禾谷镰孢荧光定量检测体系的建立及其在玉米苗枯病上的应用

为实现对田间土壤中禾谷镰孢Fusarium graminearum的定量检测,本研究构建了土壤含菌量与玉米苗枯病病情指数的回归模型。基于禾谷镰孢甾醇14α-去甲基化酶基因CYP51C序列,设计特异性引物HQ1-F/HQ1-R,利用引物建立实时荧光定量PCR(RT-q PCR)体系,选取4个玉米自交系品种进行室内苗枯病接种试验,调查其病情指数,利用RT-qPCR体系检测土壤禾谷镰孢含菌量,并对病情指数和土壤禾谷镰孢含菌量进行回归。结果表明,仅禾谷镰孢扩增出目的条带并且可从多种病原菌土壤中检测出。RT-qPCR的熔解曲线具有单一吸收峰,扩增曲线的循环阈值与模板浓度呈良好的线性关系,扩增效率为104.7%,标准曲线为y=-3.2137x+34.9560(R~2=0.9968),最低可检测到1 pg/μL的DNA。随着土壤禾谷镰孢接菌量的增加,单位土壤禾谷镰孢含菌量呈线性增加,即y=13.603x-85.370(R~2=0.9998)。4个玉米品种的病情指数与土壤禾谷镰孢含菌量的回归曲线分别为y=0.0789x+22.0590(R~2=0.7949)、y=0.0304x+7.8686(R~2=0.9579)、y=0.0458x+23.7540(R~2=0.5420)、y=0.0471x+32.0760(R~2=0.6753)。     

湖北省小麦赤霉病菌对多菌灵、戊唑醇和咪鲜胺的敏感性

对2014年采自中国湖北省7个县、市的206株小麦赤霉病菌样品进行单孢分离与鉴定,并采用菌丝生长速率法随机测定了其中100株菌株对多菌灵、戊唑醇和咪鲜胺的敏感性,建立了其敏感基线。结果表明:多菌灵、戊唑醇和咪鲜胺对湖北省小麦赤霉病菌的EC50值范围分别为0.115~0.705、0.006~1.356和0.002~0.370 μg/mL,平均值分别为0.248、0.181和0.040 μg/mL;供试100株小麦赤霉病菌对3种药剂的敏感性频率均呈单峰拟正态分布,因此可将所得各EC50平均值分别作为湖北省小麦赤霉病菌对3种药剂的敏感基线参考值。以各药剂EC50平均值的10倍作为敏感性鉴别浓度,对2015及2016年湖北省小麦赤霉病菌的敏感性进行了监测,在该鉴别浓度下,多菌灵、戊唑醇和咪鲜胺对2015年小麦赤霉病菌的平均抑制率分别为100%、85.14%和82.35%,对2016年小麦赤霉病菌的平均抑制率分别为100%、76.67%和73.62%。研究表明,虽然戊唑醇和咪鲜胺对2016年小麦赤霉病菌的抑制率略有下降,但整体而言,湖北省小麦赤霉病菌对多菌灵、戊唑醇及咪鲜胺仍具有较高的敏感性。     

基于链特异性RNA-seq的禾谷镰刀菌全生活史转录组分析

为明确植物病原真菌禾谷镰刀菌Fusarium graminearum全生活史的转录组特征和基因表达模式,采用生物信息学技术对其生活史不同阶段15个时期或组织的链特异性RNA-seq数据进行分析。结果表明,共有8 106个基因在所有时期均有表达,为禾谷镰刀菌生活史核心基因,占总基因数的47.2%;有性生殖和侵染过程中表达的基因数相对较多,其中有性生殖后期表达的基因数最多,达15 221个;无性产孢和侵染小麦穗过程中基因表达水平整体较高,而分生孢子中基因表达水平最低。在燕麦培养基上禾谷镰刀菌气生菌丝的基因表达模式与侵染过程中的基因表达模式较为相似,而与营养生长菌丝的基因表达模式差异较大。该菌次生代谢物合成特征酶基因和分泌蛋白基因的表达模式均分成3类,即分别在侵染、营养生长和有性生殖过程上调表达,暗示其在生活史不同阶段的特异性功能。     

禾谷镰孢菌对氟唑菌酰羟胺敏感性基线的建立及药剂田间防效

为明确琥珀酸脱氢酶抑制剂类新型吡啶酰胺杀菌剂氟唑菌酰羟胺在中国小麦赤霉病防治中的应用潜力,分别采用菌丝生长速率法和孢子萌发法,测定了氟唑菌酰羟胺对湖北省6个地区106株禾谷镰孢菌的室内毒力、田间防效及其与多菌灵和氰烯菌酯的交互抗性。结果显示:氟唑菌酰羟胺对106株禾谷镰孢菌菌丝生长的EC₅₀值为 (0.018 0 ± 0.209 0) mg/L,平均值为 (0.072 8 ± 0.025 9) mg/L;对分生孢子萌发的EC₅₀值为 (0.052 7 ± 0.473 2) mg/L,平均值为 (0.176 0± 0.059 6) mg/mL;且其EC₅₀值频率分布均呈单峰曲线,因此可分别将菌丝生长和孢子萌发的平均EC₅₀值作为禾谷镰孢菌对氟唑菌酰羟胺的敏感性基线。初步的交互抗性测定结果表明,抗多菌灵或氰烯菌酯的菌株对氟唑菌酰羟胺均未表现出抗性。田间试验显示,氟唑菌酰羟胺有效剂量200 g/hm2处理的防效 (超过90.0%) 显著高于对照药剂氰烯菌酯600 g/hm2的防效 (78.0%),与空白对照相比增产效果在127%~135%之间。经氟唑菌酰羟胺处理后,小麦籽粒中由禾谷镰孢菌产生的毒素脱氧雪腐镰刀菌烯醇 (DON) 的含量比空白对照降低了55.09%。研究表明,氟唑菌酰羟胺对禾谷镰孢菌呈现出较高的室内活性且田间防效优越,同时还能降低小麦籽粒中DON毒素的含量及提高小麦产量,因此可作为生产中防治小麦赤霉病的替代或后备药剂,同时也可考虑用作为禾谷镰孢菌对多菌灵抗性治理的替代药剂。     

枯草芽胞杆菌与氟环唑联用对禾谷镰孢霉的增效作用机制

为验证枯草芽胞杆菌与氟环唑联用对禾谷镰孢霉的增效作用,用平板倒扣法和固体平板扩散法—打孔法测定菌药联用后枯草芽胞杆菌挥发性物质、抗真菌蛋白对禾谷镰孢霉的抑菌活性,观察抗真菌蛋白对禾谷镰孢霉菌丝形态的影响,并用刚果红染色法测定菌药联用后枯草芽胞杆菌纤维素酶活性。结果表明:菌药联用对禾谷镰孢霉的最高抑菌活性可达74.44%,表现增效和持效作用;枯草芽胞杆菌挥发性物质和抗真菌蛋白分别与氟环唑联用均可增强禾谷镰孢霉抑菌活性;菌药联用后枯草芽胞杆菌纤维素酶活性提高。研究表明枯草芽胞杆菌与氟环唑联用的增效机制是增强枯草芽胞杆菌挥发性物质对禾谷镰孢霉的抑菌活性,抗真菌蛋白活性的提高加强了禾谷镰孢霉菌丝溶解的程度,纤维素酶活性的增强提高了禾谷镰孢霉空间竞争作用和生物防治潜能。     

FgLEU1在禾谷镰刀菌亮氨酸生物合成及产毒致病中起关键作用

为挖掘新型药剂的潜在靶标,利用靶向基因敲除和互补技术研究赤霉病病原菌禾谷镰刀菌Fusarium graminearum中必需氨基酸亮氨酸合成酶编码基因FgLEU1的功能,并测定禾谷镰刀菌的生物学表型。结果表明,FgLEU1编码亮氨酸合成途径中的3-异丙基苹果酸脱水酶,其敲除突变体表现亮氨酸营养缺陷。生物学表型测定结果显示,与野生型菌株相比,FgLEU1敲除突变体的产孢量和孢子萌发率显著下降,产孢量仅为野生型菌株的20.96%,培养4 h后孢子萌发率下降了49.45%,且合成脱氧雪腐镰刀烯醇（呕吐毒素）能力丧失,在麦穗上的致病力下降,仅能侵染接种小穗,赤霉病症状不能扩展。外源添加一定量的亮氨酸、FgLeu1催化产物或导入含启动子的全长FgLEU1基因可以恢复敲除突变体表型缺陷。表明FgLEU1基因在禾谷镰刀菌亮氨酸合成、菌丝孢子形成及产毒致病过程中发挥着重要作用,可作为新型安全杀菌剂的潜在研发靶标,用于持续有效控制麦类赤霉病和镰刀菌毒素。     

禾谷镰刀菌CYP51A基因对五种三唑类杀菌剂敏感性的影响

为进一步明确14α-脱甲基酶编码基因CYP51A在禾谷镰刀菌Fusarium graminearum对三唑类杀菌剂敏感性中的作用,利用Split-marker方法敲除禾谷镰刀菌CYP51A基因并获得敲除CYP51A基因的禾谷镰刀菌ΔFgCYP51A菌株,比较该菌株和野生型菌株PH-1对5种三唑类杀菌剂的敏感性变化。结果表明,ΔFgCYP51A菌株对丙环唑、戊唑醇、烯唑醇、三唑醇和三唑酮的EC₅₀分别为0.024、0.047、0.148、0.154和0.474 mg/L。ΔFgCYP51A菌株对戊唑醇、三唑醇、三唑酮和丙环唑的敏感性均显著增加,而对烯唑醇的敏感性无明显变化。表明FgCYP51A基因与禾谷镰刀菌对三唑类杀菌剂的敏感性有关。     

Ultrastructural and immunocytochemical investigation of pathogen development and host responses in resistant and susceptible wheat spikes infected by Fusarium culmorum

Pathogen development and host responses in wheat spikes of resistant and susceptible cultivars infected by Fusarium culmorum causing Fusarium head blight (FHB), were investigated by means of electron microscopy as well as immunogold labelling techniques. The studies revealed similarities in the infection process and the initial spreading of the pathogen in wheat spikes between resistant and susceptible cultivars. However, the pathogen’s development was obviously more slow in the resistant cultivars as in comparison to a susceptible one. The structural defence reactions such as the formation of thick layered appositions and large papillae were essentially more pronounced in the infected host tissues of the resistant cultivars, than in the susceptible one. β -1,3-glucan was detected in the appositions and papillae. Furthermore, immunogold labelling of lignin demonstrated that there were no differences in the lignin contents of the wheat spikes between susceptible and resistant cultivars regarding the uninoculated healthy tissue, but densities of lignin in host cell walls of the infected wheat spikes differed distinctly between resistant and susceptible cultivars. The lignin content in the cell walls of the infected tissues of the susceptible wheat cultivar increased slightly, while the lignin accumulated intensely in the host cell walls of the infected wheat spikes of the resistant cultivars. These findings indicate that lignin accumulation in the infected wheat spikes may play an important role in resistance to the spreading of the pathogen in the host tissues. Immunogold labelling of the Fusarium toxin DON in the infected lemma showed the same labelling patterns in the host tissues of resistant and susceptible cultivars. However, there were distinct differences in the toxin concentration between the tissues of the susceptible and resistant cultivars. At the early stage of infection, the labelling densities for DON in resistant cultivars were significantly lower than those in the susceptible one. The present study indicates that the FHB resistant cultivars are able to develop active defence reactions during infection and spreading of the pathogen in the host tissues. The lower accumulation of the toxin DON in the tissues of the infected spikes of resistant cultivars which results from the host’s defence mechanisms may allow more intensive defence responses to the pathogen by the host.     

Effect of nitrogen application at anthesis on Fusarium head blight and mycotoxin accumulation in breadmaking wheat in the western part of Japan

In the western part of Japan, two wheat cultivars, Nishinokaori and Minaminokaori, are currently cultivated for breadmaking. Breadmaking wheat requires a higher protein content compared to the Japanese noodle wheat (the major type of wheat in Japan). This high protein level in the grain is obtained by top-dressing with nitrogen (N) near anthesis. Because such N applications may increase levels of Fusarium head blight (FHB) and consequent mycotoxin [deoxynivalenol (DON) and nivalenol (NIV)] accumulation in the grain, the effect of N application (0, 4, and 8 g/m²) at anthesis on FHB and mycotoxin accumulation in Nishinokaori and Minaminokaori was tested in the greenhouse in 2004 and 2005 and in two fields in 2006. In the greenhouse, plants were spray inoculated at 3, 10, and 20 days after N treatment. In field experiments, colonized maize kernels, which generate ascospores during the testing season, served as inoculum. In all experiments for both cultivars, N application at anthesis significantly increased grain protein as expected, but had no significant effect on FHB and DON and NIV levels in grain. These results suggest that, at least in these cultivars, N can be applied close to anthesis without increasing the risk of FHB and mycotoxin (DON and NIV) accumulation.     

四种SBIs类杀菌剂对不同发育阶段小麦赤霉病菌的毒力及其作用方式

为明确不同发育阶段小麦赤霉病菌对甾醇生物合成抑制剂类(SBIs)杀菌剂的敏感性差异及杀菌剂的作用方式,对4种SBIs类杀菌剂进行了室内毒力测定及田间药效试验。室内毒力测定结果表明:不同发育阶段小麦赤霉病菌对4种SBIs类药剂的敏感性存在较大差异,其敏感性由高到低依次为菌丝生长阶段和产孢阶段芽管伸长阶段孢子萌发阶段;其中,戊唑醇对菌丝生长、产孢、芽管伸长和孢子萌发阶段的EC50值分别为0.686、0.191、3.532和3.825μg/m L,咪鲜胺分别为0.063、0.305、0.827和36.581μg/m L,苯醚甲环唑分别为0.873、3.659、5.687和79.465μg/m L,烯唑醇分别为1.961、33.658、41.881和54.986μg/m L。显微观察4种SBIs类杀菌剂对小麦赤霉病菌菌丝生长和孢子萌发形态的影响,发现杀菌剂处理后菌丝末端分枝增多,新生菌丝伸长受阻,菌丝体局部膨大,有时可见菌丝及芽管的消解。田间试验结果表明:用量为有效成分100 g/hm2的戊唑醇、200 g/hm2的咪鲜胺和200 g/hm2的苯醚甲环唑对小麦赤霉病的保护效果为77.15%~79.26%,治疗效果为70.64%~72.77%,于发病前后使用均可有效防治该病害。     

The influence of carotenoid biosynthesis modification on the Fusarium culmorum and Fusarium oxysporum resistance in flax

Flax engineering to yield increased resistance to pathogens is the goal of this study. Since carotenoids act as antioxidants it is thus postulated that the accumulation of a higher quantity of these compounds in the transgenic plants might improve their resistance to pathogen infection.Our approach was based on the generation of transgenic flax overproducing carotene and analysis of its susceptibility to Fusarium infection. For transformation bacterial gene – crtB was used. As expected, transgenic plants showed increased resistance against pathogen infection.The impact of carotenoids on plant resistance to infection was verified by generation and analysis of transgenic flax with decreased content of carotene. The transgenic plants were obtained by suppression of endogenous flax gene coding for lycopene β-cyclase. Plant analysis revealed decrease in carotene content, however, an unexpected increase in resistance against Fusarium infection was detected. Further analysis of metabolites in the plants revealed that an increase in accumulation of other terpenoids and tocopherols, squalene and menthol were among them. Thus, it is suggested that repression of carotene synthesis results in the redirecting of substrates to other branches of isoprenoids synthesis.We conclude that a general level of antioxidants rather than the presence of any particular compound is the most important factor in resistance of the flax plant to pathogen infection.     

中国河南省小麦纹枯病菌对苯醚甲环唑及戊唑醇的敏感性

为明确中国河南省小麦纹枯病菌对苯醚甲环唑及戊唑醇的敏感性,采用菌丝生长速率法测定了2种药剂对采自河南省15个地市分离的95株病菌的毒力。结果显示:供试菌株对苯醚甲环唑和戊唑醇的EC50值分别在0.014~1.609和0.002~0.064 μg/mL之间。敏感性频率分布图显示:在小麦纹枯病菌群体中已出现对苯醚甲环唑敏感性下降的亚群体,而对戊唑醇则不明显;可将其中呈连续单峰频次分布的大多数菌株群体的平均EC50值（0.165±0.101）和（0.019±0.008）μg/mL分别作为小麦纹枯病菌对苯醚甲环唑和戊唑醇的相对敏感基线。苯醚甲环唑的平均EC50值是戊唑醇的26倍,表明该病原菌对戊唑醇更为敏感。不同地区病原菌群体间对苯醚甲环唑和戊唑醇的敏感性均存在显著性差异。小麦纹枯病菌对2种杀菌剂的敏感性之间存在着微弱的正相关性。本研究结果可为河南省小麦纹枯病防控杀菌剂的合理使用提供理论依据。     

禾谷丝核菌环介导等温扩增检测技术体系的建立

为建立简便、快速和灵敏的小麦纹枯病菌——禾谷丝核菌Rhizoctonia cerealis的分子检测技术体系,基于环介导等温扩增(loop-mediated isothermal amplification,LAMP)技术,以核糖体DNA内转录间隔区(internal transcribed spacer,ITS)为靶标序列,设计并筛选特异性引物,建立禾谷丝核菌的LAMP检测方法,并对其特异性、灵敏度和应用效果进行测定。结果表明,基于筛选获得的4条禾谷丝核菌特异性引物建立的LAMP方法能够从15种植物病原菌中特异性地检出禾谷丝核菌,近缘种立枯丝核菌Rhizoctonia solani和引致早期症状易混淆病害的病原菌禾顶囊壳菌Gaeumannomyces graminis、麦根腐平脐蠕孢菌Bipolaris sorokiniana和禾谷镰孢菌Fusarium graminearum均未检出;该LAMP方法在日光下的灵敏度为10^-1 ng/μL,在蓝光下的灵敏度为10^-3 ng/μL;对发病组织的检测显示该LAMP方法能从人工接种的小麦发病组织中...     

河南省小麦赤霉病菌对咯菌腈的敏感性

为明确河南省小麦赤霉病菌Fusarium graminearum对咯菌腈的敏感性,采用菌丝生长速率法测定了咯菌腈对从该省11个地市分离的95株菌株的毒力,通过方差分析法及聚类分析法对测定结果进行了分析,并研究了咯菌腈与多菌灵、戊唑醇对病菌毒力的相关性。结果显示：咯菌腈对小麦赤霉病菌菌丝生长的最低抑制浓度为0.1 μg/mL;咯菌腈对供试95株菌株的EC₅₀范围在0.003~0.088 μg/mL之间,平均EC₅₀为0.011 μg/mL;敏感性频率分布图显示,病菌群体中存在着对咯菌腈敏感性较低的亚群体,但67.4%供试菌株敏感性频率呈正态分布,将此部分菌株的EC₅₀平均值0.007 μg/mL作为小麦赤霉病菌对咯菌腈的敏感性基线。方差分析及聚类结果均显示,同一县市内的菌株对咯菌腈EC₅₀的最大值和最小值之比为1.1~8.3;除周口市沈丘县的菌株外,其余县市的菌株对咯菌腈敏感性差异不明显,咯菌腈EC₅₀平均值变化范围在0.005~0.028 g/mL之间,最大值是最小值的5.6倍;小麦赤霉病菌对咯菌腈的敏感性与其对多菌灵、戊唑醇的敏感性之间无明显相关性。表明河南省小麦赤霉病菌群体中尽管存在着敏感性较低的亚群体,但可通过药剂复配进行防控。     

中国河南省小麦纹枯病菌对咯菌腈的敏感性

为明确河南省小麦纹枯病菌对咯菌腈的敏感性,采用菌丝生长速率法测定了咯菌腈对2013年从河南省15个地市分离的98株病菌的毒力。结果表明:咯菌腈对供试小麦纹枯病菌菌株的EC50值在0.009~0.079 μg/mL之间,平均EC50值为(0.025 ±0.012)μg/mL;98个菌株对咯菌腈的敏感性分布呈连续单峰曲线,未出现敏感性下降的亚群体,可将(0.025 ±0.012)μg/mL作为小麦纹枯病菌对咯菌腈的敏感性基线;方差分析(LSD法)及SPSS聚类结果均显示,同一地市内的菌株对咯菌腈的敏感性差异较大,EC50最大值和最小值之比在1.0~4.1之间,而不同地区菌株间对咯菌腈敏感性差异不明显,咯菌腈EC50平均值变化范围在0.015~0.060 μg/mL之间,后者是前者的4倍;小麦纹枯病菌对咯菌腈的敏感性与其对噻呋酰胺、苯醚甲环唑、井冈霉素、丙环唑及戊唑醇的敏感性之间无明显相关性。本研究结果可为河南省小麦纹枯病防控杀菌剂的合理使用提供理论依据。     

中国河南省小麦纹枯病菌对井冈霉素及甲基立枯磷的敏感性

为明确中国河南省小麦纹枯病菌对井冈霉素及甲基立枯磷的敏感性,采用菌丝生长速率法测定了两种药剂对采自该省15个地市的98株病原菌菌丝生长的毒力。结果显示:井冈霉素和甲基立枯磷对供试菌株的EC₅₀值范围分别在0.023~0.852和0.035~0.512 μg/mL之间;其敏感性频率分布图显示,病原菌群体中虽已出现对井冈霉素敏感性下降的亚群体,但77.6%和99.0%的菌株对井冈霉素和甲基立枯磷的敏感性频率仍呈正态分布,因此可将此部分菌株的平均EC₅₀值（0.424±0.081）和（0.124±0.043）μg/mL分别作为小麦纹枯病菌对井冈霉素和甲基立枯磷的敏感基线;井冈霉素的平均EC₅₀值是甲基立枯磷的3倍,表明该病原菌对甲基立枯磷更为敏感;不同地理来源病原菌群体间对井冈霉素和甲基立枯磷的敏感性均存在显著的地域性差异;小麦纹枯病菌对两种杀菌剂的敏感性之间无明显相关性。研究结果可为河南省小麦纹枯病的可持续防控提供理论依据。     

江苏省水稻赤霉病病原菌分离鉴定与致病力分析

为明确江苏省水稻赤霉病的病原菌组成及致病特征,于2018—2019年从江苏省13个市41个县区采集341份水稻样品,采用组织分离法对病原菌进行分离纯化,基于翻译延伸因子（translationelongation factor-1α,TEF-1α）序列分析对分离菌株进行鉴定,通过TRI11基因序列鉴定及产毒力测定对其产毒化学型进行分析,并按照柯赫氏法则对其致病力进行验证。结果显示,从水稻样品中共分离获得病原菌392株,其中亚洲镰刀菌Fusarium asiaticum为385株,禾谷镰刀菌F. graminearum为7株。亚洲镰刀菌分为3-乙酰化脱氧雪腐镰刀菌烯醇（3-acetyldeoxynivalenol,3ADON）化学型和雪腐镰刀菌烯醇（nivalenol,NIV）化学型菌株,分别占亚洲镰刀菌总菌株数的 66.8% 和 33.2%;禾谷镰刀菌均为（15-acetyldeoxynivalenol,15ADON）化学型。产3ADON的亚洲镰刀菌在全省范围内为优势群体,而产15ADON的禾谷镰刀菌仅在淮北地区被发现;产NIV的亚洲镰刀菌对水稻的致病性要显著高于产3ADON的亚洲镰刀菌。不同产毒化学型亚洲镰刀菌对水稻均有一定致病性,但NIV化学型亚洲镰刀菌对水稻的致病力最强。     

氟啶胺等杀菌剂对不同发育阶段小麦赤霉病菌的毒力及其作用方式

为筛选高效防治小麦赤霉病的药剂,采用菌丝生长速率法和孢子萌发法测定了5种杀菌剂对不同发育阶段小麦赤霉病菌的毒力,并测定了田间接种条件下对小麦赤霉病的保护和治疗效果。结果表明,氟啶胺对小麦赤霉病菌菌丝生长、孢子萌发、芽管伸长和产孢的毒力均较高,EC_(50)≤0.249μg/m L;咯菌腈可抑制该病菌菌丝生长、芽管伸长和产孢,EC_(50)≤0.881μg/m L,但对孢子萌发几乎无抑制作用;菌毒清对分生孢子萌发、芽管伸长和产孢的毒力稍低,EC_(50)在5.726~10.244μg/m L之间,但高于其对菌丝生长的毒力;甲基硫菌灵和嘧菌酯对各发育阶段病菌的毒力均较低。在田间,接种前3 d施药,咯菌腈和氟啶胺对小麦赤霉病的保护效果较好,200 g(a.i.)/hm~2处理的效果在80.01%以上,可作为保护剂在发病前使用;菌毒清在接种前3 d和接种后3 d使用时保护效果和治疗效果均较高,200 g(a.i.)/hm~2处理的防效均在69.99%以上,既可作为保护剂在发病前使用,也可作为治疗剂在发病后使用。     

小麦纹枯病菌对噻呋酰胺的敏感性及抗药性突变体的主要生物学性状

采用菌丝生长速率法,分别测定了采自河北、山东和河南未使用过酰胺类药剂地区的166株小麦纹枯病菌Rhizotonia cerealis Van der Hoeven对噻呋酰胺的敏感性,并对小麦纹枯病菌抗噻呋酰胺突变体的诱导方法及突变体的主要生物学性状进行了研究。结果表明:噻呋酰胺对166株小麦纹枯病菌的平均EC50值为(0.072±0.022)μg/mL,菌株频率呈连续单峰曲线分布,未发现敏感性明显下降的亚群体,因此可将该EC50值作为小麦纹枯病菌对噻呋酰胺的敏感基线。通过紫外诱导加菌落角变的方法获得了5株抗噻呋酰胺突变体,其抗药性水平介于5.5~18.9倍之间;药剂驯化未能获得抗药性突变体。突变体在PSA平板上的生长速率与亲本菌株无显著性差异,但其菌丝干重、菌核形成数量和菌核干重均明显低于亲本菌株;除突变体HD7-7U对石新828的致病力未发生明显变化外,其他突变体对石新828和良星99的致病力均明显减弱;5株突变体的抗药性均不能稳定遗传;噻呋酰胺与戊唑醇、井冈霉素、咯菌腈、三唑酮、丙环唑和多菌灵之间无交互抗性关系。     

江苏、河南、安徽和山东四省小麦纹枯病菌对井冈霉素的敏感性监测

采用菌丝生长速率法监测了1984、2001和2010年江苏省小麦纹枯病菌Rhizoctonia cerealis对井冈霉素的敏感性变化趋势,以及2010年河南、安徽和山东3省小麦纹枯病菌对井冈霉素的敏感性现状。结果表明:1984、2001和2010年江苏省小麦纹枯病菌对井冈霉素的EC50值范围分别为0.31~0.87、0.05~1.21和0.20~1.09 μg/mL,26年间江苏省小麦纹枯病菌对井冈霉素的敏感性未发生显著性变化;2010年河南、安徽和山东3省小麦纹枯病菌对井冈霉素的EC50值范围分别为0.15~1.16、0.43~0.90和0.09~1.33 μg/mL,4省小麦纹枯病菌对井冈霉素的敏感性相互间差异不显著。井冈霉素仍然可以作为防治小麦纹枯病的主要药剂。