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首页> 外文期刊>Biotechnology Progress >Development of auxotrophic Agrobacterium tumefaciens for gene transfer in plant tissue culture
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Development of auxotrophic Agrobacterium tumefaciens for gene transfer in plant tissue culture

机译：营养缺陷型根癌土壤杆菌在植物组织培养中的基因转移研究进展

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摘要

Auxotrophic strains of Agrobacterium tumefaciens were generated for use in liquid co-culture with plant tissue for transient gene expression. Twenty-one auxotrophs were recovered from 1,900 tetracycline-resistant insertional mutants generated with a suicide vector transposon mutagenesis system. Twelve of these auxotrophs were characterized on a nutrient matrix. lsolates were screened for growth in plant cell and root culture, and three auxotrophs were identified that had limited growth: adenine (ade-24), leucine (leu.-27), and cysteine (eys-32). Ade-24 displayed poor T-DNA delivery in a transient expression test delivering GUS from a binary vector, while cys-32 displayed the best ability to deliver DNA of these three auxotrophs. The growth yield of cys-32 on cysteine was assessed to provide a quantitative basis for co-culture nutrient supplementation. The utility of cys-32 for delivering T-DNA to plant tissues is demonstrated, where an 85-fold enhancement in GUS expression over wild-type A. tumefaciens was achieved.
机译：产生了根癌农杆菌的营养缺陷型菌株，用于与植物组织的液体共培养，用于瞬时基因表达。从自杀载体转座子诱变系统产生的1,900个四环素抗性插入突变体中回收了21个营养缺陷型。在营养基质上鉴定了十二种营养缺陷型。筛选分离株以在植物细胞和根培养物中生长，并鉴定出生长受限的三种营养缺陷型：腺嘌呤（ade-24），亮氨酸（leu.-27）和半胱氨酸（eys-32）。在瞬时表达测试中，Ade-24在从双元载体递送GUS的过程中显示出不良的T-DNA传递，而cys-32在传递这三种营养缺陷型DNA方面表现出最好的能力。评估了cys-32在半胱氨酸上的生长产量，为补充共培养营养提供了定量依据。证明了cys-32用于将T-DNA传递至植物组织的效用，与野生型根癌农杆菌相比，其GUS表达提高了85倍。

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