高粱赤霉素信号途径关键基因的鉴定及qPCR引物筛选

Identification of Gibberellin Signaling Pathway Key Genes and Screening of qPCR Primers in Sorghum bicolor

赤霉素(GA)在植物响应生物和非生物胁迫过程中扮演着重要角色。为了探究GA在高粱响应蚜虫胁迫过程中的作用,本研究采用激素外施实验,证实GA参与了高粱的抗蚜反应;在此基础上,利用NCBI数据库查找已报道参与GA合成、代谢及Della蛋白合成相关基因在高粱中的同源基因,并设计特异引物,采用PCR和qPCR的方法验证引物的特异性。通过PCR技术共筛选出7对引物,大小符合预期,且条带单一,进一步采用qPCR技术验证,最终筛选到5个基因的5对引物,扩增曲线光滑,呈现单峰,且Ct值较小,特异性良好。研究结果为下一步研究这些基因的表达模式变化,揭示GA信号途径在高粱抗蚜反应中的作用提供理论依据。

Gibberellin(GA) plays an important role in plant response to biotic and abiotic stresses.In order to explore the role of GA in Sorghum response to aphid stress,hormone application experiment was used in this study.The results showed that GA was involved in Sorghum resistance to aphids.Based on that,the NCBI database was used to find the homologous genes in Sorghum that have been reported to be involved in gibberellin synthesis,metabolic pathways and DELLAs protein synthesis.The specific primers were designed and PCR and qPCR technology have been used to verify the specificity of primers.A total of 7 pairs of primers with expected size and single band were screened out by PCR.The specificity of the primers was further verified by qPCR and five pairs of primers for five genes were finally screened.The amplification curve was smooth,showing a single peak,and the Ct value was small,indicating that these primers are specific.This study can provide a foundation basis for further study on the expression patterns of these genes and reveal the role of GA signaling pathway in Sorghum resistance response to aphid.