Establishment of a picloram-induced somatic embryogenesis system in Vitis vinifera cv. chardonnay and genetic transformation of a stilbene synthase gene from wild-growing Vitis species
Establishment of a picloram-induced somatic embryogenesis system in Vitis vinifera cv. chardonnay and genetic transformation of a stilbene synthase gene from wild-growing Vitis species
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毒莠定诱导的葡萄体细胞胚胎发生系统的建立。
DOI：
10.1007/s11240-015-0711-9
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发表时间：
2015-05-01
影响因子：
3
通讯作者：
Wang, Yuejin
中科院分区：
文献类型：
--
作者：
Dai, Lingmin;Zhou, Qi;Wang, Yuejin
为了促进遗传改良，在葡萄品种中建立了有效的胚胎培养诱导、维持和转化系统。霞多丽是用毒莠定开发的。全花在含有 Murashige 和 Skoog's (MS) 基础盐和 3.0 mg L-1 毒莠定以及不同浓度的 2,4-二氯苯氧基乙酸和 6-苄基腺嘌呤的诱导培养基上产生最多的胚性愈伤组织。在含有MS基础盐和毒莠定的维持和增殖培养基（MPM）上培养的原胚团（PEM）在2个月内具有最高的增殖效率。此外，发现含有 MS 基础盐和 2.0 mg L-1 毒莠定的 MPM 形成最佳 PEM。然后将在 MPM 上培养的 PEM 转移到含有 MS 基础盐、0.2 mg L-1 激动素和 0.1 mg L-1 2-萘氧基乙酸的胚萌发培养基中，这是正常幼苗再生的最佳培养基。将来自中国野生葡萄的二苯乙烯合酶基因VpSTSgDNA2的单拷贝通过根癌农杆菌介导的转化转移到再生霞多丽中，并通过Southern印迹分析证实。阳性转基因葡萄品系比野生型葡萄树表现出更高水平的二苯乙烯和H2O2，并且可以稍微减少白粉病的生长。
To facilitate genetic improvement, an efficient system of embryogenic culture induction, maintenance and transformation in Vitis vinifera cv. Chardonnay was developed using picloram. Whole flowers produced the most embryogenic calluses on induction media containing Murashige and Skoog's (MS) basal salts and 3.0 mg L-1 picloram with different concentrations of 2,4-dichlorophenoxyacetic acid and 6-benzyladenine. Proembryonic masses (PEM) cultured on maintenance and proliferation media (MPM) containing MS basal salts and picloram had the highest proliferation efficiency within 2 months. Additionally, the best PEM formation was found with MPM containing MS basal salts and 2.0 mg L-1 picloram. Then this PEM cultured on MPM were transferred to embryo germination medium containing MS basal salts, 0.2 mg L-1 kinetin and 0.1 mg L-1 2-naphthoxyacetic acid, the optimal medium for normal plantlets regeneration. A single copy of the stilbene synthase gene VpSTSgDNA2 from Chinese wild Vitis pseudoreticulata was transferred into regenerated Chardonnay via Agrobacterium tumefaciens-mediated transformation and confirmed by Southern blot analysis. The positive transgenic grapevine lines exhibited higher levels of stilbene and H2O2 than wild-type vines and could slightly reduced the growth of powdery mildew.