1.College of Agronomy,
Agricultural University of Hebei, Baoding 071001, China；College of Agronomy,
Northwest A&F University, Yangling 712100, China; 2.College of Agronomy,
Agricultural University of Hebei, Baoding 071001, China; 3.College of Agronomy,
Northwest A&F University, Yangling 712100, China;
Abstract：Powdery mildew, caused by Blumeria graminis f. sp. tritici, is one of the most important fungal diseases in common wheat (Triticum aestivum L.) worldwide. Wheat germplasm N9436 is resistant to powdery mildew. In the present study, a backward subtracted cDNA library was constructed with cDNA from N9436 leaves inoculated by Blumeria graminis as the driver and cDNA from uninoculated N9436 leaves as the tester. A total of 120 positive clones were randomly chosen from the SSH-cDNA library and were amplified with sp6 and t7 primers to examine the insert size. After screening the repeated and redundant sequences, 59 expressed sequence tags (EST) were acquired. Nucleic acid and protein homology search were performed using the basic local alignment search tool (BLAST) program with the default settings at NCBI website (http://www.ncbi.nlm.nih.gov). BlastX results in nr-protein database revealed that 23 ESTs were highly homologous with known proteins involved in primary metabolism, energy metabolism, transport, signal transduction, and disease resistance and defenses. BlastNr results showed that 47 and 10 ESTs had high identities with known Unigene and function-unknown ESTs, respectively, and two ESTs matched none in the nr-database. Twenty-one ESTs were both in the nucleic acid and protein databases, including seven ESTs associated with powdery mildew resistance. Among them, one was responsible for signal transduction and six for systemic acquired resistance (SAR) system.
Cao A Z, Li Q, Chen Y P, Zou X W, Wang X E, Chen P D (2006). Screening resistance-related gene to powdery mildewin Haynaldia villosa using barlygenechip and studying its mechanism of resistance. Acta Agron Sin, 32(10): 1444―1452 (in Chinese)
Chen Y P, Wang H Z, Wang X E, Cao A Z, Chen P D (2006). Cloning and expressionof peroxisomal ascorbate peroxidase gene from wheat. Mol Biol Rep, 33: 207―213 doi: 10.1007/s11033-005-4536-1
Diatchenko L, Lauy F C, Campbell A P, Chenchik A, Moqadam F, Huang B, Lukyanov S, Lukyanov K, Gurskaya N, Sverdlov E D, Siebert P D (1996). Suppressionsubtractive hybridization: A method for generating differentiallyregulated or tissue specific cDNA probes and libraries. Proceedings of National Academy Sciences of the USA, 93(12): 6025―6030 doi: 10.1073/pnas.93.12.6025
Gao Y L, Guo W Z, Wang L, Zhang T Z (2007). Cloning and characterization of one-1,3-glucanase gene cDNA in cotton(Gossypium barbadense L.). Acta Agron Sin, 33(8): 1310―1315 (in Chinese)
Gomes E, Sagot E, Gaillard C, Laquitaine L, Poinssot B, Sane jouand Y H, Delrot S, Coutos-Thevenot P (2003). Nonspecificlipid-transfer protein genes expression in grape (Vitis sp.) cells in response to fungal elicitor treatments. Mol Plant Microbe Interact, 16(5): 456―464
He D Y, Wang H G (2005). Cloningof a new phosphoserine aminotransferase in the substitution resistantto powdery mildew of wheat-Agropyron elongatum. Chin Sci Bull, 50(6): 535―539 (in Chinese)
Huang X Q, Röder M S (2004). Molecularmapping of powdery resistance genes in wheat: a review. Euphytica, 137: 203―223 doi: 10.1023/B:EUPH.0000041576.74566.d7
Ichiro K, Kazunori T, Tokuko T, Makiko K (2006). Radical scavenger can scavenge lipid allyl radicals complexed withlipoxygenase at lower oxygen content. BiochemJ, 395: 303―309 doi: 10.1042/BJ20051595
Li R, Rimmer R, Buchwaldt L, Sharpe A G, Séguin-Swartz G, Coutu C, Hegedus D D (2004). Interaction of Sclerotinia sclerotiorum with a resistant Brassica napus cultivar: expressed sequence tag analysis identifies genes associatedwith fungal pathogenesis. Fungal GenetBiol, 41(8): 735―753 doi: 10.1016/j.fgb.2004.03.001
Liao Y, Zhang Z Y, Du L P, Xu H J, Yao W L, Shi J Y, Ren Z L, Xin Z Y (2007). Isolation of RAR1gene from Thinopyrum intermedium and analysis of its function in wheat background. Sci Agric Sin, 40(8): 1667―1674 (in Chinese)
Lin C S, Lai Y H, Sun C W, Liu N T, Tsay H S, Chang W C, Chen J J W (2006). Identificationof ESTs differentially expressed in green and albino mutant bamboo(Bambusa edulis) by suppressive subtractive hybridization (SSH) andmicroarray analysis. Plant Cell Tiss OrganCult, 86: 169―175 doi: 10.1007/s11240-006-9105-3
Liu B, Cui S P, Wang X J, Huang L L, Kang Z S (2007). Cloning and prodaryoticexpression of a wheat β-1,3-glucanase gene induced by Puccinia striiformisWestend f. sp. tritici Eriks in E. coli. J Northwest A&FUniv (Nat Sci Edn), 35(6): 125―129 (in Chinese)
Liu X F, Xu J Z, Hou Y Q, Zhan S X, Ge X C, Chao K M (1999). Isolation and characterization of a lipid transfer proteingene from rice. Acta Bot Sin, 41(7): 736―740 (in Chinese)
Luo M, Kong X Y, Huo N X, Zhou R H, Jia J Z (2002a). ESTs analysis ofresistance to powdery mildew in wheat at primary infected stage. Acta Genet Sin, 29(6): 525―530 (in Chinese)
Luo M, Kong X Y, Huo N X, Zhou R H, Jia J Z (2002b). Analysis of generesistant to powdery mildew by suppression subtractive hybridization. Chin Sci Bull, 47(16): 1237―1241 (in Chinese)
Maldonado A M, Doerner P, Dixon R A, Lamb C J, Cameron R K (2002). A putativelipid transfer protein involved in systemic resistance signaling in Arabidopsis. Nature, 419(6905): 399―403 doi: 10.1038/nature00962
McIntosh R A, Brown G N (1997). Anticipatorybreeding for resistance to rust diseases in wheat. Annu Rev Phytopathol, 35: 311―326 doi: 10.1146/annurev.phyto.35.1.311
Michels A K, Wedel N, Kroth P G (2005). Diatom plastids possess a phosphoribulokinasewith an altered regulation and no oxidative pentose phosphate pathway. Plant Physiology, 137: 911―920 doi: 10.1104/pp.104.055285
Nascimento A S, Catalano-Dupuy DL, Bernardes A, Oliveira Neto M, Santos M A M, Ceccarelli E A, Polikarpov I (2007). Crystal structures of Leptospira interrogansFAD-containing ferredoxin-NADP+ reductase and its complex with NADP+. BMC Structural Biology, 7: 69 doi: 10.1186/1472-6807-7-69
Niu J S, Yu L, Chen P D, Liu D J (2001). cDNA library construction and identification of Triticum aestivum-Haynaldia villosa 6VS/6AL translocationline. J Nanjing Agric Univ, 24(1): 5―8 (in Chinese)
Ouyang B, Yang T, Li H, Zhang L, Zhang Y, Zhang J, Fei J, Ye Z (2007). Identification of early salt stressresponse genes in tomato root by suppression subtractive hybridizationand microarray analysis. J Exp Bot, 58: 507―520 doi: 10.1093/jxb/erl258
Runquist J A, Miziorko H M (2006). Functionalcontribution of a conserved, mobile loop histidine of phosphoribulokinase. Protein Science, 15: 837―842 doi: 10.1110/ps.052015606
Selesi D, Schmid M, Hartmann A (2005). Diversity of Green-Like and Red-LikeRibulose-1, 5-bisphosphate carboxylase/oxygenase large-subunit genes(cbbL) in differently managed agriculturalsoils. Applied and Environmental Microbiology, 71: 175―184 doi: 10.1128/AEM.71.1.175-184.2005
Singla-Pareek S L, Reddy M K, Sopory S K (2003). Genetic engineering of the glyoxalasepathway in tobacco leads to enhanced salinity tolerance. PNAS, 100(25): 14672―14677 doi: 10.1073/pnas.2034667100
Thomas J C, Ughy B, Lagoutte B, Ajlani G (2006). A second isoform of the ferredoxin: NADP oxidoreductase generatedby an in-frame initiation of translation. PANS, 103(48): 18368―18373 doi: 10.1073/pnas.0607718103
Van Loon L C, van Strien E A (1999). The familiesof pathogenesis-related proteins, their activities and comparativeanalysis of PR-1 type proteins. PhysiolMol Plant Pathol, 55: 85―97 doi: 10.1006/pmpp.1999.0213
Wang Y C, Li H Y, Yang C P, Zhang G D (2007). Study on expression of genes in Tamarixand rossowii under drought stressusing cDNA microarray technology. BullBot Res, 27(2): 186―194 (in Chinese)
Wong Y Y, Ho C L, Nguyen P D, Teo S S, Harikrishna J A, Rahim R A, Wong M C V L(2007). Isolation of salinity tolerant genes from the mangroveplant, Bruguiera cylindrica byusing suppression subtractive hybridization (SSH) and bacterial functionalscreening. Aquatic Bot, 86(2): 117―122 doi: 10.1016/j.aquabot.2006.09.009
Wu J H, Hu Y G, Zhang H, Wang C Y, Wang Q Y, Ji W Q (2008). Expression of Special Genes Resistant to Powdery Mildew(Blumeria graminis f. sp. tritici) in Wheat Germplasm N9436. Acta Agron Sin, 34(7): 1143―1152 (in Chinese) doi: 10.1016/S1875-2780(08)60040-1
Yoshizawa Y, Toyoda K, Arai H, Ishii M, Igarashi Y (2004). CO2-responsive expression and gene organization of threeribulose-1,5-bisphosphate carboxylase/oxygenase enzymes and carboxysomesin Hydrogenovibrio marinus StrainMH-110. Journal of Bacteriology, 186(17): 5685―5691. doi: 10.1128/JB.186.17.5685-5691.2004
Yu X D, Qu Z P, Guo J, Yu X M, Huang X L, Han Q M, Huang L L, Kang Z S (2008). Construction and preliminary analysisof the compatible SSH cDNA library between Puccinia striiformis and wheat. Sci Agric Sin, 41(5): 1267―1273
Yu X M, Yu X D, Qu Z P, Han Q M, Guo J, Huang L L, Kang Z S (2007). Constructionof wheat SSH cDNA library induced by Puccinia striiformis and analysis of expressedsequence tags. Acta Phytopathol Sin, 37(1): 50―55 (in Chinese)
