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【目的】研究香蕉枯萎病菌4号生理小种中促分裂原活化蛋白激酶基因FoHog1的结构特点及其功能【方法】通过PCR和RT-PCR的方法获得了FoHog1基因序列并进行生物信息学分析,利用PEG介导的原生质体转化法得到了FoHog1基因缺失突变体,分析敲除突变体与野生型的生物学特性差异【结果】FoHog1基因编码一个含有357个氨基酸的蛋白,该蛋白在不同种镰刀菌中高度保守。通过对敲除突变体的研究发现,该基因缺失后菌丝密度下降,产孢量与菌丝干重明显降低,对乙酸钠和氯化铵的利用率下降,对温度、pH及渗透压等外源胁迫更为敏感。通过致病力实验发现,基因敲除突变体的定殖能力有所降低【结论】尖孢镰刀菌古巴专化型4号生理小种中FoHog1基因参与调控菌丝生长、分生孢子生成、乙酸钠和氯化铵代谢、渗透压胁迫反应及致病相关过程。
【Objective】 The objective of this study was to investigate the structural characteristics and function of the mitogen-activated protein kinase gene FoHog1 in physiological races of Fusarium oxysporum f.sp. chinensis 4 [Method] The FoHog1 gene sequence was obtained by PCR and RT-PCR and bioinformatics analysis was performed. FoHog1 gene deletion mutants were obtained by PEG-mediated protoplast transformation. The biological characteristics of knockout mutants and wild-type were analyzed. 【Results】 FoHog1 gene encodes a protein of 357 amino acids, The bacteria are highly conserved. The results of knockout mutants showed that the mycelial density of the gene was decreased, the amount of sporulation and the dry weight of hyphae decreased significantly, the utilization of sodium acetate and ammonium chloride decreased, and the temperature, pH and osmotic pressure Exogenous stress is more sensitive. Through pathogenicity experiments, gene knockout mutants showed lower colonization ability. 【Conclusion】 The FoHog1 gene of Fusarium oxysporum f. Ecotype 4 was involved in mycelial growth, conidial generation, acetic acid Sodium and ammonium chloride metabolism, osmotic stress response and pathogenicity-related processes.