基于转录组测序与定量PCR技术挖掘北沙柳株型相关候选基因  

Preliminary screening of Northern Salix(Salix psammophila)architecture related genes based on RNA-Seq analysis and quantitative PCR technique

作　　者：贺嵘 赵恺 贺玉娇 阿拉腾苏和 王爱君 宁静 韩若霜 孙贵荣[2] 张国盛[1] HE Rong;ZHAO Kai;HE Yujiao;ALATENG Suhe;WANG Aijun;NING Jing;HAN Ruoshuang;SUN Guirong;ZHANG Guosheng(College of Forestry,Inner Mongolia Agricultural University,Hohhot 010019,China;Forest General Site of Erdos,Erdos 014300,Inner Mongolia,China;Inner Mongolia Forestry Scientific Research Institute,Hohhot 010013,China;Inner Mongolia Autonomous Region Forestry and Grassland Seedling General Station,Hohhot 010010,China)

机构地区：[1]内蒙古农业大学林学院,内蒙古呼和浩特010019 [2]鄂尔多斯市造林总场,内蒙古鄂尔多斯014300 [3]内蒙古林业科学研究院,内蒙古呼和浩特010013 [4]内蒙古自治区林业和草原种苗总站,内蒙古呼和浩特010010

出　　处：《浙江农业学报》2023年第10期2332-2345,共14页Acta Agriculturae Zhejiangensis

基　　金：内蒙古自治区自然科学基金项目(2018MS03013);内蒙古自治区科学技术厅项目(2021GG0075)。

摘　　要：为构建北沙柳株型分子辅助育种体系实现高效定向育种,采用水培群体转录组测序与野外训练群体定量PCR分析相结合的方法,以RNA-Seq分析中的基因表达量(FPKM值)与冠高比(株型特征值)的相关系数作为预测值,探讨北沙柳株型相关候选基因挖掘技术。结果表明,北沙柳冠高比与FPKM值间存在相关性,在水培预测群体和野外训练群体中保持基本一致的趋势。文献中已报道的株型(或者分枝)相关基因(TAC 2、LAZY 1b、ZFP 4、TB 1、SPA 2、ABF 2和PYL 1)的预测值在0~0.6,ATX 1、RFK 1和FHY 1基因(未报道与株型相关)的预测值较高(0.6~1.0),按照“基因个数最少,相关系数最高”的原则,选择双基因组合ATX1+FHY 1作为鉴别北沙柳株形的基因。将形态学、转录组学作为一个整体来挖掘株型性状相关基因是本研究特色,研究结果也可为其他植物、其他性状的分子辅助育种提供理论基础。In order to construct a molecularly assisted breeding system for efficient targeted breeding of Salix psammophila architectures,a combination of transcriptome sequencing in the hydroponic population and quantitative PCR analysis in the field training population was used to explore candidate gene mining techniques related to Salix psammophila architectures,using the correlation coefficient between gene expression(FPKM value)and ratio of crown and height in RNA-Seq analysis as predictive values.The results showed that there was a correlation between ratio of crown and height(plant architecture characteristic value)and FPKM value in Salix psammophila,and the trend remained largely consistent in the hydroponic prediction population and field training population.The predicted values of plant architecture(or branching)-related genes(TAC2,LAZY1b,ZFP4,TB1,SPA2,ABF2 and PYL 1)reported in the literature ranged from 0 to 0.6,and the predicted values of ATX1,RFK 1 and FHY 1 genes(not reported to be plant architecture-related)were higher(0.6 to 1.0).The two-gene combination(ATX1+FHY1)was selected as the gene for the identification of Salix psammophila architectures according to the principle of“the least number of genes and the highest correlation coefficient”.Morphology and transcriptomics as a whole are the characteristics of this study,and the results can also provide a theoretical basis for molecularly assisted breeding of other plants and other traits.

关 键 词：冠高比 相关系数 转录组测序技术 定量PCR技术 基因挖掘 北沙柳 

分 类 号：S793[农业科学—林木遗传育种]