Yeast transformation mediated by Agrobacterium strains harboring an Ri plasmid: comparative study between GALLS of an Ri plasmid and virE of a Ti plasmid
Yeast transformation mediated by Agrobacterium strains harboring an Ri plasmid: comparative study between GALLS of an Ri plasmid and virE of a Ti plasmid
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DOI：
10.1111/j.1365-2443.2012.01612.x
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发表时间：
2012-07-01
期刊：
影响因子：
2.1
通讯作者：
Suzuki, Katsunori
中科院分区：
文献类型：
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作者：
Kiyokawa, Kazuya;Yamamoto, Shinji;Suzuki, Katsunori
含有Ti质粒的农杆菌菌株不仅可以将T-DNA转移到植物中，还可以转移到真菌中，包括酵母酿酒酵母。然而，还没有含有Ri质粒的农杆菌菌株在真菌转化中被评估。一些Ri质粒有虫胆，而不是VirE1和VirE2。在植物转化中，GALS蛋白可以在功能上替代结构不同的蛋白VirE2。在这项研究中，我们比较了农杆菌供体之间的酵母转化能力：一株含有Ti质粒的菌株，一株含有农品型或mikimopine型Ri质粒的菌株，以及一株含有改良的Ri质粒并补充了一种钛质粒型Vre操纵子的菌株。具有虫胆的农杆菌菌株转化酵母细胞的效率远远低于含有病毒操纵子的菌株。在受体酵母细胞中产生虫胆改善了由一株既不缺乏虫胆也不缺乏病毒操纵子的农杆菌菌株介导的酵母转化。Cre重组酶融合蛋白动员的报告实验表明，在酵母细胞中比在虫胆中表达更丰富的病毒E2蛋白。根据这些结果，我们得出结论，具有Ri质粒的农杆菌菌株所介导的低酵母转化率是由于酵母细胞中动员的胆汁量低所致。
Agrobacterium strains containing a Ti plasmid can transfer T-DNA not only to plants but also to fungi, including the yeast Saccharomyces cerevisiae. However, no Agrobacterium strain harboring an Ri plasmid has been evaluated in fungal transformation. Some Ri plasmids have GALLS , instead of virE1 and virE2. GALLS protein can functionally substitute in plant transformation for a structurally different protein VirE2. In this study, we compared the yeast transformation ability among Agrobacterium donors: a strain containing a Ti plasmid, strains harboring either an agropine-type or a mikimopine-type Ri plasmid, and a strain having a modified Ri plasmid supplemented with a Ti plasmid type virE operon. Agrobacterium strains possessing GALLS transformed yeast cells far less efficiently than the strain containing virE operon. Production of GALLS in recipient yeast cells improved the yeast transformation mediated by an Agrobacterium strain lacking neither GALLS nor virE operon. A reporter assay to detect mobilization of the proteins fused with Cre recombinase revealed that VirE2 protein is much more abundant in yeast cells than GALLS. Based on these results, we concluded that the low yeast transformability mediated by Agrobacterium strains having the Ri plasmid is because of low amount of mobilized GALLS in yeast cells.