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《江苏农业科学》[ISSN:1002-1302/CN:32-1214/S]

卷:

第51卷

期数:

2023年第7期

页码:

49-58

栏目:

生物技术

出版日期:

2023-04-05

文章信息/Info

Title:

Optimization of genetic transformation technology and application of DRs genes in melon

作者:

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; 孙玉宏¹; 朱志坤⁴

1.武汉市农业科学院，湖北武汉 430065； 2新疆农业科学院哈密瓜研究中心，新疆乌鲁木齐 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； 4.湖北省武汉市蔡甸区农业农村局，湖北武汉 430199

Author(s):

Wan Lili; et al

关键词:

甜瓜; 农杆菌浸染; 遗传转化; 真空渗透; 不定芽诱导; 发育调控基因

Keywords:

-

分类号:

S652.03

DOI:

-

文献标志码:

A

摘要:

甜瓜(Cucumis melo L.)是我国重要的设施栽培作物，具有重要的经济价值。随着农作物全基因组测序以及基因编辑技术的建立，遗传改良和基因功能验证亟需建立高效稳定的遗传转化体系。遗传转化率由基因型和植物激素互作所决定，目前，甜瓜遗传转化技术普遍存在遗传转化载体递送到细胞困难、芽再生难、遗传转化效率低等诸多问题。本研究选取5个厚皮甜瓜和4个薄皮甜瓜品种的子叶为外植体，对遗传转化试验中农杆菌浸染的方法、植物生长调节剂、筛选剂浓度等多个因素进行筛选，建立适合厚皮和薄皮甜瓜的遗传转化技术体系。结果表明，不定芽诱导最佳激素组合是1 mg/L 6-BA和 1 mg/L ABA，最佳Basta筛选浓度为5 mg/L。由于甜瓜原形成层细胞位于维管组织的最深层，为提高农杆菌浸染效率，本研究采用纳米刷对子叶外植体表面均匀造伤，接着超声波和真空渗透压3个处理方式组合，其中在纳米刷造伤后进行超声波处理20 s，-1.0 kPa真空渗透压力下处理90 s后，绿色荧光蛋白(GFP)标记显示在甜瓜外植体的转化效率为82%~95%，在不定芽诱导培养基(SIM，1 mg/L 6-BA和 1 mg/L ABA)能产生再生苗的外植体率为3%~10%。研究了GROWTH-REGULATING FACTOR5 (AtGRF5)、WUSHEL(AtWUS)、PLETHORA(AtPLT5)、WOUND INDUCED DEDIFFERENTIATION 1(AtWIND1)等发育调控因子(DRs)对甜瓜遗传转化再生的作用，选择4种甜瓜优良自交系种质转化含有DRs表达组件的遗传转化载体，之后在低浓度细胞分裂素的培养基上诱导不定芽再生，试验结果显示，超表达AtGRF5和AtPLT5能够提高转化率以及接种外植体的再生芽率。因此，开发并利用DRs能够有效改良甜瓜的遗传转化率，进而促进基因组编辑等生物技术在甜瓜育种中的应用。

Abstract:

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[1]夏瑾华,喻慧荣,邹如意,等.6-BA、赤霉素和水杨酸对甜瓜种子萌发的影响[J].江苏农业科学,2015,43(12):204.
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[2]吕琛,李锋霞,马本学,等.甜瓜成熟度检测技术研究进展[J].江苏农业科学,2014,42(01):244.
　Lv Chen,et al.Research progress of detection technology for maturity of Cucumis melo[J].Jiangsu Agricultural Sciences,2014,42(7):244.
[3]艾子凌,高鹏,杜黎黎,等.利用CAPS初步定位甜瓜MR-1白粉病抗性基因[J].江苏农业科学,2016,44(06):66.
　Ai Ziling,et al.Location of powdery mildew resistant gene using CAPS makers in melon MR-1[J].Jiangsu Agricultural Sciences,2016,44(7):66.
[4]邢卫峰,于侦云,陈刘军,等.生物肥料“宁盾”对甜瓜枯萎病的防治效果[J].江苏农业科学,2014,42(03):78.
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备注/Memo

备注/Memo:

收稿日期：2022-07-03
基金项目：湖北省重点研发项目(编号：2021BBA101、2022BBA0062)；天山创新团队计划(编号：2022D14015)；武汉市农业科学院创新项目(编号：XKCX202204-1)；武汉市生物育种关键技术及新品种培育重大专项(2022021302024852)。
作者简介：万丽丽(1982—)，女，湖北武汉人，博士，副研究员，主要从事蔬菜作物生物技术育种。E-mail：wanlili13226@163.com。
通信作者：汤谧，博士，高级农艺师，主要从事西甜瓜新品种研究、选育及推广。E-mail：xtgyjs@wuhanagri.com。

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更新日期/Last Update: 2023-04-05