Direct measurement of appressorium turgor using a molecular mechanosensor in the rice blast fungus Magnaporthe oryzae
Direct measurement of appressorium turgor using a molecular mechanosensor in the rice blast fungus Magnaporthe oryzae
复制标题
使用分子机械传感器直接测量稻瘟病菌 Magnaporthe oryzae 的附着胞膨压
DOI：
10.1101/2022.08.30.505899
复制
发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Ryder L
中科院分区：
文献类型：
--
作者：
Ryder L
许多植物病原真菌强行进入宿主体内引起疾病。例如，稻瘟病菌Magnaporthe granule利用一种称为附着胞的特殊感染细胞感染植物，这种细胞产生巨大的膨胀力，驱动一个刚性的穿透钉穿过水稻叶片角质层。虽然这些巨大的内部压力是真菌侵入宿主的关键武器，但在宿主入侵期间直接探测它们仍然非常具有挑战性，这使得我们对这些极端细胞力学的理解不完整。在这里，我们结合联合收割机荧光寿命成像（FLIM）与膜靶向分子mechanoprobe量化膜张力的变化作为一个直接代理附着胞膨在M。米。我们报道成熟的黑色素M. Oryzaeappressoria显示出不均匀的低荧光寿命和高膜张力，与巨大的膨压一致。这些极端的压力导致膜力学的大规模空间异质性，远远大于以前在任何其他细胞类型中观察到的，突出了turgor-driven附着胞介导的植物感染的极端力学。相比之下，附着胞的非致病性黑色素缺陷突变体，alb 1andbuf 1，或不成熟的非黑化附着胞，表现出高荧光寿命，与低膜张力和膨压，保持空间均匀一致。为了评估该方法，我们研究了膨压动力学在一系列的突变体受损的附着胞功能。我们发现，膨胀传感器激酶突变体Δ sln 1，最近提出产生过量的附着胞膨胀，显示出显着更高的膜张力相比，同基因野生型M。水稻品系这种非侵入性的活细胞成像技术可以直接量化和可视化病原体感染期间部署的巨大膨压。
Many plant pathogenic fungi forcibly enter their hosts to cause disease. The rice blast fungusMagnaporthe oryzae, for example, infects plants using a specialised infection cell called an appressorium, which generates enormous turgor to drive a rigid penetration peg through the rice leaf cuticle. While these vast internal pressures are a critical weapon in fungal host penetration, they have remained very challenging to probe directly during host invasion, leaving our understanding of these extreme cellular mechanics incomplete. Here, we combine Fluorescence Lifetime Imaging (FLIM) with a membrane-targeting molecular mechanoprobe to quantify changes in membrane tension as a direct proxy for appressorial turgor inM. oryzae. We report that mature melanin-pigmentedM. oryzaeappressoria display a heterogeneous low fluorescence lifetime and high membrane tension, consistent with enormous turgor. These extreme pressures lead to large-scale spatial heterogeneities in membrane mechanics, much greater than observed in any other cell type previously, highlighting the extreme mechanics of turgor-driven appressorium-mediated plant infection. By contrast, appressoria of non-pathogenic melanin-deficient mutants,alb1andbuf1, or immature non-melanised appressoria, exhibit high fluorescence lifetime, consistent with low membrane tension and turgor, that remain spatially homogeneous. To evaluate the method, we investigated turgor dynamics in a range of mutants impaired in appressorium function. We show that the turgor sensor kinase mutantΔsln1, recently proposed to generate excess appressorium turgor, displayed a significantly higher membrane tension compared to an isogenic wild typeM. oryzaestrain. This non-invasive, live cell imaging technique allows direct quantification and visualization of the enormous turgor pressures deployed during pathogen infection.
登录
查看更多内容
影响因子：
14.8
作者：
Kagan VE;Mao G;Qu F;Angeli JP;Doll S;Croix CS;Dar HH;Liu B;Tyurin VA;Ritov VB;Kapralov AA;Amoscato AA;Jiang J;Anthonymuthu T;Mohammadyani D;Yang Q;Proneth B;Klein-Seetharaman J;Watkins S;Bahar I;Greenberger J;Mallampalli RK;Stockwell BR;Tyurina YY;Conrad M;Bayır H
通讯作者：
Bayır H
影响因子：
4.8
作者：
Garcia-Saez, Ana J.;Chiantia, Salvatore;Schwille, Petra
通讯作者：
Schwille, Petra
郁金香灰霉病菌核在土壤中的存活率及培养基形成条件
DOI：
--
发表时间：
2007
期刊：
茨城県病害虫研究会報 46
影响因子：
--
作者：
山本知代;ら
通讯作者：
ら
影响因子：
2.4
作者：
Akimov, Sergey A.;Kuzmin, Peter I.;Cohen, Fredric S.
通讯作者：
Cohen, Fredric S.