Engineering Research Center of Plant Growth Regulator, Ministry of Education/College of Agronomy, China Agricultural University, Beijing 100193, China.
Beijing Key Laboratory of Environment Friendly Management on Fruit Diseases and Pests in North China, Institute of Plant Protection, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China.
Int J Mol Sci. 2023 Jan 4;24(2):988. doi: 10.3390/ijms24020988.
, the causal agent of gray mold, is one of the most destructive pathogens of cherry tomatoes, causing fruit decay and economic loss. Fludioxonil is an effective fungicide widely used for crop protection and is effective against tomato gray mold. The emergence of fungicide-resistant strains has made the control of more difficult. While the genome of is available, there are few reports regarding the large-scale functional annotation of the genome using expressed genes derived from transcriptomes, and the mechanism(s) underlying such fludioxonil resistance remain unclear. The present study prepared RNA-sequencing (RNA-seq) libraries for three strains (two highly resistant (LR and FR) versus one highly sensitive (S) to fludioxonil), with and without fludioxonil treatment, to identify fludioxonil responsive genes that associated to fungicide resistance. Functional enrichment analysis identified nine resistance related DEGs in the fludioxonil-induced LR and FR transcriptome that were simultaneously up-regulated, and seven resistance related DEGs down-regulated. These included adenosine triphosphate (ATP)-binding cassette (ABC) transporter-encoding genes, major facilitator superfamily (MFS) transporter-encoding genes, and the high-osmolarity glycerol (HOG) pathway homologues or related genes. The expression patterns of twelve out of the sixteen fludioxonil-responsive genes, obtained from the RNA-sequence data sets, were validated using quantitative real-time PCR (qRT-PCR). Based on RNA-sequence analysis, it was found that hybrid histidine kinase, fungal HHKs, such as , , and probably involved in the fludioxonil resistance of , in addition, a number of ABC and MFS transporter genes that were not reported before, such as , , , , , , and were differentially expressed in the fludioxonil-resistant strains, indicating that overexpression of these efflux transporters located in the plasma membranes may associate with the fludioxonil resistance mechanism of All together, these lines of evidence allowed us to draw a general portrait of the anti-fludioxonil mechanisms for , and the assembled and annotated transcriptome data provide valuable genomic resources for further study of the molecular mechanisms of resistance to fludioxonil.
灰葡萄孢，是导致樱桃番茄腐烂的主要病原菌之一，也是导致番茄腐烂和经济损失的主要原因之一。氟啶酮是一种广泛用于作物保护的有效杀菌剂，对番茄灰霉病有很好的防治效果。然而，抗药性菌株的出现使得对灰葡萄孢的控制变得更加困难。虽然灰葡萄孢的基因组已经可用，但很少有报道涉及使用转录组衍生的表达基因对其基因组进行大规模功能注释，并且其抗氟啶酮的机制尚不清楚。本研究为三个灰葡萄孢菌株（两个对氟啶酮高度耐药（LR 和 FR），一个对氟啶酮高度敏感（S））制备了 RNA 测序（RNA-seq）文库，分别在有无氟啶酮处理的情况下，以鉴定与抗药性相关的氟啶酮反应基因。功能富集分析在氟啶酮诱导的 LR 和 FR 转录组中发现了 9 个与抗药性相关的差异表达基因（DEGs），这些基因同时上调，7 个与抗药性相关的 DEGs 下调。这些基因包括三磷酸腺苷（ATP）结合盒（ABC）转运蛋白编码基因、主要易化因子超家族（MFS）转运蛋白编码基因，以及高渗甘油（HOG）途径同源物或相关基因。从 RNA-seq 数据集中获得的 16 个氟啶酮反应基因中的 12 个的表达模式，使用定量实时 PCR（qRT-PCR）进行了验证。基于 RNA-seq 分析，发现杂交组氨酸激酶、真菌 HHKs（如 、 、 ）可能参与了灰葡萄孢的氟啶酮耐药性，此外，还发现了一些以前未报道过的 ABC 和 MFS 转运蛋白基因，如 、 、 、 、 、 和 ，在氟啶酮耐药菌株中差异表达，表明这些位于质膜上的外排转运蛋白的过度表达可能与灰葡萄孢对氟啶酮的耐药机制有关。综上所述，这些证据使我们能够对灰葡萄孢的抗氟啶酮机制有一个总体的了解，组装和注释的转录组数据为进一步研究灰葡萄孢对氟啶酮的分子机制提供了有价值的基因组资源。
