A single amino acid substitution in the SdhB protein of succinate dehydrogenase determines resistance to amicarthiazol in Xanthomonas oryzae pv. oryzae

BACKGROUND: Xanthomonas oryzae pv. Oryzae Ishiyama, a causal agent of rice bacterial leaf blight, was found to be sensitive in vitro to the systemic fungicide amicarthiazol (2‐amino‐4‐methylthiazole ‐5‐carboxanilide), which is a potent inhibitor of succinate dehydrogenase (SDH, EC 1.3.99.1). This paper aimed to determine the molecular resistance mechanism of X. oryzae pv. oryzae to amicarthiazol. RESULTS: UV‐induced resistant mutants of X. oryzae pv. oryzae to amicarthiazol were isolated. The activity of SDH in wild‐type X. oryzae pv. oryzae was strongly inhibited by amicarthiazol, while that in resistant mutants was insensitive, although their SDH activity was decreased compared with the wild‐type sensitive strain without amicarthiazol. A mutation of Histidine₂₂₉ (CAC) to Tyrosine₂₂₉ (TAC) was identified in sdhB, which encoded the iron–sulfur protein subunit of SDH. The sdhB from the mutant was ligated into a cosmid, pUFR034, to generate pUFR034RAX, which conferred resistance to amicarthiazol when transformed into the wild‐type sensitive strain. CONCLUSION: A mutation of His₂₂₉ (CAC) to Tyr₂₂₉ (TAC) in SdhB was responsible for determining amicarthiazol resistance. Copyright © 2010 Society of Chemical Industry     

竹醋液抑菌活性及对杀菌剂的增效作用

[目的]研究竹醋液抑菌活性及对杀菌剂增效作用,为竹醋液的综合利用及其化学杀菌剂复配提供理论依据。[方法]以苹果炭疽菌、苹果腐烂菌及番茄灰霉菌等3种重要植物病原真菌为供试病原菌,采用菌丝生长速率法,评价了4种竹醋液及多菌灵、异菌脲等6种常用化学杀菌剂的抑菌活性。[结果]培养48h后,咪鲜胺对苹果腐烂菌、苹果炭疽菌和番茄灰霉菌的EC50分别为0.0012、0.0088、0.1266mg/L;戊唑醇对苹果腐烂菌、苹果炭疽菌和番茄灰霉菌的EC50分别为0.0388、0.0725、0.7993mg/L;竹醋1对苹果腐烂菌、苹果炭疽菌和番茄灰霉菌的EC50分别为5199.7932（稀释192.32倍）、5701.2827（稀释175.40倍）、6232.0030mg/L（稀释160.46倍）。咪鲜胺EC50∶竹醋液1EC50=1∶5复配组合共毒系数（CTC）为261.23,戊唑醇EC50∶竹醋液1EC50=1∶5复配组合CTC为249.49。[结论]咪鲜胺、戊唑醇及竹醋液1的抑菌活性较强,竹醋液对咪鲜胺、戊唑醇2种供试杀菌剂具有增效作用。     

黄芪种子带菌检测及药剂消毒处理

采用离体平皿法,对黄芪种子进行了表面与内部的带菌检测;通过平皿培养法,分析了不同药剂对黄芪种子发芽率的影响,观察了药剂浸种的消毒效果。结果表明,黄芪种子携带的主要真菌类群为链格孢菌(Alternaria spp.)、曲霉属(Aspergillus spp.)和青霉属(Penicilliun spp.)。世高、多菌灵和甲基托布津浸种处理对黄芪种子的发芽率无影响,而高浓度三唑酮浸种处理会显著降低黄芪种子的发芽率(P0.05)。100μg/mL的世高和500μg/mL的多菌灵浸种处理对黄芪种子有较好消毒效果,达到85.0%以上,建议选择作为黄芪播种前种子处理的药剂。     

Characterization and taxonomic reassessment of the box blight pathogen Calonectria pseudonaviculata,introducing Calonectria henricotiae sp. nov.

Calonectria pseudonaviculata, the causal agent of the disease of Buxus spp. known as ‘box blight’, was first detected in the mid‐1990s in the UK and New Zealand. Since then, the geographic range of box blight has rapidly expanded to at least 21 countries throughout temperate regions of the world, causing significant losses in nurseries, gardens and wild boxwood populations. This study determined the genetic diversity in a collection of 234 Calonectria isolates from diseased Buxus plants, originating from 15 countries and four continents. Two genetic clades, G1 and G2, were identified within this sample using multilocus phylogenetic analysis. The application of genealogical concordance phylogenetic species recognition criteria using four independent nuclear loci determined that the Calonectria isolates in these two clades are separate phylogenetic species. The isolates in the G1 clade were upheld as C. pseudonaviculata sensu stricto. Based on phylogenetic distinctiveness and the lack of mating, a new species is proposed, Calonectria henricotiae sp. nov., for the Calonectria isolates in the G2 clade. A PCR‐RFLP assay and real‐time PCR assays were developed to easily and reproducibly differentiate these species. To assess the practical implications of the identification of the two species, their physiology, fungicide susceptibility and pathogenicity were compared. No differences in pathogenicity were observed. However, C. henricotiae isolates exhibited greater thermotolerance and reduced sensitivity to specific triazole as well as strobilurin fungicides. The identification of a second phylogenetic species causing box blight may have a substantial impact on the epidemiology and control of this destructive disease.     

浙江省果蔬灰霉病菌对嘧菌酯的抗药性研究

采用菌丝生长速率法,连续监测了2010—2012年间浙江省果蔬灰霉病菌对QoI类杀菌剂嘧菌酯的敏感性变化。 结果表明:病菌群体中的低敏感性亚群体的比例明显上升,EC50值>5 mg/L 菌株的比例分别为12.5%、15.8%和28.3%;在菌丝生长阶段和孢子萌发阶段,旁路氧化在灰霉病菌对嘧菌酯敏感性中的平均相对贡献值(F)分别为2.91±0.89和5.72±2.82;嘧菌酯抗药性菌株的菌丝生长速率、产孢量、产菌核数和致病力与敏感菌株相比无显著差异。抗药性分子机制研究表明,灰霉病菌中存在2种类型的cyt b基因:Ⅰ型cyt b基因在第143位密码子后紧跟内含子;Ⅱ型cyt b基因在第143位密码子后没有紧跟内含子。大多数的灰霉病菌菌株属于Ⅱ型。Ⅰ型菌株均为嘧菌酯敏感菌株,Ⅱ型菌株为嘧菌酯敏感菌株或抗性菌株。抗性菌株的cyt b 基因的第143位密码子由甘氨酸(GGC)突变为了丙氨酸(GCC),抗药性机制为G143A。     

不同药剂组合对马铃薯晚疫病防治效果及产量的影响

以马铃薯晚疫病感病品种费乌瑞它为试验对象,采用68％精甲霜锰锌可湿粉剂、70％安泰生丙森锌可湿粉剂和64％恶霜锰锌可湿粉剂3种药剂进行马铃薯晚疫病的防治试验,结果表明：交替使用其中2种或3种药剂防治马铃薯晚疫病的效果优于单一使用1种药剂;68％精甲霜锰锌可湿粉剂与70％安泰生丙森锌可湿粉剂、70％安泰生丙森锌可湿粉剂与64％恶霜锰锌可湿粉剂、68％精甲霜锰锌可湿粉剂与64％恶霜锰锌可湿粉剂3种组合交替施用共喷药4次,防效分别为65.96％、61.70％、60.34％,商品薯产量比对照分别增产为61.74％和60.07％和52.01％。生产上可选用药剂交替使用来防治马铃薯晚疫病。     

苜蓿花期三种杀菌剂对意大利蜜蜂保护酶和解毒酶的影响

为探究苜蓿生产过程中使用的杀菌剂对其传粉昆虫意大利蜜蜂(意蜂)的安全性,利用田间推荐使用浓度的菌核净、咪鲜胺和异菌脲处理意大利蜜蜂,分别测定蜜蜂体内3种保护酶:超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)和3种解毒酶:羧酸酯酶(CarE)、谷胱甘肽S-转移酶(GSTs)和细胞色素P450(P450)的活性。结果表明,不同浓度的3种杀菌剂均可诱导意蜂SOD和POD活性,其中咪鲜胺1500倍液(0.17 mg·L^-1)处理的蜜蜂体内SOD和POD活性最高,分别为对照的1.82和5.40倍。3种杀菌剂对CAT、CarE和GSTs活性表现为低浓度诱导,高浓度抑制,对P450活性表现为抑制作用(咪鲜胺)或者诱导作用(菌核净和异菌脲)。随着处理时间的延长,3种杀菌剂1000倍液(0.40、0.25和0.50 mg·L^-1)对意蜂体内3种保护酶的活性总体表现为诱导作用;而对3种解毒酶的影响各异,菌核净对3种解毒酶活性均为先激活后抑制;异菌脲对CarE和GSTs活性表现为先抑制后激活,对P450为诱导作用;咪鲜胺对CarE和GSTs表现为先激活后抑制,对P450表现为随时间逐渐降低的抑制作用。该结果表明意蜂可以通过调节体内保护酶和解毒酶活性,降低杀菌剂的负面影响,但对蜜蜂正常生理和代谢产生了影响。因此在生产中应当谨慎施用杀菌剂,保护蜜蜂安全授粉。     

Fusarium seedling blight of wheat and oats: effects of infection level and fungicide seed treatments on agronomic characters

The effects of seed-borne Fusarium spp. and Microdochium nivale infection in spring wheat, winter wheat, and oats in Sweden was investigated in field trials for the agronomic characters yield, thousand kernel weight, grain volume weight, gluten, protein, starch, straw strength, and plant density. Seed with high and low levels of infection was mixed to obtain six infection levels in the proportions of 100/0, 80/20, 60/40, 40/60, 20/80, and 0/100 percent. The seed was untreated or treated with Celest Extra Formula M (CEFM, difenoconazole + fludioxonil) or Celest Formula M (CFM, fludioxonil). In the field trials using untreated seed, there were significant differences between infection levels only for some agronomic characters and levels. Fungicide seed treatment with CEFM in spring wheat had no significant effect on most agronomic characters including yield. In winter wheat and oats, seed treatment with CFM increased yield by 7–11% and plant density by up to 33% while having no effect on other characters. The percentage discoloration of crown roots and stem bases due to Fusarium/Microdochium spp. was also investigated visually in winter wheat and oats and found to increase with higher infection level. Fungicide seed treatment thus mainly increased plant emergence in seed lots with low-to-moderate Fusarium/Microdochium spp. infection and had little or no effects on other agronomic characters.     

Colonization of red raspberry flowers and fruit by Botrytis cinerea under commercial production conditions in northwestern Washington,USA

Colonization of red raspberry flowers and fruit by Botrytis cinerea was determined during 2017–2018 growing seasons under commercial fungicide application programmes used for grey mould management in northwestern Washington, USA. Colonization of flowers and fruit was assessed qualitatively (incidence, %) and quantitatively (abundance, number of colonies) by recovering B. cinerea from surface-disinfested samples. Both incidence and abundance of flower colonization were significantly lower than fruit colonization in both untreated and fungicide-treated plots. Incidence of flower colonization did not differ significantly between untreated and fungicide-treated plots (43% vs. 45%, respectively). In contrast, significantly greater colonization incidence was detected at green fruit stage in untreated compared to fungicide-treated plots (96% vs. 77%, respectively). Ripe fruit had the greatest colonization incidence among the three stages sampled and colonization was not significantly different between untreated and fungicide-treated plots (100% vs. 92%, respectively). Similarly, colonization abundance of flowers did not differ significantly between untreated and fungicide-treated plots (1.0 colonies per colonized flower in both treatments), but colonization abundance of green and ripe fruit was decreased 2.3- and 2.1-fold, respectively, in fungicide-treated plots. DNA fingerprinting analysis of the pathogen revealed that different multilocus genotypes colonized flowers and fruit within the same inflorescence and that genotypic diversity increased through time, suggesting independent infection events. Overall, our results demonstrate that under current environmental conditions, raspberry flowers may not be the exclusive or major route of infection for grey mould of red raspberry in northwestern Washington. Implications of current findings for management and further research are discussed.