Ectopic Expression of PgF3′5′H in Commercial Gypsophila paniculata Cultivar through Optimized Agrobacterium-Mediated Transformation
Ectopic Expression of PgF3′5′H in Commercial Gypsophila paniculata Cultivar through Optimized Agrobacterium-Mediated Transformation
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DOI：
10.3390/horticulturae9030321
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发表时间：
2023-03
期刊：
影响因子：
3.1
通讯作者：
Chunlian Jin;Dan Sun;Lulin Ma;Xijun Mo;Chunmei Yang;Fan Li
中科院分区：
文献类型：
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作者：
Chunlian Jin;Dan Sun;Lulin Ma;Xijun Mo;Chunmei Yang;Fan Li
满天星是世界上最受欢迎的切花之一，其主要栽培品种为白色花。本研究以野生型银杏品种YX 4为材料，建立了银杏叶片的再生体系和遗传转化体系，并对F3′ 5′H基因在野生型银杏叶片中的表达进行了研究。将桔梗F3′5′H（PgF 3 ′ 5 ′H）成功地导入到‘YX 4’中。共获得潮霉素抗性植株281株，其中38株PCR阳性，转化效率为13.5%。无分生组织的茎尖再生能力强，更适合作为外植体，添加噻苯隆（TDZ）对不定芽的诱导效果最好，而添加6-苄基腺嘌呤（6-BA）和1-萘乙酸（NAA）对不定芽的诱导效果不明显。此外，预培养1d、共培养5d、侵染10 min、附加30 mg·L-1乙酰丁香酮（AS）和10 mg·L-1 Hyg筛选的组合形成了“YX 4”转化的优化体系。利用农杆菌介导法对我国优良农作物品种‘YX 4’进行遗传转化，不仅为优良农作物品种的创制和改良做出了贡献，而且为农杆菌介导法的推广应用奠定了基础。栽培品种的研究，同时也对重要观赏性状相关基因的功能研究具有重要意义。
Gypsophila paniculata is one of the most popular cut flowers in the world whose major cultivars are blooming white. As is well known, blue flowers could be generated via the overexpression of the gene encoding flavonoid 3′5′-hydroxylase (F3′5′H) in species that naturally lack it. In this study, we established the regeneration and a genetic transformation system for the commercial cultivar ‘YX4’ of G. paniculata and introduced the F3′5′H of Platycodon grandiflorus (PgF3′5′H) successfully into ‘YX4’ using the established protocol. A total of 281 hygromycin (Hyg)-resistant plantlets were obtained, and 38 of them were polymerase chain reaction (PCR) positive, indicating a 13.5% transformation efficiency. Shoot apex without meristem was more suitable for explant due to its high regeneration capacity, and the supplement of thidiazuron (TDZ) provided the most efficient promotion of adventitious bud induction, whereas the supplement of 6-Benzyladenine (6-BA) and 1-naphthaleneacetic acid (NAA) did not affect much. Additionally, the combination of 1 day (d) pre-culture, 5 d co-culture, 10 min infection, 30 mg·L−1 additional acetosyringone (AS) supplement, and 10 mg·L−1 Hyg selection formed the optimized system for ‘YX4’ transformation. This reliable and efficient agrobacterium-mediated transformation of the valuable commercial cultivar ‘YX4’ will contribute not only to the creation and improvement of G. paniculata cultivars, but also to the function research of genes associated with important ornamental traits.