Establishing a highly efficient Agrobacterium-mediated transformation system in sweet buckwheat

IF 1.4 3区农林科学 Q4 FOOD SCIENCE & TECHNOLOGY

Journal of Consumer Protection and Food Safety Pub Date : 2023-06-01 DOI:10.1007/s00003-023-01444-x

Binhan Zhou, Fugong Ding, Zhu Yang, Zhenzhen Song, Ji Sun, Shuping Wang, Xue Wang, Zhixiong Liu, Zhengwu Fang, Yingxin Zhang

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引用次数: 1

Abstract

Buckwheat (Fagopyrum esculentum Moench) is a healthy cereal with a short lifecycle, and an efficient transgenic method for gene function analysis of sweet buckwheat is lacking. In this study, we developed a simple Agrobacterium-mediated transformation protocol using a floral-dip. For this transformation experiment, the Agrobacterium strain GV3101 harboring binary vector pHZM69 with enhanced green fluorescent protein (eGFP) as well as a resistance gene to the herbicide Basta was used. The effect of different cell densities (OD₆₀₀ = 0.4, 0.6, 0.8 and 1.0) applied for infection was measured, and maximum transformation efficiency (12.42%) was obtained with cell density of OD₆₀₀ = 0.6, followed by OD₆₀₀ = 0.4 (8.48%), OD₆₀₀ = 0.8 (4.39%) and OD₆₀₀ = 1.0 (3.62%). Here, sweet buckwheat was successfully transformed by Agrobacterium-mediated floral dip method, which could be a useful tool for the production of transgenic sweet buckwheat and promote genomic function analysis in sweet buckwheat.

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建立高效农杆菌介导的甜荞麦转化体系

荞麦(Fagopyrum esculentum Moench)是一种生命周期短的健康谷物，缺乏一种有效的转基因方法来分析甜荞麦的基因功能。在这项研究中，我们开发了一种简单的农杆菌介导的转化方案，使用花浸渍。在本转化实验中，采用农杆菌菌株GV3101，该菌株携带具有增强型绿色荧光蛋白(eGFP)和抗除草剂Basta基因的二元载体pHZM69。测定不同细胞密度(OD600 = 0.4、0.6、0.8和1.0)对感染的影响，当细胞密度为OD600 = 0.6时，转化效率最高(12.42%)，其次为OD600 = 0.4(8.48%)、OD600 = 0.8(4.39%)和OD600 = 1.0(3.62%)。本研究通过农杆菌介导的花浸渍法成功转化了甜荞麦，为生产转基因甜荞麦和促进甜荞麦基因组功能分析提供了有益的工具。

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来源期刊

Journal of Consumer Protection and Food Safety Veterinary-Food Animals

CiteScore

3.70

自引率

4.20%

发文量

审稿时长

>12 weeks

期刊介绍： The JCF publishes peer-reviewed original Research Articles and Opinions that are of direct importance to Food and Feed Safety. This includes Food Packaging, Consumer Products as well as Plant Protection Products, Food Microbiology, Veterinary Drugs, Animal Welfare and Genetic Engineering. All peer-reviewed articles that are published should be devoted to improve Consumer Health Protection. Reviews and discussions are welcomed that address legal and/or regulatory decisions with respect to risk assessment and management of Food and Feed Safety issues on a scientific basis. It addresses an international readership of scientists, risk assessors and managers, and other professionals active in the field of Food and Feed Safety and Consumer Health Protection. Manuscripts – preferably written in English but also in German – are published as Research Articles, Reviews, Methods and Short Communications and should cover aspects including, but not limited to: · Factors influencing Food and Feed Safety · Factors influencing Consumer Health Protection · Factors influencing Consumer Behavior · Exposure science related to Risk Assessment and Risk Management · Regulatory aspects related to Food and Feed Safety, Food Packaging, Consumer Products, Plant Protection Products, Food Microbiology, Veterinary Drugs, Animal Welfare and Genetic Engineering · Analytical methods and method validation related to food control and food processing. The JCF also presents important News, as well as Announcements and Reports about administrative surveillance.