本研究旨在提高黄瓜绿斑驳花叶病毒(cucumber green mottle mosaic virus,CGMMV)介导的基因沉默(VIGS)在瓜类作物中应用效率.以黄瓜、甜瓜和西瓜优良种质为实验材料,采用不同的CGMMV病毒载体接种方式、并设置不同的培养温度和相对湿度,以测试基因沉默的有效性.结果表明,真空渗透和种子吸胀的接种方式能够在瓜类作物中产生最高的基因沉默率(FGS).在22℃和25℃的培养环境下,黄瓜、甜瓜和西瓜PDS沉默有效性(EGSL)较高.黄瓜、甜瓜和西瓜生长1个月后,EGSL达到100％,显著高于30℃的培养环境下EGSL.生长3个月后,甜瓜和西瓜在22℃培养环境下EGSL分别为89.7％和95％;在25℃培养环境下EGSL分别为85.6％和86.1％,均显著性高于30℃下EGSL.在相对湿度为30％和50％环境下,黄瓜的EGSL分别为70％和72％,甜瓜的EGSL分别为76％和73％,显著高于相对湿度为80％的培养环境下EGSL.西瓜在相对湿度为30％的培养环境下的EGSL为69％,显著高于相对湿度为50％时的EGSL(38％)和相对湿度在80％时的EGSL(33％).综上所述,通过优化了瓜类作物的CGMMV-VIGS的技术体系中接种方式和培养环境参数,提高了基因沉默率和有效性,并且能够快速获得整个植株基因沉默的种质资源,为作物优质和抗逆基因功能的研究提供有效途径.
Abstract
This study aims to improve the efficiency of cucumber green mottle mosaic virus(CGMMV)-mediated virus-induced gene silencing(VIGS)in cucurbit crops.Using germplasms from cucumber,melon and watermelon,effectiveness in gene silencing was tested by using various inoculation methods with CGMMV-VIGS vector,cultivation temperatures,and relative humidity in incubator.The results demonstrated that vacuum infiltration and seed imbibition were the most effective inoculation methods,achieving the highest frequency of gene silencing(FGS)in cucurbit crops.Cucumber,melon,and watermelon showed high effectiveness of PDS silencing EGSL(100％)at the temperatures of 22℃ and 25℃ after one month of growth,significantly higher than the EGSL at 30℃.After three months,the EGSL for melon and watermelon were 89.7％and 95％,respectively at 22℃ and 85.6％and 86.1％at 25℃,both significantly higher than that at 30℃.With 30％and 50％relative humidity in incubator,the EGSL for cucumber were 70％and 72％,respectively,which were significantly higher than the EGSL at 80％humidity.For watermelon,the EGSL at 30％relative humidity was 69％,significantly higher than the EGSL at 50％humidity(38％)and 80％humidity(33％).Overall,by optimizing the inoculation methods and environmental parameters in the CGMMV-VIGS,we have improved the gene silencing rate and effectiveness in cucurbit crops.This method will facilitate rapid acquisition of VIGS germplasm resources for research about crop quality and abiotic stress resistance.
关键词
黄瓜绿斑驳花叶病毒/基因沉默/PDS沉默有效性/瓜类作物/种子接种/温度/相对湿度
Key words
cucumber green mottle mosaic virus(CGMMV-VIGS)/gene silencing/PDS silencing EGSL/cucurbits/seed inoculation/temperature/relative humidity
维普
万方数据