丝核菌细胞核染色技术的研究

比较了5种不同的染色方法对丝核菌（Rhizoctonia)菌株细胞核的染色效果。结果表明，番红O－KOH的染色效果最好，Giemsa、苯胺蓝－甘油次之，染色效果最差的是苯胺蓝－乳酚油及曲利苯蓝－乳酚油。用番红O－KOH染色方法对分离自13种作物的57个丝核菌菌株进行细胞核染色，其中3个菌株为双核丝核菌，其余54个菌株为多核丝核菌。     

Development of Stem-base Pathogens on Different Cultivars of Winter Wheat Determined by Quantitative PCR

The progress of development of stem-base pathogens in crops of second winter wheat was plotted in nine experiments in three years. The amount of each pathogen present was determined by quantitative PCR. Where Tapesia yallundae was present in quantifiable amounts, it usually developed earlier than the other eyespot pathogen, T. acuformis. Both species were usually present in greater amounts on cultivars which are more susceptible to eyespot. The sharp eyespot pathogen, Rhizoctonia cerealis, developed more erratically than either of the Tapesia spp. and there were no consistent effects on different cultivars. Fusarium spp., the cause of brown foot rot, were rarely present in quantifiable amounts, but Microdochium nivale was usually present as one or both of the varieties nivale and majus. Late-season (after anthesis) decreases in M. nivale suggest that any brown foot rot symptoms attributable to this fungus would have fully developed earlier. Cultivar differences in amounts of M. nivale were most clear in stems during internode extension and when relatively large amounts of DNA were present. Such differences approximately reflected eyespot susceptibility, cv. Soissons containing most and cv. Lynx containing least DNA. The results emphasise the difficulty in relating diagnoses, by quantitative PCR or other means, at early growth stages when decisions to apply fungicides against stem-base disease are made, to later disease severity.     

Whole-Cell Fatty Acid Composition to Characterize and Differentiate Isolates of Rhizoctonia Species Associated with Turfgrass Diseases in Japan

Cellular fatty acids were analyzed to characterize and differentiate 34 isolates of Rhizoctonia species representing binucleate Rhizoctonia AG-D (I), AG-D (II), R. solani AG 2-2 IIIB, AG 2-2 LP, R. circinata var. circinata and var. oryzae associated with turfgrass diseases in Japan. Myristic, pentadecanoic, palmitic, palmitoleic, stearic, oleic, linoleic and linolenic acids were consistently present in varying quantities in all isolates. Heptadecanoic and 9-heptadecenoic acids were present in isolates of Rhizoctonia AG-D (I), AG-D (II), R. solani AG 2-2 IIIB and AG 2-2 LP but not in isolates of R. circinata var. circinata and var. oryzae. Palmitic, oleic and linoleic acids were the major fatty acids found, constituting 88.30-98.37% of the whole-cell fatty acid content. The remaining fatty acids were present in smaller amounts. Isolates within a single group were closely clustered, whereas isolates from different groups were clearly distinguishable based on average linkage cluster analysis of cellular fatty acids. Principal component analysis, based on all fatty acids detected, confirmed the distinct separation of isolates representing the six groups of Rhizoctonia species obtained from turfgrasses. These results suggested that fatty acid analysis is useful for the characterization and differentiation of isolates of Rhizoctonia species associated with turfgrass diseases. Received 21 May 2001/ Accepted in revised form 28 September 2001     

镧素对镰刀菌Fusarium solani及其致病酶的影响

采用琼脂平板生长速率法及液体培养基培养测定了La对镰刀菌Fusarium solani的抑制作用和毒力,并测定了其对病菌胞外的果胶酶、蛋白酶和纤维素酶等几种致病酶活性的影响。结果表明,随着La2O3浓度升高,对病菌菌丝生长的抑制作用增强,对病菌的EC50和EC95分别为278.2和552.0 mg/L;在一定浓度范围内,La提高了单位量菌丝所产生3种致病酶的活性,但由于菌丝生长受到抑制,除蛋白酶外,病菌胞外致病酶果胶酶和纤维素酶的总量或总活性受到了抑制,降低了病菌的致病力。     

茭白纹枯病发生规律及产量损失测定

茭白纹枯病发生规律与产量损失的研究结果表明:高温高湿、种植过密、偏施氮肥、缺乏钾素是病害加重的主要原因;田间发病盛期在7月中旬至8月中旬;单茭重减低率与病级之间呈极显著线性相关;始病期越早,危害越重,产量损失也越大;病害危害程度与有效分蘖数(孕茭数)减少率、单茭重减低率、产量损失率之间呈高度正相关。在产量损失构成中,有效分蘖数的减少为主要因素。     

几种除草剂对水稻纹枯病菌的影响

研究了丁草胺、环庚草醚、草甘膦、苄嘧磺隆、吡嘧磺隆和环丙嘧磺隆6种除草剂对水稻纹枯病菌菌丝生长和菌核形成的影响。结果表明,在一定浓度范围内,6种除草剂均能抑制菌丝生长,减少菌核形成数量,且浓度越高,抑制能力越强,但在较高浓度范围内,除草剂可增加菌核体积;吡嘧磺隆在5μg/ml、环丙嘧磺隆在10μg/ml时可增加菌核形成数量;在含除草剂培养基上产生的菌核都可萌发。     

Anastomosis Groups, Pathogenicity and Sensitivity to Fungicides of Rhizoctonia solani Isolates Collected on Potato Crops in France

A collection of 241 isolates of Rhizoctonia solani obtained from potato plants grown in different areas in France was characterized for anastomosis grouping, symptomatology on tubers of different cultivars and sensitivity to three fungicides. Most isolates collected belonged to (anastomosis groups (AGs)) AG 3, but 2% and 4% of the isolates were AG 5 and AG 2-1. AG 3 and AG 2-1 isolates were mostly obtained from sclerotia on tubers, but all AG 5, some AG 3 and some AG 2-1 isolates were recovered from superficial tuber alterations, like deformations, corky or scabby lesions. Sclerotia were formed on tubers produced by healthy stem cuttings grown in soil artificially infested with AG 3, but not on tubers grown in soil infested with either AG 5 or AG 2-1. No variation in susceptibility to sclerotial formation was observed among five potato cultivars. In all cases, a large proportion of tubers showed superficial corky lesions, often associated with deformations. The proportion of tubers with lesions and deformations was highest in soil infested with AG 2-1 and significantly lower on cv. Samba in all treatments. All isolates were highly sensitive to flutolanil, iprodione and pencycuron, except the AG 5 isolates, moderately sensitive to pencycuron. These results show that, although AG 3 is the most common R. solani group on potato in France, AG 5 and AG 2-1 may be present. Isolates differed for pathogenicity. In vitro sensitivity to fungicides varied among AGs.     

一株马铃薯干腐病生防菌的鉴定及其抑菌活性研究

镰孢菌引起的马铃薯干腐病是影响马铃薯贮存期间产量与品质的重要病害之一。寻找高效低毒的防治马铃薯干腐病的生防药剂对提高马铃薯的产量与质量具有重要意义。本研究以腐皮镰孢Fusarium solani为病原菌, 测定分离自马铃薯块茎的菌株2-1-9-CJK-2的抑菌活性。该菌株发酵液、无菌发酵液、菌悬液、胞内组分以及挥发性有机物对腐皮镰孢均具有抑菌活性, 抑制率分别为93.37%、61.92%、80.82%、44.01%和49.16%。结合形态学、生理生化特征和多基因系统发育树构建鉴定其为贝莱斯芽胞杆菌Bacillus velezensis。马铃薯切片试验和马铃薯块茎试验表明, 当菌株发酵液浓度为1×109 cfu/mL时, 对病原菌的抑制效果最好。除此之外, 菌株可以分泌蛋白酶、淀粉酶、纤维素酶, 具有溶磷能力和生物膜形成能力, 但不具有溶血性, 并且对供试的其他6种病原真菌有较强的抑菌活性。研究表明贝莱斯芽胞杆菌2-1-9-CJK-2抑菌谱广, 具有生物安全性, 对腐皮镰孢有较强的抑菌活性, 在植物病害防治中具有一定的应用潜力, 可为马铃薯干腐病生防制剂的开发奠定基础。     

华北地区十字花科芸薹属蔬菜上立枯丝核菌的病原生物学研究

 由丝核菌引起的十字花科蔬菜叶腐和茎基腐病在中国华北地区普遍发生,其中以河北、内蒙以及北京较为严重。2011~2018年,从华北地区不同省份具有典型叶腐和茎基腐症状的芸苔属蔬菜上分离获得95个丝核菌(Rhizoctonia spp.)分离物,大多数分离自发病植株的叶部,少数分离自茎基部。通过细胞核染色,87株菌属于多核丝核菌,另外8株属于双核丝核菌;经菌丝融合鉴定、rDNA-ITS区及TEF-1α(translation elongation factor 1-alpha, TEF-1α)序列分析,大多数多核丝核菌属于立枯丝核菌(Rhizoctonia solani)AG-2-1(74％),其他少数分别属于AG-1-IB(16％)、AG-4-HG II(2％)和双核丝核菌AG-A(8％)。温室条件下进行寄主范围致病力测定,各分离物对原寄主都表现出致病力,呈现典型叶腐或茎基腐症状;对其他作物的致病力差异较大。不同融合群(Anastomosis group,AG)的菌株对寄主致病力大小存在差异,AG-2-1致病力最强,只有AG-A对叶部没有致病力。AG-2-1对寄主叶部的致病力和对茎基部的致病力呈显著正相关,AG-1-IB对寄主叶部的致病力和对茎基部的致病力无显著相关性。     

草莓丝核菌根腐病病原菌鉴定及7种杀菌剂的抑菌作用测定

为了解草莓丝核菌根腐病的病原种类及筛选防治丝核菌根腐病的有效杀菌剂,本研究基于形态学特征、细胞核荧光染色、菌丝融合群测定以及rDNA-ITS的序列分析,对北京和河北承德地区的草莓丝核菌根腐病的病原菌进行了鉴定,并利用菌丝生长速率法测定了7种杀菌剂对丝核菌的抑菌作用。结果发现,北京地区的丝核菌为双核丝核菌(binucleate Rhizoctonia,BNR),属于融合群AG-A;河北的丝核菌为立枯丝核菌Rhizoctonia solani,属于融合群AG-4。氟啶胺、吡唑醚菌酯、噻呋酰胺、戊唑醇、咯菌腈、氟硅唑对2种丝核菌均有很强的抑制作用,EC_(50)值为0.063 9～2.485 7μg/mL,抑霉唑的抑制作用较差,EC_(50)值为9.966 8～11.236 8μg/mL。同一种杀菌剂对不同丝核菌的抑制作用存在差异,噻呋酰胺、戊唑醇、氟硅唑、咯菌腈和抑霉唑对立枯丝核菌的抑制作用强于对双核丝核菌。试验结果为生产上合理选用杀菌剂防治草莓丝核菌根腐病提供了科学依据。