芸薹根肿菌次生游动孢子侵染致病分析

为明确芸薹根肿菌Plasmodiophora brassicae Woron.在其它寄主中是否广泛存在无性短循环及次生游动孢子的侵染致病性,以不结球白菜为寄主培养3批幼苗(G1、G2和G3),用休眠孢子悬浮液接种G1,被侵染的G1接种G2,被侵染的G2接种G3,采用离心管水培法研究其侵染致病性。结果显示,无性短循环研究中,G1、G2和G3根毛均被侵染,除G3并株接种侵染率为33.33%外,其它处理侵染率均在50.00%以上,根毛里有明显的游动孢子囊;次生游动孢子能侵染不结球白菜的皮层组织,致使不结球白菜发病形成明显的肿根;G1、G2和G3水培发病率为20.00%、15.00%和6.00%,砂培发病率为22.50%、18.75%和7.50%;G3肿根病理切片中可观察到休眠孢子。表明芸薹根肿菌侵染不结球白菜时,其生活史中存在无性短循环,次生游动孢子具有侵染致病作用。     

Histological methods to detect the clubroot pathogen Plasmodiophora brassicae during its complex life cycle

The clubroot pathogen Plasmodiophora brassicae is an obligate biotrophic protist that lives in close relationship with its host cell. The roots of the host plants are colonized and the plant growth is altered upon infection. While shoots can be stunted and show wilt symptoms after longer infection periods, the root system is converted to a tumorous root tissue, called ‘clubroot’, by alterations of the plant growth promoting hormones auxin, cytokinin and brassinosteroid. Because the life cycle occurs largely within the host cells, this leads to dramatic changes in host root morphology and anatomy. Thus, the identification of the respective protist structures in the host tissue by microscopy is challenging. Different staining methods as well as fluorescence and electron microscopy of thin sections can reveal specific life stages of P. brassicae and can yield additional information on the changes in the host tissues concerning, for example, cell wall properties. In addition, promoter–reporter fusions, immunostaining methods and in situ hybridization techniques can be used to gain additional information on the changes in the host roots.     

NBF-926的生物测定及对大白菜根肿病的防效试验

十字花科蔬菜根肿病休眠孢子在1×103~1×107个孢子/m L浓度范围内均能侵染大白菜,随着休眠孢子浓度的不断增加,各处理大白菜感病率逐渐增高。1×107个孢子/m L浓度处理感病率最高,达100%。采用1×107芸薹根肿菌休眠孢子感染盆栽大白菜,测定几种不同药剂对根肿病的防治效果,结果表明,生防链霉菌NBF-926防治根肿病的盆栽试验的效果及植株鲜重均最高,显著高于其他几种药剂。     

十字花科蔬菜根肿病研究新进展

十字花科根肿病在我国的东北地区、西南地区、长江中上游地区以及山东青岛等地迅速扩大,危害十分严重,制约着我国十字花科蔬菜产业的发展。根肿病一直是国内外学者研究的热点,经过几十年的不懈努力,在根肿菌分类地位、根肿菌生理小种分化、病原菌检测技术、病原菌有效接种技术、病原菌生活史与生物学特性、根肿病防治等方面已取得突破性进展。近几年来,根肿病的研究在病原菌检测、病原菌生物防治技术、病原菌生活史等方面取得了比较突出的成绩。本文从十字花科蔬菜根肿病病原菌的分类地位、生活史、致病机理、诊断与检测技术以及生物防治技术等方面,对国外近两年取得的研究进展进行综述,并对我国根肿菌生活史研究趋势、根肿菌致病机理深入研究的必要性进行了探讨。     

基于LC-MS技术的代谢组学方法研究根肿菌对拟南芥代谢的影响

为探寻根肿菌侵染拟南芥后期寄主代谢的生物标志物,将罹患根肿病的油菜根切块进行组织培养,获得较为纯化的、单一的根肿菌接种体,与拟南芥种子在1/2MS培养基上共培养,利用液相色谱-质谱联用(LCMS)技术的非靶向代谢组学方法分析根肿菌侵染后期拟南芥根部代谢物差异。组织培养结果表明,根肿菌可以在愈伤组织中生长和繁殖;共培养结果显示45d后拟南芥侧根尖端肿大,显微观察显示根肿菌能完成侵染循环;代谢组数据PCA和PLS-DA分析结果显示试验组(接种根肿菌)和对照组(未接种根肿菌)明显分离;OPLSDA分析和t检验结果表明重要的差异代谢物包括油菜素内酯、类黄酮、木质素、萜类、甾体皂苷和磷脂等物质。提示这些物质可能在根肿菌侵染后期时拟南芥与根肿菌的相互作用中起重要作用。     

Effects of cyazofamid against Plasmodiophora brassicae Woronin on Chinese cabbage

Cyazofamid (4-chloro-2-cyano-N,N-dimethyl-5-p-tolylimidazole-1-sulfonamide) is a novel fungicide with high levels of activity against Oomycetes fungi and Plasmodiophora brassicae Woronin. The effects of cyazofamid were investigated against P. brassicae, the causal agent of clubroot disease in Chinese cabbage. Cyazofamid at 0.3 mg litre(-1) inhibited resting spore germination of this pathogen by about 80%. Cyazofamid at 3-10 mg litre(-1) exhibited fungicidal activity to resting spores of P. brassicae 1-10 days after treatment. When cyazofamid was applied to infested soil, both root-hair infections and club formation caused by P. brassicae were strongly inhibited at 1-3 mg kg(-1) dry soil. These results suggest that cyazofamid directly inhibits resting spore germination, thereby leading to the inhibition of root-hair infection and club formation. Cyazofamid at 3 mg kg(-1) dry soil also exhibited complete control of clubroot disease. The effect of broadcast soil application using a dust formulation at 2 kg AI ha(-1) (equivalent to 1.3 mg AI kg(-1) dry soil), and plug seedling tray application by a suspension concentrate formulation at 200 and 400 mg AI tray(-1) (30 x 60 x 4 cm3) against P. brassicae was also evaluated. Cyazofamid exhibited good efficacy against the pathogen. The sequential treatment including plug seedling tray application with cyazofamid and pre-plant broadcast soil application with the fungicide fluazinam also exhibited excellent levels of control. These results indicate that cyazofamid has a high potential to be an effective fungicide for the control of clubroot disease.     

盈江县高山大白菜根肿菌生理小种鉴定及抗源筛选

采用Williams鉴别系统对云南省盈江县高山大白菜产区采集的根肿菌进行鉴定,4个鉴别寄主均表现为感病,病情指数分别为21、33、28、30,说明该根肿菌为4号生理小种。采用菌土法接种盈江根肿菌,对18个抗根肿病大白菜品种进行抗性鉴定,其中6个品种发病率和病情指数均为0,对盈江根肿菌表现免疫;2个品种病情指数10,表现抗病,可以作为该病区大白菜新品种选育的抗性遗传资源。     

Inactivation of soil-borne plant pathogens during small-scale composting of crop residues

Samples of heavily infested crop residues were incorporated in static compost heaps (2.5–4.6 m³) of the Indore type. Temperature increased to 50–70°C within 6 days depending on the type of crop residues used and the location within the heap. The heat phase (>40 °C) lasted 2–3 weeks and was followed by a c. 5-months maturation phase (<40 °c).=" among=" the=" 17=" pathogens=" tested,=">Olpidium brassicae and one of the four formae speciales ofFusarium oxysporum that were tested survived composting, but also their inoculum was greatly reduced.Survival during specific phases of composting was studied by incorporation and retrieval of samples at various stages of the process.F. oxysporum f. sp.melonis was completely inactivated andO. brassicae andPlasmodiophora brassicae were almost completely inactivated during the short heat phase. The three pathogens survived the long-lasting maturation phase without loss of viability. Heat evolved during composting was found to be the most important factor involved with sanitation of crop residues. The possible involvement of fungitoxic conversion products and microbial antagonism is discussed.     

Soil management in a mountain agroecosystem and clubroot disease

Clubroot, caused by Plasmodiophora brassicae, is a disease limiting the production of Brassica species. Its severity varies according to the density of the pathogen's resting spores and environmental conditions. Although the soil environment (including its physical, chemical and biological characteristics) is determinant in the development of the disease, little is known regarding these characteristics in tropical regions, such as Brazil. The objective of this study was to investigate the relationships between soil attributes, bacterial population composition and clubroot intensity in a tropical mountain agroecosystem. Soil and plant samples were collected from 17 cauliflower-producing localities in Nova Friburgo, Brazil. The physical and chemical attributes, the bacterial population composition, the number of pathogen resting spores in the soil samples, and the clubroot severity were evaluated. Based on bacterial population composition, two distinct groups of soils were identified, which varied in attributes such as organic matter, potential acidity, pH and disease severity. The organic matter level was associated with differences in soil acidity, bacterial population composition and disease intensity, but there was not a cause and effect relationship between bacterial population composition and clubroot severity. Under the conditions of this agroecosystem, soils with increased organic matter presented higher acidity potential (H + Al), which was strongly correlated to clubroot severity. Thus, high acidity potential could be a key factor leading to clubroot development in mountain regions of Brazil.     

青花菜根肿病田间防治技术研究

2018年开展青花菜根肿病田间防治试验, 移栽后用5种药剂灌根处理?分别进行了5种药剂灌根次数试验?3种药剂灌根剂量试验和3种药剂灌根时期试验?试验结果表明, 5种药剂中500 g/L氟啶胺SC是防治青花菜根肿病的最佳药剂, 但使用不当会有药害风险, 移栽当天用2 250 g/hm2?移栽后3 d用3 375 g/hm2和4 500 g/hm2灌根, 花球成熟期推迟, 株高?最大叶面积均不同程度受抑制, 尤其在苗期使用会明显抑制生长, 表现为植株矮小?叶片黄化?无法正常盘根?建议用2 250 g/hm2分别在移栽后3~7 d和移栽后30 d各灌根1次?其次采用100 g/L氰霜唑SC, 338 g/hm2在移栽后0~3 d和30 d灌根也可取得较好防效?75%百菌清WP 8 440 g/hm2与100 g/L氰霜唑SC 338 g/hm2防效相当?20%噻唑锌SC防效较低, 62.5 g/L精甲·咯菌腈FSC防效最差, 不适宜用在根肿病防治上?