Hu Dongfang, Wang Chunsheng, Tao Fei, Cui Qian, Xu Xiangming, Shang Wenjing, Hu Xiaoping
State Key Laboratory of Crop Stress Biology for Arid Areas, College of Plant Protection, Northwest A & F University, Yangling, China.
State Key Laboratory of Crop Stress Biology for Arid Areas, College of Plant Protection, Northwest A & F University, Yangling, China; East Malling Research, East Malling, Kent, United Kingdom.
PLoS One. 2014 Jun 13;9(6):e100046. doi: 10.1371/journal.pone.0100046. eCollection 2014.
Verticillium dahliae is a fungal pathogen causing Verticillium wilt on a range of economically important crops. Microsclerotia are its main survival and dormancy structures and serve as the primary inoculum on many hosts. Studies were conducted to determine the effect of temperature (5 to 50°C), pH (2 to 12) and nutrient regimes on microsclerotia germination. The optimal condition for microsclerotium germination was 20°C with pH 8.0 whereas nutrient regimes had no significant effect on its germination. The whole genome wide expression profiles during microsclerotium germination were characterized using the Illumina sequencing technology. Approximately 7.4 million of 21-nt cDNA tags were sequenced in the cDNA libraries derived from germinated and non-germinated microsclerotia. About 3.9% and 2.3% of the unique tags were up-regulated and down-regulated at least five-fold, respectively, in the germinated microsclerotia compared with the non-germinated microsclerotia. A total of 1654 genes showing differential expression were identified. Genes that are likely to have played important roles in microsclerotium germination include those encoding G-protein coupled receptor, lipase/esterase, cyclopentanone 1,2-monooxygenase, H(+)/hexose cotransporter 1, fungal Zn(2)-Cys(6) binuclear cluster domain, thymus-specific serine protease, glucan 1,3-beta-glucosidase, and alcohol dehydrogenase. These genes were mainly up-regulated or down-regulated only in germinated microsclerotia, compared with non-germinated microsclerotia. The differential expression of genes was confirmed by qRT-PCR analysis of 20 randomly selected genes from the 40 most differentially expressed genes.
大丽轮枝菌是一种真菌病原体，可在一系列具有重要经济价值的作物上引发黄萎病。微菌核是其主要的存活和休眠结构，也是许多寄主上的主要接种体。本研究旨在确定温度（5至50°C）、pH值（2至12）和营养条件对微菌核萌发的影响。微菌核萌发的最佳条件是20°C、pH值8.0，而营养条件对其萌发没有显著影响。利用Illumina测序技术对微菌核萌发过程中的全基因组表达谱进行了表征。在来自已萌发和未萌发微菌核的cDNA文库中，对约740万个21核苷酸的cDNA标签进行了测序。与未萌发的微菌核相比，在已萌发的微菌核中，分别约有3.9%和2.3%的独特标签上调和下调至少5倍。共鉴定出1654个差异表达基因。可能在微菌核萌发中发挥重要作用的基因包括编码G蛋白偶联受体、脂肪酶/酯酶、环戊酮1,2-单加氧酶、H(+)/己糖共转运蛋白1、真菌Zn(2)-Cys(6)双核簇结构域、胸腺特异性丝氨酸蛋白酶、葡聚糖1,3-β-葡糖苷酶和乙醇脱氢酶的基因。与未萌发的微菌核相比，这些基因主要仅在已萌发的微菌核中上调或下调。通过对40个差异表达最显著的基因中随机选择的20个基因进行qRT-PCR分析，证实了基因的差异表达。
