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首页> 外文期刊>Australasian Plant Pathology >Stable transformation of Rhizoctonia solani with a modified hygromycin resistance gene
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Stable transformation of Rhizoctonia solani with a modified hygromycin resistance gene

机译：修饰的潮霉素抗性基因可稳定转化茄枯萎病

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摘要

A vector for Agrobacterium tumefaciens-mediated transformation of basidiomycetes was constructed by insertion of a modified hygromycin resistance gene into the plant binary vector pBin19. The hygromycin coding region is flanked by a basidiomycete promoter and terminator. Isolates from different anastomosis groupings (AG) of the phytopathogenic fungus Rhizoctonia solani (Kuhn) were transformed with this vector using A. tumefaciens. Hygromycin-resistant transformants were isolated from a single AG6 isolate, but not from AG3 or AG4 isolates. Of six transformants isolated, five showed enhanced growth on agar containing either 25 or 50 mg/mL hygromycin. However, as the hygromycin concentration increased, the difference between the transformants and thecontrol reduced, until at 100 mg/mL there was no difference. The resistance phenotype was retained through repeated subcultures on non-selective media. The presence of the gene in the transformants was confirmed by PCR analysis and Southern hybridisatio
机译：通过将修饰的潮霉素抗性基因插入植物二元载体pBin19中，构建了根癌农杆菌介导的担子菌转化载体。潮霉素编码区的侧面是担子菌启动子和终止子。使用根癌农杆菌用该载体转化植物致病性真菌Rhizoctonia solani（Kuhn）的不同吻合组（AG）的分离株。潮霉素抗性转化子是从单个AG6分离物中分离出来的，但不是从AG3或AG4分离物中分离出来的。在分离出的六个转化子中，五个在含有25或50 mg / mL潮霉素的琼脂上显示出增强的生长。然而，随着潮霉素浓度的增加，转化体和对照之间的差异减小，直到在100 mg / mL时没有差异。通过在非选择性培养基上重复传代培养来保留抗性表型。通过PCR分析和Southern杂交证实了转化子中该基因的存在。

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