AWN Vol 43: awn43b8.html Septoria tritici leaf blotch-resistant germplasm derived from bread wheat/D genome synthetic hexaploids.

L.I. Gilchrist and A. Mujeeb-Kazi.

F1 hybrids from T. turgidum/T. tauschii crosses were sources of synthetic hexaploids. Several hundred such SHs have been produced forming a bridging germplasm base for bread wheat improvement. Screening for S. tritici in Toluca, Mexico, led to identification of resistant SH combinations that were crossed onto susceptible bread wheat cultivars. Advanced, resistant derivatives were evaluated over three cycles using a mixture of five isolates. Disease severity was recorded in each cycle using a modified double digit 1.1 to 9.9 resistant:susceptible scale during the watery, milky, and doughy grain-filling stages. The diversity observed in the SH/bread wheat advanced lines was influenced significantly by 1,000-kernel weight. These resistant lines can be readily utilized in breeding programs (Tables 16, p. 162 and 17, p. 163), and are superior than the resistant conventional bread wheat germplasm.

Table 16. Pedigrees of ten Septoria leaf blotch-resistant spring bread wheat cultivars/T. tauschii derivatives.

Germplasm Cross and selection number 3
Seri1//Croc1/T. tauschii (224)2 CIGM90.358-1Y-1M-2Y-0B-0Y-1M-2M-0Y
Altar84/T. tauschii (224)//2*Cupe1 CIGM91.191-5B-3Y-0B-0Y-1M-2M-0Y
Yaco//Croc1/T. tauschii (205)/3/Yaco1 CIGM91.153-1B-2Y-0B-0Y-2M-0Y
Pgo//Croc1/T. tauschii (224)/3/2*Weaver CIGM91.248-2Y-0B-1Y-2M-7M-0Y
Altar 84/T. tauschii (191)//Yaco3/3*Bau1 CIGM92.337-0B-0Y-1M-2M-0Y
Altar 84/T. tauschii (191)//Opata1 CIGM90.483-4Y-1B-2Y-0B-1M-3M-0Y
Croc1/T.t auschii (205)//Kauz1 CIGM90.248-1Y-2B-11Y-0B-1M-1M-0Y
Croc1/T.t auschii (205)//Weaver1 CIGM90.250-4Y-3B-4Y-0B-1M-4M-0Y
Croc1/T.t auschii (205)//Weaver CIGM90.250-4Y-3B-4Y-0B-1M-3M-0Y
Croc1/T. tauschii (213)/Pgo1 CIGM90.412-5Y-3B-6Y-0B-2M-1M-0Y

1 bread wheat cultivars
2 synthetic hexaploid with Ae. tauschii CIMMYT accession number in parenthesis
3 B = Batan, M = Toluca, Y = Yaqui Valley

Conclusions. S. tritici-resistant SH wheats upon crossing with elite but susceptible bread wheats have yielded advanced derivatives resistant to the disease. Ten lines with desirable agronomic characteristics studied over 3 years, have maintained their resistances with scores between 1.1 to 2.1. Resistant bread wheats score 3.1 to 4.1, susceptible bread wheats 8.7 to 8.9, and all durum cultivars score between 8.7 to 9.9. Direct crosses between T. tauschii accessions contributing resistance to S. tritici and susceptible bread wheat cultivars are in progress. Monosomic analyses only are concentrated on the D genome, because T. tauschii is the donor source. DH protocols will be merged with the conventional monosomic analyses.

Amphiploids of Triticum turgidum with some A, B, and D genome Triticeae accessions resistant to Septoria tritici leaf blotch.

R. Delgado, L.I. Gilchrist, and A. Mujeeb-Kazi.

A-, B-, and D-genome Triticeae species in the primary and secondary gene pools offer a potent avenue that facilitates pyramiding of diverse stress resistances for wheat improvement. We have produced a wide array of genetic stocks using the above

genomic accessions. These amphiploids are hexaploids of AA AABB, AABB BB, and AABB DD genomic compositions. They were derived from crosses of several A-, B-, or D-genome diploid accessions with durum wheat cultivars. Screening these germplasms in Toluca, Mexico, for S. tritici for 3 years has led to the identification of several resistant entries. The diagnostic parameter was disease damage during three grain-filling stages. Amphiploid germplasm provides a unique diversity for improving both bread and durum wheat cultivars through bridge crosses as one utilization option. Some descriptors and disease scores of the above germplasms are presented (Table 18, p. 164). These amphiploids will enable breeders to exploit the diversity for durum and bread wheat improvement by amphiploid bridging crosses, direct diploid single-genome crosses, or tetraploid combinations involving accessions of two resistant genomes exemplified by resistant AADD tetraploids.

Table 17. Agronomic characteristics and disease reaction of Mycosphaerella graminis resistant spring bread wheat germplasm grown in Atizapan, Toluca, Mexico state, over three crop cycles.

GermplasmDays to
anthesis
Days to
physiological
maturity
Plant
height
(cms)
1000-kernal
weight
(gr)
Disease damage1
WSMSDS
1.CIGM90.35883132100391.11.12.1
2.CIGM91.19183138100381.12.12.1
3.CIGM91.1538313295371.11.11.1
4.CIGM92.24888142100331.11.12.1
5.CIGM92.3378313895351.11.12.1
6.CIGM90.4838013290251.12.12.1
7.CIGM90.2488314290291.11.12.1
8.CIGM90.2508313885411.11.11.1
9.CIGM90.25083138885411.11.11.1
10.CIGM90.41283138100391.11.12.1
11.Bobwhite CM33203
Resistant check
8313890311.11.14.1
12.Kauz CM67458
Susceptible check
8313585216.47.48.7
13.Seri 82 CM33027
Susceptible check
8314090212.18.48.9
1WS=watery stage of grainfill, MS=milky stage, and DS=dough stage

Table 18. Cross combination details of some AA AABB, AABB BB, AABB DD, AADD amphiplids resistant to Septoria tritici with a few descriptors.

Amphiplid combination
Durum cultivar/Diploid species
Cross
No.
Descriptors1S. tritici score2
FlowHt
(cms)
P.matWSMSDS
Sca/T. boeoticum (23)390.674881301341-11-11-1
Scoop/T. boeoticum (50)90.7001001351271-11-11-1
Scoop/T. boeoticum (87)90.7071001301271-11-11-1
Aos/T. monococcum (98)90.7911101051501-11-11-1
Aos/T. monococcum (118)90.7941151101441-11-11-1
CPI/-----44/T. boeoticum (93)93.1291001351381-11-11-1
Stn/T. monococcum (111)93.1341051151271-11-11-1
Stn/T. monococcum (112)93.1361001101341-11-11-1
CPI/-----44/Ae. speltiodes (129)94.37S1151001401-11-11-1
Ceta/Ae. speltoides (135)94.45S1201151451-11-11-1
Ceta/Ae. speltoides (140)94.254S1201151451-11-11-1
Arlin/Ae. speltoides (141)94.255S1201051501-11-11-1
Altar/Ae. squarrosa (224)86.9421101001351-11-11-1
Yar/Ae. squarrosa (518)90.8461051001401-11-11-1
Yar/Ae. squarrosa (493)89.4631051001351-11-11-1
Altar/Ae. squarrosa (502)93.3951001051301-11-11-1
Sca/Ae. squarrosa (409)93.2371051001401-11-11-1
Croc/Ae. squarrosa (879)89.4791001101351-11-11-1
Ae. squarrosa (1027)/T. boeoticum (42)94.32S1101351501-11-11-1
BW Susceptible Seri 82CM3302783901402-18-48-9
BW Resistant BobwhiteCM3320383901381-11-14-1
1Flow=days to flowering. Ht=plant height at maturity, and P.Mat=days to physiological maturity. 2WS=waitery stage of grainfill, MS=milky stage, and DS=dough stage. 3=CIMMYT wide cross program accession number. 4=CPI/Gediz/3/Goo//Jo69/Cra/

Management of an abiotic stress in wheat - salinity - through alien Triticeae germplasm, cytogenetic manipulations, and screening methodology development.

J.L. Diaz-de-Leon and A. Mujeeb-Kazi.

Abiotic stresses are static mechanisms that tend to be more durable, because of the absence of pathogen influence. Of these abiotic stresses, salinity tolerance in wheat poses an almost unsurmountable challenge. Conventional diversity is limited and significant practical outputs are few. We present here our research strategy that encompasses screening under two controlled conditions, and a field screening protocol using sea water dilution as the irrigation source. The dilutions equate variable electrical conductivity levels. Complementing this screening procedure is novel germplasm development by exploitation of the annual and perennial Triticeae species of the primary and tertiary gene pools.

Some salinity screening methodologies. In general the methodologies are separated into field evaluations and in vitro testing. Both procedures provide information on genetic diversity among the germplasms. We present here procedures that are being used in our experimentation.

Field Screening using sea water dilutions. Germplasm screening for salinity response is conducted under field conditions in La Paz, Baja California Sur, Mexico. Normal well-water for irrigation with an electrical conductivity (EC) level of 4.5 ds/m serves as the control concentration. Sea water, in close proximity to our field screening site, is kept in 1,200 liter containers. Mixing sea water with the normal well-water provides the necessary EC levels selected for the field evaluation. These levels have been kept at 8.0, 12.0, 16.0, and 20.0 ds/m and can be extended further. Plots are separated from each other on all sides by black plastic line dividers. The plots are individually flood-irrigated with 200 liters twice a week according to the treatment category. Electrical conductivity of the irrigation water is measured, and soil samples randomly taken from each plot after 24 hours. Determination of the soil EC follows the established extraction procedure with steps of soil sampling/lot at random points. After 1 week of plant growth, all plots are fertilized with 15 g of urea per week up to 8 weeks. The urea applications are made weekly with each irrigation and are discontinued after 8 weeks.

In vitro screening. These protocols utilize the hydroponics system (Gorham 1990) where NaCl levels from 50 mol/m3 up to 200 mol/m3 have been used as discrimination levels. Growth parameter assessments are made, and the procedure is strengthened by K:Na discrimination analyses.

In addition, we have used two other strategies where growth analyses are made by 15 days from the start of the experimentation, allowing a first selective cut of the germplasm, and using these initial selections for the field sea water dilution study. These strategies utilize (i) a saline agar media based upon the standard Murashige-Skoog culture medium with variable EC levels that simulate the field sea water dilution ranges and (ii) the same sea water dilutions for doing a petri dish germination/growth study, complemented by Hoagland's nutrition solution as a variable if necessary.

Genetic diversity of the salinity status in the Triticeae.

The conventional germplasm. Triticum aestivum cultivar Kharchia 65, a salt-tolerant cultivar, has been exploited by growers in Rajasthan, India, for several decades. Controlled testing in saline media unequivocally attests to the cultivar's salt tolerance potential (Mujeeb-Kazi et al. 1993). Attempts to incorporate the cultivar's salinity tolerance genetic diversity for wheat improvement have been made, and the singular success is accredited to the KRL 1-4 wheat cultivar release. Another salt-tolerant T. aestivum cultivar selected from Lu 26 is Lu 26S. Additional inputs to this conventional gene pool of Kharchia 65, KRL 1-4, and Lu 26S are some collections of promising cultivars and land races acquired from various programs and countries. These are Chinese Spring, SNH-9, WH-157, Sakha 8, Shorawaki, Candeal, and Pasban-90. Susceptible cultivars in the above tester group of diverse germplasms were a durum wheat, PBW34, and the bread wheat cultivar Oasis. More susceptible wheats are needed. We have developed this tester set in order to establish germplasm evaluation uniformity among researchers. Undoubtedly, additional entries will be added to the current set, and a revised list of germplasm formulated.

Our current status and projections for incorporating salinity tolerance through alien genetic diversity.

Interspecific hybridization or short-term product output strategy is focused on use of Ae. tauschii using the bridge cross route that exploits the various synthetic hexaploids. We have produced several hundred synthetics.

Through intergeneric hybridization, we are attempting to exploit the salt tolerance potential of two diploids (Th. bessarabicum and Th. elongatum). Thinopyrum bessarabicum focuses on direct exploitation through its 56-chromosome amphiploid for bread wheat improvement. The amphiploid form of both these diploids (Wang 1986) is also being used for durum and bread wheat and involves the Ph manipulation strategy. This manipulation avenue is an integral part both for bread wheat and durum wheat improvement and F1 hybrids have been produced. These hybrids were cytologically validated, and the ph role was elucidated for the alien amphiploid combination with Chinese Spring ph. The meiotic associations in the durum/diploid alien amphiploid also document the hybrid status and elucidate the genomic associations. Use of the Cappelli durum ph is a route that also has been used for alien introgressions. In bread wheat, the significance is exemplified in F1 hybrids with Ph showing less pairing as compared to the ph based high pairing.

The novel route of PhI incorporation introduced recently in this program holds substantial merit (Chen et al. 1994), and will be an option to pursue. In addition, the use of the molecular probes diagnostic for the Ph locus (Gill and Gill 1996) is seen as an efficient means to exploit the several Ph-based perennial F1 hybrids by creating Phph BC1 derivatives and exploiting only the ph-derived haploids, as described by Mujeeb-Kazi et al. (1993).

References.

Chen PD, Tsujimoto H, and Gill BS. 1994. Transfer of PhI genes promoting homoeologous pairing from Triticum speltoides to common wheat. Theor Appl Genet 88:97-101.

Gill KS and Gill BS. 1996. A PCR-based screening assay of Ph1, the chromosome pairing regulator gene of wheat. Crop Sci 36:719-722.

Gorham J. 1990. Salt tolerance in the Triticeae: ion discrimination in synthetic hexaploid wheats. J Exp Bot 41:623-627.

Mujeeb-Kazi A, Gorham J, and Lopez-Cesati J. 1993. Use of wild Triticeae relatives for stress tolerance. In: Inter Crop Science I (Buxton DR, Shibles R, Forsberg RA, Blad BL, Asay KH, Paulsen GM, and Wilson RF eds). Ames, IA. p. 549-554.

Wang R-C. 1986. Amphiploids of the diploid hybrid Thinopyrum bessarabicum x T. elongatum. Agron Abstr p. 86.

Karnal bunt screening of some D-genome synthetic hexaploids and germplasm derived from their crosses to bread wheat.

G. Fuentes-Davila and A. Mujeeb-Kazi.

D-genome synthetic hexaploid wheats derived from T. turgidum/T. tauschii accession combinations were evaluated for Karnal bunt resistance under field and greenhouse conditions in Mexico over three test cycles. Of 266 SH wheats tested, several demonstrate immunity (0 %) to KB. Four SH wheats have been registered. For breeding applications, crosses between KB-immune SH wheats and KB-susceptible bread wheats gave elite agronomic plant type segregants. These lines were inoculated and are further sources of elite KB-resistant germplasm. Infection of susceptible bread wheat cultivars ranged from 20.5 to 100 percent. The standard susceptible bread wheat cultivar WL711 was the check. Several `SH/bread wheat' derivatives were immune.

The interspecific crossing program at CIMMYT has produced 570 synthetic hexaploids (T. turgidum/T. tauschii accessions) over a span of 5 years. Those SHs produced during the first 3 years were seed increased and formed the KB-screening germplasm reported here.

STUDY 1. Screening of T. turgidum/T. tauschii SH wheats.

Field Screening. From the first SH wheats produced during 1989-91, 266 were tested for KB resistance. Several lines with 0 % infection were identified. Susceptibility also was present in this material, and infection levels up to 35 % were observed. The durum parents were either immune or showed up to 2.8 % infection. The bread wheat cultivar WL711 had a mean infection value of 65 %. We have registered four SH types from this immune group. The pedigrees and characteristics of these four SH wheats have been described (Villareal et al. 1996).

The SH germplasms gave 0 % infection to KB in all tests over three cycles of evaluation, compared with a 65 % mean infection of WL711, the susceptible bread wheat check cultivar. The durum wheats in the pedigrees of the four SH wheats had levels of infection from 0.3-0.84 %. Several other SH wheats also had an immune KB response with a large number exhibiting less than 3 % infection. The SH entries have acceptable agronomic characteristics that can significantly contribute to bread wheat improvement.

Greenhouse screening. The KB-screening data of the SH and durum parental germplasm under field and greenhouse regimes is in Table 19. From the field-resistant SH entries, a selected few showing a similar response under greenhouse controlled conditions are included. The durum parents in these SH lines were immune under field tests, but a differential response existed under the stringent greenhouse testing. Durum infections ranged from 2.5 to 64.2 %. Their corresponding SH amphiploids with immunity strongly endorse the contribution made by the T. tauschii accessions to these genetic stocks (Table 19).

STUDY 2. Screening of susceptible bread wheat/resistant SH wheat advanced derivatives.

Extending the utility of KB-immune SH lines, susceptible bread wheat cultivars were crossed by several SH wheats. Advanced `bread wheat/SH' F6 derivatives from these crosses were field tested for KB resistance. All F1 combinations had the base bread wheat cultivar susceptibility and SH immunity established earlier in Study 1 through field and greenhouse evaluations. These 750 advanced `bread wheat/SH' derivatives tested were selected earlier for desirable plant type and acceptable agronomic characteristics. Karnal bunt infections of the 750 F6 entries ranged from 0-51.9 % over two inoculation dates. There were 71 derivatives with immunity (0 %) on both inoculation dates, 80 had less than 1 % infection, and 66 lines had infections between 1.1 and 2.0 %. Though derivatives of all three categories will be tested further, only 12 derivatives are indicated in Table 20 (p. 169). These selections have desirable phenology traits and combine not only the resistance of the T. tauschii accessions, but also possess the genetic diversity of the A and B genomes of the respective durum wheat cultivars. Apart from Weaver, all bread wheats have a field KB susceptibility range between 7.9 and 11.98 %. These cultivars under greenhouse conditions range from 8.8 (Weaver) to 72.4 %.

Reference.

Villareal RL, Mujeeb-Kazi A, Fuentes-Davila G, and Rajaram S. 1996. Registration of four synthetic hexaploid wheat germplasm lines derived from Triticum turgidum x T. tauschii crosses and resistant to Karnal bunt. Crop Sci 36:218

Table 19. Karnal bunt screening of durum parents and their synthetic hexaploids under field and greenhouse conditions.

Entry
number
Parent and cross combinationPercent Karnal bunt infection
Field
inoculation
Greenhouse
inoculation
4Altar 107.4
52Altar/T. tauschii (198) 200
55Altar/T. tauschii (205)00
62Altar/T. tauschii (211)00
77Altar/T. tauschii (219)00
111Altar/T. tauschii (315)00
119Altar/T. tauschii (328)00
1Chen 105.8
162Chen/ T. tauschii (662)00
5Duergand 10.438.2
80Duergand/Ae. tauschii (221)00
98Duergand/Ae. tauschii (247)00
11CPI/Gediz"S"/3/Goo"S"//Jo"S"/CR"S"054.5
61CPI/Gediz"S"/3/T. tauschii(Aus 18912)00
162CPI/Gediz"S"/3/T. tauschii(Aus 18912)00
31Yar "S"1064.2
145Yar "S"/T.tauschii (493)00
174Yar "S"/T.tauschii (809)00
3268112/Ward021.8
12868112/Ward/T. tauschii (369)00

1Durum cultivar
2Number in parenthesis is the accession number in wide crosses program working collection.

Table 20. Advanced F6 derivatives from 'Karnal bunt-susceptible bread wheat/resistant synthetic hexploid' crosses screened under Obregon, Sonora, field conditions by the boot inoculation of ten spikes per entry, except for WL711 (the susceptible check) where 30 spikes were inoculated.

F6CombinationFloret
number
Seeds
infected
Percent
infection
Bread wheat Infection
greenhouse1field
1Esda//Altar/T.tauschii/3/Esda56120.3572.411.98
2Croc 1/T.tauschii//Pgo3010020.59.71
3Croc 1/T.tauschii//Pgo43100--
4Pgo//Croc 1/T.tauschii41300--
5Croc 1/T.tauschii//Kauz7420051.69.83
6Croc 1/T.tauschii//Weaver592008.81.1
7Croc 1/T.tauschii//Fct6430055.210.8
8Croc 1/T.tauschii//Kauz52600--
9Croc 1/T.tauschii//Kauz52600--
10Croc 1/T.tauschii//2*Weaver43700--
11T.tauschii(368)/Cno//2*Cno77450.6429.77.9
12T.tauschii(222)/Cno//2*Cno81900--
13WL7111,37342431.365.42 to 100.0

1Greenhouse data from the greenhouse screening of bread wheat cultivars in Study 1. Table 21. Some synthetic hexaploids for Triticum turgidum x T.Tauschii, 2n=6x=42; evaluated for Cochliobolus sativus in Poza Rica, Mexico.

T. turgidum cultivar/T. tauschii accession (SH) combination
and its CIGM cross number
Disease score
Leaves1Seed2Spike3
(1-9)
ab
Doy 1/T. tauschii (188) CIGM88.1175939423
Rabi//GS/CRA/2/T. tauschii (190)CIGM88.1178939423
Rok/Kmli//T. taschii (214)CIGM88.1335939423
Yuk/T. tauschii (217) CIGM90.561939422
Arlin_1//T. tauschii (225) CIGM88.1228989948
Gan/T. tauschii (236)CIGM88.1228939426
Mexi/Vic//Yav79/3/T. tauschii (434)CIGM88.1335929222
Doy 1/T. tauschii (447) CIGM88.1344929323
Doy 1/T. tauschii (510) CIGM88.1360939534
Doy 1/T. tauschii (511) CIGM88.1363939434
Doy 1/T. tauschii (515) CIGM90.566939434
Yar/T. tauschii (524)CIGM89.474949536
68.111//RGB-U/Ward/3/FGO/4/Rabi/5/T. tauschii (629)CIGM90.590929323
Ceta/T. tauschii (850) CIGM89.552939424
Ceta/T. tauschii (872)CIGM89.555939434
68.111//RGB-U/Ward/3/FGO/4/Rabi/5/T. tauschii (878)CIGM89.559929323
Rabi//GS/CRA/3/T. tauschii (895)CIGM90.603939433
Rabi//GS/CRA/3/T. tauschii (895)CIGM90.605929323
Croc 1/T. tauschii (518)CIGM86.944939434
PBW114/T. tauschii (Introduction from India)939432
Ceta/T. tauschii (895) CIGM89.56792922
Arlin_1//T. tauschii (308)CIGM90.811979848
LCK59.61/T. tauschii (344)CIGM90.832979944
Rabi//GS/CRA/3/T. tauschii (457)CIGM90.832929422
Ciano 79 (Susceptible Bread Wheat)979959
BH 1146 (Resistant Bread Wheat)959736
1Two-digit scoring system: first digit=height of infection;i.e, five=up to center of plant, 9=up to the flag leaf; second digit=disease severity on infected leaves, 1=low and 9=total leaf destroyed. a=score at early milk stage, b=score at dough stage.
2Grain infection scored as: 1=low and 5=high seed blemish at embryo points.
3Spike infection scored as: 1=low infection and 9=high infection.

Helminthosporium sativum evalutions of D-genome dexaploids and germplasm derived from their crosses to bread wheat.

R. Delgado and A. Mujeeb-Kazi.

Spot or leaf blotch caused by C. sativus (syn. H. sativum) limits wheat production in many nontraditional hot, humid, wheat-producing areas. Breeding for resistance is a high priority that is hampered by scarcity of adequate resistance. Intergeneric hybrid derivatives involving Th. curvifolium currently are the most promising and widely utilized. Additional superior resistant sources reside in T. tauschii accessions. This inference is based upon screening data of their SHs. The advanced progenies of T. aestivum/resistant SH wheats (T. turgidum/T. tauschii) have enabled selections with high resistant levels to C. sativus. These lines are agronomically acceptable and have satisfactory quality. The quality diversity of the selected resistant lines is equal to, or better than, the wheat parents involved in the respective crosses.

Study 1. Intergeneric sources of resistance. Five lines from our intergeneric hybridization program were identified and characterized as resistant to C. sativus and possessed satisfactory agronomic attributes. The pedigree of the intergeneric cross was `Chinese Spring/Th. curvifolium//Glen/3/Alondra/Pavon'. These five lines have been registered and described in Mujeeb-Kazi et al. (1996). Two of the lines are called Chirya and Mayoor, and all germplasms have been widely distributed. These lines are being utilized for wheat improvement in the breeding programs of CIMMYT, by several national programs, and other collaborators.

Study 2. Interspecific source of resistance. The diversity of resistance in the various SH wheats was observed from screening of several synthetic wheats. Some results highlighting a few SH wheats are provided in Table 21. The durum cultivars involved in these SH combinations were generally susceptible for the leaf infection and seed blemish parameters. These scores were 9-7 to 9-9 for leaf blotch, 8-9 for spike damage, and 4-5 for seed appearance. (see Table 21, p. 170, for scoring scales). A SH wheat is interpreted to be resistant based on the reaction of the respective T. tauschii accession parent.

Elite SH types have been identified for wheat improvement based upon agronomic evaluations (Villareal et al. 1995) and C. sativus disease-screening data (Table 21, p. 170). Synthetic hexaploid wheats with leaf scores of 9-5 or less and seed damage of 3 or less are the preferred resistant donors for wheat improvement.

Study 3. Bread wheat/synthetic hexaploid advanced derivatives. The SH bridge in breeding is advantageous for wheat improvement, because it allows not only the T. tauschii resistances to be exploited, but also incorporates the genetic diversity of the A and B genomes of the respective durum wheat cultivars present in the SH wheats. Using this approach, we have developed `bread wheat/SH' germplasms from which 30 highly-resistant C. sativus lines were identified. The resistance of this germplasm is similar to that observed for the interspecific material described in Study 2.

Reference.

Mujeeb-Kazi A, Villareal RL, Gilchrist LI, and Rajaram S. 1996. Registration of five Helminthosporium leaf blight resistant wheat germ plasm lines. Crop Sci 36:216-217.

Some applications of wheat/maize-induced polyhaploidy.

A. Mujeeb-Kazi and M. Inagaki.

Crosses between wheat and maize are an effective means of producing wheat polyhaploids. The haploid generation is independent of the maize pollen source and of recipient wheat cultivars, though some pollen source preference does exist. Applications of the haploid procedure are in breeding programs, developing mapping populations, producing cytogenetic stocks, and as an offshoot assisting wide crossing programs. Techniques that complement satisfactory hybridization product frequencies strongly favor the use of detached spikes, sucrose as a nutrient source, and sulfurous acid for overcoming contamination. A post-pollination 2,4-dichlorophenoxy acetic acid treatment remains crucial. Use of stored maize pollen gives positive results with a low frequency. Improved frequencies result by using stored pearl millet pollen. The technique gives a variable response with durum wheats and X TriticoSecale.

Diverse species crosses with maize and Tripsacum pollen. T. aestivum, T. turgidum, and synthetic hexaploid / maize or tripsacum. We obtained polyhaploid embryo recovery frequencies for 16 hexaploid wheats, 5 tetraploid wheats, and 15 synthetic hexaploids, averaging 15.6, 16.9, and 19.8 %, respectively. Mean plant regeneration frequencies for bread wheats was 68.5 %, durum wheats were 73.9 %, and the synthetic hexaploids were 74.5 %. When Tripsacum was the pollen source, regeneration frequencies were 78.5, 66.7, and 77.5 % for bread, durum, and synthetic wheats, respectively. Successful chromosome doubling with colchicine averaged 64 % for T. aestivum cultivars, 69.5 % for T. turgidum cultivars, and 63.6 % for the synthetic hexaploids. The use of Tripsacum allows for a prolonged crossing cycle under our conditions in Mexico if both maize and Tripsacum are used as pollen donors.

Production of alien chromosome disomic addition lines. In wheat wide crosses, polyhaploidy can be further exploited for the production of alien chromosome addition lines from populations that have varying chromosome numbers. Preferably, plants with 22 chromosomes (21 chromosomes of wheat plus 1 alien chromosome) are recovered and colchicine treated. The final products after colchicine treatment will be plants with 44 chromosomes (42 wheat plus an alien pair). This process simplifies the production of disomic addition lines, and resolves the constraints of paternal transmission of alien chromosomes. In addition, it reduces the analyses necessary for recovering 44-chromosome disomic derivatives following the selfing of 43-chromosome plants that have chromosomal associations of 21 bivalents plus 1 univalent.

If a wide cross program were built exclusively around the wheat cultivar Chinese Spring, the H. bulbosum procedure (40-45 % polyhaploid recovery) first used by Islam et al. (1981) for producing wheat/barley addition lines, would be satisfactory. However, in our program where commercial wheat cultivars are used, the H. bulbosum technique is ineffective and we have logically favored the `wheat x maize' methodology. We have initially applied the procedure to derivatives of `Th. elongatum x T. aestivum crosses. From 180 backcross derivatives, we obtained seed set after colchicine treatment of 62 plants with somatic chromosome numbers of 43, 44, and 45 that were crossed with maize. Several double haploids with 44 chromosomes have been developed, which form the starting phase for producing Th. elongatum disomic addition sets.

Embryo formation in wheat crosses with maize and pearl millet. Embryo formation frequencies (percent frequencies of the embryos obtained from wheat florets pollinated) in crosses with maize and pearl millet are shown in Table 22. In crosses using fresh pollen of maize and pearl millet, embryos were obtained at frequencies of 18.9-20.4 % and 19.7-35.6 %, respectively, across three crossing methods. Embryo formation frequencies indicated that crosses with stored maize pollen produced embryos at lower frequencies (2.8-8.5 %) than other crosses (18.9-35.6 %) with no significant difference presented for fresh and stored pearl millet pollen.

Use of pollen storage and detached-tiller culture did not increase the wheat polyhaploid production frequency, but these procedures do enhance production efficiency. Stored pearl millet pollen can be successfully used when fresh pollen is unavailable. Detached-tiller culture gives considerable savings in terms of labor and space required for handling wheat plants. Hot-water emasculation is a time saver and requires only 3 minutes for a large number of wheat spikes, whereas hand emasculation takes at least 3 minutes per spike. These techniques, used solely or in combination, provide greater flexibility for polyhaploid production in hexaploid wheat.

Table 22. Embryo formation frequencies (%)1 in crosses of hexaploid wheat with maize and pearl millet.

Pollen donorPollen storage
on plant
Crossing method 1
hand/detachedhand/detachedhot water
MaizeFresh20.4b19.4b18.9b
Stored 2.8d7.0c8.5c
Pearl milletFresh19.7b21.2b35.6a
Stored20.4b27.7b23.3b

1 Numbers followed by the same letter are not significantly different at the 5 % probability level.

go to next document