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利用具有6个无毒基因不同表现型且处于萌发中的静孢子时期的马铃薯晚疫病菌构建4个混 合池,通过cDNA AFLP分析,共获得85个与无毒基因相关的差异表达片段,其中与无毒基因Avr1、Avr2、 Avr3 Avr10 Avr11、Avr4相关的差异表达片段分别为20,28,16和21个。将这些差异表达片段对20个晚疫 病病菌个体进行无毒基因差异表达验证,得到1个无毒基因连锁群Avr3 Avr10 Avr11的候选片段,即 A11T13Avr3 10 11S157;无毒基因Avr4的候选片段2个,即A24T19Avr4S122和A24T24Avr4S142。Blast分析 表明:A11T13Avr3 10 11S157与马铃薯晚疫病菌萌发孢子囊(germinatingsporangium)EST(expressed sequencetag)库中的序列PG001F10.XT7同源;A24T19Avr4S122与晚疫病菌萌发中静孢子(germinating cyst)EST库中的PH051G10.XT7部分序列完全一致,而A24T24Avr4S142与PH051G10.XT7具有98%的 同源性。这些结果将促进通过电子克隆并结合RACE(RapidAmplificationofcDNAEnds)技术获得全长的无 毒基因Avr4。
Four mixed pools were constructed by using P. infestans in the period of conidial germination with six non-toxic genes and their phenotypes were germinated. A total of 85 differentially expressed fragments related to non-toxic genes were obtained by cDNA AFLP analysis. Among them, The differential expression fragments of avirulence genes Avr1, Avr2, Avr3 Avr10 Avr11 and Avr4 were 20, 28, 16 and 21, respectively. The differential expression fragments of these 20 Phytophthora infestans individuals were verified by non-toxic gene differential expression, obtained a non-toxic gene chain Avr3 Avr10 Avr11 candidate fragments, namely A11T13Avr3 10 11S157; non-toxic gene Avr4 two candidate fragments, Namely A24T19Avr4S122 and A24T24Avr4S142. Blast analysis showed that A11T13Avr3 10 11S157 was homologous to the PG001F10.XT7 sequence in the expressed sequence tag library of P. infestans, and that A24T19Avr4S122 was homologous with the EST library of germinating cyst in the germination of P. infestans The partial sequence of PH051G10.XT7 was identical, while A24T24Avr4S142 had 98% homology with PH051G10.XT7. These results will facilitate the production of the full-length avirulence gene Avr4 by electronic cloning combined with RACE (Rapid Amplification of cDNAEnds) technology.