紫花地丁VpTM_(6-2)在本氏烟草中的功能研究  

The gene function study of VpTM_(6-2) from Viola philippica in Nicotiana benthamiana

作　　者：李巧峡[1] 黄永林 张正蓉 王勃 LI Qiao-xia;HUANG Yong-lin;ZHANG Zheng-rong;WANG Bo(College of Life Science,Northwest Normal University,Lanzhou 730070,Gansu,China)

机构地区：[1]西北师范大学生命科学学院,甘肃兰州730070

出　　处：《西北师范大学学报（自然科学版）》2021年第6期81-87,共7页Journal of Northwest Normal University(Natural Science)

基　　金：国家自然科学基金资助项目(31800188);甘肃省自然科学基金资助项目(18JR3RA088)。

摘　　要：病毒诱导的基因沉默(VIGS)是研究植物基因功能的一种非常有用的技术,它可随意敲掉感兴趣的基因,耗时较短,不需建立稳定的遗传体系.本研究利用VIGS方法,以紫花地丁的VpTM_(6-2)作为探针序列,去沉默本氏烟草中的内源NbTM_(6),以探讨VpTM_(6-2)的生物功能.VpTM_(6-2)属于B类MADS-box基因,文中设计的探针序列长357 bp,与NbTM_(6)相应的序列之间具有69.0%的同源性,除一个碱基有差异外,存在一段连续长27个碱基的100%同源序列.结果表明,VpTM_(6-2)能够特异性敲除本氏烟草的内源NbTM_(6),使得NbTM_(6)的表达明显下降,花粉发育异常(其余花器官发育均正常).本氏烟草VIGS体系使得紫花地丁中的一些基因可在异源体系中得到功能验证.该研究进一步证明VpTM_(6-2)表达含量的下降可能与紫花地丁两型花发育有一定的关系.Virus-induced gene silencing(VIGS)is a particularly useful tool for plant functional genomics because it can knock down the genes of interest,the observation of elicited phenotypes needs a short time,and does not require the genetic transformation of plants.In order to investigate the function of VpTM_(6-2),the gene silencing efficiency of the corresponding orthologs NbTM_(6) is determined by using the coding region of VpTM_(6-2) as triggers in N.benthamiana through VIGS.VpTM_(6-2) belongs to the B-class MADS-box genes.The VpTM_(6-2) trigger sequence is 357 bp long and has 69.0%identity at the nucleotide level with the putative target sequences of NbTM_(6),and there is a 27-nucleotide stretch in VpTM_(6-2) and NbTM_(6) showing 100%identity with one base mismatch.The result showed that the NbTM_(6) is effectively knocked down by the VpTM_(6-2) trigger sequence in N.benthamiana.The pollen viability in N.benthamiana flowers is reduced,and the expression of NbTM_(6),which has the highest identity with VpTM_(6-2),is significantly reduced in the abnormal flowers of N.benthamiana compared to the wild-type flowers.This work provides a system that heterologous sequence-triggered VIGS is an efficient way to investigate functional conservation of orthologous genes.The study further showed that the decline of VpTM_(6-2) expression in floral tissue of V.philippica is related with the CH-CL flower development.

关 键 词：紫花地丁 本氏烟草 VIGS VpTM_(6-2) 功能研究 

分 类 号：R285[医药卫生—中药学]