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通过农杆菌介导转化生产转基因百合植株。

Production of transgenic lily plants by Agrobacterium-mediated transformation.

作者信息

Hoshi Y, Kondo M, Mori S, Adachi Y, Nakano M, Kobayashi H

机构信息

Niigata Agricultural Research Institute, 857 Nagakura, 940-0826, Nagaoka, Niigata, Japan.

出版信息

Plant Cell Rep. 2004 Jan;22(6):359-64. doi: 10.1007/s00299-003-0700-z. Epub 2003 Sep 4.

Abstract

A system for the production of transgenic plants was developed for the Oriental hybrid lily, Lilium cv. Acapulco, by Agrobacterium-mediated genetic transformation. Filament-derived calli were co-cultivated with A. tumefaciens strain EHA101/pIG121Hm, which harbored a binary vector carrying the neomycin phosphotransferase II, hygromycin phosphotransferase, and intron-containing beta-glucuronidase genes in the T-DNA region. Six hygromycin-resistant (Hyg(r)) culture lines were obtained from 200 calli by scratching them with sandpaper prior to inoculation and using NH(4)NO(3)-free medium for co-cultivation and a hygromycin-containing regeneration medium for selection. Hyg(r) culture lines regenerated shoots, which developed into plantlets following transfer to a plant growth regulator-free medium. All of these plantlets were verified to be transgenic by GUS histochemical assay and inverse PCR analysis.

摘要

通过农杆菌介导的遗传转化,为东方杂交百合‘阿卡普尔科’(Lilium cv. Acapulco)开发了一种转基因植物生产系统。将花丝来源的愈伤组织与根癌农杆菌菌株EHA101/pIG121Hm共培养,该菌株携带一个双元载体,其T-DNA区域含有新霉素磷酸转移酶II、潮霉素磷酸转移酶和含内含子的β-葡萄糖醛酸酶基因。通过在接种前用砂纸刮擦200个愈伤组织,并使用无NH(4)NO(3)的培养基进行共培养以及含潮霉素的再生培养基进行筛选,从这些愈伤组织中获得了6个抗潮霉素(Hyg(r))培养系。Hyg(r)培养系再生出芽,将其转移到无植物生长调节剂的培养基后发育成小植株。通过GUS组织化学分析和反向PCR分析,证实所有这些小植株都是转基因的。

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