摘要
【目的】突破棉花VIGS技术仅应用于棉花子叶期的时间限制瓶颈,弥补棉花生殖生长时期VIGS技术的应用缺陷。【方法】以农大棉8号为材料,以GhCLA1为验证基因,对4日龄棉苗(子叶未展开)进行茎注射,对7日龄棉苗(子叶完全展开)进行子叶注射,对开花期和结铃期的棉花植株分别在茎节间、新生果枝基部、新生叶片等组织部位进行VIGS注射,利用表型观察和RT-PCR技术验证目的基因的沉默效率和持续时间。【结果】4日龄棉花幼苗经茎VIGS注射后叶片白化率为80%,7日龄棉花幼苗经子叶VIGS注射后叶片白化率为100%,开花期和结铃期的棉花植株经新生果枝基部VIGS注射后白化率分别为90%和68.75%。【结论】获得了稳定的棉花全生长周期的VIGS应用技术体系。
【Objective】To break through the time limit bottleneck that cotton VIGS technology can only be applied in cotton cotyledon stage,and make up for the application deficiency of VIGS technology in cotton reproductive growth stage.【Method】Using Nongdamian 8 as the material and GhCLA1 as the verification gene,4-day-old cotton seedlings’stem(cotyledon folded)were injected,7-day-old cotton seedlings cotyledon(cotyledon fully unfolded)were injected,also flowering and bolling stages cotton stem and base of new fruit branches and new leaves were injected with VIGS respectively.Phenotype observation and RT-PCR were used to verify the silencing efficiency of target genes and photobleaching duration.【Result】The leaf photobleaching rate of 4-day-old cotton seedlings was 80%by VIGS injected to stems,that of 7-day-old cotton seedlings was 100%by VIGS injected to cotyledon,and that of flowering stage and bolling stage cotton was 90%and 68.75%by VIGS injection at the base of new fruit branches,respectively.【Conclusion】A stable VIGS application technology system for the whole growth cycle of cotton is obtained.
作者
丁若羲
豆硕
安叶芝
孔文慧
郭文静
张冬梅
王省芬
马峙英
吴立柱
DING Ruo-xi;DOU Shuo;AN Ye-zhi;KONG Wen-hui;GUO Wen-jing;ZHANG Dong-mei;WANG Xing-fen;MA Zhi-ying;WU Li-zhu(Hebei Agricultural University,Co-Innovation Center for Cotton Industry of Hebei Province,State Key Laboratory of North China Crop Improvement and Regulation,Baoding 071000)
出处
《生物技术通报》
北大核心
2025年第2期58-64,共7页
Biotechnology Bulletin
基金
国家自然科学基金项目(32072109)
河北省中央引导地方科技发展资金项目(236Z2908G)。
关键词
基因沉默
棉花
子叶
茎
生长期
VIGS
cotton
cotyledon
stem
growing period
