Web Blight of Fan Columbine, <Emphasis Type="Italic">Aquilegia flabellata, </Emphasis>Caused by <Emphasis Type="Italic">Rhizoctonia solani </Emphasis>AG1-IB : A New Disease Record

A new disease, found on fan columbine in August of 1997, first appeared as necrotic spots on leaves and within a week caused wilting of all the leaves. Fungal mycelia bound aerial parts of the plants together, formed mats of mycelia and eventually killed the plants. The pathogen, isolated from the infected leaves and stalks, was identified as Rhizoctonia solani AG1-IB in respect to hyphal anastomosis and culture types. The common name of web blight is proposed for this new occurrence on fan columbine (Kumonosu-byo in Japanese). Received 24 November 1999/ Accepted in revised form 4 February 2000     

A broad diversity survey of Rhizoctonia species from the Brazilian Amazon reveals the prevalence of R. solani AG-1 IA on signal grass and the new record of AG-1 IF on cowpea and soybeans

Leaf blight, sheath blight, and web blight are major diseases caused by Rhizoctonia species on both Fabaceae and Poaceae plant hosts in the Brazilian Amazon agroecosystem. To determine the diversity of Rhizoctonia species associated with foliar diseases on fabaceous (cowpea and soybean) and poaceous (rice and signal grass [Urochloa brizantha]) hosts, a broad survey was conducted in Pará, Rondônia, Roraima, and Mato Grosso, in the Amazon, from 2012 to 2013. We extended our survey to Cerrado areas of Mato Grosso, and the lowlands of Paraíba Valley, in São Paulo, where these Rhizoctonia foliar diseases have not been reported so far. Our findings revealed that these diseases are caused by a diversity of Rhizoctonia solani AG-1 complex. We detected that R. solani AG-1 IA (sexual phase Thanatephorus cucumeris) was the predominant pathogen associated with signal grass leaf blight and collar rot diseases in the Amazon, especially in Rondônia and northern Mato Grosso. In addition, a subgroup of R. solani (AG-1 IF), not previously reported in Brazil, was associated with leaf blight on cowpea and soybean, in Roraima. Another subgroup (AG-1 ID) was also detected in Roraima. In Mato Grosso Cerrados we did not find any of the major Rhizoctonia foliar pathogens. Instead, R. oryzae (Waitea circinata) was the predominant species associated with a collar rot on U. brizantha. In the lowlands of São Paulo, R. oryzae-sativae (Ceratobasidium oryzae-sativae) was the predominant pathogen detected causing the rice sheath spot disease.     

Efficacy of Hypovirulent Binucleate Rhizoctonia sp. to Control Banded Leaf and Sheath Blight in Corn

Water agar, artificially infested soil and leaf sheath inoculation methods were used to assess the suitable time of application, varietal host response and persistence of Rhv7, a hypovirulent, binucleate Rhizoctonia collected from soil in the Philippines, to effectively control virulent Rhizoctonia solani AG1-IA isolate RS35 on corn. With the water agar method, prior inoculation (2 to 3 days) with the biocontrol agent is essential to achieve maximum control of the pathogen. Disease protection was increased from 23 to 70% as the pre-incubation time of Rhv7 prior to the challenge inoculation with virulent isolate was lengthened from 0 to 3 days. Disease severity and incidence of banded leaf and sheath blight (BLSB) on the three corn hybrids were also suppressed in artificially infested soil. In leaf sheath inoculation, the suppressive ability of Rhv7 against BLSB on corn persisted during lesion expansion. This suppression was expressed as slower disease progress in plants with Rhv7 than in plants without Rhv7. Early and timely brace root formation that detached infected sheaths, also aided in reducing the number of diseased plants in Rhv7-treated plots at the final observation. Mycelial growth activity of RS35 was reduced when corn plants were pre-inoculated with Rhv7 before challenge even if there was no contact detected between Rhv7 and RS35, which suggests that Rhv7 protects corn against BLSB by induced resistance. The hypovirulent, binucleate Rhizoctonia Rhv7 strain effectively controlled R. solani AG1-IA isolate RS-35 in corn. Received 30 August 1990/ Accepted in revised form 25 October 1999     

Comparison of rDNA-ITS Sequences between Potato and Tobacco Strains in <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> AG-3

Genetic diversity among 51 isolates of Rhizoctonia solani AG-3, representing potato and tobacco populations, was inferred from the sequences of the internal transcribed spacer (ITS) and 5.8S ribosomal RNA (rRNA) gene. The 5.8S rDNA sequence was completely conserved not only in AG-3, but across all the AG isolates examined, whereas the rDNA-ITS sequence was found to be variable among the isolates. The nucleotide sequence similarity in the ITS 1 region was high (96-100%) for isolates within each of the two populations, but was 91-92% for isolates from different populations. The AG-3 isolates had 56 to 91% sequence similarities in the ITS 1 region with R. solani isolates of the other AGs. Phylogenetic analysis based on the ITS-5.8S rDNA sequence data indicated that the different populations in AG-3 are distantly related to each other. Genetic divergence between the two populations was also supported by the results of DNA-DNA hybridization studies. This study suggests that AG-3 consists of two genetically isolated groups corresponding to separate subgroups: AG-3 PT (potato type) and AG-3 TB (tobacco type). Specific primer sets for the detection of the two AG-3 subgroups were developed from the aligned rDNA-ITS sequences. Received 22 April 1999/ Accepted in revised form 2 July 1999     

筛选水稻纹枯病农药活性试验方法的改进

水稻纹枯病 ( sheath blight on rice)是我国稻区主要病害之一 ,其致病菌为 Rhizoctoniasolani[1 ] ,在高温、高湿条件下 ,该病不易控制 ,会造成不同程度的减产 ,所以此病一直是药剂筛选的主要模式之一。多年的试验证明 :在北方冬季用常规的“植株法”对水稻纹枯病进行药剂筛选试验难度很大 ,因为植株法占用空间大 ,难以满足其发病条件 (最佳温度 2 8℃± 1℃ ,相对湿度 97%以上 [2 ] )。笔者经多年的试验 ,探索出了用水稻离体“叶片法”代替“植株法”。“叶片法”占用空间小 ,发病条件容易控制[3] 。不足之处是每年 1 1月至翌年 4月间 ,…     

Occurrence of foliar rot of pak choy and Chinese mustard caused by Rhizoctonia solani AG1-IB in China

During December 2003, leaf rot was frequently observed on leaves of pak choy and Chinese mustard in Xishuangbanna district of Yunnnan Province, China. Isolates of Rhizoctonia solani, consistently obtained from the diseased leaves, were identical to anastomosis group (AG)-1 IB based on cultural characteristics and analysis of the ITS-5.8s rDNA region. This is the first report of foliar rot of pak choy and Chinese mustard caused by R. solani AG-1 IB in China.     

影响水稻纹枯病菌营养体亲和性分化的因子

 为了探明影响水稻纹枯病菌营养体亲和性分化的因子,将水稻纹枯病菌菌株cx-2在不同水稻品种继代接种,并在含不同杀菌剂、具不同pH及不同温度条件的PDA平板上继代培养,用对峙法测定继代菌株与原始接种菌株的营养体亲和性。此外,对营养体亲和性分化菌株与原始菌株的AFLP指纹图谱进行比较。结果显示,在供试的30个水稻品种上连续接种4次后,从9个水稻品种中分离出与原始菌株营养体不亲和的菌株。菌株在不同pH值的PDA平板继代培养4次后,在pH偏碱性端(pH 10、pH 11)开始出现营养体亲和性分化的菌株。在不同农药和温度条件下继代培养10次的菌株中没有分离到营养体亲和性分化的菌株。营养体亲和性分化菌株与原始菌株的AFLP指纹图谱没有差异。     

Distribution of Vegetatively Compatible Populations of Rhizoctonia solani AG1-IA in a Field Planted with Different Host Species

To understand the distribution pattern and divergence of Rhizoctonia solani in a field over a 4-year period, R. solani AG1-IA isolates were collected from diseased tissues of several crops. Pairing tests between isolates to detect hyphal anastomosis and vegetatively compatible population (VCP) groupings were done on 2% water agar and potato dextrose agar. A single VCP of R. solani AG1-IA dominated a large upland crop field at the Institute of Plant Breeding, University of the Philippines at Los Ba?os. The VCP changed more slowly and at a lower frequency as compared to other reports. Received 27 September 1999/ Accepted in revised form 3 February 2000     

温度对玉米纹枯病菌生长与发病的影响

室内控制性测定结果表明,玉米纹枯病菌(RhizoctoniasolaniAG1鄄IA)菌丝生长的温度10～35℃,最适温度25～30℃;菌核萌发温度18～35℃,最适温度25～30℃。在幼苗和离体叶鞘上由菌丝引起侵染和发病的温度15～35℃,以25～30℃最适宜,潜育期最短,病斑较大。田间分期播种观察分析,气温和地面温度与病害始见的历期(播种期至病害初见期)分别呈显著负相关和极显著负相关。在田间自然条件下,始病的温度为20℃左右。这些结果为了解玉米纹枯病的发生规律,开展预测预报提供了科学依据。     

Resistance of Sorghum Line CS 621 to Rhizoctonia solani AG1-IA and Other Sorghum Pathogens

Sorghum line CS 621 was evaluated along with other lines and cultivars for resistance to Rhizoctonia sheath blight, tar spot and gray leaf spot for 3 years. CS 621 was consistently resistant to these diseases even under a heavy natural outbreak of tar spot in the breeding nursery. It was also found to be more resistant to Rhizoctonia sheath blight than the resistant lines from Japan during the 1993 to 1994 screening tests. Resistance to Rhizoctonia solani AG1-IA was expressed in the form of reduction in the severity of disease and rate of infection. Evaluation of progenies from crosses involving CS 621 and a susceptible variety, UPL Sg5, indicated that additive and dominant gene effects are important in the expression of quantitative resistance to R. solani. CS 621 could therefore serve as a source of multiple resistance genes in a breeding program for high yield and stability against sorghum diseases. Received 6 August 1999/ Accepted in revised form 18 October 1999     

Pathotypic and genetic diversity in the population of Rhizoctonia solani AG1‐IA causing rice sheath blight in China

Sheath blight, caused by Rhizoctonia solani AG1‐IA, is one of the most serious diseases of rice. In this study, a total of 175 isolates of R. solani AG1‐IA were collected from five rice‐growing regions in China. Pathogenicity tests revealed that all isolates were virulent to five cultivars with different levels of resistance at the rice seedling stage in the greenhouse. There was considerable variation in aggressiveness, and the isolates were classified into three pathotypes based on disease severity, with moderately virulent isolates prevalent in the population. Forty‐three haplotypes were identified based on ITS sequencing, and 39 haplotypes were distinct among isolates. There were high levels of haplotype diversity and nucleotide diversity within the populations of R. solani AG1‐IA. High gene flow (Nm = 1·63–5·22) was detected, consistent with relatively low differentiation between pairs of populations. Five populations were divided into two distinct clusters by the unweighted pair group method with arithmetic mean (UPGMA), and no spatial population differentiation was discernible. The majority (97·8%) of genetic diversity was distributed among isolates within populations, with only 2·2% of the genetic diversity attributed to differences among populations. The star‐like shape of the haplotype network provided evidence of signatures of population expansion in recent history. No significant relationships were found between the genetic diversity and aggressiveness or geographic origin among populations of R. solani AG1‐IA. These results highlight that the population characteristics of R. solani AG1‐IA should be taken into account in evaluating the germplasm resistance of rice cultivars to sheath blight.     

井冈霉素A对水稻纹枯病菌的毒力和作用机理研究

根据水稻纹枯病菌 Rhizoctonia solani AG- 1IA在含系列浓度井冈霉素 A马铃薯葡萄糖琼脂培养基 (PDA)上的菌丝生长速率 ,计算出药剂的毒力回归方程为:Y=3.2 30 9+0 .872 0 X,r=0 .9910 ,药剂的 EC50 =10 6 .9μg/ m L,EC75=6 34.5 μg/ m L。在含 5 0 0 μg/ m L井冈霉素 A的 PDA上水稻纹枯病菌的菌丝生长抑制率为 74 .1% ,而田间水稻植株上人工接菌的药效试验结果表明 ,5 0 μg/ m L药液喷洒后 7d和 14 d的防效即可分别达到 77.4 %和 76 .7% ,即药剂在 PDA平板上的理论抑制作用仅是田间活体植株上对病菌实际作用效果的 1/ 10。比较室内毒力和田间药效试验的结果可以看出 ,井冈霉素 A具有对病原菌和寄主植物双重作用的特性。研究首次发现 ,在室内培养基上不能有效抑制病菌生长的 1μg/ m L的井冈霉素 A可以在水稻植株未喷药部位产生防御水稻纹枯病的作用 ,且能够持续诱导植物防御反应相关酶——过氧化物酶 (PO)和苯丙氨酸解氨酶 (PAL)的活性增高,表明该药剂可以激发水稻抗性防卫反应的表达。     

Effect of various crop establishment methods practised by Asian farmers on epidemics of rice sheath blight caused by Rhizoctonia solani

Establishment methods for rice crops in tropical Asia are very diverse, leading to variation in the structure of rice canopies. Differences in canopy structure can in turn affect the spread of the rice sheath blight pathogen, Rhizoctonia solani . Rice sheath blight epidemics were compared during two seasons in crops established by different methods: direct broadcasting of pregerminated rice seeds, and transplanting of rice seedlings at spacings of 20 × 20 cm, 13 × 25 cm and 25 × 25 cm between hills (i.e. along and between rows, respectively). In both years, the apparent infection rate based on incidence data and the terminal severity of sheath blight were lower in the direct-seeded crops than in any of the transplanted ones, regardless of spacing. The frequency of leaf-to-leaf contacts ( CF ) between hills (or plants) was highest in direct-seeded rice, and lowest in rice transplanted at a spacing of 25 × 25 cm. Larger CF is known to favour rice sheath blight epidemics. The apparent contradiction between higher incidence and lower CF in the transplanted stands than in the direct-seeded stands is interpreted in terms of accessibility of healthy host tissues to the spread of the pathogen in the canopy, and accounts for within-host (rice hill or plant) and between-host (hill or plant) disease spread. The analysis of incidence-severity relationships indicated a less aggregated distribution of the disease in direct-seeded rice, which was related to the spatial distribution of the tillers. These findings have direct implications for the management of the disease.     

嘧菌酯对扁豆纹枯病的物理作用方式及其生物动力学特性

以多菌灵和福美双为对照药剂,测定了嘧菌酯对立枯丝核菌Rhizoctonia solani的抑制活性,并用离体叶片法测定了嘧菌酯对该病原菌引起的扁豆纹枯病的物理作用方式及其生物动力学特性。嘧菌酯、多菌灵和福美双对立枯丝核菌菌丝生长的EC50值分别为0.072 4、1.134 4和1.202 6 μg/mL。施药后立即接种, 250 μg /mL嘧菌酯、500 μg /mL多菌灵和1 000 μg /mL福美双对扁豆纹枯病的保护效果分别为95.61％、99.88％和81.45％;施药3 d后再接种,嘧菌酯、多菌灵和福美双对扁豆纹枯病的保护效果分别为92.27％、100％和46.96％;接种36 h后再施药,3种药剂对扁豆纹枯病的治疗效果分别为90.40％、95.75％和61.94％。表明嘧菌酯对扁豆纹枯病具有很好的保护作用、持效性和治疗作用。在叶片基部施药后在顶部接种,嘧菌酯、多菌灵和福美双对扁豆纹枯病的防治效果分别为87.81％、42.09％和7.24％;在叶片背面施药后在正面接种,3种药剂对扁豆纹枯病的保护效果分别为87.30％、37.00％和16.15％。表明嘧菌酯在扁豆叶片中具有很好的木质部输导和跨层转移活性。     

水稻纹枯病菌PG的分离纯化及其理化性质研究

水稻纹枯病菌产生的多聚半乳糖醛酸酶(Polygalacturonase,PG)是其重要的致病因子之一。用丙酮法提取PG粗蛋白,分别经DEAE-Sepharose Fast Flow离子交换柱、Phenyl-Sepharose 6 Fast Flow疏水柱、Sephadex G-75凝胶柱和DE52离子交换柱层析纯化得到一种具有较高活性的PG纯蛋白。该蛋白分子量为39.81 kD;等电点为4.58;含有糖基,含糖量为1.48%;含有α氨基酸,但不含芳香族氨基酸;不含脂基。这种蛋白在pH4~12范围内均具有活性,pH5时活性最大;对热不稳定,100℃下水浴20 min,活性完全丧失;对胰蛋白酶和蛋白酶K敏感,酶处理后其活性只有对照的35.0%和35.2%;对紫外线和氯仿亦敏感,处理后活性仅为对照的40.0%和51.7%。     

微生物杀菌剂及其混配剂防治水稻纹枯病的田间药效试验

对微生物杀菌剂及其混配剂对水稻纹枯病进行田间防治试验,结果表明枯草芽孢杆菌10亿/g可湿性粉剂、井冈霉素·蜡质芽孢杆菌12.5%可湿性粉剂对水稻纹枯病有较好的防治效果。在发病初期施药,枯草芽孢杆菌10亿/g可湿性粉剂1500g/hm2防效为75.4%,产量增幅21.5%;井冈霉素·蜡质芽孢杆菌12.5%可湿性粉剂2880g/hm2防效为83.89%。可以作为井冈霉素的替代产品,进行推广应用。     

玉米衰老相关蛋白基因ZmSAP的克隆与表达分析

利用立枯丝核菌(Rhizoctonia solani)诱导胁迫下玉米高耐纹枯病自交系幼苗叶片所构建的cDNA文库中获得的EST序列,通过电子克隆和RT-PCR方法,结合cDNA末端快速扩增技术,从玉米叶片中分离克隆到衰老相关蛋白基因的全长cDNA序列,命名为ZmSAP(GenBank登录号:GU253311)。序列分析表明,ZmSAP全长1156bp,包含一个完整的603bp的开放阅读框(ORF),具有连续的Poly A尾和典型的加尾信号AATTAA。ProtParam程序预测表明该基因编码200个氨基酸,相对分子量为21.92kD,等电点为10.38。该氨基酸序列与豌豆、挪威云杉、寄生藻等有不同程度的同源性且在不同物种间具有一个保守结构域,染色体定位发现该基因位于玉米第1条染色体上。荧光定量PCR分析表明,在高耐纹枯病自交系R15和高感纹枯病材料478中ZmSAP基因在病菌胁迫初期呈诱导表达,可能参与了病原菌诱导的细胞程序性死亡过程,说明该基因与纹枯病菌胁迫响应机制密切相关。     

Biological diversity of Rhizoctonia solani (AG-3) in a northern potato-cultivation environment in Finland

A total of 119 isolates of Rhizoctonia were collected from stem canker lesions, stolon and root lesions, hymenia on stems, or from black scurf on tubers of potato plants ( Solanum tuberosum ) in Finland (latitudes 60–67°N). All isolates except three belonged to anastomosis group 3 (AG-3) of R. solani , as determined by phylogenetic analysis of the internal transcribed spacer sequences (ITS1 and ITS2) of ribosomal RNA (rRNA) genes. Sensitivity of the 119 isolates to the fungicide flutolanil was tested in vitro (EC₅₀ values 0·14–0·75  µ g active ingredient mL⁻¹). The isolates also varied considerably in growth rate (5·1–14·8 mm day⁻¹). The severity of disease caused by 99 isolates was determined based on the proportion of potato sprouts affected by lesions, discoloration or death, which was c . 1–60%. Only two isolates that were able to cause severe symptoms showed particularly low sensitivity to the fungicide and rapid growth rate. One isolate each of anastomosis groups AG-2-1 and AG-5 and an unknown, binucleate Rhizoctonia sp. were detected. The AG-5 isolate and the binucleate isolate caused mild symptoms on potato sprouts, whereas the AG-2-1 isolate was not pathogenic. Taken together, AG-3 of R. solani was the predominant causal agent of the stem canker and black scurf diseases of potato in Finland and showed considerable variability in disease severity, fungicide sensitivity and growth rate in vitro .     

Bacterial Blight of Welsh Onion : A New Disease Caused by Xanthomonas campestris pv. allii pv. nov.

In June of 1998, a new bacterial disease was observed on Welsh onion in Okinawa Prefecture, Japan. Infected plants in nursery boxes were stunted with tip dieback, and heavily infected plants died. In fields, the disease appeared on leaves as irregular gray spots or elliptical spots with creases in the center. These spots enlarged and spread rapidly continued cloudy or rainy weather, and formed blight lesions on outer leaves. Yellow mucoid bacterial colonies were consistently isolated from these lesions. The causal bacterium was identified as a pathovar of Xanthomonas campestris on the basis of bacteriological properties. The bacterium was pathogenic to Welsh onion, onion, but nonpathogenic to chive, Chinese chive and hyacinth. Of Liliaceae plants, which contain Welsh onion and onion, only hyacinth has been reported as a host for the genus Xanthomonas, namely X. campestris pv. hyacinthi. However, strains of X. campestris pv. hyacinthi were not pathogenic against either Welsh onion or onion. From these results, the bacterium isolated from Welsh onion is considered to be a new pathovar of X. campestris, and the name of X. campestris pv. allii pv. nov. is proposed. A strain MAFF 311173 is designated as the pathotype strain. Received 29 March 2000/ Accepted in revised form 4 July 2000