Morphological and molecular characterization of Neopestalotiopsis vitis associated with leaf blight disease of Manilkara zapota—a new record from India

Sapota is an important horticultural crop grown in India, and Karnataka is a major producer of sapota. A characteristic leaf blight disease was observed in Southern Karnataka during field surveys conducted in 2019 with an incidence of 13–22% in approximately 45 ha of sapota field. The leaf blight-associated pathogen was isolated on the potato dextrose agar medium. A total of 12 isolates obtained from each location were identified culturally and morphologically. Based on the morphological and cultural features, the pathogen was identified as Pestalotiopsis or Neopestalotiopsis, which was further confirmed by molecular identification using a representative isolate (MZ03). The ITS rDNA and β-tubulin genes were amplified and sequenced using ITS1/ITS4 and T1/T22 primer pairs respectively. nBLAST search analysis and concatenated (ITS-rDNA and TUB2 loci) phylogenetic analysis confirmed the pathogen identity as Neopestalotiopsis vitis. Pathogenicity tests conducted on detached leaves by inoculation with a conidial suspension of N. vitis produced typical blight symptoms after 4–5 days and progressed to cover the entire leaf lamina after 10–12 days. The pathogen’s identity was confirmed after re-isolation by cultural and morphological features. Although Pestalotiopsis clavispora and Pestalotiopsis versicolor causing diseases on sapota seedlings and trees have been reported, no reports are available for the occurrence of N. vitis to sapota from India. This is the first report of N. vitis associated with leaf blight disease of sapota from India.     

Relationship between onion thrips (Thrips tabaci) and Stemphylium vesicarium in the development of Stemphylium leaf blight in onion

Stemphylium leaf blight caused by Stemphylium vesicarium and onion thrips (Thrips tabaci) are two common causes of leaf damage in onion production. Onion thrips is known to interact synergistically with pathogens to exacerbate plant disease. However, the potential relationship between onion thrips and Stemphylium leaf blight is unknown. In a series of controlled laboratory and field trials, the relationship between thrips feeding and movement on the development and severity of Stemphylium leaf blight were examined. In laboratory assays, onions (“Avalon” and “Ailsa Craig”) with varying levels of thrips feeding damage were inoculated with S. vesicarium. Pathogen colonisation and leaf dieback were measured after 2 weeks. In pathogen transfer assays, thrips were exposed to S. vesicarium conidia, transferred to onion and leaf disease development was monitored. In field trials, insecticide use was examined as a potential indirect means to reduce Stemphylium leaf blight disease and pathogen colonisation by reducing thrips damage. Results from laboratory trials revealed that a reduction in thrips feeding decreased S. vesicarium colonisation of onion leaves by 2.3–2.9 times, and decreased leaf dieback by 40–50%. Additionally, onion thrips were capable of transferring S. vesicarium conidia to onion plants (albeit at a low frequency of 2–14% of plants inoculated). In field trials, the symptoms and colonisation of Stemphylium leaf blight were reduced by 27 and 17%, respectively with the use of insecticide to control thrips. These results suggest that onion thrips may play a significant role in the development of Stemphylium leaf blight, and thrips control may reduce disease in commercial onion fields.     

Identification of the New Pathogen (Stemphylium lycopersici) Causing Leaf Spot on Pepino (Solanum muricatum)

Pepino (Solanum muricatum var. pepino) plants were found affected by an extensive leaf spot caused by plant pathogenic fungi during a survey in the Cameron highlands, Pahang state, Malaysia. Symptomatic leaf samples were collected from infected pepino plants and cultivated on PDA medium, and the pathogen was isolated and purified; then, consequently, all isolates were identified as Stemphylium lycopersici on the basis of their cultural and morphological characteristics and combined sequences of the internal transcribed spacer (ITS) and glyceraldehyde‐3‐phosphate dehydrogenase (gpd) regions. A pathogenicity assay on detached leaves further confirmed that S. lycopersici causes leaf spot disease. To the best of our knowledge, this is the first report of S. lycopersici causing leaf spot on pepino in Malaysia and worldwide.     

Septoria Leaf Spot of Stevia rebaudiana in Canada and Methods for Screening for Resistance

The herb Stevia rebaudiana is a potential source of low-calorie sweeteners. In 1995, a severe leaf spot and blight was observed in stevia production fields and research plots in the provinces of Ontario and British Columbia, Canada. The disease was characterized by angular, shiny, olive-grey lesions that rapidly coalesced and were often surrounded by a chlorotic halo. Leaves quickly became necrotic and often dropped off the plant. The disease progressed upwards in the foliage during the growing season. A Septoria sp. was isolated from diseased leaves. Ten isolates (five from each of the two provinces) of the Septoria sp. were compared with respect to conidial size. Across isolates, conidia lengths and widths overlapped (grand means for length and width were 71.4 μm and 1.4 μm, respectively). Conidiogenesis was holoblastic. Morphological characteristics and disease symptoms were similar to those of Septoria steviae, previously reported only from Japan. It was concluded that the Canadian isolates belonged to S. steviae. Isolates from Canada did not differ significantly from one another with respect to effects of temperature on colony growth or germination of conidia. Optimum temperatures for these parameters were between 20 and 25°C. In field trials, the pathogen was shown to successfully over-winter in diseased leaf tissue. In order to develop procedures for identification of resistant germplasm and greenhouse screening of candidate fungicides, effects of leaf wetness period, inoculum concentration, and plant age on disease development were determined. Thirty-six hours of leaf wetness were required for consistent development of leaf spots. Inoculum concentrations of 5 × 10⁵ conidia/ml or more were required to produce high disease severities; 6-week-old plants were more susceptible than older plants. In the growth chamber, greenhouse, and field trials, germplasm selections with high levels of resistance to S. steviae were identified. This is the first report of resistance to this disease in S. rebaudiana.     

Loop-mediated isothermal amplification assay for the detection of Plasmopara viticola infecting grapes

Grapes downy mildew caused by obligate oomycete plant pathogen Plasmopara viticola is a devastating disease worldwide, resulting in significant yield and quality losses. A field survey was conducted in two major grapes cultivated areas of Tamil Nadu for the incidence of grapevine downy mildew. The disease incidence was 43.42%–76.69%, and the highest disease incidence of 76.69% was observed in the Theni district. Totally eight P. viticola isolates were collected from different places in Coimbatore and Theni districts. These isolates were confirmed through microscopic observation and sequencing of COX 2 gene, and the phylogenetic tree was developed to study their phylogenetic relationship among the isolates which shows 97–100% sequence similarity with other P. viticola isolates and less sequence similarity with Plasmopara species. The loop-mediated isothermal amplification (LAMP) assay was developed based on the CesA4 gene sequence of P. viticola. The assay developed was more sensitive as it detected P. viticola genomic DNA up to 20 fmg. LAMP assay specificity was proved by carrying out the assay with genomic DNA extracted from other Oomycetes and fungal plant pathogens. Finally, LAMP assay was validated by testing seventy-eight grapevine leaf samples collected from seven different locations. LAMP assay showed a positive reaction in sixty-two samples tested out of seventy-eight samples tested. Therefore, the LAMP assay described should helpful for early and specific detection of downy mildew pathogen and help in mitigating disease incidence.     

Pathogen invasion triggers an evolutionary trap for an endangered checkerspot butterfly dependent on an exotic host plant

Exotic pathogen invasions can change host eco-evolutionary interactions and possibly create an evolutionary trap when the pathogen generates mismatches between developmental phenology and reproductive cues. Taylor’s checkerspot butterfly (Euphydryas editha taylori), is an endangered species of western North America with 80 % of the extant populations dependent on an exotic host, Plantago lanceolata. Survey of occupied, recently extinct, and unsuccessful butterfly reintroduction sites spanning 4° of latitude revealed widespread disease on P. lanceolata caused by Pyrenopeziza plantaginis. This fungal pathogen, new to North America, reduces the standing crop of P. lanceolata foliage throughout the winter post-diapause larval feeding period. However, disease is absent when adult butterflies and pre-diapause larvae are active. Diseased plants were frequent in Taylor’s checkerspot populations with a history of persistence, but >90 % of the host plants in these populations had initiated new leaves within the first few weeks of post-diapause larval feeding. Conversely, host plants in recently extinct and unsuccessfully reintroduced populations were severely diseased, >66 % mean foliage necrosis/plant, and <23 % had initiated new leaves. Feeding choice trials with 25 larvae indicated that new leaves were strongly and consistently preferred by post-diapause larvae over all other available leaf types, both diseased and non-diseased. Because the influence of disease on post-diapause larval food resources is developmentally disassociated from oviposition, P. plantaginis invasion appears to have triggered an evolutionary trap for Plantago-dependent populations of Taylor’s checkerspot.     

Arbuscular mycorrhiza‐mediated resistance in tomato against Cladosporium fulvum‐induced mould disease

Root colonization with arbuscular mycorrhizal fungi (AMF) enhances plant resistance particularly against soil‐borne pathogenic fungi. In this study, mycorrhizal inoculation with Glomus mosseae (Gm) significantly alleviated tomato mould disease caused by the air‐borne fungal pathogen, Cladosporium fulvum (Cf). The disease index (DI) in local leaves (receiving pathogen inoculation) and systemic leaves (just above the local leaf without pathogen inoculation) was 36.4% and 11.7% in mycorrhizal plants, respectively, whereas DI was 59.6% and 36.4% in the corresponding leaves of AMF non‐inoculated plants, after 50 days of Gm inoculation, corresponding to 15 days after Cf inoculation by leaf infiltration. Foliar spray inoculation with Cf also revealed that AMF pre‐inoculated plants had a higher resistance against subsequent pathogen infection, where the DI was 41.3% in mycorrhizal plants vs. 64.4% in AMF non‐inoculated plants. AMF‐inoculated plants showed significantly higher fresh and dry weight than non‐inoculated plants under both control (without pathogen) and pathogen treatments. AMF‐inoculated plants exhibited significant increases in activities of superoxide dismutase and peroxidase, along with decreases in levels of H₂O₂ and malondialdehyde, compared with non‐inoculated plants after pathogen inoculation. AMF inoculation led to increases in total chlorophyll contents and net photosynthesis rate as compared with non‐inoculated plants under control and pathogen infection. Pathogen infection on AMF non‐inoculated plants led to decreases in chlorophyll fluorescence parameters. However, pathogen infection did not affect these parameters in mycorrhizal plants. Taken together, these results indicate that AMF colonization may play an important role in plant resistance against air‐borne pathogen infection by maintaining redox poise and photosynthetic activity.     

Efficacy assessment of antifungal metabolites from Chaetomium globosum No.05, a new biocontrol agent,against Setosphaeria turcica

Northern corn leaf blight (NCLB), an important and potentially destructive corn foliar disease, is caused by Setosphaeria turcica. The intent of this study was to evaluate antifungal metabolites from Chaetomium globosum (Cg) strain No.05 to suppress NCLB in maize. This strain significantly suppressed mycelial growth of numerous phytopathogenic fungi especially S. turcica on potato dextrose agar medium. The secondary metabolites of the strain inhibited mycelial growth and conidial germination of S. turcica. When co-inoculated at three droplets (5 μL/droplet) of conidial suspension (5 × 10⁴ conidia/mL) on each 8-cm-long detached leaf, 20% culture filtrates completely suppressed disease incidence of northern corn leaf blight. The application of the culture filtrates at 2 h post-inoculation (hpi) of S. turcica in greenhouse studies showed a 81.9% inhibition of NCLB on the seedlings, while culture filtrates applied before pathogen inoculation showed even higher rates of disease reduction. The application of the culture filtrates had no observed effects on the treated maize leaves or seedlings. Two active compounds, isolated from the extracts, were identified as chaetoglobosin A and chaetoglobosin C based on the spectroscopic analysis. Both in vitro and in planta bioassay experiments showed that chaetoglobosin A displayed potent biocontrol efficiency against S. turcica. To the best of our knowledge, this is the first report of the evaluation of the inhibitory effects of C. globosum and chaetoglobosin A against S. turcica both in vitro and on detached maize leaves.     

Population dynamics of Macrophomina phaseolina in relation to disease management: A review

Macrophomina phaseolina (Tassi) Goid. causes seedling blight, charcoal rot, leaf blight, stem and pod rot on over 500 plant species in different parts of the world. The pathogen survives as sclerotia formed in host tissues which are released into the soil as tissue decay. Low soil moisture is considered the more important predisposing factor for M. phaseolina-induced diseases than high temperature. The intensity of the disease on a crop is related to the population of viable sclerotia in the soil and abiotic factors. The influence of various management strategies in reducing the number of viable propagules of the pathogen in the soil has been studied in order to minimize the impact of the disease. Any management approach that reduces inoculum density in the soil may reduce disease incidence on the host. However, to reduce inoculum density, quantitative determination of viable propagules from soil is necessary in order to understand the effect of management strategies on the population dynamics of this pathogen. Considerable work has been done on organic amendments, changing crop sequences with tolerant crops, fumigants, herbicides and tillage in managing M. phaseolina populations in the soil and the resulting disease. Solarization has been used in controlling M. phaseolina in different countries where this pathogen is causing disease on economically valuable crops. However, this method of soil disinfestation was effective in eliminating viable populations at the top soil layer although by combining other approaches its effectiveness was improved at lower soil depth. Use of biological control agents with or without organic amendments or after solarization has emerged to be a practical management approach in the control of M. phaseolina. In this paper, an attempt has been made to review those research findings where the influence of various management approaches on survival of M. phaseolina mainly sclerotia have been investigated.     

Detection and Molecular Diversity of Pantoea ananatis Associated with White Spot Disease in Maize,Sorghum and Crabgrass in Brazil

Bacteria of the genus Pantoea have become important plant pathogens worldwide in recent years. Pantoea ananatis was reported as the cause of maize white spot, a serious maize disease in Brazil, causing significant yield losses. However, very little information is available about how to detect this pathogen, its genetic variability and the putative alternative hosts in maize‐growing areas. To address these issues, we implemented a rapid and efficient PCR‐based method to identify P. ananatis isolated from leaves showing white spot symptoms and evaluated its genetic diversity in maize, sorghum and crabgrass. Of the 29 bacteria isolated from typical water‐soaked lesions of white spot disease that produced yellow colonies, 15 isolates were identified as P. ananatis by 16S rDNA sequencing and correctly detected by the PCR reaction, amplifying a specific fragment of the ice nucleation gene (ina). These P. ananatis isolates included 13 from maize, one from sorghum and one from crabgrass, while the other 14 yellow colony isolates were from other bacterial species, including two Pantoea species (Pantoea dispersa and Pantoea agglomerans) that were not amplified by the ina primers. These results indicate that the optimized PCR assay can be used to detect P. ananatis isolated from white spot lesions and could be used as a large‐scale and cost‐effective method of detecting this pathogen in leaf lesions on maize and other grasses. All isolates were evaluated for hypersensitive response (HR) on tobacco, revealing that some P. ananatis were able to induce HR. The high genetic variability revealed by rep‐PCR did not differentiated the P. ananatis isolates based on their hosts or HR reaction. The detection, characterization and diversity of P. ananatis from maize, sorghum and crabgrass in our study can be applied in understanding epidemiology and designing control strategies for maize white spot disease in Brazil.     

Isolation of Trichoderma and Evaluation of their Antagonistic Potential against Alternaria porri

Nine isolates of Trichoderma were collected from Assiut Governorate, Egypt, as leaf surface and endophytic fungi associated with onion flora stalks. Four isolates were identified as Trichoderma harzianum, while five isolates were belonging to Trichoderma longibrachiatum. The antagonistic activity of these isolates against onion purple blotch pathogen Alternaria porri was studied in vitro using dual culture assay. All tested Trichoderma isolates showed mycoparasitic activity and competitive capability against the mycelial growth of A. porri. Mycoparastic activity of Trichoderma was manifested morphologically by the overgrowth upon the mycelial growth of the pathogen and microscopically by production of coiling hyphae around pathogen hyphae. Isolates of T. harzianum exhibited high ability to compete on potato dextrose agar (PDA) medium causing the maximum rate of pathogen inhibition (73.12%), while isolates of T. longibrachiatum showed inhibition rate equalling 70.3%. Chitinase activity of Trichoderma was assayed, and T. harzianum Th‐3013 showed the maximum value contributing 2.69 U/min. Application of T. harzianum Th‐3013 to control purple blotch disease in vivo under greenhouse conditions caused disease reduction up to 52.3 and 79.9% before and after 48 h of pathogen inoculation, respectively, while the fungicide Ridomil Gold Plus caused disease reduction comprising 56.5 and 71.7%, respectively. This study proved that T. harzianum Th‐3013 as a biocontrol agent showed significant reduction in onion purple blotch disease compared with the tested fungicide.     

Pantoea stewartii Causing Stewart's Wilt on Dracaena sanderiana in Korea

In April 2010, a severe occurrence of Stewart's wilt on Dracaena sanderiana plants was observed in greenhouses in Seongnam, Gyeonggi Province, South Korea, with an incidence of 35‐50%. Being imported plants, little was known about the pathogens associated with D. sanderiana. Symptoms included chlorosis, wilting and leaf blight on the leaf surfaces. Physiological analysis, pathogenicity tests, sequencing and phylogenetic analysis of the 16S rRNA gene revealed that the pathogen was the bacterium Pantoea stewartii. To the best of our knowledge, this is the first report on bacterial wilt caused by P. stewartii on D. sanderiana.     

Biological control of common blight of bean (Phaseolus vulgaris) causedby Xanthomonas axonopodis pv. phaseoli by using the bacteriumRahnella aquatilis

The aim of this study was to evaluate the bacterium Rahnella aquatilis (Ra) for protection of bean plants against common blight disease caused by Xanthomonas axonopodis pv. phaseoli (Xap). Xap isolates were isolated from a naturally blighted leaves of bean plants grown in Assiut governorate. The blight symptoms were produced by all three isolates, but the isolates differed in their degree of the pathogenicity. Xap1 was the most virulence one against bean plants. The effect of Ra against common blight of bean plant was tested. In vitro studies, Ra exhibited inhibitor effect against the pathogen. Under greenhouse and field conditions, beanvariety “Giza 6” treated by Ra resulted in marked disease suppression. Ahigh decrease of the disease was correlated with a reduction of the bacterial multiplication. In physiological studies, bean plants treated by Ra exhibited higher phenolic compounds contents and higher activity of peroxidase (PO) enzyme than untreated plants. In conclusion, application of Ra was effective and could be recommended for controlling the bean common blight disease.     

Evidence of Alternaria alternata Causing Leaf Spot of Aloe vera in Iran

A leaf spot and leaf blight disease was observed on Aloe vera plants as small, circular to oval dark brown necrotic sunken spots on leaves. Infected tissues collected from different sites in diseased fields were cultured on potato carrot agar medium, and the pathogen was identified as Alternaria alternata on the basis of morphological and cultural characteristics. The conidiophores were branched, straight, golden brown, smooth‐walled, measuring up to μm long by 3 μm wide with one conidial scar. The conidia were golden brown in colour and produced in long branched chains, obclavate in shape and in short conical flask. Pathogenicity tests conducted on healthy potted aloe plants in a glasshouse showed typical leaf spot symptoms after 4–7 days. The optimal temperature for the growth of A. alternata was 25°C.     

Analysis of Pathogenic Diversity of the Rice Bacterial Blight Pathogen (Xanthomonas oryzae pv. oryzae) in the Andaman Islands and Identification of Effective Resistance Genes

Bacterial blight (BB) caused by Xanthomonas oryzae pv. oryzae (Xoo) is a major disease of rice in the tropics for which genetic resistance in the host plants is the only effective solution. This study aimed at identification of resistance gene combinations effective against Xoo isolates and fingerprinting of the Xoo isolates of Andaman Islands (India). Here, we report the reaction of 21 rice BB differentials possessing Xa1 to Xa21 genes individually and in different combinations to various isolates of pathogen collected from Andaman Islands. Pathological screening results of 14 isolates revealed that among individual genes tested across 2 years, Xa4, Xa7 and Xa21 conferred resistance reaction across all isolates, whereas among combinations, IRBB 50 (Xa4 + xa5), IRBB 52 (Xa4 + Xa21) and IRBB 60 (Xa4 + xa5 + xa13 + Xa21) conveyed effective resistance against tested isolates. The nature of genetic diversity among four isolates selected on the basis of geographical isolation in the islands was studied through DNA finger printing. The RAPD primers S111, S119, S1117, S1109, S1103, S109 and S105 were found to be better indicators of molecular diversity among isolates than JEL primers. The diversity analysis grouped 14 isolates into three major clusters based on disease reaction wherein isolate no. 8 was found the most divergent as well as highly virulent. The remaining isolates were classified into two distinct groups. The importance of the study in the context of transfer of resistance gene(s) in the local cultivars specifically for tropical island conditions is presented and discussed.     

A role for β‐sitosterol to stigmasterol conversion in plant–pathogen interactions

Upon inoculation with pathogenic microbes, plants induce an array of metabolic changes that potentially contribute to induced resistance or even enhance susceptibility. When analysing leaf lipid composition during the Arabidopsis thaliana–Pseudomonas syringae interaction, we found that accumulation of the phytosterol stigmasterol is a significant plant metabolic process that occurs upon bacterial leaf infection. Stigmasterol is synthesized from β‐sitosterol by the cytochrome P450 CYP710A1 via C22 desaturation. Arabidopsis cyp710A1 mutant lines impaired in pathogen‐inducible expression of the C22 desaturase and concomitant stigmasterol accumulation are more resistant to both avirulent and virulent P. syringae strains than wild‐type plants, and exogenous application of stigmasterol attenuates this resistance phenotype. These data indicate that induced sterol desaturation in wild‐type plants favours pathogen multiplication and plant susceptibility. Stigmasterol formation is triggered through perception of pathogen‐associated molecular patterns such as flagellin and lipopolysaccharides, and through production of reactive oxygen species, but does not depend on the salicylic acid, jasmonic acid or ethylene defence pathways. Isolated microsomal and plasma membrane preparations exhibited a similar increase in the stigmasterol/β‐sitosterol ratio as whole‐leaf extracts after leaf inoculation with P. syringae, indicating that the stigmasterol produced is incorporated into plant membranes. The increased contents of stigmasterol in leaves after pathogen attack do not influence salicylic acid‐mediated defence signalling but attenuate pathogen‐induced expression of the defence regulator flavin‐dependent monooxygenase 1. P. syringae thus promotes plant disease susceptibility through stimulation of sterol C22 desaturation in leaves, which increases the stigmasterol to β‐sitosterol ratio in plant membranes.     

Biocontrol potential of phylloplane bacterium Ochrobactrum anthropi BMO‐111 against blister blight disease of tea

Aims

The present study was carried out to screen the phylloplane bacteria from tea for antagonism against grey blight caused by Pestalotiopsis theae and blister bight caused by Exobasidium vexans and to further evaluate the efficient isolates for disease control potential under field condition.

Methods and Results

A total of 316 morphologically different phylloplane bacteria were isolated. Among the antagonists, the isolates designated as BMO‐075, BMO‐111 and BMO‐147 exhibited maximum inhibitory activity against both the pathogens under in vitro conditions and hence were selected for further evaluation under microplot field trial. Foliar application of 36‐h‐old culture of BMO‐111 (1 × 10⁸ colony‐forming units ml^?1) significantly reduced the blister blight disease incidence than the other isolates. The culture of BMO‐111 as well as its culture filtrate effectively inhibited the mycelial growth of various fungal plant pathogens. The isolate BMO‐111 was identified as Ochrobactrum anthropi based on the morphological and 16S rDNA sequence analyses.

Conclusions

It could be concluded that the biocontrol agent O. anthropi BMO‐111 was effective against blister blight disease of tea.

Significance and Impact of the Study

Further study is required to demonstrate the mechanism of its action and formulation for the biocontrol potential against blister blight disease of tea.     

Detection of Cochliobolus heterostrophus races in South China

Cochliobolus heterostrophus is the causal pathogen of the southern corn leaf blight (SCLB). There are three known races: race O, race C and race T. To determine which C. heterostrophus races comprise the field population in southern China and to assess diversity of these strains in terms of virulence, 200 isolates from diseased plants were collected in nine provinces/municipalities. All were race O, that is, no race T or race C isolates were found. Sixty race O isolates that sporulated well were chosen for further analysis. Virulence was measured using the integral optical density (IOD) of leaf lesions on four maize inbred lines. UPGMA cluster analysis of AFLP markers was applied to the 60 race O isolates plus control race O, T and C strains. Phylogenetic distribution, geographic location and virulence were not correlated. These results can provide valuable information for guidance in early warning and disease control.     

First Report of Sclerotinia nivalis Causing White Mold Disease on Sedum sarmentosum in China

In 2010 and 2011, a disease exhibiting characteristics of white mold was found on Sedum sarmentosum, a crassulaceous weed under canopies of tea trees, in Zhushan County, Hubei Province, China. Based on the cultural and morphological characteristics, the pathogen was identified as Sclerotinia nivalis Saito. In the phylogenetic tree inferred from the internal transcribed spacer (ITS)‐rDNA sequences, the pathogen was clustered with five previously characterized isolates of S. nivalis, forming a unique clade, thus confirming the morpho‐cultural identification. Koch’s postulates were fulfilled by pathogenicity tests using the isolate SsSn‐24 and Let‐19 of S. nivalis on plants of S. sarmentosum. To our knowledge, this is the first report of S. nivalis on S. sarmentosum in the family Crassulaceae.     

Symptomology and etiology of a new disease,yellow stunt,and root rot of standing milkvetch caused by <Emphasis Type="Italic">Embellisia</Emphasis> sp. in Northern China

An Embellisia sp. has been established as the cause of a new disease of the herbaceous perennial forage legume, ‹standing milkvetch’ (Astragalus adsurgens Pall.) in Northern China, which severely reduces plant density and degrades A. adsurgens stands. The disease was common at an experimental location in Gansu Province where it was recognized by the occurrence of stunted plants with reddish-brown stems and yellow and necrotic leaf blades. An Embellisia sp. was isolated from symptomatic stem, leaf blade, petiole, and root tissues at varying frequencies of up to 90%. Single-spore isolates grew very slowly on PCA, PDA, V-8 and, wheat hay decoction agar. Pathogenicity was confirmed by inoculation of seeds, dipping 2-day-old pre-germinated seedlings in inoculum and spraying inoculum on 6-month-old plants. Symptoms on test plants included yellow leaf lesions, brown lesions on stems and petioles, stunted side-shoots with yellow, small, distorted and necrotic leaves, shoot blight, bud death, crown rot, root rot, and plant death. The disease is named as ‹yellow stunt and root rot’ of A. adsurgens to distinguish it from diseases caused by other known pathogens. Embellisia sp. is also pathogenic to A. sinicus but not to 11 other tested plant species.