陆地棉GhNFD4在棉花干旱响应中的功能  

Functional Studies of the Gossypium hirsutum GhNFD4 Gene in Response to Drought in Cotton

作　　者：马利花 侯梦娟 朱新霞[1] MA Li-hua;HOU Meng-juan;ZHU Xin-xia(School of Life Sciences,Shihezi University,Key Laboratory of Oasis Town and Mountain Basin System Ecology Corps,Shihezi 832003)

机构地区：[1]石河子大学生命科学学院绿洲城镇与山盆系统生态兵团重点实验室,石河子832003

出　　处：《生物技术通报》2025年第3期104-111,共8页Biotechnology Bulletin

基　　金：国家自然科学基金项目(32360457)。

摘　　要：【目的】分析GhNFD4在陆地棉响应干旱胁迫中的生物学功能,为陆地棉抗逆分子育种提供基因资源和理论依据。【方法】使用实时荧光定量PCR (RT-qPCR)技术检测棉花幼苗在PEG模拟的自然干旱胁迫下的GhNFD4表达模式;利用烟草脆裂病毒(TRV)诱导的基因沉默技术(VIGS)探究GhNFD4在干旱胁迫中的调控作用,克隆GhNFD4干扰序列和构建基因沉默载体,并通过瞬时转染技术侵染棉花幼苗进一步获取GhNFD4沉默植株,观察沉默植株与对照植株在自然失水15 d的表型差异,采用NBT和DAB染色法检测ROS产物的积累,并检测离体叶片的失水率和抗旱相关生理指标等。【结果】GhNFD4的表达受PEG模拟的自然干旱胁迫诱导,并在胁迫12 h表达量达到了顶峰。自然干旱15 d后,与阴性对照植株相比,GhNFD4沉默植株叶片萎蔫较为严重,且叶片在NBT和DAB染色后着色更深,其离体叶片失水率及干旱胁迫后叶片的相对电导率(REL)、丙二醛(MDA)和过氧化氢(H_(2)O_(2))含量与对照组相比均显著升高,总抗氧化物酶(T-AOC)活性与对照组相比显著下降。【结论】GhNFD4受干旱胁迫诱导,在干旱条件下通过调节抗氧化酶活性参与棉花对干旱胁迫的正向调控,改善棉花对干旱胁迫的抵抗能力。【Objective】To analyze the biological function of GhNFD4 in response to drought stress in terrestrial cotton,and to provide genetic resources and theoretical basis for molecular breeding of terrestrial cotton for stress tolerance.【Method】Real-time fluorescence quantitative PCR(RT-qPCR)was used to detect the expression pattern of GhNFD4 in cotton seedlings under PEG-mimicked natural drought stress;tobacco crisped virus(TRV)-induced gene silencing(VIGS)was utilized to explore the regulatory role of GhNFD4 in drought stress,and the interfering sequences of GhNFD4 were cloned and the gene silencing vectors were constructed.The GhNFD4 silencing plants were further obtained by infiltrating cotton seedlings through transient transfection technology,observing the phenotypic differences between silencing plants and control plants at 15 d of natural water loss,detecting the accumulation of ROS products by using NBT and DAB staining,and detecting the water loss rate and drought-related physiological indexes of isolated leaves.【Result】The expression of GhNFD4 was induced by natural drought stress simulated by PEG and peaked at the 12th h of stress.After 15 d of natural drought,the leaves of GhNFD4-silenced plants showed more severe wilting and darker coloring after NBT and DAB staining compared with the negative control plants,and the water loss rate of the isolated leaves and the relative electrical conductivity(REL),malondialdehyde(MDA),and hydrogen peroxide(H2O_(2))contents of the leaves after drought stress were significantly higher compared with the control,and the total antioxidant enzyme(T-AOC)activity was significantly lower compared with the control activity was significantly decreased compared with the control group.【Conclusion】GhNFD4 is induced by drought stress and actively participates in the positive regulation of drought stress in cotton by regulating the activity of antioxidant enzymes under drought conditions,which improves the resistance of cotton to drought stress.

关 键 词：棉花 GhNFD4 干旱 基因表达 VIGS 

分 类 号：S43[农业科学—农业昆虫与害虫防治]