苏麦3号小麦穗部病毒诱导的基因沉默(VIGS)体系的建立及验证

Construction and validation of virus-induced gene silencing(VIGS) system in spike of wheat variety Sumai 3

为了探讨利用病毒诱导的基因沉默(VIGS)体系在小麦穗部进行基因沉默的可行性,用大麦条纹花叶病毒诱导苏麦3号小麦穗部PDS基因的转录后沉默,再接种禾谷镰刀菌进行抗性鉴定。结果表明,与接种空载BSMV∶00的对照相比,接种BSMV∶PDS的小麦穗中PDS基因表达量下调了77%(P<0.05)。接种BSMV∶00的小麦穗受到赤霉病菌侵染后,穗部平均病小穗数和DON毒素含量分别为2.75个和4.55μg/g,与接种无菌水对照无显著差异(P>0.05)。说明在苏麦3号穗部VIGS系统中,通过接种病毒能使靶标基因沉默;接种赤霉病菌后,苏麦3号病小穗数和DON毒素含量不受接种病毒的影响,对赤霉病菌的抗性保持不变,建立的VIGS体系可以应用于小麦赤霉病抗性候选基因的功能分析。

Virus-induced gene silencing (VIGS) was performed in the spikes of wheat variety Sumai3 by constructing VIGS vectors and silencing PDS, in order to explore the feasibility of gene silencing by VIGS system in spikes of wheat.The results showed that the expression level of PDS in the spikes inoculated by barley stripe mosaic virus(BSMV)∶PDS was reduced by 77% in comparison with that by BSMV∶00 (P<0.05).After infection with Fusarium asiaticum, the average number of diseased spikelets and DON content were 2.75 and 4.55 μg/g,respectively, showing no significant difference with the control group (P>0.05).The results suggested that the VIGS system for spike enabled the target gene silencing.The number of diseased spikelets and accumulation of DON toxin in Sumai 3 were not affected by the virus inoculation, and the FHB resistance remained unchanged, indicating that the VIGS system for Sumai3 could be applied to functional analysis of candidate genes for wheat FHB resistance.