不同VIGS体系在棉花中沉默效率的研究

Silencing Efficiency of Different VIGS Systems in Cotton

【目的】比较不同病毒诱导的基因沉默(VIGS)体系在棉花中的沉默效率。【方法】本研究以35S启动子驱动的抗卡那霉素基因nptⅡ为目标基因,利用RNA病毒载体TRV和DNA病毒载体CLCrV构建基因沉默载体,研究这2套VIGS体系在棉花中的基因沉默效率和沉默持续时间的差异。【结果】TRV体系比CLCrV体系沉默效率高,TRV体系比CLCrV体系沉默持续时间短,CLCrV体系能够在棉花蕾、花、花药等生殖器官中发挥沉默作用。【结论】TRV病毒沉默体系主要适用于短期的棉花内源基因的沉默,CLCrV病毒沉默体系适用于研究棉花生长发育后期生殖生长相关的基因功能。本研究为棉花选取合适的VIGS体系提供参考,对研究棉花基因功能具有重要意义。

【Objective】The present paper aimed to compare the silencing efficiency of different virus-induced gene silencing systems in cotton.【Method】35 S promoter-driven nptⅡof kanamycin resistance gene was used as target genes to construct two gene silencing vector systems TRV-VIGS(RNA virus vectors)and CLCrV-VIGS(DNA virus vectors)to study the differences in gene silencing efficiency and silencing duration between the two VIGS systems in cotton.【Result】The TRV system was more efficient than the CLCrV system in the short term,and the TRV system was less efficient than the CLCrV system on silencing duration.The CLCrV system was able to silence the reproductive organ related genes such as flower bud,flower,anther and so on.【Conclusion】TRV virus silencing system is suitable for inducing the silencing of endogenous genes in early cotton plants,and CLCrV virus silencing system is suitable for studying the silencing of endogenous genes related to reproductive growth that play a long-term role in cotton growth and development.This study provides a reference for the selection of suitable VIGS systems for cotton,which is of great significance to the study of cotton gene function.