Comparative survival of Sclerotia of Sclerotinia minor and S. sclerotiorum

Survival of sclerotia of Sclerotinia minor and S. sclerotiorum was compared in irrigated fields during the summer in two major lettuce production areas in California. More than 50% sclerotia of S. sclerotiorum compared with 4 and 35% of S. minor remained viable after 24 weeks of burial at 15 and 5 cm depths, respectively, in the San Joaquin Valley while >80% of sclerotia survived in the Salinas Valley for both species. The results explain in part the lower infections from S. minor in the San Joaquin Valley. To identify factors that contribute to the rapid decline in the viability of sclerotia, the effects of soil moisture, temperature, and oxygen levels were studied in laboratory. More than 90% of sclerotia of both species survived for at least 3 months in sterilized dry soils at temperatures between 15 and 40 degrees C. Soil moisture did not affect survival at 15 and 25 degrees C. At 35 degrees C, however, survival rates were significantly lower at high (-0.3 to -0.01 MPa) water potential than at low (<-1.0 MPa) water potential. Incubation under ultralow oxygen concentration (0.01%) significantly reduced survival of sclerotia in nonautoclaved moist soils at 25 degrees C, with less than 2% sclerotia surviving over 4 weeks compared with about 45% sclerotia surviving at the ambient oxygen level (21%). The combination of high temperature, high soil moisture, and reduced oxygen in irrigated fields contribute to the lower survival of both Sclerotinia species and the responses of the two species to these conditions shape their relative geographical distribution.     

Control of lettuce drop disease, caused by Sclerotinia sclerotiorum, with metham-sodium soil treatment and foliar application of benomyl

The effects of metham-sodium (MES) soil treatment, a varying number of benomyl foliar sprays (two, three and six) and combined treatments on control of lettuce drop disease were studied at two sites in Israel naturally infested with the pathogen Sclerotinia sclerotiorum. MES killed 85% of the S. sclerotiorum sclerotia in the top 10 cm of soil where initial populations were 0.6 and 1.6 viable sclerotia per kg soil at the two sites. Of the remaining viable sclerotia. only 30% produced apothecia. MES treatment alone reduced numbers of apothecia to 5% of those counted in the unsprayed control treatment. Numbers of ascospores deposited on the crop in the MES-treated plots were 7-20% of those deposited on control plots. Lettuce drop decreased the total yield by 30% in unsprayed control plots; MES application reduced the loss to 4% and also increased the number of marketable plants. Benomyl sprays alone also significantly reduced numbers of apothecia and disease development and consequently increased yield, although not to the extent observed with the MES treatment alone. Combined treatments produced the best disease control, but this improvement was not accompanied by any major increase in yield compared to that with MES treatment alone.     

Interspecific Transmission of Double-Stranded RNA and Hypovirulence from Sclerotinia sclerotiorum to S. minor

ABSTRACT Interspecific transmission of a hypovirulence-associated double-stranded RNA (dsRNA) and hypovirulent phenotype was attempted from hypovirulent isolate Ss275 of Sclerotinia sclerotiorum to five virulent isolates of S. minor. dsRNA and the hypovirulent phenotype were successfully transmitted to one of the five isolates, Sm10. Three putative converted isolates of Sm10 were slow growing and developed atypical colony morphologies characteristic of the hypovirulent phenotype. These isolates were assayed for virulence and produced significantly smaller lesions than isolate Sm10 on detached leaves of Romaine lettuce. One of these putative converted isolates, designated Sm10T, was tested to confirm interspecific transmission of dsRNA. In northern hybridizations, dsRNA isolated from Sm10T hybridized with a digoxigenin-labeled cDNA probe prepared from dsRNA isolated from Ss275. Random amplified polymorphic DNA analysis confirmed that isolate Sm10T was derived from Sm10 and not from Ss275 or a hybrid of the two species. The dsRNA and hypovirulent phenotype were subsequently transmitted intraspecifically from Sm10T to Sm8. To our knowledge, this is the first report of interspecific transmission of dsRNA and an associated hypovirulent phenotype between fungal plant pathogens by hyphal anastomosis.     

Bacillus thuringiensis C25 which is rich in cell wall degrading enzymes efficiently controls lettuce drop caused by Sclerotinia minor

European Journal of Plant Pathology - Sclerotinia is a phytopathogenic genus of fungi that form sclerotia: black, hard, seed-like functioning bodies. Sclerotinia sclerotiorum and Sclerotinia minor...     

Analyses of Lettuce Drop Incidence and Population Structure of Sclerotinia sclerotiorum and S. minor

ABSTRACT To understand the geographical distribution of lettuce drop incidence and the structure of Sclerotinia minor and S. sclerotiorum populations, commercial lettuce fields were surveyed in the Salinas, San Joaquin, and Santa Maria Valleys in California. Lettuce drop incidence, pathogen species, and mycelial compatibility groups (MCGs) were determined and analyzed using geostatistic and geographical information system tools. Lettuce drop incidence was lowest in the San Joaquin Valley, and not significantly different between the other two valleys. Semivariogram analysis revealed that lettuce drop incidence was not spatially correlated between different fields in the Salinas Valley, suggesting negligible field-to-field spread or influence of inoculum in one field on other fields. Lettuce drop incidence was significantly lower in fields with a surface drip system than in fields with furrow or sprinkler irrigation systems, suggesting that the surface drip system can be a potential management measure for reducing lettuce drop. In the San Joaquin Valley, S. sclerotiorum was the prevalent species, causing drop in 63.5% of the fields, whereas S. minor also was identified in 25.4% of the fields. In contrast, in the Salinas Valley, S. minor was the dominant species (76.1%) whereas S sclerotiorum only observed in only 13.6% fields, in which only a few plants were infected by S. sclerotiorum. In the Santa Maria Valley, both species frequently were identified, with S. minor being slightly more common. Although many MCGs were identified in S. minor, most of them consisted of only one or two isolates. In all, approximately 91.4% of the isolates belonged to four MCGs. Among them, MCG-1 was the most prevalent group in all three valleys, accounting for 49.8% of total isolates. It was distributed all over the surveyed areas, whereas other MCGs were distributed more or less locally. Populations of S. sclerotiorum exhibited greater diversity, with 89 isolates collected from the Salinas and San Joaquin Valleys belonging to 37 different MCGs. Among them, the most recurrent MCG-A contained 16 isolates, and 30 MCGs contained only 1 isolate each. Many MCGs occurred within only one or a part of the two valleys. Potential reasons for this abundant diversity are discussed.     

Germination of Sclerotinia minor and S. sclerotiorum Sclerotia Under Various Soil Moisture and Temperature Combinations

ABSTRACT Sclerotial germination of three isolates each of Sclerotinia minor and S. sclerotiorum was compared under various soil moisture and temperature combinations in soils from Huron and Salinas, CA. Sclerotia from each isolate in soil disks equilibrated at 0, -0.03, -0.07, -0.1, -0.15, and -0.3 MPa were transferred into petri plates and incubated at 5, 10, 15, 20, 25, and 30 degrees C. Types and levels of germination in the two species were recorded. Petri plates in which apothecia were observed were transferred into a growth chamber at 15 degrees C with a 12-h light-dark regime. All retrievable sclerotia were recovered 3 months later and tested for viability. Soil type did not affect either the type or level of germination of sclerotia. Mycelial germination was the predominant mode in sclerotia of S. minor, and it occurred between -0.03 and -0.3 MPa and 5 and 25 degrees C, with an optimum at -0.1 MPa and 15 degrees C. No germination occurred at 30 degrees C or 0 MPa. Soil temperature, moisture, or soil type did not affect the viability of sclerotia of either species. Carpogenic germination of S. sclerotiorum sclerotia, measured as the number of sclerotia producing stipes and apothecia, was the predominant mode that was affected significantly by soil moisture and temperature. Myceliogenic germination in this species under the experimental conditions was infrequent. The optimum conditions for carpogenic germination were 15 degrees C and -0.03 or -0.07 MPa. To study the effect of sclerotial size on carpogenic germination in both S. minor and S. sclerotiorum, sclerotia of three distinct size classes for each species were placed in soil disks equilibrated at -0.03 MPa and incubated at 15 degrees C. After 6 weeks, number of stipes and apothecia produced by sclerotia were counted. Solitary S. minor sclerotia did not form apothecia, but aggregates of attached sclerotia readily formed apothecia. The number of stipes produced by both S. minor and S. sclerotiorum was highly correlated with sclerotial size. These results suggest there is a threshold of sclerotial size below which apothecia are not produced, and explains, in part, why production of apothecia in S. minor seldom occurs in nature.     

Effects of soil temperature, moisture, and burial depths on carpogenic germination of Sclerotinia sclerotiorum and S. minor

Extensive studies have been conducted on the carpogenic germination of Sclerotinia sclerotiorum, but carpogenic germination in S. minor has not been studied adequately. It remains unclear why apothecia of this pathogen have seldom been observed in nature. In this study, a new method was developed to produce apothecia in the absence of soil or sand, and carpogenic germination without preconditioning was recorded for 95 of the 96 S. sclerotiorum isolates tested. Carpogenic germination of the two species was compared under a variety of temperature, soil moisture, burial depths, and short periods of high temperature and low soil moisture. The optimal temperatures for rapid germination and for maximum germination rates were both lower for S. minor than for S. sclerotiorum. The temperature range for carpogenic germination was also narrower for S. minor than for S. sclerotiorum. A 5-day period at 30 degrees C, either starting on the 10th or 20th day of incubation, did not significantly affect carpogenic germination of S. sclerotiorum. For both S. minor and S. sclerotiorum, the percentage of carpogenically germinated sclerotia increased as soil water potential increased from -0.3 to -0.01 MPa. In the greenhouse, a 10- or 20-day dry period completely arrested carpogenic germination of S. sclerotiorum, and new apothecia appeared after an interval of 35 days following rewetting, similar to the initial carpogenic germination regardless of when the dry period was imposed. In naturally infested fields, the number of sclerotia in 100 cc of soil decreased as depth increased from 0 to 10 cm before tillage, but became uniform between 0 and 10 cm after conventional tillage for both species. Most apothecia of S. minor were, however, produced from sclerotia located at a depth shallower than 0.5 cm while some apothecia of S. sclerotiorum were produced from sclerotia located as deep as 4 to 5 cm. These results provide the much needed information to assess the epidemiological roles of inoculum from sexual reproduction in diseases caused by the two Sclerotinia species in different geographical regions. However, more studies on effects of shorter and incompletely dry periods are still needed to predict production of apothecia of S. sclerotiorum in commercial fields under fluctuating soil temperature and moisture.     

Adjustment of soil-surface pH and comparison with conventional fungicide treatments for control of lettuce drop (Sclerotinia minor)

The use of soil-surface applications of finely powdered calcium hydroxide (Ca(OH)₂) to inhibit Sclerotinia minor sclerotial germination and infection at the collar region of lettuce plants is described. In the laboratory, a pH > 8·0 reduced sclerotial germination of the three S. minor isolates tested. In the glasshouse, surface applications of 2–10 t Ca(OH)₂ ha⁻¹ raised the pH of the top 1–2 cm of a duplex sandy loam soil above 8·5 for at least 8 weeks without affecting soil pH within the transplant root zone. There was a linear relationship between the rate of Ca(OH)₂ applied and disease control, with complete disease suppression at 10 t Ca(OH)₂ ha⁻¹. In field trials on two soil types (duplex sandy loam, pH 6·0; and red ferrosol, pH 6·9), a rate of 2·5 t Ca(OH)₂ ha⁻¹, maintained soil-surface pH above 8·5 for 1–3 weeks and provided up to 58% reduction in lettuce drop. Application of polyvinyl alcohol (a soil-conditioning polymer) over the Ca(OH)₂ layer appeared to reduce Ca(OH)₂ loss by wind, but did not improve retention of raised soil-surface pH or disease suppression. Ca(OH)₂ treatment gave similar disease control to the industry standard treatment of a procymidone-based fungicide seedling drench. A combined treatment of Ca(OH)₂ and fungicide drench gave greater control than either individual treatment, and equivalent control to fungicide drench and three procymidone foliar sprays, offering integrated management options. The use of soil-surface-applied Ca(OH)₂ with fungicides, rotation and drip irrigation offers an opportunity for enhanced and sustainable control of lettuce drop.     

Population structure of Sclerotinia sclerotiorum in crop and wild hosts in the UK

Sclerotinia sclerotiorum is an important pathogen of many crop plants which also infects wild hosts. The population structure of this fungus was studied for different crop plants and Ranunculus acris (meadow buttercup) in the UK using eight microsatellite markers and sequenced sections of the intergenic spacer (IGS) region of the rRNA gene and the elongation factor 1‐alpha (EF) gene. A total of 228 microsatellite haplotypes were identified within 384 isolates from 12 S. sclerotiorum populations sampled in England and Wales. One microsatellite haplotype was generally found at high frequency in each population and was distributed widely across different hosts, locations and years. Fourteen IGS and five EF haplotypes were found in the 12 populations, with six IGS haplotypes and one EF haplotype exclusive to buttercup. Analysis of published sequences for S. sclerotiorum populations from the USA, Canada, New Zealand and Norway showed that three of the IGS haplotypes and one EF haplotype were widely distributed, while eight IGS haplotypes were only found in the UK. Although common microsatellite and IGS/EF haplotypes were found on different hosts in the UK, there was evidence of differentiation, particularly for one isolated population on buttercup. However, overall there was no consistent differentiation of S. sclerotiorum populations from buttercup and crop hosts. Sclerotinia sclerotiorum therefore has a multiclonal population structure in the UK and the wide distribution of one microsatellite haplotype suggests spatial mixing at a national scale. The related species S. subarctica was also identified in one buttercup population.     

小核盘菌菌丝生长和乙二酸合成影响因素研究

小核盘菌菌丝在V8溶液培养基和酵母浸膏溶液培养基中生长显著优于其他供试培养基,菌丝干重分别为6.666 mg/mL和6.632 mg/mL。在葡萄糖土豆浸汁培养基中产生的乙二酸量显著高于其他培养基,为1.981 mg/mL。在蔗糖液培养基中,培养7 d菌丝干重达到最大,为4.455 mg/mL。培养6 d乙二酸产生量达到最大,为0.966 mg/mL。培养基初始pH4.5最适合菌丝生长,干重为4.678 mg/mL。乙二酸含量随初始pH增加而增加,pH7.0的培养基中乙二酸含量为1.835 mg/mL。氮源含量增加可以促进乙二酸的合成,当蔗糖与大豆水解蛋白之比为25 g/10 g时,乙二酸含量为1.897 mg/mL。添加琥珀酸钠乙二酸含量显著增加,为3.741 mg/mL。     

核盘菌对菌核净的抗药性机制初探

经药剂筛选获得对菌核净不同表型的核盘菌(Sclerotinia sclerotiorum ) 抗药性突变体。与敏感亲本菌株相比, 抗药突变体MN 61 (MR ) 和MN 91 (HR) 在含1% 和8% 葡萄糖的PDA 上生长受到抑制,MN 113 (LR) 只对1% 葡萄糖敏感。通过测定抗药突变体MN 61 和野生敏感菌株PN 061 的电导率, 发现抗药突变体能在更短的时间里渗出更多的电解质。抗药突变体苯丙氨酸解氨酶活性比敏感亲本菌株高出1 倍以上, 当用不同浓度菌核净处理或饥饿处理时, 抗药突变体和敏感亲本菌株PAL 活性均上升, 但抗药突变体的酶活始终高于敏感亲本菌株。     

Factors affecting the production of apothecia and longevity of sclerotia of Sclerotinia sclerotiorum

Sclerotia of Sclerotinia sclerotiorum produced naturally on winter oilseed rape and by an isolate from this host on sterile wheat grain were placed in two different soil types in pots, either on the soil surface or 1. 2, 3, 4 or 6 cm deep. Nitrogen (as TN 34, Thames Nitrogen Co.) was applied to one set of pots in autumn 1982 and spring 1983. Production of apothecia was recorded in this set in 1983 only. In pots without added N apothecia were counted in 1983 and 1984 and sclerotia were recovered from these pots at the end of the experiment.
Overall, production of apothecia was not affected by soil type. More apothecia formed from cultured sclerotia and they appeared earlier than from natural sclerotia. More apothecia were produced from sclerotia placed at 0–2 cm than from those buried deeper. Fewer apothecia were produced in pots with added N and their production was delayed. Fewer sclerotia were recovered from the upper layers of soil with a higher clay content, and when sclerotia were obtained from oilseed rape rather than from cultures.     

不同栽培措施对制种生菜菌核病的影响

本文研究不同栽培措施对制种生菜菌核病的影响。采用不同覆膜栽培方式试验结果表明,黑色和白色膜全覆盖栽培制种生菜对制种生菜菌核病有极显著的防治效果;其中黑色膜全覆盖对制种生菜菌核病的防效高,为76.7%,白色膜全覆盖防效为66.8%,黑膜与白膜全覆盖防效之间没有显著差异。不同钾肥施用量试验结果表明,最经济合理的硫酸钾用量为150～225kg/hm2;其中追肥施钾150～225kg/hm2对生菜菌核病的防治效果为44.6%～47.3%,基肥施钾150～225kg/hm2防治效果为43.2%～45.9%。不同栽培密度试验结果证明:合理的栽培密度对制种生菜菌核病有极显著的防治效果,酒泉市针对绿叶半结球型生菜最经济合理的栽培密度为35cm×50cm～40cm×50cm。     

油菜内生生防菌BY-2在油菜体内的定殖与对油菜菌核病的防治作用

 从油菜植株体内分离出的内生细菌BY-2,经过生物学鉴定为枯草芽孢杆菌(Bacillus subtilis)。用BY-2回接油菜,重新分离到具有BY-2相同形态特征和抑制病原真菌能力的内生菌株。油菜接种后的第10 d,体内的BY-2菌数达(2.24~9.02)×10³ cfu/g鲜植株,25 d仍然保持在(3.13~8.59)×10³ cfu/g鲜植株。BY-2与油菜核盘菌[Sclerotinia sclerotiorum(Lib.) de Bary]对峙培养可以形成直径为3.1 cm的抑菌圈;可使油菜核盘菌菌丝细胞浓缩变短,细胞壁破裂,原生质外溢,从而抑制真菌生长发育;同时还能抑制菌核的萌发,抑制率达60%~70%;在油菜离体叶片试验中,BY-2对菌核病的防治效果达100%。     

BTH诱导花椰菜对菌核病的抗性研究

 利用苯并噻二唑BTH处理菌核病抗性不同的花椰菜品种幼苗, 采用营养生长期活体叶片菌丝块接种鉴定法评价菌核病抗性诱导效果,结果表明经BTH处理的植株菌核病病情指数明显下降, 对感病品种和抗病品种的诱抗效果分别达到81.5%和63.8%。对于花椰菜重要的防御酶活性变化研究结果表明,BTH诱导处理的花椰菜植株过氧化物酶（POD）、抗坏血酸酶( SOD )、过氧化氢酶（CAT）、苯丙氨酸解氨酶（PAL）和多酚氧化酶( PPO)的活性均有所提高。同时病程相关蛋白几丁质酶和β-1,3-葡聚糖酶的活性也增加。 利用半定量RT-PCR方法检测防御反应基因表达,结果表明BTH诱导首先激发了植株 PR-1等基因参与的水杨酸信号传导防御反应途径的发生,同时PDF1.2 基因的上调表达说明BTH诱导也影响了茉莉酸信号传导途径。     

Glasshouse trials of Coniothyrium minitans and Trichoderma species for the biological control of Sclerotinia sclerotiorum in celery and lettuce

Coniothyrium minitans, Trichoderma harzianum (HH3) and Trichoderma sp. (B1) were tested for ability to control disease caused by Sclerotinia sclerotiorum in a sequence of a celery crop and two lettuce crops in the glasshouse. In control plots, over 80% of celery and 90 and 60% of lettuce in first and second crops, respectively, were infected at harvest. Only the C. minitaris treatment in the first lettuce crop decreased disease and increased marketable yield. Nevertheless, C. minitans reduced the number of sclerotia recovered at harvest in the celery and first lettuce crops and decreased sclerotial survival over the autumn fallow periods following the celery and second lettuce crop. C. minitans survived in soil for over 1 year and spread to infect sclerotia in virtually all other plots. C. minitans infected sclerotia at all times of the year but sclerotia still failed to degrade during the summer months when the soil was dry. The Trichoderma species tested had no effect on disease and almost no effect on the survival of the sclerotia. even though they could be recovered from soil for the duration of the experiments.