北沙柳株型候选基因表达量与株型相关性的验证  

Validation of the correlation between the expression levels of candidate genes and plant architecture traits in Salix psammophila strains

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] 王瑞平 张国盛[1] He Rong;Zhao Kai;He Yujiao;Suhe Alateng;Wang Aijun;Ning Jing;Han Ruoshuang;Sun Guirong;Wang Ruiping;Zhang Guosheng(Forestry College of Inner Mongolia Agricultural University,Hohhot 010019,China;Forest General Site,Erdos,Erdos 017000,China;Inner Mongolia Forestry Scientific Research Institute,Hohhot 010013,China;Inner Mongolia Autono-mous Region Forestry and Grassland Seedling General Station,Hohhot 010010,China;Ordos Forestry and Grassland Bu-reau,Ordos 017010,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,内蒙古树林召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 [5]鄂尔多斯市林业和草原局,内蒙古鄂尔多斯017010

出　　处：《山西农业大学学报（自然科学版）》2023年第2期82-91,共10页Journal of Shanxi Agricultural University(Natural Science Edition)

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

摘　　要：[目的]构建北沙柳株型MAS体系实现高效定向育种。[方法]以不同株型北沙柳作为研究材料,构建预测群体、训练群体及验证群体。采用RNA⁃seq与RT⁃qPCR分析相结合的方法,以测序结果中FPKM值与北沙柳冠高比的相关系数作为预测值,并在大群体和不同时间采样群体中进行验证,最后通过基因组合法筛选与株型相关的最佳基因组合,探讨了北沙柳株型候选基因挖掘技术。[结果]冠高比是体现株型性状较理想的特征值。RNA⁃seq结果中不同基因的预测值不同,预测值的变化范围是0.0~0.9。目标基因(选择已验证与分枝有关SpsLAZY1b、SpsTAC2基因,与调控株型有关的差异基因ZFP4、TB1、SPA2、ABF2、PYL1以及预测值较大的ATX1、FHY1、RFK1基因作为目标基因)表达量与冠高比的相关性在预测群体、训练群体以及验证群体中具有相同的变化趋势,且ATX1、FHY1、RFK1在训练群体和验证群体中相关系数较高。按照“基因个数最少,相关系数最高”的原则,选择(ATX1+FHY1)为鉴别北沙柳株型的最佳基因组合。[结论]可利用RNA⁃seq结果筛选北沙柳株型候选基因,基因表达量与目标性状相关系数越高,功能基因与性状的关系越密切,且水培实验不会改变北沙柳的株型。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,利用性状与相关基因建立北沙柳株型后备基因的筛选是本研究的创新之处。研究结果也可为其他植物、其它性状的MAS提供理论基础。[Objective]The research purpose of current study was to construct a Market Assisted Selection(MAS)for efficient and targeted breeding of Salix psammophila strains.[Methods]Different plant architecture types of S.psammophila were used as research materials to construct a predictive population,training population,and validation population.A combination of RNA-seq and RT-qPCR analysis was used to predict the FPKM value and crown height ratio in the sequencing results and vali⁃dated them in large populations and populations sampled at different times.[Results]The results showed that the crown height ratio was the ideal value for plant architecture traits.The predicted values of different genes in the RNA-seq results varied from 0.0 to 0.9.The expression of target genes(SpsLAZY1b and SpsTAC2 genes,ZFP4,TB1,SPA2,ABF2,PYL1,and ATX1,FHY1,and RFK1 genes)with higher predicted values were selected.The correlation between the expression of ZFP4,TB2,SPA2,ABF2,PYL1 and ATX1,FHY1 and RFK1 genes,which had higher predicted values,had the same trend in the predictive,training,and validation populations.The correlation coefficients of ATX1,FHY1 and RFK1 were higher in the training and validation populations.According to the principle of"the lowest number of genes and the highest corre⁃lation coefficient",(ATX1+FHY1)was selected as the best gene combination for the identification of S.psammophila plant architecture.This study was characterized by the use of morphology and transcriptomics as a whole to explore genes related to strain traits,and the use of traits and related genes to establish the screening of back-up genes for S.psammophila plant archi⁃tecture was an innovative feature of this study.[Conclusion]The RNA-seq results can be used to screen the candidate genes of plant architecture of S.psammophila,and the higher the correlation coefficient between gene expression and target traits,the closer the relationship between functional genes and traits.The hydroponic experiment will not change the plant architecture of S.ps

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：S722.33[农业科学—林木遗传育种]