山东小麦纹枯病菌致病性与遗传分化关系的研究

 对山东麦区被鉴定为双核丝核菌的35个菌株进行致病性测定的基础上,选用15个随机引物对上述菌株进行了RAPD分析,共标记出171条DNA片段,其中多态性片段161个,多态率为94.15%。用UPGMA法构建系统树,以遗传距离0.33为阈值,被鉴定为AG-D融合群的33个菌株隶属于同一个RAPD组,而3个未定融合群菌株(WK-6、WK-37和WK-13)均为独立的RAPD组。以遗传距离0.25为阈值,将属于同一个融合群的33个AG-D群菌株划分为7个亚组,说明受试小麦纹枯病菌株间存在丰富的遗传多样性。综合分析受试菌株的致病力测定结果与RAPD分析发现,供试菌株的RAPD组与菌株致病力的强弱无明显的相关性,但少数菌株的致病力强弱与亚RAPD组有一定的相关性。     

小麦纹枯病菌中的dsRNA及其与菌株生物学特性的相关性

对采集自我国山东、河南、江苏、安徽4省的21株小麦纹枯病菌菌丝中的dsRNA进行了检测,研究了dsRNA条带与菌株菌落形态、生长速度、对小麦的致病力等生物学特性之间的相关性。结果表明,从16个菌株中检测到dsRNA条带,dsRNA的大小和数量存在丰富的多样性。致病力测定结果表明这21个小麦纹枯病菌菌株的致病力差异显著。目前暂未发现小麦纹枯病菌中dsRNA的类型和数量与菌株的菌落形态、生长速度及致病力等性状存在明显的相关性,但是致病力较弱的菌株中dsRNA条带的多样性更丰富。本研究为今后发掘小麦纹枯病菌中的低毒真菌病毒作了初步的探索。     

江苏省几种作物丝核菌交互致病性的聚类分析

 本文在鉴定江苏省几种作物病原丝核菌所属种及菌丝融合群的基础上,进行六种作物病原丝核菌菌株与其寄主的人工交互接种试验,结果表明六种作物病原丝核菌对各自的原分离作物均表现强致病性。将该六种病菌在六种作物上的病情指数经聚类分析可划分为三类,第一类为水稻纹枯病菌,在人工接种条件下可侵染六种作物,综合致病力最强,平均病指为42、38;第二类为玉米、大豆纹枯病菌及棉花立枯病菌,其致病特性及症状与水稻纹枯病菌相似,但致病力较弱,平均病指分别为31、0,25、80和18、95:第三类为大、小麦纹枯病菌,其致病特性与前两者截然不同,不侵染大豆和棉花,平均病指为28、1和26、53。据致病力划分的类群与菌株所属的分类地位基本吻合。另外,本文还讨论了病原丝核菌在自然界中相互侵染的可能性,以及各种作物丝核菌种群的多样性等问题。     

新疆北疆棉田立枯丝核菌不同菌丝融合群致病力的研究

从新疆北疆棉区采集了典型的棉花立枯病病苗及棉田土标样686份,按常规分离方法分离得到399个分离物,从中鉴定出272个纯化的立枯丝核菌(Rhizoctonia solani Kühn)菌株。用标准菌株,通过载玻片菌丝融合试验测定,将纯化的272个菌株划归为3个菌丝融合群:即AG-2、AG-4和AG-5,分别占总菌株的6.24%、84.2%和1.1%。另有23个菌株不与任何标准菌株融合,占8.46%,说明新疆北疆棉田立枯丝核菌的优势菌系是多核丝核菌的AG-4融合群。通过从10种不同配方培养基中筛选效果好的麦芽蛋白胨(MPDA)配方培养基(Ⅱ)进行对峙培养,将纯化获得的272个丝核菌菌株,划分为6个不同的营养亲和群。将多核的不同菌丝融合群及其各营养亲和群代表菌株在3种不同主栽棉花品种上(每品种都加不接菌的对照)进行温室盆栽致病力测定,结果以AG-4对棉花的致病力最强,其次是非融合类,AG-2和AG-5虽致病,但并不致死苗,其平均病指数分别为94.9、81.4、53.1、43.5。     

河南省小麦纹枯病菌的群体组成及其致病力分化研究摇

纹枯病已成为我国小麦生产的主要障碍之一.许多学者研究表明,我国小麦纹枯病菌优势菌群为禾谷丝核菌(Rhizoctonia cerealis),其中主要为AG-D融合群,其次还包括AG-A、AG-R、AG-E、AG-Bo融合群和立枯丝核菌(R.solani)的AG-2、AG-4及AG-5融合群等[1].     

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。     

江苏省大、小麦纹枯病病原学的初步研究

 1984至1986年间,采集江苏各地大、小麦纹枯病标样,分离获得丝核菌属菌株分别为23个和50个。经比较鉴定其培养特性,菌丝形态、隔膜孔器以及细胞核数目,结合菌丝融合测试,50个小麦纹枯丝核菌菌株中除2个属于立枯丝核菌(Rhizoctonia solani Kühn)AG₅群外,其余均为禾谷丝核菌(R.cetea-lis Vander Hoeven),其中CAG₁、CAG₃、CAG₆、AGC₁分别为44、1、1和2个菌株。在23个大麦纹枯丝核菌菌株中,除2个属于立枯丝核菌AG₅群外,其余均为禾谷丝核菌,其中CAG₁、CAG₂、AGE的菌株分别为18、1、1、另有一菌株不属所测融合群。
人工交互接种试验表明:来源于大麦和小麦的禾谷丝核菌CAG₁群菌株对于大、小麦均有极强的致病力;立枯丝核菌AG₅群菌株次之,也有一定致病力;其他融合群的菌株致病力较弱。因此,引致江苏省大小麦纹枯病的主要病原菌为禾谷丝核菌(R.cerealis)的CAG₁群。     

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。     

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。     

东北地区烟草靶斑病菌(Rhizoctonia solani)融合群、致病力分化及品种抗病性研究

2013-2014年吉林省、黑龙江省烟区首次大面积发生烟草靶斑病,损失严重。采集吉林省柳河县、黑龙江省林口县和宾县的烟草靶斑病病叶进行分离纯化,得到了12个菌株。将所获菌株分别与立枯丝核菌标准融合群菌株AG-2、AG-3、AG-4进行对峙培养,结果表明:此12个菌株均与立枯丝核菌AG-3标准菌株发生融合反应,不与其他融合群发生融合;随机选取3个县各1个代表性菌株,提取菌丝基因组DNA并对其ITS序列进行分析与比对,菌株HLJ-2、JL-1、HLJ-7的ITS序列与辽宁省烟草靶斑病菌菌株LF-2、LJT-8(AG-3融合群)的同源性达99%~100%,因此推断,吉林省、黑龙江省烟草靶斑病菌与辽宁省烟草靶斑病菌应属于同一个融合群,即立枯丝核菌AG-3融合群(Rhizoctonia solani AG-3)。根据各菌株在鉴别寄主上表现的抗性不同,可将吉林省、黑龙江省所有菌株分为3个致病类型,即致病类型Ⅰ、致病类型Ⅱ和致病类型Ⅲ。同一省份不同菌株间致病力有差异,黑龙江省菌株3种致病类型均有,以致病类型Ⅱ为主;吉林省菌株只有致病类型Ⅰ和致病类型Ⅱ。分别选取3个省的强致病力菌株,用离体叶片接种法鉴定了我国21个主栽烟草品种对靶斑病的室内抗病性,结果表明:没有免疫或抗病品种,‘龙江981’、‘NC297’对供试的3个强致病力菌株均表现中抗,‘中烟202’、‘云烟97’、‘NC55’、‘KRK26’和‘G28’分别对不同菌株表现中抗。     

Identification and pathogenicity of<Emphasis Type="Italic">Rhizoctonia solani</Emphasis> and binucleate<Emphasis Type="Italic">Rhizoctonia</Emphasis> anastomosis groups isolated from forage legumes in Erzurum,Turkey

Abstrast  Three-hundred-twenty-five isolates ofRhizoctonia (215R. solani and 110 binucleateRhizoctonia) were obtained from roots and crowns of alfalfa, sainfoin and common vetch grown in Erzurum, Turkey. The isolates were assigned to five anastomosis groups (AG) ofR. solani (AG-2-1, AG-3, AG-4, AG-5, and AG-10) and two anastomosis groups of binucleateRhizoctonia (AG-I and AG-K). In pathogenicity tests on alfalfa, sainfoin and common vetch, the highest disease severities were caused by isolates of AG-4 and AG-5. Isolates of AG-10 and AG-I were not pathogenic on the three tested forage legumes, whereas isolates of AG-K on alfalfa and sainfoin, and of AG-2-1 on sainfoin, were moderately virulent. Alfalfa isolate AG-3 was moderately virulent on sainfoin. This is the first report ofR. solani AG-3, AG-5, AG-10 and binucleateRhizoctonia AG-I on alfalfa. In addition, all theR. solani and binucleateRhizoctonia groups isolated from sainfoin and common vetch were recovered from these crops for the first time in Turkey. http://www.phytoparasitica.org posting Dec. 16, 2002.     

Characterization ofRhizoctonia solani isolates from potatoes in turkey and screening potato cultivars for resistance to AG-3 isolates

A total of 304Rhizoctonia solani isolates and 60 binucleateRhizoctonia-like fungi were recovered from stems and tubers of infected potato plants over a 2-yr period in northeast Turkey.R. solani isolates were identified to 11 anastomosis groups (AGs): AG-1 (0.66%), AG-2-1 (5.6%), AG-2-2 (0.99%), AG-3 (83.9%), AG-5 (4.6%), AG-6 (0.66%), AG-8 (1.32%), AG-9 (0.33%), AG-10 (1.32%), AG-12 (0.33%), and AG-13 (0.33%). In the greenhouse tests, most of the AG-3 isolates were significantly more virulent than isolates belonging to other AGs on potato cv. Batum. Isolates of other anastomosis groups differed in their virulence. Results indicated that AG-3 is an important pathogen on potatoes grown in the study area. Five of 22 commercial and local potato cultivars evaluated for their reaction toR. solani AG-3 isolates (TP-2) under greenhouse conditions were highly resistant; the remaining cultivars exhibited different levels of susceptibility to the pathogen isolate. http://www.phytoparasitica.org posting July 14, 2005.     

Genetic diversity,anastomosis groups and virulence of <Emphasis Type="Italic">Rhizoctonia</Emphasis> spp. from strawberry

Virulent Rhizoctonia spp. isolated from strawberry in Israel belonged to anastomosis groups (AG) of: binucleate Rhizoctonia (BNR) AG-A, AG-G, AG-K and AG-F, and to multinucleate Rhizoctonia (MNR) AG 4 subgroup HG-I. In addition, a soil isolate of AG 4 subgroup HG-III was also found to be virulent on strawberry. None of the Israeli isolates obtained in the present study belonged to BNR AG-I, or other MNR AGs. In the cluster analysis of rDNA-ITS sequences, all of the isolate sequences consistently clustered according to their known AGs and subgroups. One AG-F cluster included sequences of 10 strawberry isolates, while another AG-F cluster included sequences of two isolates submitted to GenBank. Additional work is needed to determine whether the isolates of these two clusters may belong to different AG-F subgroups. The current virulence bioassay used for Rhizoctonia spp. isolates on strawberry is based on inoculation of stolon-derived daughter plants with the isolates and estimation of the reduction in plant biomass, rather than on specific distinct disease severity symptoms. The duration of this test is relatively long (ca. 5 weeks or more) and the availability of daughter plants from runners is naturally limited to a certain season. Among the possible alternative methods evaluated in the present study (inoculation of fruits or seedlings developed from germinated strawberry seeds), the method based on seedlings was best. This method has a potential to replace the currently used stolon-daughter plant inoculation bioassay for testing virulence of strawberry root pathogens. This is the first report indicating that Rhizoctonia spp. isolates that belong to AG-F, AG-K, AG 4 HG-I and AG 4 HG-III are virulent to strawberry.     

我国小麦纹枯病病原学的初步研究

从北京、江苏、浙江、安徽、山东、河南、江西、四川、贵州、陕西及甘肃等11个省市采集到的小麦纹枯病病株上共分离出43个丝核菌分离物,分别属于双核丝核菌CAG-1、CAG-2、CAG-3、CAG-5、CAG-6,以及多核丝核菌AG-2-1和AG-5等7个不同的菌丝融合组;各组分离物的数目依次为27、2、1、3、1、2和4个。致病性测定结果表明,引起我国小麦纹枯病的主要病原物是禾谷丝核菌(Rhizoctonia cerealis)中的第一菌丝融合组(CAG-1),但立枯丝核菌(R.solani)中的第五菌丝融合组(AG-5)亦可致病。文中讨论了小麦纹枯病病原的复杂性,复合侵染,微侵染,菌丝融合组中致病力差异和生物防治方法的应用前景等问题;对小麦纹枯病病原学研究在理论和实践上的意义进行了阐述。     

Differentiation of Rhizoctonia AG-D Isolates from Turfgrass into Subgroups I and II Based on rDNA and RAPD Analyses

Binucleate Rhizoctonia anastomosis group (AG) D is the cause of rhizoctonia-patch and elephant-footprint diseases of zoysiagrass, and winter-patch disease of bentgrass. Rhizoctonia AG-D is also known as the causal pathogen of other diseases such as sharp-eye-spot of cereals, foot-rot of cereals and winter-stem-rot of mat rush. Isolates of AG-D have been divided into the two subgroups AG-D (I) and AG-D (II), based on the results of cultural characteristics and pathogenicity tests. Isolates obtained from zoysiagrass exhibiting symptoms of rhizoctonia-patch disease, from bentgrass with winter-patch disease, from wheat with foot-rot disease, and from mat rush with winter-stem-rot disease were reported to belong to subgroup AG-D (I). On the other hand, isolates obtained from zoysiagrass with elephant-footprint disease were assigned to subgroup AG-D (II). To confirm the existence of these two subgroups in AG-D, the genetic structure of AG-D isolates from turfgrass and other crops was compared. RFLP analysis of the ITS region from rDNA after digestion with the restriction enzymes EcoRI, HaeIII, HhaI, HinfI, and MboI separated AG-D isolates into two groups corresponding to AG-D (I) and AG-D (II). Furthermore, other AGs except AG-Q (AGs-A, Ba, Bb, C, E, F, G, I, K, L, O, P, and R. solani AG1-IC) did not have the same patterns that were seen for the two AG-D subgroups. AG-Q isolates from bentgrass showed the same patterns as AG-D (I). The results of the RAPD analysis also revealed the existence of two groups that corresponded to AG-D (I) and AG-D (II). These analyses revealed that Rhizoctonia AG-D isolates from turfgrass could be divided into two subgroups consistent with those based on cultural characteristics and pathogenicity. In addition, isolates of foot-rot disease of wheat and isolates of winter-stem-rot disease of mat rush whose cultural characteristics were the same as those of AG-D (I) also showed similar RFLP and RAPD patterns to those of AG-D (I) isolates from turfgrass.     

Characterization of Rhizoctonia Species Associated with Foliar Necrosis and Leaf Scorch of Clonally-propagated Eucalyptus in Brazil

The objective was to identify and characterize the causal agent of foliar necrosis and leaf scorch of Eucalyptus spp. in Brazil. Nineteen putative isolates of Rhizoctonia obtained from Eucalyptus plants during clonal propagation were compared with isolates from other hosts and with tester strains of anastomosis groups of Rhizoctonia solani. Features compared were morphological characteristics of anamorphs and teleomorphs, numbers of nuclei per cell in the vegetative hyphae, anastomosis of hyphae, and ability to produce necrotic lesions on cuttings and damping-off of E. grandis×E. urophylla hybrid seedlings. Rhizoctonia solani AG1 (AG1-IB like) was the most frequent causal agent isolated from Eucalyptus plants and cuttings with symptoms of leaf scorch and foliar necrosis respectively. These isolates were highly virulent on Eucalyptus cuttings and presented naturally epiphytic growth on Eucalyptus shoots. Binucleate isolates and isolates of R. solani AG4 were also virulent on cuttings and were most virulent on Eucalyptus seedlings causing pre- and post-emergence damping-off. Virulence on Eucalyptus cuttings and seedlings was not restricted to a single species or anastomosis group of Rhizoctonia.     

Isolation and characterization of lectins from the AG-D group of binucleate <Emphasis Type="Italic">Rhizoctonia</Emphasis> species

Isolates from 18 anastomosis groups (AGs) of binucleate Rhizoctonia were screened for lectin activity. Eight AGs (AG-B, AG-D, AG-F, AG-G, AG-H, AG-I, AG-R and AG-U) had low to moderate lectin activities. Among these, members of AG-D and AG-I had the highest activity. Partially purified lectins from AG-D preferentially agglutinated human blood type A to type B and O. Mucin and galactose were the most potent inhibitors among the tested carbohydrates. The molecular masses of these lectins ranged from 12.7 kDa for the monomer to 62 kDa for the pentamer type. Proline, alanine, glutamic acid, aspartic acid, leucine, threonine, serine and tyrosine were the major amino acid components of these lectins. Lectins from AG-D were stable at 4–50°C and from pH 6.0 to 10.0. When assayed with isoelectric focusing, these lectins gave bands at pI 9.30. Specificity of lectins from AG-D to galactose and its derivatives suggest a possible recognition role in this fungal species.     

Effect of cropping system on composition of the <Emphasis Type="Italic">Rhizoctonia</Emphasis> populations recovered from canola and lupin in a winter rainfall region of South Africa

Rhizoctonia spp. anastomosis groups (AGs) associated with canola and lupin in the southern and western production areas of the Western Cape province of South Africa were recovered during the 2006 and 2007 growing seasons and identified using sequence analyses of the rDNA internal transcribed spacer regions. The effect of crop rotation systems and tillage practices on the recovery of Rhizoctonia spp. was evaluated at Tygerhoek (southern Cape, Riviersonderend) and Langgewens (western Cape, Moorreesburg) experimental farms. Isolations were conducted from canola planted after barley, medic/clover mixture and wheat, and lupin planted after barley and wheat, with sampling at the seedling, mid-season and seedpod growth stages. In the 2006 study, 93.5% of the Rhizoctonia isolates recovered were binucleate and 6.5% multinucleate; in 2007, 72.8% were binucleate and 27.2% were multinucleate. The most abundant AGs within the population recovered included A, Bo, I and K, among binucleate isolates and 2-1, 2-2 and 11 among multinucleate isolates. Crop rotation sequence, tillage and plant growth stage at sampling all affected the incidence of recovery of Rhizoctonia, but certain effects were site-specific. The binucleate group was more frequently isolated from lupin and the multinucleate group from canola. AG-2-1 was only isolated from canola and AG-11 only from lupin. This study showed that important Rhizoctonia AGs such as AG-2-1, 2-2 and 11 occur in both the southern and the western production areas of the Western Cape province and that crop rotation consistently influences the incidence and composition of the Rhizoctonia community recovered from the cropping system.     

Population structure of the rice sheath blight pathogen <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> AG-1 IA from India

The population structure of Rhizoctonia solani AG-1 IA causing rice sheath blight from India was evaluated for 96 isolates using seven RFLP loci. Nineteen of the isolates did not hybridise to R. solani AG-1 IA RFLP probes and rDNA analyses subsequently confirmed that they were either Ceratobasidium oryzae-sativae isolates or another Rhizoctonia sp. The population structure of the remaining 77 R. solani AG-1 IA Indian isolates was similar to that of a previously characterized Texas population. Clonal dispersal of R. solani AG-1 IA in India was moderate within fields and no clones were shared among field populations. Low levels of population subdivision and small genetic distances among populations were consistent with high levels of gene flow. Frequent sexual reproduction was indicated by the fact that most populations were in Hardy–Weinberg equilibrium (HWE). The two loci (R68 and R111) that deviated significantly from HWE showed an excess of heterozygosity. Although Texas and Indian populations were geographically very distant, they exhibited only moderate population subdivision, with an F_ST value of 0.193.