ABSTRACTS OF POSTERS
P - 01
CHARACTERIZATION OF A AND C GENOME-SPECIFIC DISPERSED AND CLUSTERED
REPETITIVE DNA SEQUENCES FROM AVENA SATIVA
Evgueni V. Ananiev, M. Isabel Vales, Ronald L. Phillips, and
Howard W. Rines
Department of Agronomy and Plant Genetics, University of Minnesota
and USDA-ARS, St. Paul, MN 55108
The genomic DNAs from eight species of oat with A, C, AB, AC, and ACD
genomes were used as probes in Southern blot hybridization to a blot panel
of cosmid clones from A. sativa (ACD) genomic DNA cut with different
restriction enzymes. Several DNA subfragments were identified which specifically
hybridized with labeled A or C genomic DNA. Those subfragments were recloned
and characterized. Among them, eight A/D-genome and five C-genome specific
DNA sequences have been identified as members of different families of
dispersed repetitive elements. In situ hybridization of an A/D genome-specific
repeat, termed As-A1, to metaphase chromosomes of A. sativa showed
that it is dispersed along 14 out of 21 A. sativa chromosome pairs,
representing the A and D genomes. One of the C genome-specific repeats,
termed As-C1, is dispersed along 7 of the 21 pairs of A. sativa
chromosomes. Six of the seven C genome chromosome pairs have chromosomal
segments of different length at their termini which gave hybridization
with the As-A1 but not with the As-C1 repeats while termini of six out
of fourteen AD genome chromosome pairs gave hybridization with the As-C1
but not with the As-A1 repeats. Thus, these two probes, As-A1 and As-C1,
enabled the detection of presumably ancient reciprocal terminal translocation
segments of the C chromosomes and A or D chromosomes. A second C genome-specific
repeat, termed As-C2, revealed clusters on six of the seven C chromosome
pairs. Size, number and location of those clusters are specific for each
chromosome pair of the C genome. These three genome-specific repetitive
DNA sequences were used to screen the A. sativa genomic cosmid library
to selectively isolate C or A/D genome-specific cosmid clones. Analysis
of these new clones revealed that they contain additional genome- or chromosome-specific
DNA probes.
P - 02
EARLY DETECTION OF CHROMOSOME RETENTION/ELIMINATION IN OAT X
MAIZE HYBRIDS USING GENOMIC IN SITU HYBRIDIZATION
Evgueni Ananiev, Gang Chen, Howard W. Rines, and Ronald L. Phillips
Department of Agronomy and Plant Genetics, University of Minnesota
and USDA-ARS, St. Paul, MN 55108
Interspecific hybridization between different species of cereals is
widely used for the production of haploid plants or chromosome addition
lines as the result of preferential elimination or occasional retention
of chromosomes of one of the parents. The factors which affect chromosome
elimination/retention are poorly understood and difficult to monitor. In
most cases, cytological and molecular characterization requires well developed
plants. We applied genomic fluorescence in situ hybridization for
the early detection of chromosome elimination/retention in developing embryos,
endosperm and early seedlings in oat x maize hybrids. Developing embryos
and endosperm were isolated for analysis from caryopses at 3 and 6 days
after pollination. In addition, root tips of young seedlings from rescued
embryos were cut off, fixed in 45% acetic acid, squashed and made into
permanent preparations. Maize genomic DNA or a multiprobe ( a collection
of dispersed highly repetitive elements of the maize genome) were labeled
with fluorescein and used for in situ hybridization. Maize chromosomal
material was detected in metaphase plates as well as in interphase nuclei
using this technique. All embryos from oat-maize crosses containing maize
chromosomes were chimeric. Different cells in a developing embryo can have
different numbers of retained maize chromosomes from zero to ten. In older
embryos it is possible to detect sectors of cells without maize chromosomes.
Most endosperm cells in contrast contain practically a full set of maize
chromosomes. The cell division in endosperm usually ceases early in development.
Different levels of chromosome retention/elimination were detected in developing
young seedlings. The process of maize chromosome elimination appears to
begin during the first cell divisions after fertilization, occurs with
different rates in different tissues, and is highly asynchronous.
P - 03
THE TRANSFER OF CROWN RUST RESISTANCE GENE PC94 FROM
A WILD DIPLOID TO CULTIVATED HEXAPLOID OAT
Taing Aung1*, James Chong1 and Mike Leggett2
1 Cereal Research Centre, Agriculture and Agri-Food
Canada, 195 Dafoe Road, Winnipeg, MB, Canada, R3T 2M9; 2Institute
of Grassland & Environmental Research, Aberystwyth, Wales
Crown rust Puccinia coronata Cda. f.sp. avenae Eriks.
is the major foliar disease of cultivated oats in Canada and the United
States. Estimates of the reduction of the grain yield, based on yearly
reports of Cereal Rust Laboratories U.S. Department of Agriculture, caused
by the pathogen vary from 10% to 35% depending on the severity of the infection
and the cultivar used. The ability of the crown rust pathogen to produce
new biotypes capable of overcoming previously resistant cultivars usually
within a few years has led to a continuous search for further sources of
resistance. Several wild diploid species of oat particularly, Avena
strigosa, confer high degree of resistance to crown rust. Highly
effective crown rust resistance gene(s) was identified in a genotype of
diploid oat A. strigosa Schreb and the genotype was desiganated
as (RL1697) at CRC, Winnipeg. The resistance diploid genome of RL1697
was combined with the hexaploid genome of oat cultivar SunII in the
form of a synthetic octaploid (2n = 56). Resistant disomic addition lines
were generated in the subsequent backcross progenies from the crosses between
the octaploid and SunII recurrent parent. The resistance from the RL1697
chromosomes in the addition lines was introgressed into the SunII genome
through induced chromosome pairing procedure. We report on the successful
transfer of crown rust resistance from A. strigosa (RL1697)
to cultivar SunII.
The diploid oat A.strigosa (RL1697) resistant to crown rust race
CR152 was obtained from the Institute of Grassland and Environmental Research,
Wales, UK. in 1991. The F1 plants resistant to CR152 were produced from
the reciprocal crosses between (RL1697) and A. strigosa {Saia(Wpg)}
which is susceptible to CR152. The selfed progeny of the diploid F1 plants
segregated in a ratio of 3 R : 1 S, indicating that the RL1697
resistance is inherited as a single dominant gene. Oat cultivar SunII (pistillate
parent) was crossed with RL1697 (pollen parent) and the F1 hybrids
were treated with 0.2% colchicine to produce three synthetic octaploids
(2n = 56). Resistance reaction observed in the synthetic octaploids (2n
= 56) indicated that the resistance confer by RL1697 genome when
combined with SunII genome is expressed. Resistant plants with 2n = 44,
45 chromosome (disomic addition) were selected from the selfed progenies
of the BC1 plants produced from a cross between SunII and the octaploid.
The selected resistant chromosome addition lines were cytologically unstable
due to addition decay (loss of added RL1697 chromosomes resulted
in the loss of the resistance) which indicate that breed-true resistance
line cannot be establish from the disomic RL1697 chromosome addition
line. . The disomic addition lines were then crossed to another synthetic
octaploid (2n=56) derived from a cross between SunII (hexaploid) and A.
longiglumis,CW57 (diploid) and the resistant F1 plants so produced
were backcrossed to SunII as pistillate parent for five times.
The homozygous resistant plants which breed true for resistance in the
selfed progenies of BCV populations from heterozygous resistant lines which
segregated in a monofactorial ratio of 3 resistant : 1 susceptible
were produced. The homozygous and heterozygous plants with 42 chromosomes
were selected from the F3 generation of the BCV progenies. The selected
resistance lines exhibited no cytological abnormality (no loss or gain
of chromosome, no unusual chromosome pairing at meiosis) all the heterozygous
resistant lines segregated in 3R:1S ratio and all the homozygous lines
breed true for resistance. This confirms the transfer of crown rust resistance
gene from A. strigosa (RL1697) to cultivar SunII. One homozygous
line which is cytologically stable (2n=42 and meiosis with regular 21 bivalents)
and with agronomically desirable characteristics was selected, the line
was designated as S42, the resistance gene transferred from
RL1697 to SunII is designated as Pc94. Subsequent
evaluations showed that Pc94 is highly resistant (0 ; reaction
type) to crown rust tested in the green house and under field conditions
in Manitoba and in New Zealand.
Introgression of genes from species of lower ploidy (diploids and tetraploids)
into the genome of hexaploid oat cultivars is very difficult to achieve
by means of regular backcrossing procedures. This is mainly due to the
low frequency with which the chromosomes of the wild species pair with
those of cultivated oat, consequently reducing the chance of recombination
and gene transfer. It is well documented that a genotype of A. longiglumis
(CW57) suppress the activity of the gene or genes controlling regular bivalent
pairing in A. sativa and induced pairing between non homologous
chromosomes that resulted in translocation between two homoeologous chromosomes
as well as between unrelated chromosomes. The ability of CW57 gene(s) in
inducing pairing between A.strigosa (RL1697) chromosome and A.sativa
chromosome probably have affected the transfer of the RL1697 resistance
into SunII genetic background.
S42 was used as pollen parent and crossed to 11 North
American oat cultivars (AC Belmont, AC Marie, AC Preakness, Caliber, Dal,
Derby, Dumont, Jasper, Riel, Robert, Steele). The resistance gene
Pc94 is inherited as dominant gene in five oat cultivar (SunII,
Calibre, Dal, Derby and Jasper) genetic backgrounds. However, Pc94
is inherited as a recessive gene in seven cultivar (AC Belmont, AC
Marie, AC Preakness, Dumont, Riel, Robert and Steele) genetic backgrounds.
Since crown rust resistance gene Pc38 is present in all the
above seven cultivars and the expression of Pc94 is modified
in all these cultivars it is logical to conclude that Pc94
expression is suppressed by Pc38 (J. Chong & T. Aung
, see the following article). However, the modification of Pc94
gene expression could also be due to reversal of dominance to recessive
nature of the gene in some genetic backgrounds similar to the phenomena
reported in wheat with reference to leaf rust resistance genes. While Pc94
is proven to be a highly effective gene, there is still a need to deploy
this gene in combination with other Pc genes with the aim
to provide adequate protection to the oat crop from the ravages of the
disease and to prolong the life of the resistance gene. The S42
genotype with the Pc94 gene has provided a valuable new source
of resistance to crown rust for oat improvement programs in Canada.
*Corresponding author (Email: TAUNG@EM.AGR.CA)
*Corresponding author (Email: TAUNG@EM.AGR.CA
P - 04
OAT GERMPLASM RESOURCES IN THE USDA-ARS NATIONAL SMALL GRAINS
COLLECTION
Harold E. Bockelman
USDA-ARS, National Small Grains Collection, Aberdeen
The USDA-ARS National Small Grains Collection (NSGC) is a large working
collection of wheat, barley, oat, rye, triticale, and rice. The origins
of these small grain collections date back to the beginning of the 20th
century with the first USDA plant explorers. For the first half of the
century the collections were maintained individually by USDA Cereal Investigators.
The NSGC was officially organized in 1948, bringing together the various
small grain collections. The oat (Avena) collection presently consists
of 20,781 accessions representing 17 species originating in 95 countries
and regions of the world. Important components of the oat collection include
9,904 accessions of the hexaploid cultivated species, A. sativa,
and 8,264 accessions of the wild hexaploid species, A. sterilis.
More than 225,000 evaluation data records are available, including important
disease, insect, quality, and agronomic characters. Passport and evaluation
data are maintained on the Germplasm Resources Information Network (GRIN),
available online at http://www.ars-grin.gov/npgs.
P - 05
AGRONOMIC AND GRAIN QUALITY CHARACTERISTICS OF HULLESS OATS
D. E. Burrup1, D.M. Wesenberg1, D.M. Peterson2,
and J.C. Whitmore3.
1National Small Grains Germplasm Research Facility,
USDA, ARS, 1691 S. 2700 W., P.O. Box 307, Aberdeen, Idaho 83210, USA; 2Cereal
Crops Research Unit, USDA, ARS, 501 Walnut St., Madison, Wisconsin 53705,
USA; 3University of Idaho, Tetonia Research & Extension
Center, 888 West Highway 33, Newdale, Idaho 83436, USA
Grower interest in hulless oats in Idaho and adjacent states is being
maintained, but most planted acreages remain small and largely constitute
on-farm trials. Inquiries received by the authors frequently involve growers
interested in specialty markets, dairy producers, or horse trainers. Hulless
oat improvement currently accounts for about 25% of the oat program effort
at Aberdeen. One of the highest yielding selections in the program continues
to be 86Ab1616, which averaged 5622 lbs/A in groat yield in irrigated trials
conducted at Aberdeen, Idaho in 1994-97. The well adapted high yielding
cultivar 'Monida' averaged 5275 lbs/A in groat yield in the same trials.
The best Aberdeen hulless oat selections relative to expression of the
hulless trait (e.g., 90Ab1500) averaged about 98% hulless kernels and some
of the hulless oat selections developed by Agriculture and Agri-Food Canada,
Ottawa, Ontario (e.g., NO51-1) averaged over 98% hulless kernels in these
trials. These selections represent marked improvement over some of the
earlier hulless oat cultivars relative to the expression of the hulless
trait. Major problems frequently associated with hulless oats that continue
to impede grower and user acceptance include lower yield, shattering, poor
expression of the hulless trait, a severe itch factor, germ damage, and
grain handling problems in elevators. Data that may impact on the latter
issue are presented here for review. During the conduct of test weight
determinations it was observed that hulless oat selections vary significantly
in their rate of flow in test weight equipment. The time required for a
standard amount of grain (one quart in this instance) to flow from a standard
test weight device was measured. Trichome characteristics, including trichome
dehiscence, were also studied. Among 11 oat cultivars grown in an elite
trial for four years, three hulled cultivars ('Ajay', Monida, and 'Ogle')
were very similar in flow time, averaging 2.4 seconds. The eight hulless
cultivars averaged 2.8 seconds and ranged from 2.6 to 3.6 seconds. At least
in some cases the presence of prominent, long trichomes appeared to be
associated with the reduced rate of flow. In a larger series of 37 hulless
oat entries grown in the same trial at Aberdeen in 1997, the flow time
ranged from 2.6 to 3.6 seconds, with the hulled entries again averaging
2.4 seconds. In a second trial grown under irrigation at Aberdeen in 1997,
the flow time for 23 hulless cultivars ranged from 2.7 to 4.1 seconds.
This newly studied characteristic might be an additional useful criterion
to consider in the development of hulless oat cultivars.
P - 06
RECURRENT SELECTION FOR INCREASED PARTIAL RESISTANCE TO OAT
CROWN RUST>
C. Castell1, L. Savin2, L. Federizzi3,
G. Ochocki4, and D. Stuthman1
Dep. of Agronomy and Plant Genetics, University of Minnesota; 2ENSAT,
Toulouse France; 3 UFRGS, Porto Legre, Brazil; and 4USDA-ARS
Cereal Disease Laboratory, St. Paul, MN
Major gene crown rust resistance has not ever been long lasting. Because
it is usually race-specific, it can be overcome as new virulences emerge
in the pathogen population. Partial resistance, especially if it is race
non-specific, is likely longer lasting, or durable. This alternative type
of resistance, also called incomplete, rate-reducing, slow rusting, or
late rusting among other names, is often under polygenic control. Therefore
using recurrent selection (RS) to accumulate genes with smaller effects
should theoretically be effective. Another important difference between
major gene and partial resistance is that partial resistance can not be
measured in absolute terms. Rather, it is a relative measure comparing
a possibly partially resistant test candidate to a fully susceptible genotype.
Our objectives were to 1) practice RS to increase the level of partial
resistance, 2) evaluate the parents from each cycle of selection for their
relative level of partial resistance, and 3) determine which mechanisms
of partial resistance (infection efficiency, latent period, or spore production),
if any, were modified during the RS. F5 lines from cycle 7 of our original
Closed Recurrent Selection System (CRSS) were used as the base population
for this study. Twenty to 25 lines from 63 families were planted at Rosemount,
MN, in May, 1996, and evaluated for partial crown rust resistance after
exposure to natural inoculation. Based on family average, the 21 best families
(least rust at maturity) were chosen as parents for the next cycle. These
21 parents were crossed in a circulant partial diallel in fall 1996 and
the resulting 63 families of F1 plants were grown in the spring 1997 greenhouse.
In May 1997, these resulting F2's were planted at Rosemount to initiate
another cycle of RS for partial resistance after exposure to natural infection.
For this cycle, selection was first for the 21 best families and then for
the best plant in each selected family. This procedure was repeated for
the next cycle and on May 5, 1998, F2 families were planted at Rosemount
to initiate the third cycle of selection for partial crown rust resistance.
Simultaneously, the 12 C0 parents from the CRSS, the 21 parents from cycle
one RS for resistance, the 21 parents from cycle 2 and 'Starter', a susceptible
check, were planted in a RCB design to determine the putative genetic progress
for partial crown rust resistance. Using adult plant inoculation in the
greenhouse on the same three sets of parents and Starter, possible resistance
mechanisms of slow-rusting are being studied.
P - 07
CHARACTERIZATION OF
MAIZE CHROMOSOME RETENTION IN DEVELOPING EMBRYOS AND PLANTS FROM OAT X
MAIZE CROSSES
Gang Chen1,
Evgueni Ananiev1, M. Isabel Vales1, Oscar Riera-Lizarazu1,
Howard Rines2, and Ronald Phillips1
1University
of Minnesota; 2USDA-ARS, St. Paul, MN 55108
When the oat cultivar Starter
was pollinated with maize hybrid Seneca 60, about 20% of the florets developed
embryos. However, only 12% of the rescued embryos germinated and only about
half of those survived to maturity. Maize chromosome identification with
maize microsatellite (SSR) probes showed that all plants that ceased growth
at an early stage retained more than one maize chromosome, 35% of the surviving
plants retained one to two maize chromosomes and the remaining 65% surviving
plants had no maize chromosomes. The retention of multiple maize chromosomes
apparently retards embryo germination and seedling growth. In attempts
to obtain faster and more complete maize chromosome elimination during
embryo development, the effects of different temperatures were tested on
the chromosome constitution of young embryos in oat x maize hybridization.
Oat x maize crosses were made with the traditional emasculation and pollination
method followed by a treatment of 10 mg/L 2,4-D solution to florets two
days after pollination. After 2,4-D application, sets of plants were put
in growth chambers under temperatures of 16°C/14° C (day/ night),
20°C/18°C, or 25°C/23°C. Six days after pollination embryos
were harvested and analyzed for chromosome constitution by an in situ
hybridization technique using fluorescein-labeled maize total genomic DNA
as the probe. All of the embryos from the different temperature treatments
at this stage were chimeric in terms of maize chromosome retention. In
this initial test of 5 embryos per treatment, maize chromatin was detected
in 93.1% of the embryo cells in the high temperature treatment, 47.9% in
the middle temperature treatment, and 27% in the low temperature treatment.
These results indicate that low temperature leads to less maize chromosome
retention during embryo development.
P - 08
EVALUATION OF PARTIAL
RESISTANCE TO OAT CROWN RUST FOR QTL IDENTIFICATION
Gang Chen1,
Howard Rines2, Kurt Leonard2, Jerry Ochocki2,
Deon Stuthman1, and
Ronald Phillips1
1University
of Minnesota and 2USDA-ARS, St. Paul, MN 55108
The overall objective of this
research is to identify molecular (RFLP, AFLP) markers associated with
genes for crown rust partial resistance in oat to be able to transfer these
genes to elite germplasm using marker assisted selection. A population
of 158 F6:8 recombinant inbred lines (RILs) derived from a cross between
MN841801, a partially resistant line, and Noble, a susceptible cultivar,
were evaluated for their resistance to oat crown rust in 1997 and 1998
in both greenhouse and field tests. In the greenhouse experiments, a single
rust isolate MNB236 was chosen based on its virulence to the parents in
the seedling stage and used to inoculate the flag leaf at the floret emergence
stage. The pustule numbers per leaf area were hand-counted every 2 days
for 5 dates starting from 12 days after inoculation. Digital pictures of
the inoculated leaves were taken for image analysis of pustule numbers
per leaf area, percentage of diseased area vs. total leaf area, and pustule
size. For the field experiments, a natural crown rust population was used
to inoculate adult plants at floret emergence and a computer simulated
scale was used as a reference to read the percentage of diseased area vs.
total leaf area. The number of pustules per leaf area obtained by digital
image analysis and by hand counting matched well , the correlation coefficient
was 0.91. The values for the different progeny lines for the area under
the disease progress curve (AUDPC) from the greenhouse test had the same
patterns as those from the field. Tests of selected RILs with four other
isolates gave no indication of race specificity in the resistance responses.
Restriction enzymes BamHI,DraI,EcoRI,EcoRV and HindIII
are being used to survey parents and progeny for RFLP polymorphisms for
marker analysis.
P - 09
MOLECULAR CLONING,
CHARACTERISATION, MAPPING AND FUNCTIONAL ANALYSIS OF DISEASE RESISTANCE
GENES FROM OAT
Davis W Cheng and K
C Armstrong
Eastern Cereal and Oilseed
Research Centre, Agriculture & Agri-Food Canada, Ottawa, ON
Molecular cloning, characterisation,
mapping and functional dissection of oat disease resistance genes will
elucidate the mechanisms of gene mutation and reorganisation involved in
the evolution of new forms of disease resistance. Cloning of the
disease resistance genes would offer the possibility to use differences
between the resistant and susceptible alleles as markers for selection
in breeding and to engineer crop varieties with a higher level of resistance.
Resistance genes cloned from
flax (L6, M), Arabidopsis (RPS2, RPM1, RPP5), tomato
(prf, I2, Cf-9, Cf-2), sugar beet (HS1), tobacco (N),
rice (Xa21), and others barley (b1-b9),rice (rz1-15
), share conserved domains from which primers can be designed to amplify
disease resistance gene analogues. We have used degenerate primers to these
conserved domains and primers designed to other isolated genes eg wheat
(Lr10), barley (mlo), peroxidase from wheat etc. We have
cloned two peroxidase genes which may be induced by powdery mildew; two
NBS-LRR type genes from primers for b1 of barley; three resistance
gene analogues which contain a typical LRR motif from degenerant primers
designed from L6. We have also cloned related sequences with primers
from Lr10 of wheat, three related sequences to mlo from barley,
two sequences related to rice pi-2(t), and thirteen related sequences
to flax L6 and tobacco N. Blast search results indicate that
several of these genes contain domains with high homology to domains from
different genes from other species.
Crown and stem rust are probably
the two most important diseases of oat. Our efforts will focus on cloning
and identifying rust resistance gene analogues. In particular we will focus
on the Pg3, Pg9, Pc68 etc gene cluster by amplifying sequences
from the telo-VIII cytogenetic stock using the microdissection protocol.
We have confirmed that the genes are probably located on this chromosome
arm. Mapping the amplified products to the known map positions of rust
genes will imply function and northern analysis after pathogen challenge
will imply induction by the pathogen.
P - 10
MOLECULAR CLONING AND
CHARACTERIZATION OF PEROXIDASE GENES FROM OAT ( AVENA SATIVA L.
VAR. OGLE ) WHICH MAY BE INVOLVED IN INDUCED RESISTANCE TO ERYSIPHE
GRAMINIS F. SP. TRITICI
Davis W. Cheng and
Ken C. Armstrong
Eastern Cereal and Oilseeds
Research Center, Agriculture & Agri-Food Canada, Bldg 50, Central Experimental
Farm, Ottawa, Ontario K1A 0C6, Canada
Two peroxidase genes, temporarily
designated pxc2 and pxc6, were cloned and characterized from
oat based on a peroxidase gene sequence from common wheat. They are 1191
bp and 1197 bp in length and contain 3 exons and 2 introns in almost the
same position . The mRNA splicing site is typical of eukaryotic genes.
Their exon sequences are more conserved ( 79.4% - 88.7% ) than their intron
sequences ( 42.1% - 45% ). Blast search results indicates that the gene
sequences reported here are very similar to the barley mRNA for peroxidase
induced by powdery mildew ( 85% - 91% ), the wheat pox2 gene induced
by powdery mildew with haem-binding activity ( 85% - 89% ), and the wheat
pathogen peroxidase gene induced to powdery mildew ( 86% - 100% ). The
putative proteins encoded by oat pxc2 and pxc6 genes consist
of 313 aa ( pI 7.98 ) and 314 aa ( pI 8.18 ) respectively compared to the
wheat peroxidase gene which consists of 310 aa ( pI 7.98 ). The alignment
of amino acid sequences indicates that the structural homologies between
oat peroxidase PXC2 and PXC6 and wheat and barley peroxidases are still
very high (64.3% - 75.4% ). Experiments is to study the developmental expression
and chromosomal localization of these peroxidase genes from oat which will
also be outlined.
P - 11
IDENTIFICATION OF OAT
SPECIES AND VARIETIES USING MINISATELLITE AND MICROSATELLITE DNAs
ChengDao Li, Brian
G. Rossnagel, and Graham J. Scoles
Plant Sciences Department
& Crop Development Centre, University of Saskatchewan, 51 Campus Drive,
Saskatoon, SK S7N 5A8
The detection and analysis
of DNA polymorphisms in crops is an essential component of marker-assisted
selection and cultivar identification in plant breeding. We have explored
microsatellite DNA primers and the direct amplification of minisatellite
DNA to identify oat species and varieties.
Five primers were designed
based on minisatellite DNA core sequences. The PCR products of primers
from A longiglumis, A wiestii, A canarien, A strigosa,
A clauda, A abyssinica, A barbata, A maroccana,
A murphyi, A sterillis, A fatua, A byzantina
and A sativa demonstrated a high degree of polymorphism and the
various genomes could be identified. However, the level of polymorphism
among cultivated varieties was low.
Microsatellite DNA enrichment
libraries have been constructed. Genomic DNA of oat (Avena sativa
cv. Ogle) was digested with a combination of restriction enzymes DraI,
AluI and RsaI, and 200 to 900 bp fragments were excised and
purified from the gel. The SSR fragments were enriched with 3'-biotinilated
oligos and cloned into a pCR 2.1 vector. More than 90% of the white clones
showed positive hybridization with the synthesized SSR oligos. Sequence
analysis demonstrated that the most enriched SSR library is a (GAA/CTT)n
repeat, with 92.3% of the white clones containing that sequence. The (TC/AG)n
and (TG/AC)n libraries have 69% and 51% of the SSR clones respectively.
However, no (AT/TA)n repeat clone was identified in the present study,
although it has been reported to be the most abundant SSR in plants. These
libraries could provide enough SSR clones for constructing a saturated
SSR linkage map for genetic analysis of oat. Twenty pairs of primers were
designed and synthesized based on the above sequences. These primers were
used to characterize 11 Avena species and 20 cultivated oat varieties.
P - 12
QUANTITATIVE TRAIT
LOCI IDENTIFIED IN A HULLESS BY COVERED OAT (AVENA SATIVA
L.) POPULATION
David De Koeyer1,
Nick Tinker1, Jitka Deyl1, Vern Burrows1,
Corinne Chenier1, Steve Molnar1, Ken Armstrong1,
George Fedak1, Charlene Wight1, Darrell Wesenberg2,
Brian Rossnagel3, Deon Stuthman4, Charles Brown5,
Fran Webster5, and Art McElroy1
1 Eastern Cereal
and Oilseed Research Centre, Agriculture and Agri-Food Canada, Ottawa,
ON K1A 0C6; 2 USDA-ARS, National Small Grains Germplasm Research
Facility, Aberdeen, ID; 3 Dept. of Crop Science and Plant Ecology,
U. of Saskatchewan, Saskatoon, SK; 4 Dept. of Agronomy and Plant
Genetics, U. of Minnesota, St. Paul, MN; 5 The Quaker Oats Company,
Barrington IL
Quantitative trait loci (QTLs)
detected in adapted populations may ultimately be useful for improving
germplasm via marker-assisted selection. An F6 recombinant inbred
oat (Avena sativa L.) population, "T/M", was developed from
a cross between a hulless line and an elite covered seeded line. One-hundred
and one T/M lines (59 covered and 42 hulless) were grown in several environments
since 1992. Fourteen agronomic and quality traits were evaluated in trials
at Aberdeen, ID, Rosemount and St. Paul, MN, Ottawa, ON, and Saskatoon,
SK. The linkage map used for QTL analysis includes 193 RFLP, RAPD, and
AFLP loci. These are organized into 35 linkage groups and 40 unlinked loci.
For some traits, QTL analysis was performed separately on the covered and
hulless portion of the population or only on the covered lines. Simple
interval mapping was utilized for the QTL analyses (LOD =2.9). In total,
27 QTLs were identified affecting heading date, plant height, lodging,
tertiary kernels, grain yield, kernel weight, test weight, thin and plump
kernels, and groat beta-glucan, oil, and protein content. Two QTL regions
affecting plant height and grain yield appear to correspond to QTLs or
major genes identified in other oat populations. Many of the QTLs affecting
kernel weight, % thin and % plump kernels were concentrated in the same
genomic regions within T/M. There were several lines in this population
with improved groat percentage compared to the covered parent. This would
indicate that groat percentage QTLs exist in this population and further
mapping efforts may identify these loci. Marker-trait relationships identified
in this QTL study are being applied to marker-assisted selection projects
to evaluate this strategy for developing high quality oat lines with superior
agronomic performance.
This research was made possible
by generous financial support from the Quaker Oats Company, and by collaborative
dialogue and technical assistance from a consortium of Quaker funded institutions.
P - 13
DIFFERENCES AMONG PRIMARY,
SECONDARY, AND TERTIARY OAT (AVENA SATIVA L.) KERNELS FOR MORPHOLOGICAL
PARAMETERS AND GROAT PERCENTAGE
David De Koeyer, Jitka
Deyl, and Art McElroy
Eastern Cereal and Oilseed
Research Centre, Agriculture and Agri-Food Canada, Ottawa, ON, Canada,
K1A 0C6
Improving groat percentage
is a high priority for developing superior oat cultivars for both feed
and milling markets. A sample of oat grain is comprised of sub-populations
of primary, secondary, and sometimes tertiary kernels. The objectives of
this study were to determine the variation that exists for the "kernel
population" characteristics in elite breeding material and to assess the
relationship between these parameters and groat percentage. Twenty cultivars
and breeding lines were grown at Ottawa, ON in 1997. Panicles were randomly
selected and the primary, secondary, and tertiary kernels were manually
separated, counted, and weighed. Groat percentage was obtained for samples
of the three kernel types. The proportion of each kernel type was significantly
different among the lines tested, based on number of kernels and total
weight. Germplasm with higher groat percentage was characterized as having
high primary groat percentage and a smaller difference in groat percentage
between the primary and secondary kernels. Only two lines had greater than
80% groat values for secondary kernels, a plateau that appears to exist
for the other lines in this study. Improved selection efficiency may be
obtained by preferentially selecting for groat percentage of primary kernels,
the largest (61.0%) component on a total weight basis. Only 3 lines in
this study exceed 75% for primary groat percentage, whereas, only 3 lines
did not exceed 75% for secondary groat percentage. Increasing the proportion
(weight) of secondary kernels relative to the primary kernels may also
be an effective way of increasing groat percentage. This study has helped
to identify some important factors that may be used to help develop varieties
with higher groat percentage.
P - 14
ANTIOXIDANT ACTIVITY
AND PHENOLIC ANTIOXIDANT CONTENTS
OF OAT FRACTIONS
Cheryld L. Emmons1,
David M. Peterson1,2, Greg Paul3, Jeffrey B. Blumberg4,5,
Garry J. Handelman4,5,
Ronald L. Prior5, and Guohua Cao5
1Cereal Crops
Research Unit, Agricultural Research Service, USDA, Madison, Wisconsin;
2Department of Agronomy, University of Wisconsin, Madison,
Wisconsin; 3The Quaker Oats Company, Barrington, Illinois; 4Tufts
University, Boston, Massachusetts; and 5Jean Mayer USDA Human
Nutrition Research Center on Aging, Boston, Massachusetts
The potential of oat fractions
to contribute to dietary antioxidants was evaluated. Antioxidant activity
was measured in methanol extracts of flour, pearlings, bran fines, and
trichomes from groats by 1) inhibition of coupled autoxidation of linoleic
acid and -carotene (AOA), 2) inhibition of copper and peroxyl radical-induced
low-density-lipoprotein oxidative damage (LDL-C, LDL-P, respectively) (Trolox
equivalents), and 3) oxygen radical (peroxyl and hydroxyl) absorbance capacity
(ORAC-P and ORAC-H, respectively) (Trolox equivalents). Pearlings had greater
AOA (62-78% inhibition) than flour (53%), bran fines (57%), or trichomes
(47%). Pearlings had greater ORAC-P (5.81) and ORAC-H (6.34) than flour
(3.40, 3.70), bran fines (2.71, 1.80), and trichomes (2.74, 1.11). LDL-P
and LDL-C were greatest for pearlings (3.82 and 3.42, respectively), followed
by trichomes (1.54, 1.87), flour (1.26, 0.55), and bran fines (1.28, 0.46).
AOA was correlated to LDL-C (r=0.75).
Total phenolic content (TPC),
determined using the Folin-Ciocalteau phenol reagent, was expressed as
gallic acid equivalents. Pearlings contained the greatest TPC (318 mg/kg),
followed by flour (156 mg/kg), trichomes (139 mg/kg), and bran fines (107
mg/kg). TPC was correlated to AOA (r=0.85), ORAC-P (r=0.80), and LDL-C
(r=0.93). Phenolic compounds were separated by HPLC on a C18
column using a linear 1-40% acetonitrile gradient (pH 2.8). Peaks were
identified by comparing retention times and UV spectra with known standards.
Tannic acid, p-hydroxy-benzoic acid, vanillic acid (VA), caffeic
acid (CA), syringic acid, vanillin, p-coumaric acid, and ferulic
acid were identified and quantified in each sample. AOA was correlated
to VA and CA contents (r=0.84 and r=0.79, respectively). LDL-C was correlated
to CA (r=0.60). Three peaks correlated to AOA and LDL-C have spectra similar
to hydroxy cinnamic acids and retention characteristics similar to avenanthramides
(AV). They have been tentatively identified as AV1, AV3, and AV4 (r>0.83
and r>0.54 for AOA and LDL-C, respectively). A fourth peak, with
a spectrum similar to catechin (a flavan-3-ol), has been denoted F3OL (r=0.80
and r=0.56 for AOA and LDL-C, respectively). Removing trichomes from the
correlation analysis increased coefficients for some of the identified
compounds, suggesting 1) oat fractions contain many phenolic compounds
that contribute to antioxidant activity, and 2) trichomes have potential
for good activity, but may contain interfering pro-oxidative compounds.
These data indicate a potential
for oat products, especially those enriched in the outer layers of the
kernel, to significantly contribute to dietary intake of antioxidant phytonutrients.
Analysis of a wider selection of genotypes, fractions, and products is
planned to determine the full potential of these materials as sources of
dietary antioxidants.
P - 15
PARACENTRIC INVERSIONS
IN F1 HYBRIDS OF OAT CULTIVARS
George Fedak, Nick
Tinker, L. O'Donoughue, Vern Burrows, Ken Armstrong, Steve Molnar
Eastern Cereal and Oilseed
Research Centre, Agriculture and Agri-Food Canada, Ottawa, Ontario, Canada
K1A 0C6
The presence of duplicated
regions in the oat genome as detected by RFLP markers and other anomalies
in segregation patterns of parental genotypes in segregating populations
has prompted a study of meiotic chromosome behaviour in F1 hybrids
that gave rise to the mapping populations. A population obtained from a
Hinoat × Newman F1 hybrid showed an excess of Newman type
parental gametes (L. O'Donoughue, p.c. N. Tinker p.c.)
Chromosome behaviour was examined
at meiosis in F1 hybrids derived from cultivars Tibor ×
Marion, Hinoat × Newman, Marion × Tibor and Hinoat ×
Marion. A very low frequency of quadrivalent configurations was observed
in most plants of most hybrid combinations. A-I bridges were observed at
frequencies of 30 - 49% in Hinoat × Newman and 7% in some but not
all Tibor × Marion F1 hybrids. In meiocytes where bridges
occurred, single bridges were most common but up to five bridges were seen
in some cells. The single A-I bridge configurations were the result of
single crossovers or 3 strand double crossovers in the inverted segments
in inversion heterozygotes. Double bridges were the result of four strand
double crossovers in the inversion loops. The expected free fragments that
accompany a bridge configuration were rarely seen at A-I but were frequently
present in the cytoplasm of telophase I. There was some decline in proportion
of stainable pollen in plants containing inversions.
The above observations indicate
that the cultivars Hinoat and/or Newman contain paracentric inversions.
Some of the consequences of the presence of inversions include asynapsis
and nonhomologous pairing in regions adjacent to the inversion, thus causing
reduced recombination in those regions. Chromatids with duplication and
deficiencies would result from the recombination process leading to the
production of like gametes. The viability of certain gametes could thus
be affected. This concept requires additional study in other cultivar combinations.
P - 16
AN UPDATE OF THE MOLECULAR
LINKAGE MAP OF CULTIVATED OAT
Susanne Groh1,
Nick Tinker2, Louise O'Donoughue3, Shahryar Kianian4,
Stephen Fox5, Suzanne Livingston1, Charlene Wight2,
Howard Rines6, Mark Sorrells7, Steve Molnar2,
George Fedak2, Ken Armstrong2, Ronald Phillips1.
1Dept. of Agronomy
and Plant Genetics, University of Minnesota, St. Paul, MN 55108, USA; 2Agriculture
and Agri-Food Canada, Research Branch, Eastern Cereal and Oilseed Research
Centre, Ottawa, ON K1A 0C6, Canada; 3DNA Landmarks Inc., St.
Jean Sur Richelieu, Quebec J3B 6Z1, Canada; 4Dept. of Plant
Sciences, North Dakota State University, Fargo, ND 58105, USA; 5Crop
Development Centre, University of Saskatchewan, Saskatoon, SK S6N 5A8,
Canada; 6Plant Science Research Unit, USDA-ARS, and Dept. of
Agronomy and Plant Genetics, University of Minnesota, St. Paul, MN 55108,
USA; 7Dept. of Plant Breeding, Cornell University, Ithaca NY,
14853, USA
A molecular genetic linkage
map of cultivated oat based on the cross Kanota (Avena byzantina)
× Ogle (A. sativa) has been developed by a consortium effort.
This map has provided the foundation for additional genetic studies and
is a pivotal component of ongoing comparative-mapping efforts. We have
continued to update this map since it was first reported by O'Donoughue
et al. (Genome 38:368-380, 1995). Here, we report an updated version of
the Kanota × Ogle linkage map. The former map was based on 561 loci
mapped in 71 recombinant inbred lines (RILs). We have now mapped additional
RFLP markers detected by cDNA clones from oat, barley, and wheat as well
as probes from oat genomic DNA. After checking for duplicates and severe
segregation distortion, data were available for 703 marker loci. The population
has been extended to 137 RILs, in which 108 of the loci have been mapped
to date. Markers were assigned to linkage groups at LOD 7. Some unlinked
groups have been joined based on chromosome assignments determined by aneuploid
studies (reported elsewhere), resulting in a total of 32 linkage groups.
Marker ordering within linkage groups was performed for a subset of 259
loci providing a robust framework with a uniform marker density of approximately
5 to 10 cM. Most ordering within this framework was performed at LOD 3.
Markers not included in the framework have been placed relative to this
framework. The current size of this base map is 1770 cM excluding unlinked
markers. Future work will include mapping of additional markers on the
extended population, with emphasis on markers that will allow for comparisons
with maps of other crop species. Updated versions of the map will be made
available on the GrainGenes database (http://wheat.pw.usda.gov/).
P - 17
TERTIARY KERNEL EVALUATION
IN OAT
Cristine Handel1,
Melissa Huhn1, Deon Stuthman1 and Gary Fulcher2
1Department
of Agronomy and Plant Genetics, University of Minnesota; 2Dept.
of Food Science and Nutrition, University of Minnesota
Seed physical traits, such
as seed size and shape, are very important for oat processing industry.
It is known that the number of kernels per spikelet is variable in oat
(Avena sativa L.). Commonly there are only two kernels, but three and even
four can be present. In most cases the primary kernel is bigger than the
secondary one, and it is bigger than the tertiary kernel (TK). In studying
TKs, a precise phenotypic rating of this trait is needed, especially if
the data are used for molecular marker development. One way of rating TK
is by looking at the top of the panicle of 5 plants in a row, and scoring
zero (absence) or one (presence). This method is very fast but genotypes
that are intermediate or even low for TK's might be misscored in some environments.
Another way of rating TKs is by counting the number of florets and TKs
in a panicle. This method is precise, but very time consuming. In our study
these two methods were compared. In addition, the panicles were divided
into three parts and TK numbers determined for each third of the panicle.
The genetic material used for the study was composed of seven F6 pairs
of iso-lines (high and low TK) planted in St. Paul MN (regular TK expression)
and Aberdeen ID (high TK expression). Genotypes, locations, types of rating
and the genotypeXlocation interaction were significant sources of variation.
In both locations, the rating of the second third (middle of the panicle)
gave the same results as rating the whole panicle for TKs, making the rating
process faster but still adequately precise.
P - 18
AMPLIFIED DNA FRAGMENT
LENGTH POLYMORPHISM BETWEEN OAT AND BARLEY MAPPING PARENTS
D. L. Hoffman and J.
E. McNeil
USDA-ARS Small Grains
Germplasm Research Facility, Aberdeen, Idaho 83210 USA
Genetic maps have become an
important tool for understanding the basic genetics of crop plants. Detailed
molecular maps for oat and barley have been constructed with the use of
DNA restriction fragment length polymorphism (RFLP) markers. The use of
amplified DNA fragment length polymorphism (AFLP) markers is an efficient
method for mapping additional populations and adding more markers to existing
maps. Before mapping with AFLP markers, it is essential to know which primer
pairs yield numerous DNA amplification differences between parental cultivars.
The objectives of this study
were to 1) identify which AFLP primer pairs will yield the most DNA amplification
differences among oat mapping parents and 2) compare this information with
that found for barley. Total genomic DNA was extracted from plants of oat
mapping cultivars Kanota, Ogle, and TAM O-301, and from plants of barley
mapping cultivars Steptoe and Morex. AFLP was conducted using protocols
established by Gibco BRL Life Technologies, and the five cultivars were
surveyed for potential AFLP markers with 62 EcoRI-MseI primer pairs.
The results indicated that
some primer pairs are better than than others in producing AFLPs in both
oat and barley, and that the conduct of a preliminary screen is helpful
for efficient mapping. AFLP was more readily found in barley. Some adjustments
to the procedure may need to be made to find more AFLP for oat. The next
step will be to determine the actual number of AFLP markers with segregating
populations. This information will be used to help construct an oat molecular
genetic map in Ogle/TAM O-301 and increase the number of non-RFLP markers
for barley.
P - 19
NINE CYCLES OF RECURRENT
SELECTION FOR INCREASED GROAT OIL CONTENT IN OAT
J.B. Holland and K.J.
Frey
Dep. of Agronomy, Iowa
State University, Ames, IA 50010
Nine cycles of recurrent phenotypic
selection for high groat (caryopsis) oil content were conducted in a genetically
broad-based oat (Avena sativa L.) population including germplasm
from the wild relative A. sterilis. The objectives of the experiment
were to determine if selection for high groat oil content was effective
throughout all nine cycles and if selection reduced genetic variation for
oil content; to identify agronomically acceptable lines with high groat
oil content; and to document the effects of selection on grain yield, groat
fraction, or oil yield. We evaluated 100 random lines from the base (C0)
population and each of the nine selection cycle populations in three environments.
Mean groat oil content increased linearly over cycles of selection at a
rate of 6.6 g kg-1 per cycle (equivalent to 6.7% of C0 population
mean per cycle) from a mean of 98.2 g kg-1 in C0 to 158.5 g
kg-1 in C9. The rate of gain from selection did not decrease
in later cycles. Oil yield increased at a rate equivalent to 1.9% of C0
population mean per cycle, while mean groat fraction did not change and
mean grain yield decreased at a rate equivalent to 3.2% of C0 population
mean per cycle. For all traits, genetic variation did not decrease over
cycles. Selected S0-derived lines from the population with groat
oil contents greater than 150 g kg-1 exhibited agronomic deficiencies
such as lower yield and greater lodging and disease susceptibility compared
to the best check cultivar.
P - 20
COMBINED AFLP AND RFLP
MAPPING IN TWO HEXAPLOID OAT RECOMBINANT INBRED POPULATIONS
H. Jin1,
L. L. Domier1,2, F. L. Kolb1, and C. M. Brown1
1Department
of Crop Sciences, University of Illinois, 1102 South Goodwin Ave., Urbana,
IL 61801 USA; 2United States Department of Agriculture, Agricultural
Research Service, Crop Protection Research Unit, 1102 South Goodwin Ave.,
Urbana, IL 61801 USA
The segregation of AFLP markers
was analyzed in two different hexaploid oat recombinant inbred populations.
The first population was derived from a cross of Kanota x Ogle and consisted
of 71 recombinant lines, which were previously used to produce an RFLP
map of the hexaploid oat genome. The second population was derived from
a cross of Clintland64 and IL86-5698, barley yellow dwarf virus sensitive
and tolerant oat lines, respectively. One hundred twenty-six lines of this
population were used to identify loci contributing to tolerance to barley
yellow dwarf virus infection. More than 300 AFLP markers were scored in
each of the populations, of which 85 could be scored in both populations.
These markers were linked to RFLP markers in 20 of 38 Kanota x Ogle RFLP
linkage groups. The addition of the 306 AFLP markers to the Kanota x Ogle
RFLP data set reduced the number of linkage groups to 33 at a LOD of 8.0
and a recombination fraction of 0.3. Most of the markers that showed distorted
segregation in one population were more normally distributed in the other.
All but eight of the AFLP markers scored in the Kanota x Ogle population
were linked to previously mapped RFLP markers. Thirty six linkage groups
were observed in the Clintland64 x IL86-5698 population. Two of the linkage
groups from this population were consolidated by comparing the segregation
of markers in both populations. Markers were linked in similar orders in
the two populations and showed a very similar distribution to that of the
RFLP markers.
P - 21
GENETIC ENGINEERING
OF MODEL AND ELITE OAT VARIETIES WITH ANTIFUNGAL GENES USING A NOVEL SELECTION
SYSTEM
Heidi F. Kaeppler,
Geeta K. Menon, Ronald W. Skadsen, and Anna M. Nuutila.
Department of Agronomy,
University of Wisconsin, Madison, WI, 53706, and USDA-ARS Cereal Crops
Quality Unit, Madison WI, 53706
Genetic engineering can be
utilized in oat as a complimentary tool to breeding to incorporate genes
encoding novel or altered traits for improved grain quality or agronomic
performance. Limitations in oat transformation include low transformation
frequencies, difficulty in application of current transformation systems
to elite oat varieties, and few choices of marker genes for efficient selection
of transformants. The objectives of our research were to: 1. develop a
new selection system based on visual selection for expression of the green
fluorescent protein gene, and 2. transform the highly culturable oat genotype,
GP-1, and elite oat varieties Belle, Dane and Gem, with transgenes encoding
antifungal proteins using the new gfp-based selection system. The gfp expression
construct and the antifungal transgene constructs were codelivered into
friable, embryogenic oat tissue cultures via microprojectile bombardment.
Transgenic cell lines of GP-1, Belle, Dane and Gem were recovered following
visual selection for gfp-expressing cells. Fertile, transgenic plants were
regenerated from all selected cell lines of GP-1. Transgenic cell lines
of Belle, Dane and Gem are currently on regeneration medium. Strong expression
of gfp was observed in callus, regenerated plant tissues and transgenic
seedlings. Southern blot and PCR analysis confirm integration of the transgenes
in primary regenerants and seedling progeny. The period of time between
initiation of selection and placement of cells on regeneration medium was
4-5 weeks, compared to 8-12 weeks when selection was based on antibiotic
resistance. Analysis of transgenic plants and progeny for expression of
the antifungal proteins is underway.
P - 22
NUCLEAR MAGNETIC RESONANCE
IMAGING FOR DETERMINING THE DISTRIBUTION OF OIL IN OAT (AVENA SATIVA
L.) GROATS
S. Kibite, H. Grondey
and A. Blazek
Agriculture & Agri-Food
Canada, Lacombe Research Centre Lacombe, Alberta, Canada.
Nuclear magnetic resonance
(NMR) imaging is an extension of NMR spectroscopy where the NMR signal
is resolved by the application of 3-dimensional magnetic field gradient.
NMR imaging was used in the present study to determine the feasibility
of using this technology to examine the distribution of oil in the oat
groat, and also to elucidate any differences that may exist among high-
and low-oil genotypes. This information would be useful to plant breeders
who may be interested in breeding new cultivars for various end-use applications.
Three oat genotypes, Waldern,
LAO-505-001, and N326-7 were selected to give a range of oil contents from
less that 6% to more than 16% on a dry groat-weight basis. The three genotypes
were grown at the same location and in the same growing season to eliminate
environmental differences, which have been shown to affect oil content
in oat. A Brunker mico-imaging probe equipped with a 10-mm horizontal coil
was used for imaging the groats. When a short TE (time to echo) was used,
both the oil and water components of the groats were visible, and the resulting
images were blurred due to the fact that the oil and water signals were
offset slightly from each other. A longer TE dephased the signals from
the water component and produced sharper images. The length of TE required
to dephase the water signals was 23 ms for Waldern, 29 ms for N326-7 and
50 ms for LAO-505-001. These TE values appeared to be inversely related
to the moisture content of the samples since the Waldern, N326-7 and LAO-505-001
groats contained 13.1, 10.1 and 9.5% moisture, respectively. In all three
genotypes, the oil component appeared to be localized in the germ as well
as in the periphery (presumably the bran fraction) of the groat, with very
little oil present in the endosperm. Although the amounts of oil in the
germ and bran fractions of each genotype were not quantified, it was evident
from visual observation of the images that the major differences among
high, medium- and low-oil genotypes were due to the amounts of oil that
were present in the germ. These observations indicate that NMR imaging
can be a useful indicator of the distribution of oil in the groat, and
can aid plant breeders in cultivar selection for specific end-uses.
P - 23
GENETIC DIVERSITY FOR
ALUMINUM TOLERANCE IN AVENA SPP
S. Kibite.
Agriculture & Agri-Food
Canada, Lacombe Research Centre, Lacombe, Alberta, Canada.
Aluminum (Al) toxicity is
a major factor limiting oat (Avena sativa L.) production in the
Grey-Wooded soils of western Canada. Progress in breeding for Al tolerant
oat cultivars has been limited by a lack of suitable sources of genes for
Al tolerance. In an effort to find new genes for Al tolerance, ~3,500 germplasm
accessions for the USDA World oat Collection were evaluated for Al response
in the greenhouse using a modified hydroponics system. Accession lines
were grown in Al-toxic (300 M of Al, pH 4.5) and Al-free (0 M of Al, pH
6.5) nutrient culture solutions. Ten seedlings from each treatment combination
were harvested 14 days after seeding, and the roots measured to assess
the level of Al tolerance in each accession. Using this method, 11 Al tolerant
lines (5 A. strigosa, 4 A. brevis, 1 A. fatua and
1 A. abyssinica) were identified. Subsequent greenhouse experiments
revealed that these 11 accession lines were more tolerant than previously
reported Al tolerant genotypes, and suggested that they may carry new genes
for Al tolerance.
P - 24
THE INHERITANCE OF
ALUMINUM TOLERANCE IN THREE OAT CROSSES
S. Kibite
Agriculture & Agri0Food
Canada, Lacombe Research Centre, Lacombe, Alberta, Canada
Aluminum (Al) toxicity is
a major problem in oat (Avena sativa L.) grown in acid soils. Breeding
for Al tolerance (AT) is hampered by a lack of information on the inheritance
of this trait. Three experiments were conducted in a greenhouse to determine
the mode of inheritance of AT in oat. The results showed that AT is a heritable
trait and that it is governed by nuclear genes. Data from two experiments
exhibited inheritance patterns consistent with those expected for a single
gene with complete dominance for the tolerant reaction. Data from the third
experiment showed a more complex inheritance suggesting that several genes
may be involved in the expression of AT. Broad-sense heritability estimates
in the three experiments ranged from 0.23 to 0.81.
P - 25
QTL MAPPING OF GROAT QUALITY TRAITS IN OAT: ACETYL-COA CARBOXYLASE,
A CANDIDATE GENE FOR OIL CONTENT
Shahryar F. Kianian1, Susanne Groh2, Margaret
A. Egli2, Ronald L. Phillips2, Howard W. Rines3,
David A. Somers2, Burle G. Gengenbach2, Francis H.
Webster4, Suzanne M. Livingston2, Louise S. O'Donoughue5,
Mark E. Sorrells6, Darrell M. Wesenberg7, Deon D.
Stuthman2, and R. Gary Fulcher8
1Dept. of Plant Sciences, North Dakota State University,
Fargo ND 58105, USA
2Dept. of Agronomy and Plant Genetics, University of Minnesota,
St. Paul MN 55108, USA
3Plant Science Research Unit, USDA-ARS, and Dept. of Agronomy
and Plant Genetics, University of Minnesota, St. Paul MN 55108, USA
4The Quaker Oats Co., John Stuart Research Laboratories,
Barrington IL 60010, USA
5DNA Landmarks Inc., St. Jean Sur Richelieu, Quebec, J3B
6Z1, Canada
6Dept. of Plant Breeding, Cornell University, Ithaca NY
14853, USA
7USDA-ARS, National Small Grains Germplasm Research Facility,
Aberdeen ID 83210, USA
8Dept. of Food Science and Nutrition, University of Minnesota,
St. Paul MN 55108, USA
Genetic manipulation of groat quality traits such as oil and beta-glucan
content is desired for optimizing the value of oats in human and livestock
diets. The localization and characterization of quantitative trait loci
(QTLs) for these traits is necessary for identifying genetic markers to
be used for marker-assisted selection (MAS) to improve the quality of oat
groats. Two populations of recombinant inbred lines derived from the crosses
Kanota × Ogle (K×O) and Kanota × Marion (K×M) were
genotyped using RFLP markers and tested for oil and beta-glucan content
across several locations and years. QTL analysis was conducted using composite
interval mapping with 350 RFLP loci in K×O and single factor ANOVA
with 60 loci in K×M. A major QTL affecting oil content was found
consistently across environments in both populations on linkage group 11,
explaining up to 43% of the phenotypic variance. A second locus was found
in each population, explaining an additional 10 to 20% of the phenotypic
variance. A cDNA clone encoding oat acetyl-CoA carboxylase (ACCase) mapped
to the same region as the major QTL on linkage group 11 in both populations.
ACCase, which catalyzes the first step in de novo fatty acid synthesis,
is therefore likely to play a major role in determining groat oil content
and provides an excellent marker for MAS. Furthermore, the genomic region
around ACCase also showed an effect on beta-glucan, indicating that selection
for this locus could simultaneously influence oil and beta-glucan content.
However, the allele from Kanota increased the numerical value of both traits
in K×O, while in K×M the allele from Kanota increased oil and
the allele from Marion increased beta-glucan content. Additional research
will be needed to gain a better understanding of the role of ACCase in
determining groat oil content and possible effects on correlated traits.
P - 26
THE EFFECT OF RIPENESS
AND HARVESTING TECHNIQUE ON THE GERMINATION OF NAKED OAT IN NORTHERN GROWING
CONDITIONS
Anna-Maija Kirkkari1,
Ari Rajala2, and Pirjo Peltonen-Sainio3
1Department
of Agriculture, TTS-Institute; 2Department of Plant Production,
University of Helsinki; 3Department of Plant Production, University
of Helsinki
In Finland, oat is the most
cultivated cereal crop after barley (370 000 hectares, 34 % of the grain
crop cultivation area in 1996). Presently, all of the cultivated oat in
Finland is conventional hulled oat. The interest in naked oat is however
increasing, due to its high quality grain material for various industrial
purposes and also due to possibilities to utilise naked oat in novel healthy
food production. In Scandinavian growing conditions, grain is most often
harvested at moisture content between 15 - 30 % . Combine harvesting at
high moisture content, exposes the naked grain to mechanical damages. After
harvesting grain is dried in grain circulating, batch type dryers. Heavy
mechanical handling of naked grain during harvesting and drying especially
at high moisture levels often result in reduction in germination. Poor
germinating grain material is unacceptable for seeding or for industrial
processing that requires high germination.
The aim of this study was
to optimise the harvesting procedure of naked oat in order to minimise
grain damage and loss of germination. A practical guide, i.e. 'harvesting
recipe' will be produced for the benefit of farmers interested in this
new crop. In farm scale trials, supported with laboratory tests, naked
oat will be harvested and dried with conventional machinery to establish
the effect of ripeness and various grain moisture on the germination. Hand
threshed grain material is used as reference. The trials are carried out
in 1998-2000.
In preliminary trials, highest
germination has been achieved when harvested at 19% moisture content. In
field trials, the optimal moisture range will be established in relation
to ripening and weather conditions.
In conclusion, naked oat is
an attractive new crop in Finland for food and feed processing industry.
Prior to introduction to the farmers, the especially problems related to
harvesting and drying need to be solved.
P - 27
DEVELOPMENT OF BARLEY
YELLOW DWARF VIRUS TOLERANT OAT GERMPLASM LINES
F.L. Kolb1,
C.M. Brown1, L.L. Domier
2,1, and N.J. Smith1
1Department
of Crop Sciences, University of Illinois at Urbana-Champaign; 2
USDA-ARS
A population from a four-way
cross was used to develop barley yellow dwarf virus (BYDV) tolerant oat
germplasm lines. Our objective was to combine genes for tolerance to BYDV
into germplasm lines with a higher level of tolerance to BYDV than the
parents. The four-way cross involved four BYDV tolerant parents: IL86-1156,
IL86-5698, IL86-6404 and Ogle. The F3 population was space-planted
in the field in 1993 and inoculated with BYDV-PAV-IL when the plants were
at Feekes GS 1. Plants that exhibited BYDV symptoms were destroyed. About
780 of the most tolerant plants (based on lack of symptoms) were harvested
individually. In 1994, a single hill of each line was evaluated for BYDV
tolerance in a BYDV-PAV-IL inoculated nursery, and 139 lines were selected.
The 139 lines (plus the parents and checks) were evaluated in 1995 and
1996 using three replications of paired control and BYDV-PAV inoculated
hills. Because these lines exhibit little or no BYDV symptoms, we evaluated
BYDV tolerance based on virus titer using ELISA, percent stunting (height
difference between control and inoculated hills), and percent yield loss
(grain yield difference between control and inoculated hills). In 1996,
62 lines were evaluated for agronomic performance in a replicated experiment
using 6 row plots at one location. In addition to BYDV tolerance, lines
were selected for further evaluation based on grain yield per se, kernel
morphology and absence of awns. In 1997, 42 lines were evaluated further
in replicated trials at two locations. Based on all of the criteria used
for selection, 7 lines are being increased for release as BYDV tolerant
germplasm lines. These lines differ somewhat in height, maturity and ELISA
virus titer. All of the lines have excellent tolerance to BYDV-PAV.
P - 28
NIRS PREDICTION OF
GROAT PROTEIN AND OIL CONTENT USING WHOLE OAT (AVENA SATIVA L.)
SAMPLES
A.R. McElroy and D.T.
Grégoire
Eastern Cereal and Oilseed
Research Centre, Agriculture and Agri-Food Canada, Ottawa, Ontario. K1A
0C6
Near infrared reflectance
spectroscopy (NIRS) is routinely used for quality analysis of grains and
oilseeds. Quality analyses are made on whole oat samples for feed purposes
and on groats for milling purposes. Therefore, milling oat samples must
be dehulled before quality analysis. NIRS prediction equations were developed
to determine the protein and oil content of the groat using whole oat samples.
Groat samples, prepared by
Codem dehulling, were evaluated for protein (Kjeldahl N x 6.25) and acid
hydrolysis fat. Initial equations were developed from a 100 sample population
from Ottawa and Elora, Ontario, and then enhanced with a 154 sample set
from Saskatchewan (11 cultivars x 14 sites). The per cent groat ranged
from 60 - 79 % (Ontario) and from 63 - 80% (Saskatchewan). The mean groat
protein was approximately 15% for both locations (11 - 23% overall), while
the mean groat oil content was 8.5 (Ontario) and 9.9 % (Saskatchewan) with
an overall range from 5 - 13.5%.
Comparisons of the standard
error of prediction (SEP), slope and ratio of standard deviation of predicted
values to SEP (RPD) were made between two techniques: 1) whole oat prediction
equations developed here, and 2) bias-corrected predictions using Quaker
Oat universal groat equations.
The SEP using whole oat and
groat equations were similar for protein (0.51 vs 0.68) and oil (0.27 vs
0.31) for the Ontario samples. Slope and RPD values were similar for both
techniques. The SEP for the Saskatchewan samples, using the enhanced quation,
was 0.72 for protein and 0.49 for oil. It is concluded that the protein
and oil content can be predicted as accurately using whole oat equations
as with traditional groat equations, despite a wide variation in the groat
content of the samples.
P - 29
CURRENT STATUS AND
PERSPECTIVES OF OAT BIOTECHNOLOGY IN SOUTHERN BRAZIL
S.C.K. Milach
UFRGS, Depto. Plantas
de Lavoura, C.P. 776, Porto Alegre, RS, Brazil, 915011-970, milach@vortex.ufrgs.br
There are two main groups in southern Brazil that
have been conducting research in oat (Avena sativa L.) biotechnology.
One is at Federal University of Rio Grande do Sul (UFRGS) and the other
at University of Passo Fundo. In the later, the main effort has been in
using tissue culture to generate somaclonal variants. Several lines regenerated
in vitro have shown variation for agronomic traits and one of these
has been selected and tested in the official yield nursery; but has not
been promoted to a variety. At UFRGS until recently the main effort was
also in using tissue culture to select for new variation. In the last few
years, however, this program has been expanded. In this paper we will present
the current efforts and perspectives of oat biotechnology at UFRGS. Thirty-five
oat genotypes grown in the Southern Brazilian environments are being characterized
at the molecular level using RAPD markers. These genotypes were chosen
because they represent a broad range of variability and have traits of
interest to be mapped with markers in the future. Partial resistance (PR)
to oat crown rust has being identified in the germplasm of UFRGS. Through
a collaborative research project with Dr. Greg Penner from the Cereal Research
Centre of the Agriculture and Agri-Food Canada Institute, current work
is being conducted on mapping PR to crown rust using AFLP markers in one
population from a cross between two UFRGS lines. The perspective on this
project is to convert some of the AFLP markers identified as associated
to PR to Amplicon Specific Alleles to screen oat lines for PR in the UFRGS
breeding program. Finally, the development of a system to transform Brazilian
oat lines is underway with the objective to obtain resistance to spot blotch,
which is not available at desirable levels in the oat germplasm.
P - 30
VARIABILITY FOR OAT KERNEL MORPHOLOGY IN
BRAZILIAN GENOTYPES AND ITS RELATION TO MILLING YIELD
S.C.K. Milach, C.R.A. Bothona, L.M. Tisian,
M.C.C. Teixeira
UFRGS, Depto. Plantas de Lavoura, C.P. 776,
Porto Alegre, RS, 915011-970, Brazil
Kernel morphology of oat (Avena sativa L.)
Brazilian genotypes has been studied in the last few years at the Federal
University of Rio Grande do Sul, in attempt to identify germplasm with
superior quality. The objective of this paper is to present the main results
of the studies carried out to characterize the genetic variability, to
access the effect of years in the expression of kernel morphology, and
to identify kernel traits possibly associated to milling yield. Nine genotypes
were grown in 1995 and 1996 in a complete randomized experiment with four
replications. Five panicles per replication were harvested and primary,
secondary and tertiary kernels per panicle were separated and counted.
A sample of 50 kernels of each type per genotype and replication was prepared
and evaluated using digital image analysis (DIA). Kernel traits evaluated
were area, length, width, perimeter and shape factor. In 1996, the 5 meters
long and wide plots were harvested after the five panicles were taken;
then test weight, milling yield, weight of a thousand kernels and grain
yield were evaluated. Variability among oat genotypes and the interaction
genotype x year were significant for all kernel morphology traits. However,
the top (UFRGS911747) and bottom (UFRGS7) genotypes have been the same
in both years for all traits. Significant correlation was not found between
kernel morphology and milling yield traits. However, the genotype with
the best milling yield, CTC3, did not have tertiary kernels and had high
test weight. The results obtained so far indicate that morphology per se
is not enough to select genotypes with high milling yield in the Brazilian
oat germplasm.
P - 31
STRATEGIES TO CHARACTERIZE PARTIAL RESISTANCE
TO OAT CROWN RUST OCCURRING IN GENOTYPES GROWN IN THE SOUTHERN BRAZILIAN
ENVIRONMENTS
S.C.K. Milach1, G.C.H Thome1,
G.O. Mellos1, L.C. Federizzi1, J.A. Martinelli1,
G.A. Penner2
1 UFRGS, Depto. Plantas de Lavoura,
Porto Alegre, RS, C.P. 776, 91501-970, Brazil; 2 Cereal Research
Center, Agriculture and Agri-Food Canada, Winnipeg, Canada
Oat (Avena sativa L) crown rust (Puccinia
coronata f. sp. avenae) encounters favorable environments and
has large variability in Southern Brazil. For this reason, a major challenge
for the Oat Breeding Program at the Federal University of Rio Grande do
Sul has been to breed genotypes with more durable resistance to crown rust.
Thus, we have been searching for alternatives to identify genotypes with
partial resistance (PR). The objective of this paper is to outline our
approach in identifying and characterizing PR to crown rust in oat Brazilian
genotypes and to present the main results obtained so far. The strategies
used include: identification of traits and methods to evaluate PR for the
Southern Brazilian environments; identification and characterization of
oat lines for PR; verification that PR will hold through different years;
determination of the PR genetic basis in selected lines; and molecular
mapping of quantitative trait loci (QTL) associated to PR. A digital image
analysis procedure was developed to evaluate leaf area infected with rust.
Other traits evaluated were pustule size and area under the disease progress
curve. The results indicated that a more precise analysis was obtained
by using the data of all the PR components. Twenty-seven oat lines were
evaluated in 1995 using this criteria. Six of them were selected for the
genetic studies and 15 populations developed from crosses among them. The
diallel analysis for pustule size and leaf area affected by crown rust
revealed that additive gene action was most important for both traits.
The genotype UFRGS91906 was identified as one with the best General Combining
Ability for components of PR and its phenotypic data was consistent for
three years. The segregating population from the cross UFRGS7 x UFRGS91906
is being used to construct a molecular map for PR QTL identification.
P - 32
SCAR MARKERS FOR RUST RESISTANCE GENES PC68,
PG3 AND PG9 DESIGNED FOR MARKER ASSISTED SELECTION IN OATS
W. Orr, D. De Koeyer, C. Chenier, N. Tinker
and S. J. Molnar
Eastern Cereal and Oilseed Research Centre,
Agriculture and Agri-Food Canada, Ottawa, Ontario K1A 0C6
Crown rust, caused by Puccinia coronata and
stem rust, caused by P. graminis, are important diseases of oat
(Avena sativa L.). The resistance genes Pc68, Pg3
and Pg9 form a single tightly linked cluster. To facilitate introgression
of these genes into other germplasm, we converted strategic RAPD markers
to PCR based Sequence Characterized Amplified Region (SCAR) markers. One
RAPD marker (UBC195) has been linked to Pg3 in Rodney O-Pg3
and to Pg9 in OT328 x Dumont. The diagnostic 0.4kb fragments were
sequenced. A primer pair designed from the Pg3 sequence consistently
amplifies a band on the susceptible line (Rodney O) but not on the resistant
line (Rodney O-Pg3). The resultant 0.4 kb fragment is digested by
Rsa I to 0.24kb in some lines (example OT966-1 which carries Pc68)
and to 0.2kb in others (example OT974-1 which carries Pc48) such
that homozygotes and heterozygotes of such crosses can be distinguished.
A second primer pair was designed from the sequence of the fragment produced
by UBC195 from Dumont (Pg9). A single 0.4kb band is produced on
Dumont (resistant) but not on OT328 (susceptible). This fragment is also
amplified in lines known to carry Pc68 (OA966-1, AC Francis and
OT274) and in OA974-1 (Pg9?) but not in genotypes with neither gene
(OT328, OA971-2 and OA971-8). These "user-friendly" PCR based SCAR markers
are currently being employed in breeding programs to expedite selection
for rust resistance.
This research was made possible by generous financial
assistance from Quaker Oats and by collaborative dialogue and technical
assistance from a consortium of Quaker funded institutions.
P - 33
DEVELOPMENT OF A GENOMIC MAP OF CULTIVATED
OAT IN TAM-0-301 X OGLE RIL POPULATION
V. Portyanko1, D. Hoffman2,
M. Lee1, and J. Holland1
1Department of Agronomy, Iowa State
University, Ames, IA 50011; 2 USDA-ARS Small Grains Research
Facility, Aberdeen, ID 83210
Complete genetic maps have become an essential component
of progress in crop improvement and basic genetics. The first RFLP map
of the cultivated oat genome (n=21) involved 38 linkage groups and was
based on Kanota x Ogle RIL population (O'Donoughue et al., 1995).
The major objectives of this study are 1) to develop
a cultivated oat RFLP map with the number of linkage groups corresponding
to the basic number of chromosomes; 2) to connect this map to genome maps
of related grass genera; 3) to use this map for experiments to map QTLs
affecting vernalization and photoperiod responses, disease resistance,
and grain yield under diverse conditions.
To date 292 cDNA clones (including a core set of
175 anchor probes being mapped in other cereals) were screened in combinations
with 6 restriction enzymes to identify polymorphisms between mapping parents,
Avena byzantina, cv. TAM-0-301 and Avena sativa, cv. Ogle.
About two thirds (or 215) of a total set of probes screened were
found to be informative in combination with at least one enzyme. Mapping
of these clones inTAM-0-301 x Ogle population of 136 F6:7 RILs
is underway.
A number of important developmental and agronomic
traits were scored in the mapping population to map underlying QTLs/genome
regions. These include heading date and plant height under different vernalization
and photoperiod regimes; heading date, grain yield, and test weight in
the field under different planting dates and environments.
P - 34
OAT PLANTS WITH INTROGRESSED MAIZE CHROMOSOME
SEGMENTS FROM GAMMA IRRADIATION OF PROGENY OF OAT X MAIZE CROSSES
O. Riera-Lizarazu1, M. I. Vales1,
E. V. Ananiev1, H. W. Rines2, and R. L. Phillips1
1University. of Minnesota; 2USDA-ARS,
St. Paul, MN.
Crosses of oat (Avena sativa L., 2n = 6x
= 42) by maize (Zea mays L. 2n = 2x = 20) have resulted in the recovery
of both haploid oat plants (2n = 21) and plants with 21 oat chromosomes
plus one or more retained maize chromosomes. From these, self-fertile disomic
addition lines (2n = 42 + 2) with a full complement of 21 oat chromosome
pairs plus an added maize chromosome pair were recovered for 6 of the 10
maize chromosomes. The phenotypes of these oat-maize chromosome addition
lines ranged from that of near normal oat plants (e.g., maize chromosome
4 additions) to quite marked effects on vigor and plant morphology (e.g.,
maize chromosome 3 addition). The added maize chromosome represents a potential
source of new traits for oat such as disease resistance or seed composition
modification. In addition, for maize genomics analyses, the addition lines
represent a valuable source for mapping or isolating maize DNA sequences
specific to a particular maize chromosome. Irradiation of these whole chromosome
addition lines to induce the transfer of maize chromosome segments into
the oat genome was seen as a way to further reduce the amount and complexity
of maize chromatin present. For this purpose, we treated oat maize chromosome
9 monosomic seed with gamma rays (30, 40, and 50 k rad). Progeny plants
from self-fertilization of plants from irradiated seed were screened for
the presence of maize DNA by Southern analysis using a multiprobe of a
mixture of maize-specific repeated sequences. The maize chromatin present
was characterized by fluorescence genomic in situ hybridization
of root tip cells and by analyzing DNA extractions with a series of 34
maize RFLP and SSR markers specific for chromosome 9. Various maize chromosome
9 rearrangements in oat were identified including translocation introgressions
of maize chromosome segments onto oat chromosomes.
P - 35
INDEPENDENT TESTS AND VALIDATION OF PUTATIVE
-GLUCAN QTLS IN OAT
V. JM. Robert1, R. L. Phillips1,2,
and D. D. Stuthman1
1 Department of Agronomy and Plant
Genetics; 2Plant Molecular Genetics Institute, University of
Minnesota, 411 Borlaug Hall, 1991 Buford Circle, St. Paul, MN 55108, USA
Several studies have demonstrated the hypocholesterolemic
effect associated with the consumption of oat soluble fiber [(1-3), (1-4)--D-Glucans].
This effect can reduce the risk of heart disease in humans. The development
of a genetic map in cultivated oat (Avena byzantina X Avena sativa)
has permitted the study and mapping of quantitative trait loci (QTLs) associated
with inheritance of soluble fiber. This study revealed a complex inheritance
as many QTLs were detected and it appeared that further validation would
be useful prior to initiating a marker assisted selection (MAS) breeding
program for soluble fiber. The objective of this research was to test the
effect of each QTL previously identified, independently of other QTLs.
This research should allow validation and selection of QTLs with large
effects on soluble fiber inheritance.
Recombinant inbred lines (RILs) of the mapping population
with residual heterozygosity at specific soluble fiber QTLs were identified.
Sets of near isogenic lines (NILs) were generated by isolating a series
of selfed progenies from each RIL. This material was grown for two years
and at several locations. Fiber contents were obtained through near infrared
technology and chemical analysis. These NIL sets were segregating for specific
molecular markers in the vicinity of specific QTLs in an otherwise homogeneous
genetic background. Therefore, allelic substitutions through regression
analysis were tested at each independent QTL. Moreover, pleiotropy and/or
genetic linkage with seed or plant characteristics were sometimes detected
as correlated responses were observed in some sets of segregating NILs.
This method allowed validation of the soluble fiber QTLs and provided furhter
understanding of soluble fiber inheritance.
P - 36
GROAT BREAKAGE
Brian G. Rossnagel
Crop Development Centre, University of Saskatchewan,
51 Campus Drive, Saskatoon, SK,
S7N 5A8 Canada
Groat breakage during dehulling, which leads to
increased mill loss and lower mill yield, has been a problem for millers
from time to time for many years. Traditionally it has been associated
with certain environmental conditions around crop harvest, especially frost.
However, when reported as a major problem with Canadian shipments to the
US in 1993 and 1994, since this also coincided with an increase in the
portion of those shipments which would be the variety Derby, we became
suspicious that a genetic component might be involved. As a result a project
was set up to evaluate genetic differences for groat breakage.
Several studies have been undertaken to evaluate
methodology to evaluate groat breakage and to determine if genetic differences
do exist. One study comparing samples from 11 varieties over 14 locations
in 1996 demonstrated that significant differences between varieties exist,
with Derby demonstrating greater and Elvy less breakage than others. Environment
was also a factor. Unfortunately, it has been determined that hand separation
of broken from whole groats is the only accurate method to evaluate this
trait, since sieving to separate broken from whole groats is influenced
by original groat size. One year of data from a 90 genotype SSD population
segregating for groat breakage indicates that the trait is normally distributed
and thus probably under multi-genic control. While there is some association
of breakage with lower fat, greater plumpness and thin hulls, these relationships
are not absolute and with careful monitoring of breeding materials breeders
should be able to select good milling quality oat with minimal groat breakage.
Special project funding from the Saskatchewan
Agriculture Development Fund for this groat breakage evaluation project
is gratefully acknowledged.
P - 37
THE AMMI MODEL FOR GROUPING ENVIRONMENTS
AND OAT GENOTYPES
Jose J. Salmerón
INIFAP-Campo Experimental Sierra de Chihuahua,
Cd.Cuauhtémoc, Chihuahua, Mexico 31500
A study was performed in ten environments under rainfall
conditions during the oat growing seasons of 1992, 1993 and 1994. Ten oat
genotypes (six lines and four varieties) were seeded in the above environments.
The AMMI model was used to obtain homogenous groups of genotypes and locations
and identify stable and high yielding genotypes. Genotypes were arranged
in three categories: One of them with three oat varieties clasiffied as
instable due to their high and negative PCA1 scores Papigochi and Chihuahua
are late, and Páramo is an early cultivar, also this group was susceptible
to stem and crown rust. A second group formed by five lines had a high
and positive PCA1 scores, these were the high yield potential genotypes,
intermediate maturity and resistant to oat rust and a third group constituted
by two genotypes with PCA1 scores of nearly 0 has small interaction effects
and hence classified as stable genotypes, these were a variety Cusi an
early resitantat to one rust and other advanced intermediate maturity and
resistant line. According to grain yield, two groups of environment were
identified: High interaction genotype-environment Tepatitlán, Jal.,
and Pedernales 92, Chih. with high yield potential and Sto. Tomas, Chih
with intermediate yield potential. Another group were locations with low
interaction effects: Campo 1A, Cuauhtémoc, Chih. in 1993 with high
yield potential, Calera and Sombrerete, Zac. with intermediate yield potential.
The low yield potential locations were: Campo 1A(1994) , Campo 38(1993
and 1994), and Pedernales(1994) all within Chihuahua State. The AMMI model
represents another technique to visualize the genotype by environment interaction.
P - 38
DEVELOPMENT OF A "SUN II" OAT ANEUPLOID SERIES
FOR RFLP MARKER ASSOCIATIONS TO CHROMOSOME.
M.A. Start1, H.W. Rines2,
and R.L. Phillips1
1University of Minnesota; 2USDA-ARS,
St. Paul, MN.
The molecular genetic linkage map for cultivated
hexaploid oat (2n=6x=42) constructed from an Avena byzantina cv.
Kanota by A. sativa cv. Ogle cross using principally RFLP markers
consists of at least 532 loci assigned to 38 linkage groups. Additional
mapping efforts and the use of aneuploids have lowered the number of linkage
groups to 33. The objective of this research is to reduce the number of
linkage groups to 21, the expected number for hexaploid oat, by assigning
linkage groups and unlinked markers to physical chromosomes using a nearly
complete Sun II monosomic series. Twenty oat monosomics each missing a
different chromosome have been identified based on C-banding analysis.
Fourteen of the monosomics were newly derived from Sun II oat haploids
generated in oat by maize crosses. Three of the monosomics were derived
by Hacker and Riley in a different selection of Sun II and have a 3C-14
interchange. Backcrosses have been made in an attempt to eliminate the
interchange and to standardize the background of the Sun II monosomics.
Three monosomics have only been recovered in the cultivar Kanota, as described
by Morikawa. Backcrosses with Sun II have been made with these three Kanota
monosomics. Each monosomic line available was crossed with Kanota and Ogle.
Monosomic F1s from these crosses were identified by mitotic chromosome
counts of root tips, and leaf tissue was collected for DNA extraction.
RFLP autoradiograms will be analyzed and missing bands will allow assignment
of sequences to the monosomic chromosomes.
P - 39
CONSTRUCTION OF A GENETIC LINKAGE MAP IN
HEXAPLOID OAT USING AFLP: SEARCHING FOR GENETIC MARKERS FOR PARTIAL RESISTANCE
TO CROWN RUST
G.C.H. Thome1, G.A. Penner2,
A. Zacharias2, G.O. Mellos1, S.C.K. Milach1,
L.C. Federizzi1
1 Federal University of Rio Grande
do Sul - Brazil; 2 Cereal Research Centre, Agriculture and Agri-Food
Canada, Winnipeg
Crown rust (Puccinia coronata f. sp. avenae)
is the most important fungal disease of oat (Avena sativa L) in
southern Brazil. In susceptible cultivars, severe infections prior to the
seed development can result in reduced quality and yield losses as high
as 50%. Since the pathogen has been able to easily overcome the major resistance
genes, more attention is currently being paid to other forms of resistance,
such as partial resistance (PR). Screening for PR to oat crown rust and
the analysis of its components in Brazilian oat genotypes started in the
Oat Breeding Program at the Federal University of Rio Grande do Sul (UFRGS)
in 1995. The oat line UFRGS 910906 was observed to have components of PR
in the field and it has maintained its PR for over 3 years under high rust
inoculum pressure. The number of genes controlling PR in UFRGS 910906 is
not known. This is currently under study at UFRGS. Due to the difficulty
of screening for PR in the field and the effects of environment on its
expression, selection for PR by conventional methods is not always reliable.
The ability to identify individuals with PR at the seedling stage using
molecular markers is desirable since it would increase the speed and reliability
of selection in breeding programs. This poster presents preliminary results
of a study under way to develop molecular markers for PR to crown rust
using Amplified Fragment Length Polymorphism (AFLP) analysis in a cross
between UFRGS 910906 and the susceptible cultivar UFRGS 7. In a field trial
of the F2 generation (200 individuals), continuous variation of PR to crown
rust was observed. Different primer combinations are being tested on this
F2 population in order to develop a genetic map providing markers linked
to QTLs or genes that control PR. To date, 20 different primer combinations
have been tested yielding a total of 150 polymorphisms segregating across
the population.
P - 40
HOMEOLOGOUS OR DUPLICATED GENOMIC REGIONS
AFFECTING GROAT OIL CONCENTRATION
Nick Tinker, David De Koeyer, Fran Webster,
George Fedak, Steve Molnar
Eastern Cereal and Oilseed Research Centre,
Agriculture and Agri-Food Canada, Ottawa ON, Canada K1A 0C6
Oil concentration of oat groats has important feed
and nutritional implications. Low oil is considered desirable in most food
products, while high oil contributes to greater feed energy. We used a
genetic linkage map to identify quantitative trait loci (QTLs) in a naked-by-covered
population of cultivated oat (see companion poster). Here, we report and
discuss a more detailed analysis of QTLs affecting groat oil concentration.
Based on simple interval mapping (LOD=3), three genomic regions were associated
with oil concentration. A fourth region, which showed marker homology to
the other three QTL regions, was significant based on single factor analysis
(P<0.01). The allelic effects at all four regions were approximately
equal, with each pair of alleles showing substitution effects of up to
0.5% oil. In two regions, alleles from the covered parent gave increased
oil, while in the other two regions, alleles from the naked parent gave
increased oil. These four QTL effects were primarily additive (no significant
epistasis), accounting for more than 50% the variation in oil concentration,
and explaining much of the transgressive segregation seen in this population.
Based on commonality of cDNA sequences mapped in these regions, it is possible
that all four QTLs result from genetic variation at a homeologous or duplicated
locus (i.e. a common point in a biochemical pathway). The above hypothesis
is supported by the additivity of allelic effects shown by these QTL: variation
at different points in a biochemical pathway would be more likely to have
produced epistatic effects.
This research was made possible by generous financial
support from the Quaker Oats Company, and by collaborative dialogue and
technical assistance from a consortium of Quaker funded institutions.
P - 41
AN RFLP / RAPD / AFLP LINKAGE MAP OF CULTIVATED
OAT FROM A HULLESS-BY-COVERED CROSS
Nick Tinker, Charlene Wight, Louise O'Donoughue,
George Fedak, Ken Armstrong, Vern Burrows, Art McElroy, David De Koeyer,
Corinne Chenier, Anissa Lazreg-Lybaert, Shan He, Steve Molnar
Eastern Cereal and Oilseed Research Centre,
Agriculture and Agri-Food Canada, Ottawa, ON, Canada K1A 0C6
Molecular linkage maps provide important tools for
gene discovery and genomic research. In oat, previously reported RFLP linkage
maps have been developed in two diploid populations and in the interspecific
hexaploid population Kanota (Avena byzantina) x Ogle (A. sativa)
("K/O"). Here, we report a linkage map of RFLP, RAPD, and AFLP markers
developed in an intraspecific (A. sativa) hexaploid oat population.
The mapping population consisted of 102 recombinant-inbred lines derived
by single-seed descent (to the F6) from a hulless-by-covered cross ("T/M").
Approximately 170 RFLP markers, 24 RAPD markers, and 250 AFLP markers were
mapped to 35 linkage groups (LOD 6). Genome coverage in the 35 linkage
groups is approximately 750 cM. An additional 45 unlinked markers probably
account for a large portion of the remaining genome. The AFLP markers appear
to be randomly distributed on most linkage groups, with five linkage groups
containing exclusively AFLPs. Most of the RFLP markers were based on cDNA
clones mapped in K/O. Since most clones map to multiple homeologous loci,
definitive locus identity between two maps requires an allele that is common
to both populations. Thus, it was possible to confirm the identity of only
46 RFLP loci common to both the T/M and the K/O map. These loci were used
as anchors to align linkage groups in T/M with the K/O map. Other loci
were presumed to be allelic when the same clone was mapped within a linkage
group containing a confirmed allelic relationship. Homology to the K/O
map was determined for 19 of the T/M linkage groups and several of the
unlinked T/M markers. As with the K/O map, it has not been possible to
reconcile all linkage groups to 21 oat chromosomes. Future comparisons
among these and other oat maps may eventually make this possible. Presently,
the T/M map provides a useful tool for the discovery of QTL, and for comparative
genome mapping.
P - 42
TRANSFORMATION OF ELITE OAT CULTIVARS
K.A. Torbert1, H.W. Rines2,
H.F. Kaeppler3, G.K. Menon3,
and D.A. Somers1
1Department of Agronomy
and Plant Genetics, University of Minnesota, 411 Borlaug Hall, 1991Upper
Buford Circle, St. Paul, MN 55108; 2Plant Science Research Unit,
USDA-Agricultural Research Service, St. Paul, MN 55108; 3Department
of Agronomy, University of Wisconsin, Madison, WI 53705
Mature embryos of a specific oat (Avena
sativa L.) have shown to be a reliable explant for initiating tissue
cultures that can be genetically engineered to produce transgenic, fertile
plants (Torbert et al. 1998). The purpose of this research was to investigate
mature embryos from 16 elite North American spring oat cultivars for tissue
culture initiation and the capability to produce transgenic plants from
elite cultivars. Fifteen of the genotypes tested yielded at least some
embryogenic callus that was similar in appearance to transformable callus
based on our previous work. One elite cultivar, 'Belle' was utilized in
determining transformation efficiency. From 30 microprojectile bombardments,
17 independent genetically engineered tissue cultures produced transgenic,
fertile plants. DNA sequences designed to influence a variety of traits
have been introduced into Belle. These traits include aluminum tolerance,
antisense for starch composition modification, and wheat high molecular
weight glutenin in oat kernels to investigate the dough making properties
of oats by these proteins. These results demonstrate that mature embryo-derived
tissue cultures will be useful for transforming elite oat cultivars.
P - 43
BREEDING FOR LEAF RUST RESISTANCE
IN OATS
L. Song and Weining Song
Leslie Research Centre, Farming Systems
Institute, Department of Primary Industries, Toowoomba, Qld 4350, Australia
The constant emergence of new and virulent
races of leaf rust has long been recognised as a threat to the grazing
oat industry in Australia, where over 400,000 hectares are grown annually
between southern Queensland and northern New South Wales. The breeding
program addresses this problem by pyramiding rust resistance genes with
the aid of molecular markers. Almost all the rust resistance genes were
introduced from programs overseas. These genes have been identified and
backcrossed to adapted cultivars. In this process, recombinants have been
selected for other agronomic characters which include foliage yield, growth
habit and recovery after cutting. This breeding program also looks into
other sources of rust resistances, including slow rusting. Good collaboration
has been established with other oat improvement programs in Australia as
well as programs overseas. Some of the more advanced materials have reached
the F7 or F8 stage and are now ready for commercialisation.
P - 44
BETA-GLUCAN CONTENT IN ELITE OAT
GERMPLASM AND BREEDING POPULATIONS
D.M. Wesenberg1, D.M.
Peterson2, L. Herrin2, and D.E. Burrup1
1National Small Grains
Germplasm Research Facility, USDA, ARS, 1691 S. 2700 W., P.O. Box 307,
Aberdeen, Idaho 83210, USA; 2Cereal Crops Research Unit, USDA,
ARS, 501 Walnut St., Madison, Wisconsin 53705, USA.
Beta-glucan content determinations are
routinely completed along with evaluations of other biochemical and agronomic
traits for elite oat germplasm grown in irrigated and dryland trials in
southern Idaho. In irrigated trials grown at Aberdeen, ID in 1994-97, beta-glucan
content for 26 oat cultivars ranged from 3.74% ('Derby') to 6.22% (92Ab300).
The beta-glucan content for the same entries grown under irrigation at
Tetonia, ID in 1994-97 ranged from 3.91% ('Newdak') to 5.91% (92Ab300).
In dryland trials involving the same entries at Tetonia during the same
time period beta-glucan content ranged from 4.38 % (Derby) to 5.83% (92Ab300).
Data from these trials indicate that environment influences beta-glucan
content to a modest degree, with beta-glucan content averaging 4.73% at
Aberdeen (Aberdeen), 4.88% at Tetonia (irrigated), and 5.11% at Tetonia
(dryland). The relative performance of genotypes was similar over locations
and apparently influenced little by environment. No association of beta-glucan
content with other traits evaluated was apparent in these trials. In a
study involving F3 and F4 bulk selections from a
cross of NPB 871754/'Monida' grown in hills at Aberdeen in 1996 and 1997
and Tetonia in 1997, beta-glucan content of 164 F3 selections ranged from
4.74% to 7.32% at Aberdeen in 1996 with the parents averaging 6.91% and
5.49%, respectively. Monida averaged 189% of NPB 871754 in yield at Aberdeen
in 1996, but despite the yield contrast protein content was similar for
the parental lines, averaging 19.1% for NPB 871754 and 18.3% for Monida.
The relationship of beta-glucan to protein content and yield in these early
generation studies indicated that selection for high or low beta-glucan
content should not be significantly impeded by protein or yield considerations.
Correlations between beta-glucan and protein content were positive. Correlations
between beta-glucan content and yield were negative, but relatively low
ranging from -0.15 to -0.40 in the three environments. Standard unit heritability
estimates for beta-glucan content based on F3 populations grown
at Aberdeen and F4 populations grown at Aberdeen and Tetonia
were less than 0.40.