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     

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。     

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.     

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。     

中国东北地区水稻纹枯病病原菌种类及融合群的分子鉴定

为明确中国东北地区水稻纹枯病病原菌种类及融合群的归属情况, 2015－2017年从黑龙江省、吉林省和辽宁省的17个水稻主产区采集水稻纹枯病标样, 分离获得水稻纹枯病菌214株, 运用水稻纹枯病菌的不同病原菌及融合群的特异性引物对214株水稻纹枯病菌进行病原菌种类和融合群鉴定, 并利用rDNA内转录间隔区(ITS)序列, 对供试水稻丝核菌的融合群归属进行了分析。结果表明:供试214株水稻纹枯病菌分属于茄丝核菌Rhizoctonia solani和水稻丝核菌Rhizoctonia oryzae-sativae, 菌株数分别为198株和16株, 占比分别为92.52%和7.48%。茄丝核菌菌株分属于2个融合群, 分别为AG1-IA和AG4, 菌株数分别为191株和7株, 占比分别为96.46%和3.54%。水稻丝核菌菌株均属于AG-Bb融合群, 菌株数为16株。不同年份水稻纹枯病的病原菌种类及融合群出现的频率和地域分布无明显变化, 而不同地域间水稻纹枯病病原菌的种类及融合群具有明显的分化特征, AG1-IA融合群在中国东北三省各个水稻产区均有分布且均为优势融合群, AG4融合群在辽宁省盘锦市出现频率最高, 水稻丝核菌AG-Bb融合群在吉林省吉林市、通化市和梅河口市出现频率最高。     

中国北纬33度地区小麦纹枯病菌的群体组成及致病力研究

为明确中国北纬33度地区小麦纹枯病病原菌的组成和致病力,对分离自江苏、安徽、河南、湖北4省的157株丝核菌,通过Hoechst 33258荧光染色法进行细胞核数量观测,利用菌丝融合的方法测定其所属融合群,并测定了其中111株丝核菌株对3个小麦品种(CI12633、陕229、扬麦158)麦苗的致病力.结果表明,在供试菌株中,150株为双核丝核菌,占95.54%,都属于AG-D融合群;7株为多核丝核菌,占4.46%,其中3株属于AG-1-IB融合群,3株属于AG11融合群,1株属于AG5融合群.供试菌株的致病力存在明显差异,双核丝核菌的致病力显著高于多核丝核菌,AG-D融合群中两种不同融合频率的菌株致病力无明显差异.不同地区的菌株间致病力无明显差异,菌株对3个小麦品种的致病力有显著差异,对CI12633致病力最强,对陕229致病力最弱,对扬麦158的致病力居中.     

浙江省水稻纹枯病菌的遗传分化与致病力研究

鉴定了从浙江省各地采集的60株水稻纹枯病菌株的菌丝融合群归属情况。该60个菌株均与AG-1融合群的3个亚群AG-1-ⅠA、AG-1-ⅠB、AG-1-ⅠC的标准菌株融合,当属AG-1融合群。其中,有58株均与标准菌株AG-BⅠ融合,所有菌株与其他标准菌株均无融合现象。通过离体接种对菌株致病力进行鉴定,发现各菌株间致病力差异显著。致病力最强的TTZF-1与其中52个菌株差异极为显著。进一步的转录间隔区(ITS)序列分析显示,系统发育树的结果与融合群鉴定的结果一致,与菌株间致病力差异无相关性。