通过辅助方法优化农杆菌介导的棉花胚尖遗传转化

doi:10.19586/j.2095-2341.2024.0201

生物技术进展 ›› 2025, Vol. 15 ›› Issue (3): 495-501.DOI: 10.19586/j.2095-2341.2024.0201

通过辅助方法优化农杆菌介导的棉花胚尖遗传转化

丁璇¹^,²(), 张祖英¹^,², 苏秀娟¹(), 胡文冉²()

^1.新疆农业大学农学院，乌鲁木齐 830052
^2.新疆维吾尔自治区农业科学院生物育种实验室，乌鲁木齐 830091

收稿日期:2024-12-19 接受日期:2025-02-12 出版日期:2025-05-25 发布日期:2025-07-01
通讯作者: 苏秀娟,胡文冉
作者简介:丁璇 E-mail：dx_0201@163.com
基金资助:
新疆维吾尔自治区自然科学基金项目(2022D01A88);新疆维吾尔自治区重点实验室开放课题(2021D04002)

Optimization of Agrobacterium-mediated Transformation in Cotton Embryonic Tips Through Auxiliary Method

Xuan DING¹^,²(), Zuying ZHANG¹^,², Xiujuan SU¹(), Wenran HU²()

^1.Agricultural College，Xinjiang Agricultural University，Urumqi 830052，China
^2.Biological Breeding Laboratory，Xinjiang Uygur Autonomous Region Academy of Agricultural Sciences，Urumqi 830091，China

Received:2024-12-19 Accepted:2025-02-12 Online:2025-05-25 Published:2025-07-01
Contact: Xiujuan SU,Wenran HU

摘要/Abstract

摘要：

棉花遗传转化过程中因受基因型限制、转化周期长等诸多因素制约，导致转化效率较低。以陆地棉TM-1的胚尖作为受体实验材料，利用GhTCP4基因编辑载体，通过农杆菌介导法对其进行侵染，对转化植株恢复培养基中添加的外源激素6-芐氨基嘌呤（6-benzylaminopurine, 6-BA）与萘乙酸（naphthalene acetic acid, NAA）的浓度进行探究，最终确定恢复培养基中不添加外源激素。在此基础上分别采用超声波、真空渗透法、真空渗透法和超声波相结合的辅助方式，研究辅助手段对棉花胚尖遗传转化效率的影响。结果发现超声频率40 kHz，额定功率300 W处理15 min时转化效率最高，可达到26.86%。研究旨在为建立高效棉花胚尖转化体系提供思路，为其他功能基因研究和遗传转化奠定基础。

关键词: 棉花胚尖, 遗传转化, 辅助手段, 超声波, 真空渗透

Abstract:

The genetic transformation of cotton is constrained by multiple limiting factors， including genotype restrictions and protracted transformation cycles， resulting in low transformation efficiency. In this study， embryonic tips of upland cotton TM-1 were selected as recipient material for Agrobacterium-mediated transformation using a GhTCP4 gene editing vector. We systematically investigated the effects of different concentrations of exogenous hormones 6-benzylaminopwrine （6-BA） and naphthalene acetic acid （NAA） in plant regeneration media， ultimately determining that hormone-free medium yielded optimal recovery. Furthermore， three auxiliary treatments， ultrasonication， vacuum infiltration， and their combination， were evaluated for their impacts on transformation efficiency. The results demonstrated that 15 minute ultrasonic treatment at 40 kHz frequency with 300 W rated power achieved the highest transformation efficiency of 26.86%. This study provided methodological insights for establishing an efficient cotton embryonic tip transformation system and offered a foundation for future functional gene research and genetic transformation applications in cotton.

Key words: cotton embryonic tips, genetic transformation, complementary method, sonication, vacuum infiltration

中图分类号:

Q789

丁璇, 张祖英, 苏秀娟, 胡文冉. 通过辅助方法优化农杆菌介导的棉花胚尖遗传转化[J]. 生物技术进展, 2025, 15(3): 495-501.

Xuan DING, Zuying ZHANG, Xiujuan SU, Wenran HU. Optimization of Agrobacterium-mediated Transformation in Cotton Embryonic Tips Through Auxiliary Method[J]. Current Biotechnology, 2025, 15(3): 495-501.

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通过辅助方法优化农杆菌介导的棉花胚尖遗传转化

Optimization of Agrobacterium-mediated Transformation in Cotton Embryonic Tips Through Auxiliary Method

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