Fungal plant pathogen "mutagenomics" reveals tagged and untagged mutations in Zymoseptoria tritici and identifies SSK2 as key morphogenesis and stress-responsive virulence factor.
Fungal plant pathogen "mutagenomics" reveals tagged and untagged mutations in Zymoseptoria tritici and identifies SSK2 as key morphogenesis and stress-responsive virulence factor.
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真菌植物的病原体“诱变”揭示了Zymoseptoria tritici中标记和未标记的突变，并将SSK2鉴定为关键的形态发生和应激响应性毒力因子。
DOI：
10.3389/fpls.2023.1140824
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发表时间：
2023
影响因子：
5.6
通讯作者：
中科院分区：
文献类型：
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作者：
“突变基因组学”是随机诱变、表型筛选和全基因组重测序的组合，以揭示与生物体中的表型变化相关的所有标记和未标记突变。在这项研究中，我们进行了诱变组学筛选小麦病原真菌Zymoseptoria triumphium改变形态发生开关和应力敏感性表型，使用农杆菌介导的“随机”T-DNA诱变（ATMT）。生物筛选鉴定出4个突变体，这些突变体对小麦的毒力大大降低。全基因组重新测序确定了T-DNA插入事件的位置，并揭示了几个可能影响基因功能的非连锁突变。值得注意的是，两个独立的降低毒力的突变株，具有类似的改变应力敏感性和异常菌丝生长表型，被发现有ZtSSK 2 MAPKKK基因的功能突变的明显损失。一个突变株有一个直接的T-DNA插入影响预测的蛋白质的N-末端，而另一个具有一个不相关的移码突变的C-末端。我们使用遗传互补来恢复两种菌株的野生型（WT）功能（毒力、形态发生和应激反应）。我们证明，ZtSSK 2具有非冗余的功能，ZtSTE 11的毒力通过生化激活的应激激活HOG 1 MAPK途径。此外，我们目前的数据表明，SSK 2在激活这一途径响应特定的压力具有独特的作用。最后，WT和SSK 2突变株的基于双重RNAseq的转录组分析揭示了在早期感染期间真菌中的许多HOG 1依赖性转录变化，并表明宿主反应在该早期阶段不区分WT和突变株。这些数据共同定义了与病原体毒力有关的新基因，并强调了全基因组测序步骤在诱变基因组发现管道中的重要性。
“Mutagenomics” is the combination of random mutagenesis, phenotypic screening, and whole-genome re-sequencing to uncover all tagged and untagged mutations linked with phenotypic changes in an organism. In this study, we performed a mutagenomics screen on the wheat pathogenic fungus Zymoseptoria tritici for altered morphogenetic switching and stress sensitivity phenotypes using Agrobacterium-mediated “random” T-DNA mutagenesis (ATMT). Biological screening identified four mutants which were strongly reduced in virulence on wheat. Whole genome re-sequencing defined the positions of the T-DNA insertion events and revealed several unlinked mutations potentially affecting gene functions. Remarkably, two independent reduced virulence mutant strains, with similarly altered stress sensitivities and aberrant hyphal growth phenotypes, were found to have a distinct loss of function mutations in the ZtSSK2 MAPKKK gene. One mutant strain had a direct T-DNA insertion affecting the predicted protein’s N-terminus, while the other possessed an unlinked frameshift mutation towards the C-terminus. We used genetic complementation to restore both strains’ wild-type (WT) function (virulence, morphogenesis, and stress response). We demonstrated that ZtSSK2 has a non-redundant function with ZtSTE11 in virulence through the biochemical activation of the stress-activated HOG1 MAPK pathway. Moreover, we present data suggesting that SSK2 has a unique role in activating this pathway in response to specific stresses. Finally, dual RNAseq-based transcriptome profiling of WT and SSK2 mutant strains revealed many HOG1-dependent transcriptional changes in the fungus during early infection and suggested that the host response does not discriminate between WT and mutant strains during this early phase. Together these data define new genes implicated in the virulence of the pathogen and emphasise the importance of a whole genome sequencing step in mutagenomic discovery pipelines.
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影响因子：
56.9
作者：
HAN, J;LEE, JD;ULEVITCH, RJ
通讯作者：
ULEVITCH, RJ
DOI：
10.1534/g3.115.017731
发表时间：
2015-04-27
期刊：
G3 (Bethesda, Md.)
影响因子：
--
作者：
Grandaubert J;Bhattacharyya A;Stukenbrock EH
通讯作者：
Stukenbrock EH
DOI：
10.1111/j.2517-6161.1995.tb02031.x
发表时间：
1995-01-01
影响因子：
5.8
作者：
BENJAMINI, Y;HOCHBERG, Y
通讯作者：
HOCHBERG, Y
影响因子：
6.4
作者：
Habig, Michael;Quade, Jakob;Stukenbrock, Eva Holtgrewe
通讯作者：
Stukenbrock, Eva Holtgrewe
影响因子：
16.6
作者：
通讯作者：
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