Efficient plant regeneration and Agrobacterium-mediated transformation of Begonia semperflorens-cultorum
Efficient plant regeneration and Agrobacterium-mediated transformation of Begonia semperflorens-cultorum
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四季秋海棠的高效植物再生和农杆菌介导转化
DOI：
10.1007/s11240-020-01858-7
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Igawa Tomoko
中科院分区：
文献类型：
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作者：
Hirutani Sakiko;Shimomae Kazuki;Yaguchi Akira;Chin Dong Poh;Mii Masahiro;Igawa Tomoko
四花秋海棠，又称蜡秋海棠，是最受欢迎的海棠品种之一，已培育出不同的商业品种。该物种的进一步改良需要遗传转化技术，因为通过常规育种不能引入一些所需的性状，如新的花色。在这里，我们报道了利用四花秋海棠叶片进行高效植株再生和获得转基因植株的方法。外植体在MS附加1.5 mg/L噻二唑脲、0.5 mg/Lα-萘乙酸、30 mg/L蔗糖和2.5 mg/L结冷胶的培养基上能有效地诱导出不定芽。当共培养基中加入10 mM MES时，瞬时GUS表达显著增加。将侵染的外植体在10 mg/L潮霉素(HM)的选择压力下培养，获得了HM抗性的独立芽，频率为0.78个/外植体。再生植株中潮霉素磷酸转移酶(HPT)基因已整合到基因组中，GUS基因稳定表达，这是遗传转化的证据。因此，本研究建立的条件使四花秋海棠的转化成为可能。
Begonia semperflorens-cultorum, known as wax begonia, is one of the most popularBegoniaspecies in which variable commercial cultivars have been produced. The genetic transformation technique is required for further modification of this species because introduction of some desired traits such as novel flower colors cannot be achieved by conventional breeding. Here we report the procedures of an efficient plant regeneration from leaf segment and production of transgenic plants usingBegonia semperflorens-cultorum. Efficient induction of adventitious shoots was achieved when the explants were cultured on MS medium supplemented with 1.5 mg/l thidiazuron (TDZ), 0.5 mg/l α-naphthaleneacetic acid (NAA), 30 g/l sucrose and 2.5 mg/l gellan gum.Agrobacterium tumefaciensstrain EHA101 containing the plasmid pIG121-Hm was used for gene transfer. The transient GUS expression was significantly increased when 10 mM MES was added to the co-cultivation medium. By culturing the infected explants under the selection pressure with 10 mg/l hygromycin (Hm), the Hm-resistant independent shoots were obtained at a frequency of 0.78 per explant. The regenerated Hm-resistant plants showed the integration ofhygromycin phosphotransferase(HPT) gene into the genome and stable GUS expression, as the proof of genetic transformation. Consequently, the conditions established in this study enabled the transformation ofBegonia semperflorens-cultorum.