【摘要】 据灰葡萄孢菌丝发育和菌核形成时期的RNA-seq数据,挑选出在这2个时期表达量差异较大的基因BC1G_<sub>03293进行基因功能的研究。结果发现,BC1G_<sub>03293在菌核发育时期表达明显上调,表达量比菌丝生长时期上调了70倍以上。该基因编码的蛋白质含有229个氨基酸,N端包含信号肽,但不含有任何已知保守结构域。通过同源重组的方法得到了BC1G_<sub>03293的敲除转化子ΔBC1G_<sub>03293-2和ΔBC1G_<sub>03293-4,并运用农杆菌转化技术得到了互补转化子ΔBC1G_<sub>03293-2-C2和ΔBC1G_<sub>03293-2-C3。BC1G_<sub>03293基因敲除后生长、致病及菌核形成等表型无明显变化,但敲除转化子的分生孢子产量显著下降,仅为野生型菌株B05.10分生孢子产量的45%,并且BC1G_<sub>03293基因互补可使敲除转化子的产孢量得到明显恢复。结果说明BC1G_<sub>03293基因与灰葡萄孢的分生孢子形成相关。更多还原

【Abstract】 According to the RNA-Seq mycelium growth and sclerotia formation stage,was selected to study in B.cinerea.The transcript level of BC1G_<sub>03293 during sclerotial formation stage was higher than that during mycelium growth stage.The BC1G_<sub>03293 of 229 aa signal peptide on the N end and no known conserved domain.The knockout mutantsΔBC1G_<sub>03293-2 andΔBC1G_<sub>03293-4 were obtained using the homologous recombination method.The complemented transformantsΔBC1G_<sub>03293-2-C2 andΔBC1G_<sub>03293-2-C3 were also by Agrobacterium tumefaciens-mediated transformation technique.The deletion of BC1G_<sub>03293 did not affect the growth,pathogenicity and formation of sclerotia.The conidia production of the knockout mutants obviously decreased and was approximately 45% of the wild type strain.And the conidia production of the complemented transformants recovered obviously.Our research results suggested that BC1G_<sub>03293 is involved the regulation of conidiation of B.cinerea.更多还原