Stable plastid transformation in kiwifruit(Actinidia chinensis)OA
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.
Qiqi Chen;Yuyong Wu;Yanchang Wang;Jiang Zhang;Shengchun Li
Key Laboratory of Plant Germplasm Enhancement and Specialty Agriculture,Wuhan Botanical Garden,Chinese Academy of Sciences,Wuhan 430074,ChinaSchool of Pharmacy,Binzhou Medical University,Yantai 264003,ChinaKey Laboratory of Plant Germplasm Enhancement and Specialty Agriculture,Wuhan Botanical Garden,Chinese Academy of Sciences,Wuhan 430074,ChinaState Key Laboratory of Biocatalysis and Enzyme Engineering,School of Life Sciences,Hubei University,Wuhan 430062,China Shenzhen Branch,Guangdong Laboratory of Lingnan Modern Agriculture,Key Laboratory of Synthetic Biology,Ministry of Agriculture and Rural Affairs,Agricultural Genomics Institute at Shenzhen,Chinese Academy of Agricultural Sciences,Shenzhen 518000,ChinaState Key Laboratory of Biocatalysis and Enzyme Engineering,School of Life Sciences,Hubei University,Wuhan 430062,China
农业科技
Actinidia chinensisWoody vineTransplastomic technologyaadAGreen fluorescent protein
《aBIOTECH》 2025 (1)
P.72-80,9
supported by grants from the National Natural Science Foundation of China(32271912,32071477).
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