Stable plastid transformation in kiwifruit (Actinidia chinensis)

IF 5 4区农林科学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY aBIOTECH Pub Date : 2024-11-28 DOI:10.1007/s42994-024-00186-0

Qiqi Chen, Yuyong Wu, Yanchang Wang, Jiang Zhang, Shengchun Li

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Abstract

Plastid transformation offers valuable benefits in plant biotechnology, such as high-level transgene expression and the absence of gene silencing. Here we describe the first protocol of a plastid transformation system for a woody vine (liana) kiwifruit (Actinidia chinensis). The transgenic DNA carries a spectinomycin-resistance gene (aadA) cassette and a green fluorescent protein (GFP) reporter gene cassette, flanked by two adjacent kiwifruit plastid genome sequences, thereby allowing targeted insertion between the trnfM and trnG genes. Six spectinomycin-resistant shoots were obtained out of 12 plates subjected to bombardment, and two were positive events, confirmed through PCR and Southern blot analyses. The GFP was localized to plastids as monitored by confocal laser scanning microscopy and reached 2.5% of leaf total soluble protein. Success in kiwifruit extends transplastomic technology of woody species beyond poplar, and will provide an attractive biosynthetic chassis for molecular farming.

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猕猴桃稳定质体转化的研究

质体转化为植物生物技术提供了宝贵的优势，如高水平的转基因表达和基因沉默的缺失。本文描述了木质藤本猕猴桃（猕猴桃）质体转化系统的第一个方案。该转基因DNA携带一个大霉素耐药基因（aadA）盒和一个绿色荧光蛋白（GFP）报告基因盒，两侧是两个相邻的猕猴桃质体基因组序列，从而可以在trnfM和trnG基因之间进行靶向插入。经PCR和Southern blot分析证实，12个靶板中有6个对观菌素有抗性，其中2个为阳性。通过共聚焦激光扫描显微镜检测，GFP定位于质体中，占叶片可溶性蛋白总量的2.5%。猕猴桃的成功推广了杨树以外木本植物的转质体技术，并将为分子农业提供一个有吸引力的生物合成基础。

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来源期刊

aBIOTECH

CiteScore

7.70

自引率

2.80%

发文量