Triticeae species of origin Genotype or cultivar or accession of origin Tissue Developmental stage Condition (environmental stress; disease challenge; etc.) Vector Size of library (est. no. of clones) Status (prescreened?; any sequencing done? confirmation of insert presence? indication of insert size? etc.) Contact (if library is available) ITEC contact if source of ESTs Comment
Libraries from which sequences will be contributed to ITEC
Aegilops speltoides . anthers immature; at meiosis normal conditions . . . . Jan Dvorak .
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Hordeum vulgare Barke leaf 14 day normal growth pBS/SK- (Stratagene) 5 million prescreened for SSU, LHC, sequencing, 99% with insert, average insert size: 850 bp . Prof. R.G. Herrmann .
Hordeum vulgare Forester inflorescence immature; developing . Lambda Zap II (Stratagene) greater than 1 million clones good library + well used Dave Laurie Paul Bailey .
Hordeum vulgare Franka epidermal cells 10-day-old seedlings normal conditions Triplex 2 lambda 1 million being tested for insert size and sequencing, selecting only clones with insert size between 400 and 1,500kb. Catherine Feuillet Catherine Feuillet Preliminary sequencing and BLAST analysis indicate good value for this library.
Hordeum vulgare Nure (2-row; winter) leaf first leaf cold stress UNI-ZAP XR 1 million clones made February 1999, average insert size 1kb, range 0.7-2.0 kb; sequencing started. Nicola Pecchioni, Valeria Terzi Nicola Pecchioni .
Hordeum vulgare Sultan 5 leaf . 6 and 12 h after infection with Erysiphe graminis f. sp. hordei pBAD42 - Clontech 1 million clones, 90% have insert 1.4 kb . Pietro Piffanelli .
Hordeum vulgare Triumph whole grain mature 3-day malted Lambda Zap express 40,000 good insert size in over 500 sequenced . Gordon C. Machray .
Hordeum vulgare . culm . etiolated . . . . Andreas Graner .
Hordeum vulgare . leaf . etiolated . . . . Andreas Graner .
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Lophopyrum elongatum Accession D root seedling salt-stressed with 250 mM NaCl for 6 hours under hydroponic conditions. PBluescript SK-, converted from original cDNA library in Lambda zap about 500,000 80-90% inserts with average size about 700 bp P. Gulick P. Gulick Limited copies of the Lambda zap library are available.
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Secale cereale Blanco root tip young roots stressed for aluminum in the process not yet known not yet Perry Gustafson Perry Gustafson .
Secale cereale Blanco root tip young roots nonstressed for aluminum in the process not yet known not yet Perry Gustafson Perry Gustafson .
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Triticum aestivum Atlas (Al tolerant) root 8 day old grown in vermiculite with 0.6 mM CaCl2 in water pYES2 (Invitrogen - yeast expression plasmid vector) 8.7x10 ^6 size selected for 500 - 1500bp, not sure about frequency of inserts . Marilyn Warburton .
Triticum aestivum Bobwhite endosperm mature seed nonstressed and insect-infected (Russian wheat aphid) . . screening underway, insert size >500 Anna-Maria Botha-Oberholster Anna-Maria Botha-Oberholster .
Triticum aestivum Catoctin (USDA-ARS, Beltsville MD) leaf four fully developed leaves after seed germination normal greenhouse conditions pSport-P Primary library 1.12x10^7 cfu/ml in progress J.-J. Lin J.-J. Lin Contact Dr. Lin for information about the library. With sufficient interest, it could become an LTI product.
Triticum aestivum Cheyenne endosperm 5 - 30 days post anthesis control lambda-Zap II 1,000,000 initial sequencing without prescreening - later sequencing after some normalization; about 50% of cDNAs of coding regions of 2000-2500 bp are full-length. Olin Anderson Olin Anderson .
Triticum aestivum Chinese Spring anther early meiosis . . . . . Peter Langridge .
Triticum aestivum Chinese Spring root . . . . made in 1995, well used Dave Lonsdale Paul Bailey .
Triticum aestivum Chinese Spring scutelum callus undifferentiated . . . made in 1995, well used Dave Lonsdale Paul Bailey .
Triticum aestivum Chinese Spring spikelet immature, developing . . . in preparation Graham Moore Paul Bailey .
Triticum aestivum Chinese Spring spikes immature . pT-Adv 100 cDNAs are differential display products. Sizes of inserts are known and in some cases have been assigned to chromosomes. . Bikram Gill .
Triticum aestivum Gamtoos-R (1RL.1BL translocation) leaf & stem 12- to 15-day-old seedlings infected with Russian Wheat Aphid pCMV Sport 6.0 not complete, est. at 3 million . . Nora Lapitan,
Dennis Knudson
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Triticum aestivum Germplasm line P29 leaf & stem 9-day-old seedlings unstressed Lambda Zap Express 1 million average insert size is 0.9 kb with 85% of clones having an insert . Joseph M. Anderson .
Triticum aestivum Mustang leaf 10-day-old seedlings heat stress Lambda Zap II (Stratagene) 2 million five full-length HSP clones isolated and sequenced (clone lengths of ca. 1 kb and ca. 3kb) Henry Nguyen Henry Nguyen Could produce cDNA libraries of other conditions and genotypes.
Triticum aestivum Norin 26 spikelet Feeke's scale 6 field grown . 782,000 fpu nonscreened, confirmed the insert, average insert size ca. 1.2kbp Yasunari Ogihara Yasunari Ogihara Norin 26 is spring-sown Japanese cv., no vernalization requirement, early flowering, shows photoperiod cytoplasmic male sterility in combination with cytoplasm of Aegilops crassa.
Triticum aestivum Novosibirskaya 67, red grain isoline pericarp/testa 3:1 mix, respectively of 10+20 days post anthesis . Lambda Zap II (Stratagene) 2 million made Dec. 1998 Paul Bailey, John Flintham Paul Bailey .
Triticum aestivum Odeon root seedling unstressed pUC18 819 screened from original library of 2104 clones by size, which inserts ranging from 0.3 to 2kb, most around 1 kb. J.M. Jacquemin J.M. Jacquemin 29 clones from this library have been mapped to the Opata x Synthetic wheat reference population (clones gbx).
Triticum aestivum Palmiet leaf 4-leaf stage nonstressed and insect-infected (Russian wheat aphid) . . screening underway, insert size >500 Anna-Maria Botha-Oberholster Anna-Maria Botha-Oberholster .
Triticum aestivum Soleil endosperm 1:1 mix of 10+20 days post anthesis . Lambda Zap II (Stratagene) 2 million made Dec. 1998 Kay Denyer Paul Bailey .
Triticum aestivum Sumai #3 spikes at anthesis challenged with head blight fungus pGemT 200 . . Bikram Gill .
Triticum aestivum TAM 101 root 10-day-old seedlings water deficiency stress Lambda Zap II (Stratagene) 400,000 cDNA clones isolated so far are full length. Henry Nguyen Henry Nguyen Could produce cDNA libraries of other conditions and genotypes.
Triticum aestivum Thatcher Lr1 leaf young leaf 24 h after challenge with LR-AVR1 Lambda Zap 1 million screened with resistance gene analogs; only three full sequences obtained; insert size est. 1 kb. Sylvie Cloutier Sylvie Cloutier .
Triticum aestivum Tugela endosperm mature seed nonstressed and insect-infected (Russian wheat aphid) . . screening underway, insert size >500 Anna-Maria Botha-Oberholster Anna-Maria Botha-Oberholster .
Triticum aestivum Wyuna endosperm mid-development . Lambda Zap . . . Rudi Appels .
Triticum aestivum . embryo . . . . . . Mike Holdsworth .
Triticum aestivum . endosperm . . . . . . Mike Holdsworth .
Triticum aestivum . leaf seedling challenged with an avirulent isolate of wheat yellow rust Lambda Zap Express (Stratagene) . in preparation Lesley Boyd, Rob Koebner Paul Bailey .
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Triticum monococcum DV92 shoot apex and young leaves 5-week-old plants not induced for flowering Phagemid pBK-CMV (4518 bp) Primers T3 and T7 764,000 Two empty colonies from 24. Average size 1000 bp. 50 clones sequenced . Jorge Dubcovsky .
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Triticum turgidum ssp. durum Siliana root 3-day-old seedling drought stress pSPORT1 (BRL) about 50,000 before amplification screened for dehydrins; clones between 0.3 and 2 kb. Philippe Joudrier Philippe Joudrier An additional 2000 sequences will be contributed, possibly from other libraries, during INRA's participation.
Libraries available; but use for ITEC sequences not yet confirmed.
Hordeum vulgare Morex shoot; etiolated 5-day-old seedling no stress; dark grown Uni-ZAP XR vector; excised pBluescript phagemids; directionally 6x10^5 pfu; mass excised to yield 1.3x10^6 cfu/ml on SOLR (Stratagene) cells; amplification=148; nonrecombinant clones=3%; blue colonies=5%; but 75% of these had inserts. insert size ranges from 0.6 to 2.2 with peak at 1.6 kb; no sequencing done up to now. Phagemids: Tim Close,
Arrayed library (later): Rod Wing
Andy Kleinhofs We do not recommend sequencing from this library at this time. Once it is arrayed and the abundant mRNA clones removed; it will be an excellent source for random EST sequencing.