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芒果胶孢炭疽病菌对土槿皮乙酸抗性突变菌株的诱导及其生物适合度分析
引用本文:文小东,朱发娣,谷梦萱,叶火春,冯岗,张静.芒果胶孢炭疽病菌对土槿皮乙酸抗性突变菌株的诱导及其生物适合度分析[J].热带作物学报,2022,43(8):1654-1662.
作者姓名:文小东  朱发娣  谷梦萱  叶火春  冯岗  张静
作者单位:1.贵州大学农学院,贵州贵阳 5500252.中国热带农业科学院环境与植物保护研究所,海南海口 5711013.贵州大学精细化工研究开发中心,贵州贵阳 550025
基金项目:国家自然科学基金项目(31872001)
摘    要:土槿皮乙酸(pseudolaric acid B, PAB)是一个结构独特的天然二萜酸,前期发现该化合物对芒果胶孢炭疽菌等多种植物病原真菌具有广谱的抑菌活性,但其杀菌作用机制未知。本研究以芒果胶孢炭疽菌菌株JB-Q为供试对象,采用药剂驯化、紫外照射和钴辐射等方法诱导产生抗PAB的突变体,并分析抗性菌株的生物适合度。结果表明,仅药剂驯化法得到了18株不同抗性水平且可稳定遗传的抗PAB的胶孢炭疽菌,抗性频率为1.69×10-7。适合度研究表明,所有供试菌株在15~35℃范围内均能生长,且在25℃时菌丝生长速率达到最大值;而不同抗性水平的突变菌株的生长速率存在一定差异,其中低抗菌株>亲本菌株>中抗菌株>高抗菌株;渗透压测试结果表明,亲本菌株对0.02% 十二烷基硫酸钠(sodium dodecyl sulfate, SDS)表现更为敏感,其抑制率高达70%;在金属离子(NaCl、KCl和CaCl2)胁迫下,抗性突变菌株则表现更为敏感。因此,研究认为PAB不易导致胶孢炭疽菌的抗性产生,但诱导产生的抗性突变菌株的生存竞争力与敏感菌株相当。此外,交互抗性结果显示,土槿皮乙酸与吡唑醚菌酯、咪鲜胺和戊唑醇之间不存在交互抗性,但与苯并咪唑类杀菌剂多菌灵存在正交互抗性,其皮尔逊相关系数为0.7729(P<0.01),这表明PAB可能与微管抑制剂多菌灵有相似的作用机制,但仍需开展进一步的研究验证。

关 键 词:胶孢炭疽菌  土槿皮乙酸  抗性突变菌株  生物适合度  
收稿时间:2022-02-21

Induction and Biological Characterization of Mutants of Colletotrichum gloeosporioides to Pseudolaric Acid B
WEN Xiaodong,ZHU Fadi,GU Mengxuan,YE Huochun,FENG Gang,ZHANG Jing.Induction and Biological Characterization of Mutants of Colletotrichum gloeosporioides to Pseudolaric Acid B[J].Chinese Journal of Tropical Crops,2022,43(8):1654-1662.
Authors:WEN Xiaodong  ZHU Fadi  GU Mengxuan  YE Huochun  FENG Gang  ZHANG Jing
Institution:1. College of Agricultural, Guizhou University, Guiyang, Guizhou 550025, China2. Institute of Environment and Plant Protection, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan 571101, China3. Center for Rearch and Development to Fine Chemicals, Guizhou University, Guiyang, Guizhou 550025, China
Abstract:Pseudolaric acid B (PAB) is a natural diterpenoid acid with unique structure. Previous studies found that PAB has antifungal activity with broad-spectrum against a variety of plant pathogenic fungi such as Colletotrichum gloeosporioides. However, its antifungal mechanism is unknown. In this study, C. gloeosporioides strain JB-Q was selected for resistance induction by three methods, including chemical acclimation, UV irradiation and cobalt radiation. The biological characteristics of the stable resistant mutants were analyzed in order to provide research materials and clues for exploring the mode of actions of PAB. The results showed that 18 strains of C. gloeosporioides with stable inheritance and different resistance levels were obtained by chemical acclimation, and the resistance frequency was 1.69 × 10-7. The evaluation of biological characteristics showed that both the parent strain JB-Q and the resistant mutants could grow in the range of 15-35℃, and the mycelium grew fastest at 25℃. The growth rate of mutants with different resistance levels was different, and the resistance level was negatively correlated with the growth rate. The results of osmotic pressure test showed that JB-Q was more sensitive to 0.02% SDS, and its inhibition rate was as high as 70%. Under metal ion stress, the resistant mutants were more sensitive. The above results suggested that PAB could not be easy to cause the resistance of C. gloeosporioides, but the survival competitiveness of the induced resistant mutant strains is equivalent to that of the parent strain. In addition, we found that there was no cross resistance between PAB and pyraclostrobin, prochloraz and tebuconazole, while there was positive cross resistance with carbendazim, and its Pearson correlation coefficient was 0.7729 (P<0.01), indicating that PAB may have a similar action mechanism with microtubule inhibitor carbendazim, then further experiments are needed to verify this.
Keywords:Colletotrichum gloeosporioides  pseudolaric acid B  resistant mutants  biological characterization  
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