中国油料作物学报2011，Vol.33Issue(2)：157-161,5.
核盘菌诱导下甘蓝型油菜宁RS-1的SSH文库构建
Construction of SSH cDNA library from Ning RS - 1 after inoculation of Sclerotinia sclerotiorum
摘要
Abstract
Total RNA was extracted from the leaves of Ning RS - 1 before and after inoculation by Sclerotinia sclerotiorum ( Lib.) de Bary.Leaves'mRNA were purified and transcribed into cDNA.An adaptor was added on to the cDNA by enzymatic hydrolysis, followed by two rounds of hybridization.Finally differential expression of suppression subtracted cDNA library of the induced Ning RS - 1 was built.In subtractive hybrid library, the induced Ning RS - 1 cDNA was used as a tester and the normal Ning RS - 1 cDNA was used as a driver.In reverse library,they were used in opposite way.The subtractive library had a recombination rate of 96.2％ with a base capacity of 2 190.And the reversed one had a recombination rate of 94.5％ with capacity of 2 780.It indicated that the library quality could meet the requirements of screening resistance genes to Sclerotiorum.Genes related to metabolism of resistance to the disease were identified by sequencing of the clones picked randomly from the cDNA library.They included γ- glutamylcysteine synthetase gene, anti- fungal protein, iron protein and ACC oxidase, etc.关键词
甘蓝型油菜/核盘菌/抑制消减杂交/cDNA文库Key words
Brassica napus L./ Sclerotinia sclerotiorum / SSH/ cDNA library分类
农业科技引用本文复制引用
周晓婴,陈松,戚存扣..核盘菌诱导下甘蓝型油菜宁RS-1的SSH文库构建[J].中国油料作物学报,2011,33(2):157-161,5.基金项目
江苏省支撑计划(BE2009304) （BE2009304）
江苏省自主创新基金(cx(09)636) （cx(09）
国家948项目(2006-G04) （2006-G04）
