mCherry-Labeled Verticillium dahliae Could Be Utilized to Investigate Its Pathogenicity Process in Nicotiana benthamiana.
mCherry-Labeled Verticillium dahliae Could Be Utilized to Investigate Its Pathogenicity Process in Nicotiana benthamiana.
复制标题
mCherry 标记的大丽轮枝菌可用于研究其在本塞姆氏烟草中的致病过程
DOI：
10.3390/genes9100508
复制
发表时间：
2018-10-18
期刊：
影响因子：
3.5
通讯作者：
Cheng H
中科院分区：
文献类型：
--
作者：
Su X;Lu G;Rehman L;Li X;Sun L;Guo H;Cheng H
大丽轮枝菌（Verticillium dahliae）是一种土传植物病原真菌，可引起植物枯萎病，但其致病性的分子机制尚不十分清楚。本研究通过原生质体转化的方法获得了一株红色荧光分离物，并利用共聚焦显微镜研究了其在本氏烟草（Nicotiana benthamiana）中的定殖、入侵和扩展等致病机制。优化mCherry基因的核苷酸序列，将其克隆到pCT-HM质粒中，转化大丽轮枝菌原生质体。该菌株（Vd-m）显示出强烈的红色荧光，其表型、生长速率和致病性与野生型大丽轮枝菌（Vd-wt）没有显著差异。接种后1 - 3天，Vd-m成功地定殖并侵入根的表皮细胞。4 ~ 6 dpi时，菌丝在根伤口和侧根原基上生长，进入木质部导管。7 ~ 9 dpi时，菌丝沿着细胞壁表面伸展，大量生长在根的木质部导管中。在10 dpi时，在叶柄和叶脉中发现了Vd-m。研究结果清楚地揭示了大丽轮枝菌侵染和定殖的途径。benthamiana的表达，优化表达可以加深我们对致病性分子机制的理解。
Verticillium dahliae is a soil-borne phytopathogenic fungus that causes a destructive vascular wilt, but details of the molecular mechanism behind its pathogenicity are not very clear. Here, we generated a red fluorescent isolate of V. dahliae by protoplast transformation to explore its pathogenicity mechanism, including colonization, invasion, and extension in Nicotiana benthamiana, using confocal microscopy. The nucleotide sequences of mCherry were optimized for fungal expression and cloned into pCT-HM plasmid, which was inserted into V. dahliae protoplasts. The transformant (Vd-m) shows strong red fluorescence and its phenotype, growth rate, and pathogenicity did not differ significantly from the wild type V. dahliae (Vd-wt). Between one and three days post inoculation (dpi), the Vd-m successfully colonized and invaded epidermal cells of the roots. From four to six dpi, hyphae grew on root wounds and lateral root primordium and entered xylem vessels. From seven to nine dpi, hyphae extended along the surface of the cell wall and massively grew in the xylem vessel of roots. At ten dpi, the Vd-m was found in petioles and veins of leaves. Our results distinctly showed the pathway of V. dahliae infection and colonization in N. benthamiana, and the optimized expression can be used to deepen our understanding of the molecular mechanism of pathogenicity.
登录
查看更多内容
影响因子：
3.2
作者：
Duressa, Dechassa;Rauscher, Gilda;Klosterman, Steven J.
通讯作者：
Klosterman, Steven J.
影响因子：
3.2
作者：
GERIK, JS;HUISMAN, OC
通讯作者：
HUISMAN, OC
影响因子：
17.3
作者：
Kendall, JM;Badminton, MN
通讯作者：
Badminton, MN
影响因子：
2.8
作者：
GRIFFITHS, DA
通讯作者：
GRIFFITHS, DA
影响因子：
4.2
作者：
Pantelides, Iakovos S.;Tjamos, Sotirios E.;Paplomatals, Epaminondas J.
通讯作者：
Paplomatals, Epaminondas J.