果生刺盘孢Crz1基因对DMI类杀菌剂抗性、外界胁迫应答、附着胞形成和致病力的影响  

Effects of Crz1 gene on resistance to DMI fungicides,stress response,appressorium formation and pathogenesis of filamentous fungus Colletotrichum fructicola

作　　者：马梦婷 李河 Ma Mengting;Li He(Key Laboratory for Non-Wood Forest Cultivation and Conservation of Ministry of Education,College of Forestry,Central South University of Forestry and Technology,Changsha 410004,Hunan Province,China)

机构地区：[1]中南林业科技大学林学院,经济林培育与保护省部共建教育部重点实验室,长沙410004

出　　处：《植物保护学报》2023年第1期231-239,共9页Journal of Plant Protection

基　　金：国家自然科学基金(32071765)。

摘　　要：为明确钙调磷酸酶应答锌指转录因子1(calcineurin responsive zinc finger transcription factor 1,Crz1)在果生刺盘孢Colletotrichumfructicola中的生物学功能,进而有效防控油茶炭疽病,该研究构建果生刺盘孢Crz1基因敲除突变体ΔCfcrz1和回补菌株ΔCfcrz1/Crz1,并测定突变体ΔCfcrz1、回补菌株ΔCfcrz1/Crz1和野生型菌株的生长发育、附着胞形成、外界胁迫应答、致病力以及对唑类杀菌剂的敏感性。结果显示,果生刺盘孢Crz1蛋白氨基酸序列与暹罗刺盘孢C.siamese的同源性最高,支持率高达100%;在马铃薯葡萄糖琼脂(potato dextrose agar,PDA)和基本培养基上,野生型菌株、回补菌株ΔCfcrz1/Crz1和突变体ΔCfcrz1的菌落直径分别为5.7、5.5、5.1 cm和5.3、5.1、2.8 cm,突变体ΔCfcrz1菌丝直径均极显著低于其他2株菌株;在含CaCl2和二硫苏糖醇(dithiothreitol,DTT)的PDA培养基上,突变体ΔCfcrz1菌丝生长抑制率分别为100.0%和46.3%,极显著高于其他2株菌株;突变体ΔCfcrz1形成的分生孢子量和附着胞形成率分别为7.2×105个/mL和17.3%,极显著小于其他2株菌株;突变体ΔCfcrz1对油茶叶的致病力明显减弱,病斑直径仅为0.06 cm,极显著低于其他2株菌株;当抑霉唑浓度大于0.1μg/mL、苯醚甲环唑浓度大于0.5μg/mL、三唑酮浓度大于4μg/mL、戊唑醇浓度大于0.5μg/mL时,突变体ΔCfcrz1菌丝生长抑制率分别大于34%、80%、39%和22%,极显著高于野生型菌株和回补菌株ΔCfcrz1/Crz1。表明转录因子Crz 1参与调控油茶果生刺盘孢的生长、产孢、外界胁迫应答、致病力和对DMI类杀菌剂的抗性。To study the biological functions of calcineurin responsive zinc finger transcription factor 1(Crz1)in fruit rot pathogen Colletotrichum fructicola,and to control anthracnose of Camellia oleifera effectively,a gene knockout mutantΔCfcrz1 and its complementary strainΔCfcrz1/Crz1 of C.fructicola were generated,then the biological functions of the mutant strainΔCfcrz1,complementary strainΔCfcrz1/Crz1 and wild strain were determined,including growth,appressorium formation,stress response,pathogenicity and resistance to DMI fungicides.This result indicated that Crz1 protein from C.fructicola had the highest amino acid sequence homology,100%identify,with that of C.siamense;the colony diameters of wild type,complement strainΔCfcrz1/Crz1 and mutantΔCfcrz1 cultured on the potato dextrose agar(PDA)and minimal medium(MM)were 5.7,5.5,5.1 cm and 5.3,5.1,2.8 cm,respectively,significantly lower for the mutant than for the other two strains;the inhibition rates on the PDA medium supplemented with CaCl2and dithiothreitol(DDT)were 100.0%and 46.3%,respectively,for the mutant strainΔCfcrz1 of C.fructicola,which was significantly higher than those for the other two strain;the percent conidia produced appressorium and conidia volume for mutantΔCfcrz1 was 17.3%or 7.2×10~5/mL,which that was significantly lower than those for the other two strains;the mutant strainΔCfcrz1showed weaker pathogenicity,with a mean lesion diameter of 0.06 cm,on wounded tea-oil than the other two strains;when the concentration of imazolium was greater than 0.1μg/mL,the concentration of difenoconazole was greater than 0.5μg/mL,the concentrations of triazolone greater than 4μg/mL,and the concentration of tebuconazole greater than 0.5μg/mL,the inhibition rates of mutantΔCfcrz1 were34%,80%,39%and 22%,respectively,which were significantly higher than that of wild type and supplement strain.These results revealed that the transcription factor Crz1 was involved in regulating the growth,conidiation,response to external stresses,pathogenicity,and resista

关 键 词：油茶 果生刺盘孢 杀菌剂 致病力 胁迫响应 

分 类 号：S763.7[农业科学—森林保护学]