农杆菌介导的商业甜瓜（Cucumis melo L.，cv. Amarillo Oro）转化。

Agrobacterium-mediated transformation of commercial melon (Cucumis melo L., cv. Amarillo Oro).

机构信息

Departamento de Genética y Producción Vegetal, Laboratorio Asociado de Agronomía y Medio Ambiente. Estación Experimental de Aula Dei, C.S.I.C., Av Montañana 177, E-50080, Zaragoza, Spain.

出版信息

Plant Cell Rep. 1994 Jan;13(3-4):145-8. doi: 10.1007/BF00239881.

DOI:10.1007/BF00239881

PMID:24193640

Abstract

Cotyledon explants of muskmelon (Cucumis melo L., cv. Amarillo Oro) seedlings were co-cultivated with disarmed Agrobacterium tumefaciens strain LBA4404 that contained the binary vector plasmid pBI121.1. The T-DNA region of this binary vector contains the Nopaline synthase/neomycin phosphotransferase II (NPTII) chimeric gene for kanamycin resistance and the Cauliflower Mosaic Virus 35S/β-glucuronidase (GUS) chimeric gene. After infection, the cotyledon pieces were placed in induction medium containing 100 mg/l kanamycin. Putative transformed shoots were obtained, followed by the development of morphologically normal plantlets. The transgenic nature of regenerants was demonstrated by polymerase chain reaction, Southern blot analysis, plant growth on medium selective for the transgene (NPTII) and expression of the co-transformed GUS gene. Factors affecting the transformation procedure are discussed.

摘要

甜瓜（Cucumis melo L.，cv. Amarillo Oro）幼苗子叶外植体与解除武装的根癌农杆菌 LBA4404 菌株共培养，该菌株含有二元载体质粒 pBI121.1。该二元载体的 T-DNA 区域包含卡那霉素抗性的胭脂碱合成酶/新霉素磷酸转移酶 II（NPTII）嵌合基因和花椰菜花叶病毒 35S/β-葡萄糖醛酸酶（GUS）嵌合基因。感染后，将子叶块放入含有 100mg/l 卡那霉素的诱导培养基中。获得了可能转化的芽，随后发育出形态正常的小植株。通过聚合酶链反应、Southern 印迹分析、对转基因（NPTII）选择的培养基上植物的生长以及共转化的 GUS 基因的表达，证明了再生体的转基因性质。讨论了影响转化过程的因素。