Reaction of Oilseed Rape Cultivars to Sclerotinia Stem Rot in Field Environments

Sclerotinia sclerotiorum (Lib.) de Bary, as an ubiquitous phytopathogenic Ascomycete fungus capable of infecting a wide range of plants, has increased in importance for oilseed rape production worldwide in recent years. The objective of this research was to determine the effectiveness of two measurements of disease rating in predicting the field reaction of cultivars to sclerotinia stem rot. One hundred and ten cultivars or lines of oilseed rape were repeatedly evaluated the disease severity index (DSI) and lesion length for their resistance to nine isolates of S. sclerotiorum in two field stations.There were greatly significant differences between two stations, among isolates, among population of cultivars for the disease severity index and lesion length, and also significant station×isolate interaction for lesion length and population×isolate interactions for DSI and lesion length, but interactions of station×isolate for DSI were not significant.Cultivars such as Pomorzanin and Lisek consistently had the lowest disease severity ratings in the field tests. However,most of cultivars were susceptible to sclerotinia stem rot in the field test. A positive correlation between the mean DSI and lesion length was found for both isolates (Sc23 and B1). The correlations ranged from 0.23 to 0.83 at various plots, with nine of the twelve correlations significant. The correlations for disease caused by different fungal isolates were statistically significant difference. These correlations ranged from -0.44 to 0.90, with nine of the thirty-two correlations significant. It indicated that resistance to some isolates was significantly correlated and selection of some varieties for resistance to one isolate appeared to improve resistance to other isolates.     

油菜几丁质酶的特点及其与抗菌核病的关系

通过测定甘蓝型油菜西农长角几丁质酶对底物的特异性等性质,结果表明,几丁质酶以脱乙酰化几丁质7B（乙酰化程度为30％）为底物时活性最高,而对8B,9B,10B（乙酰化程度为20％,10％,0％）和菌核菌细胞壁的活性较低。几丁质酶活性的最适宜反应温度为30℃-40℃中,并在25℃以下时活性大大降低,这与菌核菌生长和入侵的最适温度15℃-25℃不一致。西农长角的种子中几丁质酶的活性很强,发芽后逐渐下降,到第5天时开始提高。研究还发现,不同类型和抗性油菜材料的几丁质酶活性和对菌核菌的抗性间存在相关关系,相关系数达显著水平,这表明几丁质酶在提高油菜的抗菌核病能力起着重要的作用。     

用等位基因特异性寡核苷酸（ASO）-PCR快速检测抗多菌灵的油菜菌核病菌

 通过3对兼并性引物扩增获得与S.sclerotiorum抗药性相关的β-微管蛋白基因，全长1 685 bp，包含4个内元，相应的编码447个氨基酸。该基因与其它6种线状真菌的β-微管蛋白基因相比，氨基酸序列同源性达95.78%～97.66%，但内元数目和大小不同。比较S.sclerotiorum敏感型和抗药性菌株的β-微管蛋白基因，发现该基因第198位氨基酸由谷氨酸突变为丙氨酸，从而导致田间抗药性的产生。为了快速、准确监测田间抗药性频率，根据S.sclerotiorumβ-微管蛋白基因的突变位点设计了2对等位基因特异性寡核苷酸（ASO），直接以菌核的基因组DNA为模板扩增，所需时间约为6 h，抗药性检出率为100%，与传统菌丝直径法的测定结果相比检测准确率为96%，而传统的检测方法至少需要1～2周。     

Excel在油菜菌核病灰色预测模型构建中的应用

灰色系统理论，为病虫害预测预报提供了新的途径与方法。灰色预测模型的计算，其中关键是计算微分方程^AX（1）（K 1)=(x^(0)(1)-u/a)e^-ak u/a的解。利用Excel计算求解，具有操作方法简便易行，结果准确可靠、实用性强的优点。本文根据洞庭湖区1980～1992年油菜菌核病发病资料，利用Microsoft Excel 200进行计算，建立灰色GM(1，1)长期预测模型，经检验该模型精确度高，预测结果可靠。     

甘蓝型油菜品种（系）对菌核病抗（耐）病性的测定

用草酸浸根和黑麦粒接种两种方法分别对来自中国和加拿大等国的１５个春性和１６个冬性、半冬性甘蓝型油菜品种（系）进行了抗（耐）菌核病的鉴定。结果没有发现高抗类型，但是甘蓝型油菜品种（系）之间存在着感染菌核病轻重程度的显著差异。     

Resistance to Rhizoctonia solani AG‐2‐2 (IIIB) in creeping bentgrass plants transformed with pepper esterase gene PepEST

A pepper esterase (PepEST) gene was introduced into creeping bentgrass (Agrostis stolonifera) by Agrobacterium‐mediated transformation. Purified recombinant PepEST proteins were sufficient to inhibit the growth of the fungal pathogens Rhizoctonia solani AG2‐2 (IIIB) (causing brown patch) and Sclerotinia homoeocarpa (dollar spot), but not the oomycete responsible for pythium blight, Pythium aphanidermatum. PepEST proteins were most effective against R. solani. After genetic transformation of creeping bentgrass with PepEST, the genomic integration of transgenes bar and PepEST was confirmed by Southern blot analysis, and their expression was also validated by northern blot and western blot analyses. Disease severity on R. solani‐inoculated leaves of transgenic plants was <10% compared to ca. 50% in non‐transgenic plants. Microscopic observation of infected leaves indicated that PepEST inhibited the growth of hyphae upon fungal infection.     

A nested‐PCR method for rapid detection of Sclerotinia sclerotiorum on petals of oilseed rape (Brassica napus)

This study established a quick and accurate method to detect petal infection of oilseed rape (Brassica napus) by Sclerotinia sclerotiorum using a nested‐PCR technique. DNA samples were extracted from each petal using a microwave method, followed by two rounds of PCR amplification. The first‐round PCR amplification was performed using the universal fungal primer pair ITS4/ITS5, and the second‐round amplification with a specific primer pair XJJ21/XJJ222, which was designed using the single‐nucleotide polymorphisms among nuclear rDNA ITS sequences of Sclerotinia spp., Botrytis spp. and other selected fungi. The established technique is rapid and inexpensive, and has a high degree of specificity and sensitivity. This assay can distinguish Sclerotinia spp. from other fungi, including Botrytis cinerea, a closely related and frequent cohabitant on oilseed rape petals, and can detect 50 fg genomic DNA, five ascospores of S. sclerotiorumin vitro or 50 ascospores of S. sclerotiorum on one petal in approximately 6 h, even in the presence of a high background of oilseed rape DNA. This technique was successfully applied in detecting natural petal infections.     

In vitro and in vivo antifungal activity of synthetic pure isothiocyanates against Sclerotinia sclerotiorum

BACKGROUND: Isothiocyanates (ITCs) released by the enzymatic hydrolysis of glucosinolates in the Brassicaceae are potentially useful for controlling fungal pathogens. In vitro activity of pure ITCs against Sclerotinia sclerotiorum (Lib.) de Bary was studied by adding them to glass filters in petri dishes and dissolving them in the growing media. RESULTS: Methyl, allyl and benzyl ITCs were the most fungitoxic of the compounds in bioassays with S. sclerotiorum isolate Ss31. In the volatile phase, mycelial growth was completely inhibited by these three compounds. Aromatic ITCs were less toxic in the petri dishes but were more toxic than aliphatic ITCs when dissolved in the agar. Benzyl ITC exhibited the highest inhibitory effect on sclerotial germination, with an EC₅₀ value of 75.1 µmol L^?1. Butyl and benzyl ITCs reduced apothecial production of S. sclerotiorum by 92.5% at the highest concentration. In in vivo assay, only allyl and 2‐phenylethyl ITCs reduced disease incidence (by 76.7 and 70% respectively) at low concentrations. CONCLUSION: Sclerotinia sclerotiorum in the soil might be suppressed by the higher concentrations of allyl and benzyl ITCs released from decomposition of Brassica juncea, B. carinata, B. nigra and Sinapis spp. Copyright © 2011 Society of Chemical Industry     

Multivariate effects of plant canopy, soil physico-chemistry and microbiology on Sclerotinia stem rot of soybean in relation to crop rotation and urban compost amendment

The effects of canopy, soil physico-chemical and microbiological variables on Sclerotinia stem rot (SSR) on soybean were assessed in two soils (clay loam and sandy loam) using multiple regression and canonical redundancy analysis (RDA) and their partial form to control for the rotation (2 or 3-y-corn/soybean monoculture) and fertilization (mineral/urban compost) or spatial variables effects. The models revealed the minimal sets of variables that best explain the variance of the survival of Sclerotinia sclerotiorum’s sclerotia, carpogenic germination, disease severity and their associations. In clay loam, the 3-y-corn rotation reduced disease severity mainly through the reduction of weed biomass that favoured carpogenic germination. Urban compost has a conducive effect explained by a better soil surface drainage. Additionally, total N was found suppressive to sclerotial survival. In sandy loam, the carpogenic germination was negatively correlated with high C mineralization quotient and aggregate stability but correlated positively with Ca. Sclerotial survival was negatively correlated with pH and Ca, and positively correlated with biological fertility index. Aggregate stability, Ca and pH were associated with the urban compost. The regression and RDA analyses allowed to identify key variables that drived SSR development and explain their relationship with the cultural practices, soil health, as well as the spatial variation of disease variables.     

向日葵菌核病菌核及子囊柄萌发峰期的预测

向日葵菌核病的流行取决于田间菌核量与气候因素的影响。试验研究得知,菌核子囊柄萌发程度在菌核经过一场透雨后与七月底,八月份的降雨量及降雨次数呈正相关,而与此时期的平均温度,降雨间隔期呈负相关。由此建立了预测菌核子囊柄萌发程度的多元回归式为y=65.25—2.58x_1 0.04x_2 2.36x_3-2.10x_4。子囊柄萌发期是进行药剂防治的最有效时期,因此预测子囊柄萌发峰期是防治菌核病的关键。通过多元回归建立预测式为y=76.52-1.38x_1 0.06x_2-1.84x_3-3.86x_4。用上述两式回测前2年的田间实测值其结果与预测值极近。