Characterization of Rhizoctonia solani AG 2 isolates causing bare patch in field grown tulips in the Netherlands

During a spring survey in 1991, 130 isolates of R. solani were collected in 25 commercial flower bulb fields from diseased plants occurring in bare patches. On the basis of hyphal fusion frequency and pathogenicity to flower bulbs, tulip isolates were provisionally assigned to AG 2-t to distinguish these isolates from AG 2-1 isolates which were non-pathogenic to bulbs. Hyphal fusion frequency of a subgroup of 7 AG 2-t isolates was highly variable when paired with 7 AG 2-1 isolates (2-75%), thus making assignment of AG 2-t isolates to AG 2-1 inconclusive. The mean hyphal fusion frequency among AG 2-t isolates was 65% (±6%) indicating AG 2-t to be a relatively homogeneous group. Hyphal fusion frequency among AG 2-1 isolates was highly variable with a mean 51% (±25%) indicating AG 2-1 to be a heterogeneous group. The optimum growth temperature for AG 2-t and AG 2-1 isolates on malt peptone agar was 20-25 °C. The host range of AG 2-t and two AG 2-1 isolates comprised tulip, iris, hyacinth and lily at both 9 and 18 °C, and cruciferous, sugarbeet and lettuce seedlings at 18 °C. Six other AG 2-1 isolates were pathogenic to cruciferous seedlings, but not to any of the bulbous crops. The tested narcissus, Tagetes patula, tomato, potato, wheat, leek and maize cultivars were not susceptible to AG 2-t and AG 2-1 isolates. Statistical analysis using a proportional-odds model revealed significant differences in aggressiveness between R. solani AG 2-t isolates and differences in susceptibility between tulip and iris cultivars. At 18 °C, but not at 9 °C, isolates representing AG 2-2, AG 4, AG 5 and AG BI were pathogenic to bulbous crops. In addition to bare patch causing AG 2-t isolates, other anastomosis groups may cause disease in field grown tulips. For the development of optimal crop rotation schedules, the impact of bulb rot causing isolates under field conditions needs further study.     

Possible Mechanisms Influencing the Dynamics of Rhizoctonia Disease of Tulips

looseness-1In two observation fields, where six sites were artificially infested with Rhizoctonia solani AG 2-t, bare patches developed. These patches did not re-occur at the site of infestation in three successive years. In fields with and without artificial infestation, natural infection of tulip bulbs by Rhizoctonia spp. occurred. The spatial distribution of infected tulip bulbs was visualised in maps after kriging. The influence of sampling intensity was evaluated by stepwise reduction obtained in the observed data set of the first year. Omnidirectional semivariogram characteristics did not change when sampling intensity was reduced down to 10%. The average maximum prediction error was minimised at sampling intensities varying from 7% to 25%. Naturally occurring bare patches slowly vanished during successive cropping of flower bulbs and did not re-appear in the fourth growing season. A high frequency of isolation of R. solani AG 2-t in one field (Lisse-2) in the fourth consecutive crop did not result in bare patches in that year. It is hypothesised that a reduction in aggressiveness may account for this observation. In contrast, bulb rot due to Rhizoctonia spp. increased during the observation period. R. solani AG 5 isolates were seldom isolated before the bulbs flowered, but were the dominant isolate from bulbs at harvest. In a growth chamber experiment, it was demonstrated that AG 5 did not account for replacement of AG 2-t. However, it was demonstrated that competition may partially explain replacement of AG 2-t isolates during the growing season. At 18 °C, but not at 9 °C, an AG 4 isolate prevented AG 2-t colonising and infecting iris bulbs when both isolates were introduced together to soil. Rhizoctonia populations develop in relation to soil temperature and plant development. It is hypothesised that a temporal niche differentiation may be one of the mechanisms affecting the dynamics of rhizoctonia bare patch of tulips.     

Characterization of Rhizoctonia solani from potato in Great Britain

One hundred and thirty five isolates of Rhizoctonia solani were obtained from British potato crops between 2001 and 2003. Isolates were assigned to anastomosis group (AG) using conventional PCR assays for AG2-1 or AG3 or through the observation of hyphal interactions, where appropriate. A previously published primer set was modified in this study to enhance specificity for AG3PT. Most of the isolates (92·6%) belonged to AG3PT whilst some (6·7%) belonged to AG2-1. Only one isolate recovered (0·7%) belonged to AG5. Isolates of AG2-1 were diverse, with variation in both the length of the rDNA intergenic spacer 1 (IGS1) region and the categories of hyphal interaction observed between pairings of AG2-1 isolates. No variation in the length of the rDNA IGS1 region was observed amongst the AG3 isolates collected. Tests carried out on potato stems with a sub-set of the isolates revealed a wide range of aggressiveness amongst AG2-1 isolates. Sequencing of the rDNA internal transcribed spacer (ITS) region of the AG2-1 isolates and construction of a neighbour joining tree with other AG2-1 sequences available indicated that AG2-1 isolates with the short IGS1 region were closely related. This is the first investigation which provides evidence of the relative AG composition of R. solani populations causing disease in potato crops in Great Britain.     

Characterization of a new cultural type (LP) of Rhizoctonia solani AG2-2 isolated from warm-season turfgrasses, and its genetic differentiation from other cultural types

Isolates of Rhizoctonia solani AG2-2 obtained from turf with symptoms of large-patch disease of warm-season turfgrasses were compared with known AG2-2 isolates belonging to cultural types IIIB and IV. Some isolates that were previously identified as type IV have been separated here and named LP isolates. Comparisons among isolates were based on cultural morphology, hyphal growth rate, pathogenicity and restriction fragment length polymorphism (RFLP) analysis in the nuclear encoded ribosomal DNA (rDNA) genes. The cultural characteristics of LP isolates varied from those of types IIIB and IV. LP isolates did not show distinct sclerotial formation and zonation, and the colour of their mycelia and pigment deposition was dark brown. LP isolates had slower hyphal growth rates than types IIIB and IV, with an optimum temperature of 25°C compared with 28°C for types IIIB and IV. LP isolates were less virulent on radish but highly virulent on zoysia grass when compared with isolates of types IIIB and IV. Genomic DNA was digested separately with Eco RI, Ban III, Xba I and Sal I, and probed with cloned rDNA from Alternaria alternata in Southern hybridizations. LP isolates had one RFLP pattern, while both IIIB and IV possessed four different patterns each. Cluster analysis of RFLPs showed that R. solani AG2-2 is divided into three genetic subgroups, consisting of the IIIB, IV and LP isolates, respectively. The polymerase chain reaction (PCR) amplified rDNA internally transcribed spacer (ITS) regions of the IIIB, IV and LP isolates had the same length but produced different restriction patterns when digested with Msp I and Taq I. These results indicate that there are three cultural types in R. solani AG2-2, namely IIIB, IV and LP.     

小麦纹枯病菌核糖体基因内转录区序列比较

 对7个从江苏省小麦纹枯病样本分离到的丝核菌菌株,进行形态学鉴定、融合群分类和致病性测定,提取病菌的DNA,采用通用引物ITS1(TCC GTA GGT GAA CCT GCG G)和ITS4(TCC TCC GCT TAT TGA TAT GC),扩增病菌的rDNA内转录区(ITS),并对扩增产物进行了测序.用这些序列在NCBI中进行BLAST分析,得到与这些菌株亲缘关系最近的菌株序列,并明确了这些菌株的分类地位.对以上的菌株序列进行Alignment分析,结果表明,病菌的5.8S rDNA序列高度保守,而ITS区的可变性则相对较高,在双核和多核丝核菌、双核丝核菌CAG1融合群和非CAG1融合群菌株间存在差异,可用于反映菌株间的进化关系和双核丝核菌种下分类.     

Characterization of AG-13, a Newly Reported Anastomosis Group of Rhizoctonia solani

ABSTRACT Rhizoctonia solani anastomosis group (AG)-13 was collected from diseased roots of field grown cotton plants in Georgia in the United States. Isolates of AG-13 did not anastomose with tester isolates of AG-1 through AG-12. Mycelium of all isolates of AG-13 were light brown but darkened as cultures aged. All isolates produced aerial mycelium. Concentric rings were visible after 3 to 4 days of growth but disappeared as cultures aged and darkened. Individual sclerotia were up to 1.5 mm in diameter, similar in color to the mycelium, and generally embedded in the agar. Clumps of sclerotia up to 5 mm in diameter were produced on the agar surface. All attempts to induce basidiospore production were unsuccessful. The 5.8S region of the rDNA from isolates of AG-13 was identical in length and sequence to isolates of all other AGs of R. solani. Length and sequence of the internal transcribed spacer (ITS) regions of rDNA from isolates of AG-13 were unique among AGs of R. solani. Similarity between AG-13 and other AGs of R. solani ranged from 68 to 85% for ITS region 1 and 85 to 95% for ITS region 2. Selected isolates of AG-13 caused minor or no damage to barley, cauliflower, cotton, lettuce, potato, and radish in laboratory or greenhouse studies.     

Genetic variability and pathogenicity of Rhizoctonia solani associated with black scurf of potato in New Zealand

Isolates (a total of 129) of Rhizoctonia solani were collected from black scurf on potato tubers from different potato‐growing regions in New Zealand. Sequence analysis of the nuclear ribosomal DNA internal transcribed spacer (rDNA–ITS) regions from these isolates identified three anastomosis groups (AGs), AG‐3PT, AG‐2‐1 and AG‐5. Isolates classified as AG‐3PT were widely distributed, whereas AG‐2‐1 and AG‐5 were confined to distinct locations. Sequence heterogeneity was identified in the ITS regions of 100 AG‐3PT and AG‐2‐1 isolates. Variation in the sequence and length of the rDNA–IGS1 region was also observed for selected isolates of AG‐3PT and AG‐2‐1. Phylogenetic studies found all AG‐2‐1 isolates belong to AG‐2Nt, a subset of AG‐2‐1 previously associated with solanaceous crops in other countries. AG‐2‐1 isolates were consistently more aggressive than those of AG‐3PT. Delayed emergence, severe infection on stolons, formation of aerial tubers and considerable yield losses were associated with AG‐2‐1, but they caused negligible black scurf. In contrast, AG‐3PT caused black scurf on progeny tubers but variable effects on stem emergence and stolons. Furthermore, AG‐2‐1 isolates caused severe tuber malformation, but isolates of other AGs did not. This is the first report on the AG composition, genetic variability and pathogenicity of R. solani isolates associated with black scurf of New Zealand potatoes.     

十字花科蔬菜黑斑病菌的PCR鉴定

 在对十字花科蔬菜黑斑病菌(Alternaria sp.)3个种及相近种的5.8SrDNA和其侧翼ITS区进行测序的基础上,分别设计合成了鉴定白菜黑斑病菌3个种的特异性引物。PCR扩增结果表明:Abre1和Abre2引物对能特异性扩增芸苔链格孢(A.brassicae)371bp的片段,Abra1和Abra2引物对能特异性扩增甘蓝链格孢(A.brassicicola)457bp的片段,Ajap1和Ajap2引物对能特异性扩增萝卜链格孢(A.japonica)411bp的片段,而且其它近源种未扩增出目标片段,说明这3个引物对可以作为十字花科蔬菜黑斑病菌3个种快速检测鉴定的分子特征标记。     

稻瘟病标样中分离真菌的分析和鉴定

 通过分离稻瘟病标样中的病原真菌,回接稻瘟病菌普感的水稻品种—丽江新团黑谷,得到多个不能引发稻瘟病的菌株。随机选取2个致病的分离菌株和4个不致病的分离菌株,以稻瘟病菌Guy11为对照,观察它们的菌落形态和色泽、分生孢子形态和大小、分生孢子梗形态等生物学特性,发现6个分离菌株与Guy11的形态均较为相近。附着胞形成试验发现,不致病的分离菌株在疏水表面不能形成附着胞,但外源添加cAMP可使部分孢子形成附着胞。PCR扩增分离菌株的ITS、β-tubulin、actin和calmodulin等序列,测序并根据比对结果绘制相应的系统发育树,鉴定表明分离的2个致病菌株9-1、214属于Magnaporthe oryzae;4个不致病菌株18-1、18-2、122和132为梨孢属真菌Pyricularia。不致病菌株的发现和鉴定可为稻瘟病菌侵染机制研究和稻瘟病防治奠定基础。     

Characterization, Genetic Structure, and Pathogenicity of Rhizoctonia spp. Associated with Rice Sheath Diseases in India

ABSTRACT Isolates of Rhizoctonia spp. were obtained from rice in India during 2000-2003. Characterization by conventional techniques and polymerase chain reaction showed that from 110 isolates, 99 were R. solani and 11 were R. oryzae-sativae. Of 99 isolates identified as R. solani, 96 were AG1-IA, 1 was AG1-IB, and 2 were AG1-IC. Amplified fragment length polymorphism (AFLP) analyzes were used to determine genetic relationships in Rhizoctonia pathogen populations collected from different geographic regions. Cluster analysis based on the AFLP data separated isolates belonging to the three different intraspecific groups of R. solani AG1 and differentiated R. solani from R. oryzae-sativae. Analysis of molecular variance (AMOVA) revealed that geographic region was the dominant factor determining population structure of R. solani AG1-1A; host cultivar had no significant effect. Pathogenicity tests on Oryza sativa cv. Zenith revealed that isolates of R. solani AG1-1A and AG1-1B were more virulent than R. solani AG1-IC and R. oryzae-sativae isolates.     

PCR-based detection of Colletotrichum acutatum on strawberry

An oligonucleotide primer ( Ca Int 2) was synthesized from the variable internal transcribed spacer (ITS) 1 region of ribosomal DNA (rDNA) from Colletotrichum acutatum . PCR with primers Ca Int2 and ITS4 (from a conserved sequence of the rDNA) amplified a 490 bp fragment from several isolates of C. acutatum but not from other members of the genus Colletotrichum . Amplification of this fragment was achieved from 100 fg of fungal DNA. These primers amplified a fragment of the same size from DNA extracted from strawberry tissues infected by C. acutatum . Southern hybridization analysis confirmed the 490 bp fragment from C. acutatum DNA and infected strawberry to be identical. The species-specific primer ( Ca Int2) developed in this work could be used for the accurate identification of C. acutatum and its detection on other host plants.     

Morphological and molecular characterization of the sudden-death syndrome pathogen of soybean in Brazil

Brazilian Fusarium isolates causing soybean sudden death syndrome (SDS) were characterized by comparing them with other Fusarium isolates associated with soybean root rot, as well as F. solani f.sp. glycines isolates associated with the disease in the USA, using molecular (mitochondrial and nuclear rDNA), morphological, cultural and pathogenic characteristics. On the basis of pathogenicity data, and restriction fragment length polymorphism and sequence analysis of the rDNA internal transcribed spacer (ITS) regions, isolates formed a group distinct from nonSDS F. solani isolates, as well as other Fusarium species. ITS sequence analysis also revealed that Brazilian isolates were distinct from the majority of SDS pathogens from the USA ( Fusarium virguliforme ) and conformed to Fusarium tucumaniae .     

Characterisation of Rhizoctonia solani Anastomosis Groups Causing Bottom Rot in Field-Grown Lettuce in Germany

Bottom rot caused by Rhizoctonia solani is an increasing problem in field-grown lettuce in Germany. During the growing seasons of 1999 and 2000, 95 isolates of R. solani from lettuce plants with bottom rot symptoms were collected from eight locations. The isolates were characterised using hyphal anastomosis, pectic zymograms and morphological characteristics. Ninety-three isolates were identified as anastomosis group (AG) 1-IB, one as AG 1-IC and one as AG 2-1. Optimum hyphal growth was measured over a temperature range of 20–30 °C with an optimum at 25 °C. Aggressiveness of the AG 1-IB isolates varied from weak to strong when tested on detached lettuce leaves. The pathogenic potential of six AG 1-IB isolates was determined on 14 plant species in comparison with lettuce under conditions favourable for the fungus. Radish, broccoli, kohlrabi, spinach and millet seedlings were as severely infected as lettuce seedlings. The same isolates caused little symptoms on maize, tomato and onion. Knowledge about the host range of AGs of R. solani are important for planning an effective crop rotation as part of a control management system.     

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     

Molecular and experimental evidence of multi-resistance of <Emphasis Type="Italic">Cercospora beticola</Emphasis> field populations to MBC,DMI and QoI fungicides

Binucleate Rhizoctonia (BNR) spp. isolates were collected from taro (Colocasia esculenta (L.) Schott) and ginger (Zingiber officinale (Willd.) Roscoe) (Yunnanxiaojiang cv.) in Yunnan province. These Yunnan (YN) isolates did not anastomose with any of the tester isolates of the known AGs of binucleate Rhizoctonia spp. The growth of YN cultures on PDA was appressed, mealy and matlike after 4 days of incubation, then turned white brown, producing brown to dark brown, irregularly shaped sclerotia were embedded in the PDA medium after 14 days. All attempts to induce basidiospore production were unsuccessful, but the length and sequence of the internal transcribed spacer (ITS1 + 5.8S rDNA + ITS2) regions of 5.8S rDNA from the YN isolates were identical in length and sequence to isolates of all the other AGs of binucleate Rhizoctonia /Ceratobasidium spp. The sequences of 5.8S rDNA-ITS from the YN isolates were unique among AGs of BNR. The YN isolates had sequence similarities of 94% with isolates of AG Fb and P, 93% with AG E, 91% with AG R, 79–94% with AG S, and 74–87% with AG A, Ba, Bb, Bo, C, DI, DII, DIII, Fa, G, H, I, K, L, O, and Q. Four isolates of AG YN caused minor virulence (lesions ≦1mm²⁾ to ginger or taro in growth chamber studies. It was concluded that the YN isolates belong to a new anastomosis group AG-V of the Ceratobasidium spp..     

广西地区葡萄黑痘病病原菌的分离与鉴定

采用常规组织分离法对广西南宁、河池两地的葡萄黑痘病菌进行分离、纯化,分别得到37和31株分离菌。经菌落形态观察及r DNA ITS序列分析,南宁37株分离菌为同一菌株,河池31株分离菌为同一菌株,以NN和HC分别代表两地菌株,对它们进行形态学、致病性鉴定及r DNA ITS区域序列分析。结果显示,两地菌株形态学与致病性存在较大差异,但都符合黑痘病菌生长形态。NN菌落为红棕色、近圆形,边缘光滑。菌落中心位置丘状凸起,表面有白色菌丝和透明粘稠的小液滴,周围边缘有较规则的褶皱。HC菌落呈浅橙色、近圆形,边缘光滑。菌落中心位置丘状凸起,表面有少量白色菌丝,无液滴,周围边缘有不规则褶皱隆起。人工接种葡萄后均能引起典型的黑痘病症状,NN致病性强于HC。使用r DNA ITS区域通用引物ITS1F/ITS4进行PCR扩增后,NN和HC分别得到1 124 bp和818 bp的片段。比对结果显示1 124 bp与Elsinoe ampelina(AY826763.1)序列覆盖率达92%,序列一致性达99%;818 bp与Elsinoe ampelina(AY826762.1)序列覆盖率达75%,序列一致性达99%。因此,NN和HC均是引起广西葡萄黑痘病的病原菌。     

Characterization and pathogenicity of Rhizoctonia isolates associated with cauliflower in Belgium

Sixty-two isolates of Rhizoctonia spp. were collected from Belgian cauliflower fields during 2005 and 2006. The majority of the isolates (60 out of 62) had multinucleate cells and were identified as Rhizoctonia solani . Characterization of anastomosis groups (AGs) was performed using pectic zymograms, PCR-RFLP and sequencing of the rDNA-ITS region. The most prevalent AG was AG 2-1 (55% of isolates), followed by AG 2-1 subset Nt (11%), AG 1-1C (8%), AG 5 (8%), AG 4 HGII (6%), AG 3 (5%) and AG 1-1B (3%). Pathogenic potential towards different vegetable crops and towards maize was determined. Damage to cauliflower and endive was caused by different AGs, with the isolates aggressive towards cauliflower belonging to AG 2-1, AG 2-1 subset Nt, AG 4 HGII, AG 1-1C, AG 1-1B and AG 2-2, and those aggressive towards endive belonging to AG 1-1B, AG 1-1C, AG 2-1 subset Nt, AG 2-2, AG 4 HGII and AG 5. The most aggressive isolates towards bean belonged to AG 2-1 subset Nt and AG 2-2, for lettuce to AG 1-1B and AG 2-1, on carrot to AG 4 HGII and towards maize to AG 2-2. Within the isolates of AG 2-1, variability was observed in PCR-RFLP pattern and in aggressiveness towards several crops, indicating this subgroup to be heterogeneous. This is the first study concerning the occurrence of R. solani AGs causing wirestem in Belgian cauliflower fields and the first report of aggressive isolates of AG 1-1C, AG 2-1 subset Nt and AG 4 HGII associated with cauliflower.     

Pathogenic, Genetic and Molecular Characterisation of Fusarium oxysporum f.sp. lilii

Isolates of Fusarium oxysporum from lily were screened for pathogenicity, vegetative compatibility and DNA restriction fragment length polymorphisms, and compared to reference isolates of F. oxysporum f.sp. gladioli and F. oxysporum f.sp. tulipae to justify the distinction of F. oxysporum f.sp. lilii. Twenty-four isolates from different locations in The Netherlands (18 isolates), Italy (4 isolates), Poland and the United States (1 isolate each) shared unique RFLP patterns with probes D4 and pFOM7, while hybridization did not occur with a third probe (F9). Except for a self-incompatible isolate, these 24 isolates all belonged to a single vegetative compatibility group (VCG 0190). Isolates belonging to VCG 0190 were highly pathogenic to lily, but not to gladiolus or tulip, except for a single nonpathogenic isolate. Six saprophytic isolates of F. oxysporum from lily were nonpathogenic or only slightly aggressive to lily, gladiolus and tulip, belonged to unique VCGs and had distinct RFLP patterns. Three pathogenic isolates previously considered to belong to F. oxysporum f.sp. lilii were identified as F. proliferatum var. minus; all three belonged to the same VCG and shared unique RFLP patterns. These three isolates were moderately pathogenic to lily and nonpathogenic to gladiolus and tulip. The reference isolates of F. oxysporum f.sp. tulipae were pathogenic to tulip, but not to lily and gladiolus; they shared a distinct RFLP pattern, different from those encountered among pathogenic and saprophytic isolates from lily, and formed a separate new VCG (VCG 0230). Reference isolates of F. oxysporum f.sp. gladioli belonging to VCG 0340 proved pathogenic to both gladiolus and lily, but not to tulip. These isolates, as well as isolates belonging to VCGs 0341, 0342 and 0343 of F. oxysporum f.sp. gladioli, had RFLP patterns different from those encountered among the isolates from lily or tulip. These findings identify F. oxysporum f.sp. lilii as a single clonal lineage, distinct from F. oxysporum f.sp. gladioli and f.sp. tulipae.     

The first report of tomato foot rot caused by <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> AG-3 PT and AG-2-Nt and its host range and molecular characterization

Foot rot of mature tomato plants was found in four cities of Hokkaido, Japan, from 2004 to 2007. Six of eight isolates obtained from damaged tissues were identified as Rhizoctonia solani anastomosis group (AG)-3, and the remaining two isolates belonged to AG-2-1. We compared these isolates with nine reference isolates including the different subgroups in AG-3 (PT, TB and TM) and AG-2-Nt (pathogen of tobacco leaf spot) within AG-2-1 in terms of pathogenicity to tomato, tobacco and potato. All eight isolates caused foot rot on tomato. The six AG-3 isolates caused stem rot on young potato plants. While, all reference isolates of AG-3 PT causing stem rot of young potato plants incited foot rot on tomato. The two AG-2-1 isolates and an AG-2-Nt reference isolate caused severe leaf spot on tobacco leaves. The sequences of rDNA- ITS region and rDNA-IGS1 region of the AG-3 isolates showed high similarity to that of AG-3 PT isolates. Phylogenetic tree based on ITS and IGS1 regions of rDNA indicated that the AG-2-1 isolates from tomato formed a single clade with AG-2-Nt isolates and that they were separate from Japanese AG-2-1 isolates (culture type II). Pathogenicity tests and DNA sequence evaluation of the causal fungi revealed that the present isolates of AG-3 and AG-2-1 belonged to AG-3 PT and AG-2-Nt, respectively. This is the first report of tomato foot rot caused by R. solani in Japan.     

Rhizoctonia spp. Recovered from Strawberry Roots in Central Coastal California

ABSTRACT Rhizoctonia spp. were commonly recovered from the roots of strawberry plants growing in nonfumigated soil in the central coastal region of California. With the exception of one multinucleate isolate of R. solani (frequency of recovery of 0.8%), all other isolates were binucleate and were in anastomosis groups (AG) A, G, or I. AGs-A and -I were recovered from all five collection sites, whereas AG-G was recovered from only two sites. AG-A was the most commonly isolated AG, followed by AGs-I and -G. Similar levels of virulence were observed among the different AGs, but differences in virulence were observed among isolates in the same AG. Evaluating anastomosis grouping by pairing isolates recovered from strawberry with known tester isolates did not always yield a positive anastomosis reaction, even though both isolates anastomosed with other members of the same AG. Subsequent investigations with multiple isolates in the same AG from the same collection location confirmed that there was a lack of anastomosis or weak anastomosis reactions for some combinations of pairings, highlighting the need for to use multiple tester isolates or molecular techniques for AG determination. Restriction fragment length polymorphism (RFLP) analysis of a polymerase chain reaction-amplified region of the rDNA was effective for differentiating AGs. Sixteen RFLP groups were observed after cluster analysis with data for the size of the amplified products and fragment sizes after digestion with four restriction enzymes. Although each AG had isolates in multiple RFLP groups, any one individual RFLP group contained isolates of only a single AG. There was no consistent correlation between RFLP group and location of isolate collection.