【摘要】 为挖掘获得新的抗性基因,培育鹰嘴豆Cicer arietinum抗壳二孢叶枯病(病原菌为Ascochyta rabiei)新品种,以项目组前期获得的102个差异表达的新基因为基础,随机选取29个基因进行同源性分析,以鹰嘴豆Actin(EU529707)和Ef-1α(AJ004960)作为参考基因,利用实时荧光定量PCR技术检测这29个基因在宿主植物鹰嘴豆抗性品种系选03中的表达规律。结果显示,基于同源性分析结果可将29个基因大致分成4类,涉及信号传导机制、细胞运输、转录和细胞拯救、防御、毒性;功能分析结果显示,功能未知的基因数目最多,达到了11个,其中多数为鹰嘴豆未定性基因。这29个基因在A. rabiei胁迫下都发生了不同程度的差异表达,表达差异时间点集中在胁迫后72 h,并在96 h恢复至正常表达水平。其中解毒相关基因474在72 h时相对表达水平最高,是对照处理0 h的19.773倍,抗氧化修复相关基因1137的相对表达水平最低,约为对照处理0 h的1/3。筛选获得的差异表达基因中,表达上调的基因有10个,表达下调的基因有16个,其余3个基因的表达差异不明显。上调表达基因可能与鹰嘴豆应对A. rabiei侵染的应答机制有关,其中与免疫应激相关的蛋白基因如谷胱甘肽S-转移酶、咖啡酰辅酶A、泛素蛋白基因等可能直接参与了鹰嘴豆应对A. rabiei的免疫识别和防御。更多还原

【Abstract】 To excavate new resources for breeding of new chickpea resistant varieties to Ascochyta rabiei, the pathogen causing Ascochyta blight, 29 genes were randomly selected for homology analysis based on 102 differentially expressed new genes obtained by the research group in the early stage. Actin(EU529707) and Ef-1α(AJ004960) were used as reference genes to detect these genes in the resistant chickpea variety Xixuan 03 by real-time fluorescence quantitative PCR. The results showed that the29 genes were roughly divided into four categories: cell rescue, defense, toxicity, signal transduction mechanism, cell transport and transcription. The results of functional analysis showed that the number of genes with unknown functions was the largest(11), most of which were undetermined genes of chickpea. The 29 genes were differentially expressed under A. rabiei stress, and the expression was concentrated at 72 h after treatment, and their normal expression level was restored at 96 h. Among them, the detoxification-related gene 474 showed the highest relative expression level at 72 h, which was 19.773 times of the control at 0 h, and the relative expression level of anti-oxidation repair-related gene 1137 was the lowest, about 1/3 of the control at 0 h. Among the differentially expressed genes obtained by screening, there were 10 up-regulated genes and 16 down-regulated genes, and the expression differences of the other three genes were not significant. In particular, up-regulated genes might be involved in the response mechanism of chickpeas to A. rabiei infection, among which protein genes related to immune stress such as glutathione S-transferase, caffeyl-CoA, ubiquitin protein gene etc. might directly participate in the immune recognition and defense of chickpeas in response to A. rabiei.更多还原