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Early development of Moniliophthora perniciosa basidiomata and developmentally regulated genes

机译：Perniciosa perniciosa basidiomata的早期发育和发育调控基因

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Background The hemibiotrophic fungus Moniliophthora perniciosa is the causal agent of Witches' broom, a disease of Theobroma cacao. The pathogen life cycle ends with the production of basidiocarps in dead tissues of the infected host. This structure generates millions of basidiospores that reinfect young tissues of the same or other plants. A deeper understanding of the mechanisms underlying the sexual phase of this fungus may help develop chemical, biological or genetic strategies to control the disease. Results Mycelium was morphologically analyzed prior to emergence of basidiomata by stereomicroscopy, light microscopy and scanning electron microscopy. The morphological changes in the mycelium before fructification show a pattern similar to other members of the order Agaricales. Changes and appearance of hyphae forming a surface layer by fusion were correlated with primordia emergence. The stages of hyphal nodules, aggregation, initial primordium and differentiated primordium were detected. The morphological analysis also allowed conclusions on morphogenetic aspects. To analyze the genes involved in basidiomata development, the expression of some selected EST genes from a non-normalized cDNA library, representative of the fruiting stage of M. perniciosa, was evaluated. A macroarray analysis was performed with 192 selected clones and hybridized with two distinct RNA pools extracted from mycelium in different phases of basidiomata formation. This analysis showed two groups of up and down-regulated genes in primordial phases of mycelia. Hydrophobin coding, glucose transporter, Rho-GEF, Rheb, extensin precursor and cytochrome p450 monooxygenase genes were grouped among the up-regulated. In the down-regulated group relevant genes clustered coding calmodulin, lanosterol 14 alpha demethylase and PIM1. In addition, 12 genes with more detailed expression profiles were analyzed by RT-qPCR. One aegerolysin gene had a peak of expression in mycelium with primordia and a second in basidiomata, confirming their distinctiveness. The number of transcripts of the gene for plerototolysin B increased in reddish-pink mycelium and indicated an activation of the initial basidiomata production even at this culturing stage. Expression of the glucose transporter gene increased in mycelium after the stress, coinciding with a decrease of adenylate cyclase gene transcription. This indicated that nutrient uptake can be an important signal to trigger fruiting in this fungus. Conclusion The identification of genes with increased expression in this phase of the life cycle of M. perniciosa opens up new possibilities of controlling fungus spread as well as of genetic studies of biological processes that lead to basidiomycete fruiting. This is the first comparative morphologic study of the early development both in vivo and in vitro of M. perniciosa basidiomata and the first description of genes expressed at this stage of the fungal life cycle.
机译：背景技术半生营养真菌百日草（Moniliophthora perniciosa）是巫婆扫帚（一种可可可可的病）的病原体。病原体的生命周期以受感染宿主的死皮组织中的紫苏果产生而告终。这种结构产生了数百万个孢子孢子，可重新感染同一或其他植物的年轻组织。更深入地了解这种真菌性相的潜在机制可能有助于发展化学，生物学或遗传学策略来控制这种疾病。结果通过立体显微镜，光学显微镜和扫描电子显微镜对担子瘤出现之前的菌丝体进行了形态学分析。进行果胶化之前，菌丝体的形态变化显示出类似于木耳菌其他成员的模式。通过融合形成表面层的菌丝的变化和外观与原基出现有关。检测菌丝结节，聚集，初始原基和分化原基的阶段。形态分析还可以得出形态发生方面的结论。为了分析参与担子菌发育的基因，评估了来自未标准化的cDNA文库的一些选定的EST基因的表达，该文库代表了多年生番茄的结实阶段。用192个选定的克隆进行了大阵列分析，并与从菌丝体形成的不同阶段的菌丝体中提取的两个不同的RNA库杂交。该分析显示了在菌丝体原始阶段的两组上调和下调的基因。疏水蛋白编码，葡萄糖转运蛋白，Rho-GEF，Rheb，延伸蛋白前体和细胞色素p450单加氧酶基因归类于上调。在下调组中，相关基因聚簇编码钙调蛋白，羊毛甾醇14α脱甲基酶和PIM1。另外，通过RT-qPCR分析了具有更详细表达谱的12个基因。一个aegerolysin基因在原基的菌丝体中有一个表达高峰，而在basidiomata中则有一个表达高峰，证实了它们的独特性。桃红色菌丝体中多菌溶菌素B基因的转录物数量增加，即使在此培养阶段也表明初始担子菌的产生被激活。应激后，菌丝体中葡萄糖转运蛋白基因的表达增加，这与腺苷酸环化酶基因转录的减少相吻合。这表明养分吸收可能是触发这种真菌结实的重要信号。结论鉴定多年生支原体生命周期此阶段表达增加的基因，为控制真菌传播以及导致担子菌结实的生物学过程的遗传研究提供了新的可能性。这是首次对球孢分枝杆菌basidiomata体内和体外早期发育的比较形态学研究，并且是在真菌生命周期此阶段表达的基因的第一个描述。

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