葡萄座腔菌Mcm基因家族全基因组鉴定及其对硫化物的响应特性

由葡萄座腔菌Botryosphaeria dothidea引起的轮纹病是严重威胁我国苹果产业健康持续发展的重要病害之一。前期研究表明,韭菜及其主要成分硫化物能显著致死病原菌并能高效防控该病的发生。为了明确微小染色体维持蛋白（Mcm）基因家族在葡萄座腔菌中的基本功能,在全基因组水平对葡萄座腔菌Mcm基因家族进行了鉴定,并分析了葡萄座腔菌Mcm基因家族对硫化物的响应特性。结果显示：葡萄座腔菌Mcm基因家族共有6个成员（BdMcm2-BdMcm7）,分别位于5条重叠群（contig）上,含有3-14个CDS区。葡萄座腔菌Mcm基因家族编码产物主要定位于细胞核,每个成员均具有Mcm-box结构域及其Walker A、Walker B及R-finger结构特征,长度为718-1 618aa,等电点为5.04-8.33,分子量为79.78-180.30kDa。RNA-seq数据和qRT-PCR分析表明硫化物三硫醚能高效抑制葡萄座腔菌Mcm基因家族的表达,与对照相比,6个葡萄座腔菌Mcm基因表达量分别下调20.56%-78.10%和10.74%-73.83%,两者呈现高度一致性（R²=0.8958）。细胞流式仪检测表明硫化物也显著阻滞了葡萄座腔菌细胞周期。该研究为进一步揭示韭菜及其主要成分防控苹果轮纹病机理提供了理论依据。

Genome-wide identification and the responses to sulfides of the Mcm gene family in Botryosphaeria dothidea

Apple ring disease caused by Botryosphaeria dothidea is one of the important diseases that seriously threaten the healthy and sustainable development of the apple industry in China. Our previous study showed the Chinese leek and its main component sulfide efficiently killed the pathogen and significantly control the occurrence of the disease. To clarify the basic functions of the minichromosome maintenance protein (Mcm) gene family, the Mcm gene family in B. dothidea was identified at the whole genome level and their expression upon sulfide treatment was analyzed. The results showed that there were six members in the Mcm gene family in B. dothidea (BdMcm2-BdMcm7). They were located on five contigs and contain 3-14 CDS regions. The encoded proteins are mainly located in the nucleus and all have Mcm-box domain and Walker A, Walker B, and R-finger structure characteristics. The BdMcm proteins were 718-1 618aa in length, with 5.04-8.33 isoelectric point, and 79.78-180.30kDa in molecular weight. The RNA-seq and qRT-PCR analysis showed that sulfide dimethyl trisulfide efficiently inhibited the expression levels of the Mcm gene family in B. dothidea. Compared with that of the control, the expressions of the BdMcm2-BdMcm7 were down-regulated by 20.56%-78.10% and 10.74%-73.83%, respectively, with a high consistency (R²=0.8958). The flow cytometer detection revealed that sulfide significantly blocked the cell cycle of B. dothidea. The findings provided a theoretical basis for further revealing the mechanism that the apple ring disease was reduced by Chinese leek and its main components.