Construction and Preliminary Analysis of the SSH Library from the Foxtail Millet Rust Resistant Variety of Shilixiang

doi:10.7668/hbnxb.2014.06.022

Abstract

Abstract: In order to understand key process controling rust resistance and molecular mechamism,a suppression subtractive hybridizaion(SSH) library was constructed from Shilixiang with young leaves collected from seedling inoculated with Uromyces setariae after 12,24,48,72,96 h.To identify differentially express gene in response to Uromyces setariae,differentially express gene were screened and putative functions were assigned by analysing comparing sequence in non-redundance database from GenBank.The exprssion pattern of partially screened gene were checked by real-time-quantitative PCR.Positive clone were randomly picked out and the fragment were 200-750 bp in length.368 ESTs were obtained and only 32 ESTs were related to defence response by blast analysis in GenBank.By analysing the defence related EST,it was conclued that WRKY transcription factor,mitogen-activated protein kinase cascades,calcium signal transduction,glutathione S-transferase,cytochrome P450,pathogen-related protein,were supposed to involved in the process of incompatible interaction between Shilixiang and Uromyces setariae.Four genes from the SSH library were selected to measure their expression levels by real-time-quantitative PCR and the results showed the four genes were up-regulated in foxtail millet leaves inoculated with Uromyces setariae.Through the construction of SSH library,rust resistant genes were identified,which laid the foundation for the molecular breeding of rust resistant foxtail millet.

Key words: Suppression subtractive hybridization, Uromyces setariae, Foxtail millet, EST

CLC Number:

LI Zhi-yong, JIA Li-xia, DONG Li, WANG Nan, BAI Hui, QUAN Jian-zhang, LIU Lei, DONG Zhi-ping. Construction and Preliminary Analysis of the SSH Library from the Foxtail Millet Rust Resistant Variety of Shilixiang[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2014, 29(6): 126-130. doi: 10.7668/hbnxb.2014.06.022.

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References

[1] 梁克恭,武小菲,王雅儒,等.谷子品种对粟锈病的抗性鉴定研究[J].植物保护,1995,21(6):17-18.
[2] Yamamizo C,Kuchimura K,Kobayashi A,et al.Rewiring mitogen-activated protein kinase cascade by positive feedback confers potato blight resistance[J].Plant Physiology,2006,140(2):681-692.
[3] 于秀梅,喻修道,屈志鹏,等.条锈菌诱导的小麦抑制差减杂交文库构建及其表达序列标签研究[J].植物病理学报,2007,37(1):50-55.
[4] 李星,于秀梅,李亚宁,等.叶锈菌诱导的小麦叶片抑制差减杂交文库构建及其分析[J].中国农业科学,2008,41(12):4069-4076.
[5] Rinaldi C,Kohler A,Frey P,et al.Transcript profiling of poplar leaves upon infection with compatible and incompatible strains of the foliar rust Melampsora larici-populina[J].Plant Physiology,2007,144(1):347-366.
[6] Choi J J,Alkharouf N W,Schneider K T,et al.Expression patterns in soybean resistant to Phakopsora pachyrhizi reveal the importance of peroxidases and lipoxygenases[J].Functional & Integrative Genomics,2008,8(4):341-359.
[7] 赵立强,潘文嘉,马继芳,等.一个谷子新抗锈基因的AFLP标记[J].中国农业科学,2010,43(21):4349-4355.
[8] 董志平,李志勇,马继芳,等.谷子抗病基因同源序列的克隆与分析[J].植物病理学报,2011,41(1):93-97.
[9] Li J,Brader G,Palva E T.The WRKY70 transcription factor:A node of convergence for jasmonate-mediated and salicylate-mediated signals in plant defense[J].Plant Cell,2004,16(2):319-331.
[10] Qiu D Y,Xiao J,Ding X H,et al.OsWRKY13 mediates rice disease resistance by regulating defense-related genes in salicylate-and jasmonate-dependent signaling[J].Molecular Plant-Microbe Interactions,2007,20(5):492-499.
[11] 王凯,杜丽璞,张增艳,等.中间偃麦草 SGT1 基因的克隆及其抗病功能的分析[J].作物学报,2008,34(3):520-525.
[12] 崔艳红,贾芝琪,李颖,等.利用VIGS技术研究马铃薯抗晚疫病基因 R3a 和 RB 的信号传导[J].园艺学报,2009,36(7):997-1004.
[13] Ferrari S,Galletti R,Denoux C,et al.Resistance to botrytis cinerea induced in arabidopsis by elicitors is independent of salicylic acid,ethylene,or jasmonate signaling but requires Phytoalexin deficient3[J].Plant Physiology,2007,144(1):367-379.
[14] Du L,Ali G S,Simons K A,et al.Ca(2+)/calmodulin regulates salicylic-acid-mediated plant immunity[J].Nature,2009,457(7233):1154-1158.

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