‘陕麦139’受条锈菌抑制表达相关基因的分析

Study on Gene Expression Inhibited by Innoculation of Puccinia striiformis West. in 'Shaanmai 139'

以‘条中32’接种前后的小麦抗条锈病种质‘陕麦139’幼苗叶片为材料,利用抑制消减杂交技术,构建了条锈菌接种48 h小麦抗病品种叶片的抑制表达SSH-cDNA文库,通过测序以了解条锈菌侵染后被抑制表达的相关基因。从构建的文库中随机选取48个阳性克隆,进行测序,经过序列拼接等,获得高质量EST序列34条(Gen-Bank序列号为EL930132-EL930165),序列比对分析表明其中31条EST序列与已知功能的基因序列同源性较高,主要涉及植物的能量代谢途径、膜转运、信号传导及核酸加工等方面的基因,包括光合系统Ⅱ抗性蛋白D1、碳酸酵素、线粒体磷酸盐转运子、苹果酸脱氢酶、过敏性诱导反应蛋白和rRNA核酸内切酶等相关基因。此外,获得未知功能的EST序列3条。研究表明,小麦在条锈病菌侵染初期,许多基因的正常表达调控受到影响,如能量代谢和寄主细胞结构木质化,阻止小麦抗病过敏性反应等。

In order to study the effect of stripe rust pathogen on seedling leaves of wheat germplasm with reistance,＇Shaanmai 139＇ were inoculated with Puccinia striiformis West.as driver and not inoculated as tester.Subtractive hybridization was adopted to construct the inhibited SSH-cDNA library of the leaves in their early with DH5α.Plasmids were extracted from 48 positive clones,randomly selected from the library,and sequence and homology searches were conducted using the BLAST program.34 EST were achieved and GenBank Accession No.is from EL930132 to EL 930165.The information show that 31 EST sequences with known functions,related to energy and metabolism,membrane and transport,signal transduction,transcription and translation,including PSⅡ inhibitor resistant D1 protein,carboxylase,mitochondrial phosphate transporter,glyoxysomal malate dehydrogenase,hypersensitive-induced reaction protein and rRNA intron-encoded homing endonuclease.The result indicated that stripe rust pathogen may be influencing the expressi on of regulatory genes,the energy metabolism,the lignifications and impacting hypersensitive reaction of tissue.