摘要
【目的】筛选小麦抗麦长管蚜基因,以揭示小麦抗蚜虫的分子机制。【方法】利用mRNA差异显示反转录PCR(Differential display reverse transcription PCR,DDRT-PCR)技术,以小麦抗蚜种质98-10-35、感蚜种质1376及其F3代和BC1F1代(98-10-35/1376//1376)为材料,分析了在抗蚜材料与感蚜材料之间差异表达基因的序列及功能。【结果】从24对引物组合中,共筛选到8对引物可以在98-10-35/1376的F3代抗性群体中扩增出74条差异条带,平均每对引物扩增条带数为9.3,并对其中差异明显的20条条带进行了回收、克隆、测序和序列比对分析,其中1条在所有抗性样品池中特异表达,将其经电子克隆延长后获得了长度为2 260bp的cDNA序列,经ORF finder软件翻译后,获得了一个含642个氨基酸的蛋白质,将其命名为TA642,该蛋白与其他植物黏连蛋白质复合体亚基——STAG域蛋白质高度同源,推测其主要通过参与真核生物的姊妹染色单体黏连作用而影响植物的生理生化代谢。【结论】获得了一些与小麦抗蚜性相关的差异表达基因序列,鉴定的TA642蛋白质可能与小麦麦长管蚜抗性分子机理有关。
The objective of this paper was to screen wheat genes to resist aphid,Sitobion avenae and to reveal the resistance mechanism.【Method】Sequences and functions of differential expressed genes between resistant and susceptible population were analyzed by mRNA differential display reverse transcription PCR（DDRT-PCR）with resistant parents of 98-10-35,susceptible parents of 1376,and their F3 and BC1F1（98-10-35/1376//1376）population.【Result】8 pairs specific primers out of 24 were selected were able to amplify 74fragments with differential resistances.A mean of 9.3 amplification products were detected from above cDNA templates.20 bands with significant differences were cloned,sequenced and compared.A cDNA sequence with 2 260bp,encoding 642 amino acids,was obtained and named as TA642.TA642 was highly homologous to proteins in caderin complex subunit-STAG domain of other plants are highly homologous.It was assumed that it would affect physiological and biochemical metabolism by pla-ying a role in adhesion of sister chromatid in eukaryotes.【Conclusion】An expressed sequence tags（ESTs） sequence of wheat resistance gene,namely TA642,was obtained and it would participate in the resistance to Sitobion avenae.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2013年第8期195-201,共7页
Journal of Northwest A&F University(Natural Science Edition)
基金
国家"863"计划重大专项(2009AA101102)
中德农业合作资助项目(2008/2009(04))
河南科技大学人才基金项目(09001595)
关键词
小麦
麦长管蚜
候选基因
差异表达
同源性分析
wheat
Sitobion avenae F.
candidate genes resistant on aphid
difference expression
homologous analysis
