Host‐induced gene silencing in the necrotrophic fungal pathogen Sclerotinia sclerotiorum

Sclerotinia sclerotiorum is a necrotrophic fungus that causes a devastating disease called white mould, infecting more than 450 plant species worldwide. Control of this disease with fungicides is limited, so host plant resistance is the preferred alternative for disease management. However, due to the nature of the disease, breeding programmes have had limited success. A potential alternative to developing necrotrophic fungal resistance is the use of host‐induced gene silencing (HIGS) methods, which involves host expression of dsRNA‐generating constructs directed against genes in the pathogen. In this study, the target gene chosen was chitin synthase (chs), which commands the synthesis of chitin, the polysaccharide that is a crucial structural component of the cell walls of many fungi. Tobacco plants were transformed with an interfering intron‐containing hairpin RNA construct for silencing the fungal chs gene. Seventy‐two hours after inoculation, five transgenic lines showed a reduction in disease severity ranging from 55·5 to 86·7% compared with the non‐transgenic lines. The lesion area did not show extensive progress over this time (up to 120 h). Disease resistance and silencing of the fungal chs gene was positively correlated with the presence of detectable siRNA in the transgenic lines. It was demonstrated that expression of endogenous genes from the very aggressive necrotrophic fungus S. sclerotiorum could be prevented by host induced silencing. HIGS of the fungal chitin synthase gene can generate white mould‐tolerant plants. From a biotechnological perspective, these results open new prospects for the development of transgenic plants resistant to necrotrophic fungal pathogens.     

The control of sclerotinia stem rot on oilseed rape (Brassica napus): current practices and future opportunities

Sclerotinia stem rot (SSR) caused by the phytopathogenic fungus Sclerotinia sclerotiorum is a major disease of oilseed rape (Brassica napus). During infection, large, white/grey lesions form on the stems of the host plant, perturbing seed development and decreasing yield. Due to its ability to produce long‐term storage structures called sclerotia, S. sclerotiorum inoculum can persist for long periods in the soil. Current SSR control relies heavily on cultural practices and fungicide treatments. Cultural control practices aim to reduce the number of sclerotia in the soil or create conditions that are unfavourable for disease development. These methods of control are under increased pressure in some regions, as rotations tighten and inoculum levels increase. Despite their ability to efficiently kill S. sclerotiorum, preventative fungicides remain an expensive gamble for SSR control, as their effectiveness is highly dependent on the ability to predict the establishment of microscopic infections in the crop. Failure to correctly time fungicide applications can result in a substantial cost to the grower. This review describes the scientific literature pertaining to current SSR control practices. Furthermore, it details recent advances in alternative SSR control methods including the generation of resistant varieties through genetic modification and traditional breeding, and biocontrol. The review concludes with a future directive for SSR control on oilseed rape.     

向日葵菌核病接种方法及品种抗病性鉴定

为建立有效的向日葵菌核病田间接种鉴定方法,以核盘菌菌丝体悬浮液和孢子悬浮液作为接种物,分别对抗、感向日葵品种在现蕾期、始花期和盛花期进行人工接种,并对接种后保湿材料和保湿时间进行比较。试验结果表明:两种接种物均可使向日葵抗、感品种产生盘腐症状。用菌丝体悬浮液和孢子悬浮液接种时,浓度分别为10.0~15.0g/L和200~500个/mL,始花期接种,牛皮纸袋保湿2~4d,即可区分出向日葵品种间抗感性差异。用此方法鉴定出13个对盘腐型菌核病表现抗病的向日葵品种。     

不结球白菜感染菌核病后4个生理生化指标分析

选用4个对菌核病具有不同抗性的不结球白菜品种003-R-6、Y3-R-4、109-S-9和003-S-7为试验材料,于四叶一心期接种核盘菌,采用苗期琼脂块叶片接种法测定各品种对菌核病的抗性,并分别测定接种前后叶片中4个生理生化指标。结果表明,苗期琼脂块叶片接种法与田间自然诱发鉴定结果相一致,可有效反映抗、感品种的差异。在核盘菌影响下,4个抗、感病品种中不结球白菜的丙二醛(MDA)含量、游离氨基酸(FAA)总量、几丁质酶活性和β-1,3-葡聚糖酶活性均有所变化。其中丙二醛(MDA)含量、游离氨基酸(FAA)总量表现为感病品种增加幅度大于抗病品种,而几丁质酶活性、β-1,3-葡聚糖酶活性均表现为抗病品种增加幅度大于感病品种,说明各指标与品种抗病性强弱存在一定的相关性。     

油菜菌核病致病机理的研究 Ⅶ.抗(耐)病性的组织病理学

本文切片分析了油菜菌核病抗(耐)病性的组织病理学。结果表明:在不同感病程度的甘蓝型油莱品种(系)上菌核病菌的侵染和扩展特性包括附着胞类型、病菌在组织内的扩展以及寄主的能动反应诸方面均存在显著不同。     

防治油菜菌核病的复配农药赤霉清

１９９１－１９９４年进行了赤霉清对油菜菌核病菌的离体毒力测定及田间试验示范．离体毒力测定结果表明，赤霉清对油菜茎、叶菌核病菌的ＥＣ５０为１３１．９８和１２７．４７ｐｐｍ，共毒系数分别为４７１．０８和２８７．８０，其毒力与保护作用显著高于多菌灵和防霉宝．应用２５％赤霉清可湿粉每亩７０克（ａ．ｉ．１７．５ｇ），在油菜开花始盛期或盛花期喷一至二次．其防病效果显著高于多菌灵（ａ．ｉ．５０ｇ／亩）和防霉宝（ａ．ｉ．４２ｇ／亩）。     

Transformation of Arabidopsis thaliana via Agrobacterium tumefacience with an endochitinase gene from Trichoderma, and enhanced resistance to Sclerotinia sclerotiorum

Sclerotinia sclerotiorum is an important pathogen to many crops and is especially damaging to rape in China. As a model plant Arabidopsis thaliana (Col0) was transformed by spraying Agrobacterium tumefacience with Trichoderma endochitinase gene ThEn-42 at initial bud stage. Eleven seedlings (corresponding to about 0.22 percent transformation) exhibited resistance to hygromycin. The DNA fragment unique to endochitinase (ThEn-42) was amplified by Arabidopsis leaf-PCR or genomic DN…     

油菜菌核病菌丝转移过程中致病力变化研究

 油菜菌核病菌Sclerotinia scleroitorum Lib de Bary在PDA连续转移时,菌落形态和致病力会发生改变.本文采用离体叶片菌丝块接种法.测定菌丝体的致病性,菌丝经不同次数连续转移后,菌丝间的致病力强弱发生明显变化,部分菌丝致病力减弱或丧失.低温下保存的原始菌丝体和菌核致病力最强.     

内蒙蔬菜菌核病发生和病原种类研究初报

菌核病是呼伦贝尔盟和呼和浩特市郊蔬菜生产上的重要病害。通过1987～1988年的调查和研究,明确该地区蔬菜菌核病的蔬菜寄主分属6科16种。通过对菌核形态,菌核萌发和子囊盘产生、子囊、子囊孢子和子囊盘尺度,病菌培养性状的比较,肯定呼盟和呼市郊区蔬菜菌核病的病原均为Sclerotinia sclerotiorum(Lib.)de Bary。该病在呼市郊区保护地黄瓜上发病株率常年可达0.24～40.0%,在呼盟保护地青椒上发病株率常年可达10.0——15.0%