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中国农学通报

中国农学通报 ›› 2024, Vol. 40 ›› Issue (30): 40-47.doi: 10.11924/j.issn.1000-6850.casb2024-0328

• 林学·园艺·园林 • 上一篇    下一篇

瓜类作物CGMMV-VIGS体系的优化研究

万丽丽1(), 王转茸1, 汤谧1(), 张学军2,3, 任俭1, 曾红霞1, 张娜1, 卫佳琪1, 熊建顺1   

  1. 1 武汉市农业科学院，武汉 430345
    2 新疆农业科学院哈密瓜研究中心，乌鲁木齐 830091
    3 新疆农业科学院海南三亚农作物育种试验中心，海南三亚 572014
  • 收稿日期:2024-05-17 修回日期:2024-07-15 出版日期:2024-10-25 发布日期:2024-10-23
  • 通讯作者:
    汤谧，1981年出生，湖北武汉人，正高级农艺师，博士，研究方向：西甜瓜新品种选育及推广。通信地址：430345 武汉市黄陂区武湖农业生态园农科院作物研究所，Tel：027-61805671，E-mail：。
  • 作者简介:

    万丽丽，女，1982年出生，湖北武汉人，高级农艺师，博士，研究方向：西甜瓜生物技术育种。通信地址：430345 湖北武汉黄陂区武湖农业生态园武汉市农业科学院北部园区，作物研究所西甜瓜研究室，E-mail：。

  • 基金资助:
    湖北省重点研发项目“瓜果类蔬菜作物CRISPR基因编辑技术研发”(2022BBA0062); 武汉市生物育种关键技术及新品种培育重大专项(2022021302024852); 湖北省农业农村厅湖北省支持种业高质量发展资金项目“地方特色经济作物新品种选育与推广”(HBZY2023B004-5); 武汉市农业科学院2024年创新项目“西甜瓜抗病优异种质资源创制的研究”(XKCX202404-1); 天山创新团队计划“哈密瓜生物技术育种创新团队”(2022D14015)

Optimization of CGMMV-VIGS System in Cucurbits

WAN Lili1(), WANG Zhuanrong1, TANG Mi1(), ZHANG Xuejun2,3, REN Jian1, ZENG Hongxia1, ZHANG Na1, WEI Jiaqi1, XIONG Jianshun1   

  1. 1 Wuhan Academy of Agricultural Sciences, Wuhan 430345
    2 The Research Center of Hami-melon, Xinjiang Academy of Agricultural Sciences, Urumqi 830091
    3 Key Laboratory of Crop Genetic Breeding in Hainan, Xinjiang Academy of Agricultural Sciences, Sanya, Hainan 572014
  • Received:2024-05-17 Revised:2024-07-15 Published:2024-10-25 Online:2024-10-23

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摘要：

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。

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

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.

Key words: cucumber green mottle mosaic virus (CGMMV-VIGS), gene silencing, PDS silencing EGSL, cucurbits, seed inoculation, temperature, relative humidity