JIA-2021-2108 富平楸子高效再生体系和遗传转化体系的建立

doi:10.1016/j.jia.2022.07.023

摘要/Abstract

摘要：

本研究评估了影响富平楸子再生及遗传转化体系的因素，发现富平楸子最佳再生体系为使用叶片为外植体，对叶片进行横切，近轴面贴近包含有MS、30 g L^-1蔗糖、8 g L^-1琼脂、5 mg L^-1 6-BA、2 mg L^-1 TDZ 和1 mg L^-1 NAA，pH为5.8的培养基上暗培养三周后见光培养，最佳遗传转化体系为在根癌农杆菌重悬液中侵染8min，然后进行4天共培养及3天延迟培养，并且使用150 mg L^-1头孢噻肟和15 mg L^-1卡那霉素作为筛选压。本研究建立了一套有效的富平楸子再生及遗传转化体系，通过此体系可快速获得富平楸子转基因材料，对苹果生物学研究具有重要意义。

Abstract:

Malus prunifolia Borkh. ‘Fupingqiuzi’ has significant ecological and economic value and plays a key role in germplasm development and resistance research. However, its long juvenile phase and high heterozygosity are barriers to the identification of ‘Fupingqiuzi’ progeny with excellent traits. In-vitro regeneration techniques and Agrobacterium-mediated genetic transformation systems can efficiently produce complete plants and thus enable studies of gene function. However, optimal regeneration and genetic transformation systems for ‘Fupingqiuzi’ have not yet been developed. Here, we evaluated the factors that affect the in-vitro regeneration and transformation of ‘Fupingqiuzi’. The best results were obtained when transverse leaf sections were used as explants, and they were grown in dark culture for three weeks with their adaxial sides contacting the culture medium (MS basal salts, 30 g L⁻¹ sucrose, 8 g L⁻¹ agar, 5 mg L⁻¹ 6-benzylaminopurine (6-BA), 2 mg L⁻¹ thidiazuron (TDZ), and 1 mg L⁻¹ 1-naphthlcetic acid (NAA), pH 5.8). A genetic transformation system based on this regeneration system was optimized: after inoculation with A. tumefaciens solution for 8 min, 4 days of co-culture, and 3 days of delayed culture, the cultures were screened with cefotaxime (150 mg L⁻¹) and kanamycin (15 mg L⁻¹). We thus established an efficient regeneration and genetic transformation system for ‘Fupingqiuzi’, enabling the rapid production of transgenic material. These findings make a significant contribution to apple biology research

Key words: Malus. prunifolia Borkh , ‘Fupingqiuzi’ , regeneration system , Agrobacterium-mediated transformation system

. JIA-2021-2108 富平楸子高效再生体系和遗传转化体系的建立[J]. Journal of Integrative Agriculture, 2022, 21(9): 2615-2627.

LIU Yu-song, WANG Hong-ying, ZHAO Yong-juan, JIN Yi-bo, LI Chao, MA Feng-wang. Establishment of an efficient regeneration and genetic transformation system for Malus prunifolia Borkh. ‘Fupingqiuzi’[J]. Journal of Integrative Agriculture, 2022, 21(9): 2615-2627.

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