Conditional promoters to investigate gene function during wheat infection by Zymoseptoria tritici.
Conditional promoters to investigate gene function during wheat infection by Zymoseptoria tritici.
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DOI：
10.1016/j.fgb.2020.103487
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发表时间：
2021-01
期刊：
影响因子：
--
通讯作者：
Steinberg G
中科院分区：
文献类型：
--
作者：
Fantozzi E;Kilaru S;Cannon S;Schuster M;Gurr SJ;Steinberg G
对小麦黑穗病叶片中5个条件启动子的活性进行了分析。启动子Pnar1和Picl1在植物中被抑制。启动子PlaraB在植物侵染的所有阶段都在菌丝中被诱导。启动子Pgal7和Pex1A不紧密，但在感染后期被诱导。条件启动子响应植物细胞壁源性糖的外质体。小麦黑穗病真菌引起小麦黑穗病叶斑病，对温带小麦造成严重威胁。最近，我们描述了大量的分子工具来研究这种真菌的体外生物学。其中有5个条件启动子（Pnar1, Pex1A, Picl1, Pgal7, PlaraB），它们允许在液体培养的细胞中控制靶基因的过表达或抑制。然而，它们在植物宿主-病原体相互作用中的应用尚未得到检验。在这里，我们通过绿色荧光蛋白表达细胞的定量活细胞成像研究了这些启动子在植物侵染过程中的6个阶段的行为。我们发现Pnar1和Picl1在植物中被抑制，并证明它们适合研究植物定植过程中必需基因的表达和功能。启动子Pgal7和Pex1A在植物中不被完全抑制，但在丙酮化过程中被诱导。这表明存在诱导半乳糖或木糖和/或阿拉伯糖，通过真菌水解酶的活性从植物细胞壁释放。相反，通常控制α-l-阿拉伯糖醛酸苷酶B表达的PlaraB启动子在叶片内部被强烈诱导。这表明真菌在叶肉外质体中暴露于l -阿拉伯糖。综上所述，本研究建立了2个可抑制启动子（Pnar1和Picl1）和3个可诱导启动子（Pgal7、Pex1A、PlaraB）用于植物分子研究。此外，我们还提供了在小麦偃麦草感染生物营养期植物细胞壁降解的间接证据。
We analyze the activity of 5 conditional promoters in Z. tritici-infected wheat leaves. The promoters Pnar1 and Picl1 are repressed in planta. The promoter PlaraB is induced in hyphae during all in planta infection stages. The promoters Pgal7 and Pex1A are not tight, but induced during late infection. Conditional promoters respond to plant cell wall-derived sugars in the apoplast. The fungus Zymoseptoria tritici causes Septoria tritici leaf blotch, which poses a serious threat to temperate-grown wheat. Recently, we described a raft of molecular tools to study the biology of this fungus in vitro. Amongst these are 5 conditional promoters (Pnar1, Pex1A, Picl1, Pgal7, PlaraB), which allow controlled over-expression or repression of target genes in cells grown in liquid culture. However, their use in the host-pathogen interaction in planta was not tested. Here, we investigate the behaviour of these promoters by quantitative live cell imaging of green-fluorescent protein-expressing cells during 6 stages of the plant infection process. We show that Pnar1 and Picl1 are repressed in planta and demonstrate their suitability for studying essential gene expression and function in plant colonisation. The promoters Pgal7 and Pex1A are not fully-repressed in planta, but are induced during pycnidiation. This indicates the presence of inducing galactose or xylose and/or arabinose, released from the plant cell wall by the activity of fungal hydrolases. In contrast, the PlaraB promoter, which normally controls expression of an α-l-arabinofuranosidase B, is strongly induced inside the leaf. This suggests that the fungus is exposed to L-arabinose in the mesophyll apoplast. Taken together, this study establishes 2 repressible promoters (Pnar1 and Picl1) and three inducible promoters (Pgal7, Pex1A, PlaraB) for molecular studies in planta. Moreover, we provide circumstantial evidence for plant cell wall degradation during the biotrophic phase of Z. tritici infection.
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DOI：
10.1016/j.fgb.2015.04.013
发表时间：
2015-06
期刊：
Fungal genetics and biology : FG & B
影响因子：
--
作者：
Cairns TC;Sidhu YS;Chaudhari YK;Talbot NJ;Studholme DJ;Haynes K
通讯作者：
Haynes K
影响因子：
15
作者：
HOFFMANN, RA;VANWIJK, J;VLIEGENTHART, JFG
通讯作者：
VLIEGENTHART, JFG
影响因子：
--
作者：
Duncan, KE;Howard, RJ
通讯作者：
Howard, RJ
DOI：
10.1007/bf00282651
发表时间：
1991-01-01
期刊：
MOLECULAR & GENERAL GENETICS
影响因子：
--
作者：
DOSHI, P;BOSSIE, CA;MORRIS, NR
通讯作者：
MORRIS, NR
影响因子：
16.6
作者：
Bielska, Ewa;Higuchi, Yujiro;Schuster, Martin;Steinberg, Natascha;Kilaru, Sreedhar;Talbot, Nicholas J.;Steinberg, Gero
通讯作者：
Steinberg, Gero