利用病毒介导基因沉默方法研究小麦抗光氧化相关基因

doi:10.3724/SP.J.1006.2014.01905

摘要/Abstract

摘要：

为了鉴定可能的小麦抗光氧化基因，利用大麦条斑病毒(barley stripe mosaic virus，BSMV)介导的基因沉默(virus-induced gene silencing, VIGS)系统，对6个小偃54响应强光的基因进行了沉默表达研究。以BSMV:GFP植株为对照，分析了这些基因的减量表达植株在低温强光、DCMU、MV及H₂O₂等处理下的PSI最大光化学效率(F_v/F_m)和光合性能指数(P.I.)、MDA含量及整株生物量变化。结果显示，Ta23008和Ta92165均参与小麦对低温强光、DCMU、MV和H₂O₂等胁迫的响应过程；Ta106078参与小麦对DCMU、MV和H₂O₂等胁迫的响应过程；Ta27787参与小麦对低温强光、DCMU和H₂O₂等胁迫的响应过程；Ta24695参与小麦对低温强光和H₂O₂胁迫的响应过程；而Ta119251仅参与小麦对DCMU的响应过程。此外，Ta23008、Ta92165、Ta106078、Ta119251四个基因被抑制表达后，其转化株系的生物量比对照显著降低，推测其可能参与调控小麦生物量的积累。

关键词: 小麦, 大麦条斑病毒(BSMV), 病毒介导的基因沉默, 基因功能, 光氧化

Abstract:

Functional analysis of photo-oxidative stress responsive genes in wheat (Triticum aestivum L.) may benefit wheat improvement for high radiation use efficiency. A Chinese variety Xiaoyan 54 developed from distant hybridization between common wheat (T. aestivum, 2n=42) and tall wheatgrass (Thinopyrum ponticum, 2n=70) shows significant tolerance to high light induced photo-oxidative stress. Based on previous transcriptome analysis of Xiaoyan 54 in response to high light stress, six genes were selected in this study to assess their possible roles in photo-oxidative stress response using barley stripe mosaic virus (BSMV) mediated virus-induced gene silencing (VIGS) system in Xiaoyan 54. The BSMV induced silencing of the targeted genes together with the BSMV:GFP control plants were exposed to low temperature and high lightII (F_v/F_m), the photosynthetic performance index (P.I.), malondialdehyde (MDA) content, and biomass were evaluated. The results showed that Ta23008 and Ta92165 were involved in the responses of wheat to LTHL, DCMU, MV, and H₂O₂, respectively. Ta106078 was responsible for wheat tolerance to DCMU, MV, and H₂O₂ while Ta27787 was responsible for LTHL, DCMU, and H₂O₂ stress. Ta24695 participated in the response of wheat to both LTHL and H₂O₂. However, Ta119251 seemed to be only responsible for the DCMU stress in wheat. Additionally, four genes, Ta23008, Ta92165, Ta106078 and Ta119251, were likely to regulate biomass accumulation because the biomass was significantly reduced when they were silenced in wheat. These results provided new hints toward understanding the molecular mechanism of tolerance to photo-oxidative stress in Xiaoyan 54.(LTHL), N-(3,4-dichlorophenyl)- N’,N’-dimethylurea(DCMU), methylviologen (MV), and hydrogen peroxide (H₂O₂) stress, respectively. The maximum photochemical efficiency of photosystem

Key words: Triticum aestivum, Barley stripe mosaic virus, VIGS, Genomic function, Photo-oxidative stress

陈坤梅,李宏伟,林凡云,陈耀锋,李滨,郑琪,李振声. 利用病毒介导基因沉默方法研究小麦抗光氧化相关基因[J]. 作物学报, 2014, 40(11): 1905-1913.

CHEN Kun-Mei,LI Hong-Wei,LIN Fan-Yun,CHEN Yao-Feng,LI Bin,ZHENG Qi,LI Zhen-Sheng. Functional Analysis of Photo-Oxidative Stress Responsive Genes in Wheat Using Virus-Induced Gene Silencing System[J]. Acta Agron Sin, 2014, 40(11): 1905-1913.

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