玉米青枯病病原腐霉对其伴生镰刀菌的影响

 本试验以玉米青枯病病原肿囊腐霉(Pythium inflatum Malthews)与禾生腐霉(Pythium gramlnicola Subram)为材料,着重研究了两种腐霉的生长与代谢对其伴生病原禾谷镰刀菌(Fusarium graminearum Schw.)的影响,结果表明:1.腐霉的定殖生长能力弱于禾谷镰刀菌。在腐霉菌落上,禾谷镰刀菌仍可生长,而在禾谷镰刀菌菌落上,腐霉则不能生长。2.在伴生条件下,腐霉被镰刀菌覆盖的时间长短随接种间隔而有变化,接种间隔愈短,则覆盖愈快。3.两种腐霉的培养滤液均对禾谷镰刀菌的孢子萌发与芽管伸长及菌落扩展有明显的促进作用,表明滤液中含有对镰刀菌生长有利的活性成分。4.腐霉培养滤液对禾谷镰刀菌红色色素的产生有一定的抑制作用,其原因可能与腐霉的嗜糖性有关。5.田间结果表明,腐镰复合接种的发病率接近禾谷镰刀菌单菌接种,但低于腐霉单菌接种。     

黄瓜绵腐病病原菌鉴定及防治药剂的筛选

为有效防控黄瓜绵腐病,本研究对自江西省南昌市南昌县武阳镇和八一乡采集的黄瓜绵腐病病样进行分离,基于致病性测定、形态学特征和分子生物学方法对其进行鉴定,并采用菌丝生长速率法测定甲霜灵、霜霉威、霜脲氰、烯酰吗啉4种常用杀菌剂对该病原菌和辣椒疫霉Phytophthora capsici的抑制效果。结果表明,自武阳镇和八一乡采集的病样经分离、纯化培养后分别获得12株和10株菌株;自武阳镇分离的病原菌的藏卵器柄较直,藏卵器大小为18.1~32.9 μm;自八一乡分离的病原菌的藏卵器柄较弯曲,藏卵器大小为15.1~25.3 μm;结合致病性测定及rDNA-ITS测序结果,将自武阳镇分离的病原菌鉴定为瓜果腐霉Pythium aphanidermatum,将自八一乡分离的病原菌鉴定为德里腐霉Py.deliense。室内毒力测定结果表明,甲霜灵对瓜果腐霉和德里腐霉抑制效果最好,EC₅₀分别为0.71 μg/mL和0.25 μg/mL;霜脲氰次之,EC₅₀分别为3.68 μg/mL和3.36 μg/mL,甲霜灵、霜霉威、霜脲氰3种杀菌剂对瓜果腐霉、德里腐霉和辣椒疫霉的抑制效果差别不大,但烯酰吗啉对瓜果腐霉、德里腐霉和辣椒疫霉抑制效果差异明显,EC₅₀分别为18.06、33.58和0.18 μg/mL,表明防治黄瓜绵腐病优先选择甲霜灵。     

北京和浙江地区玉米青枯病病原菌的分离与鉴定

 1982-1986年，对北京、浙江地区的玉米青枯病病原菌进行分离和鉴定。在1985和1986两年分离的200株病株中，腐霉菌（Pythium）占66.5%，镰刀菌（Fusarium）占37%，丝核菌（Rhizoctonia）占10.5%，细菌（Bacteria）占4.5%，蠕孢菌（Helminthosporium）占3%。回接成株期玉米试验表明，肿囊腐霉菌（P.inflatum Malthews）和禾生腐霉菌（P.graminicola Subram）是玉米青枯病的病原菌，而禾谷镰刀菌（F.graminearum Schw.）则不是，至少不是主要病原菌。从玉米上分离出肿囊腐霉菌，过去未见报道。     

蔬菜土传病原菌三重PCR检测体系的建立与应用

【目的】 针对生产中危害严重的3种蔬菜土传病原菌瓜果腐霉（Pythium aphanidermatum）、尖镰孢（Fusarium oxysporum）、大丽轮枝菌（Verticillium dahliae）,建立三重PCR（triplex PCR）检测体系,为蔬菜土传真菌病害的早期诊断和鉴定提供技术与方法。【方法】 筛选3种病原菌特异性引物组合,通过设定不同的PCR退火温度、引物浓度、循环次数以及延伸时间,探索影响三重PCR扩增的因素,建立蔬菜土传病原菌三重PCR检测体系,并对体系灵敏度进行检测。为了验证建立体系的稳定性,选择2×Taq Master PCR Mix（北京博迈德生物技术有限公司）和TaKaRa Taq酶（大连宝生物工程有限公司）2个不同公司的试剂,分别利用C1000 Touch ^TM型（赛默飞世尔科技有限公司）与Aeris ^TM型（新加坡艺思高科技有限公司）热循环仪进行三重PCR反应,比较检测结果的可重复性。对田间采集的35份病害样本和149份土壤样本,分别进行三重PCR和病原菌分离检测,以确定所建立得三重PCR检测体系的适用性。 【结果】 三重PCR反应体系中引物对AsAPH2B/AsPyF、FOF1/FOR1、VActF/VActR可分别扩增出瓜果腐霉、尖镰孢、大丽轮枝菌长度为163、328和530 bp的特异性目的片段。反应体系（25 μL）：0.12 μmol·L ^-1 AsAPH2B/AsPyF,0.16 μmol·L ^-1 FOF1/FOR1,0.24 μmol·L ^-1 VActF/VActR,2×Taq Master PCR Mix 12.5 μL,退火温度为60.8℃,35个循环。该体系对病原菌纯培养物的检测灵敏度为10 ^-1ng·μL ^-1,对土壤中大丽轮枝菌、尖镰孢和瓜果腐霉的检测灵敏度分别为10 ⁵、10 ⁶个孢子/g土壤和10 ^-2mg菌丝/g土壤。分别采用2×Taq Master PCR Mix和TaKaRa Taq酶,利用C1000 Touch ^TM型和Aeris ^TM型热循环仪进行三重PCR,扩增结果一致,说明三重PCR体系检测稳定性好。对田间采集的病害和土壤样本进行检测,25份病害样本和71份土壤样本中检测出带菌,三重PCR检测与病原菌分离培养结果一致。 【结论】 本研究建立的瓜果腐霉、尖镰孢、大丽轮枝菌三重PCR检测体系具有灵敏度高、稳定性和重复性好的特点,能够快速、准确地检测田间病株及其根围土壤中的瓜果腐霉、尖镰孢、大丽轮枝菌,为蔬菜土传病害的早期预防和流行监测提供了有效的技术手段。     

玉米茎腐病研究进展

系统综述了玉米茎腐病的发病症状、分级标准、致病菌、致病机理、抗性遗传规律、抗性基因定位和抗病机理的研究进展,并提出了抵御玉米茎腐病的综合防治措施。     

寄生玉米的6种腐霉及其致病性研究

 从国内12省、自治区、直辖市玉米植株上分离出腐霉菌菌株,依据形态特征、培养性状、生长抑制温度,以及产生的有性与无性器官,鉴别出6个腐霉种:棘腐霉(Pythium acanthicum Drechsler),瓜果腐霉(P.aphanidermatum(Edson)Fitzpatrick),强雄腐霉(P. arrhenomanes Drechsler),禾生腐霉(P.graminicola Subramaniam),肿囊腐霉(P.inflatum Matthews),寡雄腐霉(P.oligandrum Drechsler),其中肿囊腐霉出现频率高,分布广泛。回接玉米表明,腐霉能够侵染玉米并引起病害,肿囊腐霉、禾生腐霉、强雄腐霉、棘腐霉是玉米青枯病的重要致病菌;瓜果腐霉能够引起玉米中部茎节腐烂;寡雄腐霉虽寄生玉米,但对玉米成株无致病力。     

黄瓜腐霉菌苗期猝倒病致病机制研究

 本文明确了Pythium aphanidermatum主要靠生产以PG(多聚半乳糖醛酸酶)为主的一系列细胞壁降解酶的协调作用侵染黄瓜幼苗。植株胚轴细胞壁随苗龄增加,果胶物质增加最明显。病菌可在健胚轴及其细胞壁上培养产生果胶酶,其中细胞壁对PG的诱导作用比胚轴明显。随病菌致病力增强或病菌果胶酶浓度加大,胚轴的浸解和释放还原糖愈加明显。健株外渗物中虽也含有细胞壁降解酶,但比病菌的产生量少,活性低。病菌在寄生体内产生的细胞壁降解酶总酶活性,尤其是PG总酶活性与病菌致病力关系最为密切。这类细胞壁降解酶易受培养基成分和酸度的影响。经染色、显微及超微观察证实了该菌侵入黄瓜苗的机制是以酶解作用为主。     

Development and evaluation of an enzyme-linked immunosorbent assay for the serodiagnosis of pythiosis in dogs

Pythiosis (caused by the aquatic oomycete Pythium insidiosum) is a devastating and often fatal cause of either severe transmural gastroenteritis or locally invasive subcutaneous disease in dogs living in the southeastern United States. Although early diagnosis is essential for successful treatment, tools available for this task are limited. Therefore, we developed and evaluated an enzyme-linked immunosorbent assay (ELISA) for the detection of anti-P insidiosum antibodies in canine serum. A soluble mycelial extract of P insidiosum was utilized as antigen in the ELISA, which was used to evaluate serum from 43 dogs with pythiosis, 8 dogs with lagenidiosis (another canine oomycosis), 16 dogs with nonoomycotic fungal or algal infections, 22 dogs with nonfungal gastrointestinal or skin disease, and 55 healthy dogs. Results were expressed as percent positivity (PP) relative to a strong positive control serum run on each plate. Medians and ranges for each of the 5 groups were as follows: pythiosis (81.7%, 50.6-98.5%), lagenidiosis (17.3%, 11.3-29.2%), other fungal or algal infections (8.2%, 4.7-15.4%), nonfungal gastrointestinal or skin disease (6.2%, 3.9-20.7%), and healthy dogs (6.7%, 3.0-15.2%). When using a cutoff value of 40% PP, the sensitivity and specificity of the ELISA both were 100%. In addition, ELISA values measured after successful surgical therapy in 2 dogs showed a decrease of anti-P insidiosum antibody concentrations into the normal range as early as 2 months after treatment. We conclude that the ELISA is a sensitive and specific test for the diagnosis of canine pythiosis, and may be a useful tool for monitoring response to medical or surgical therapy.     

Assessment of the response of carrot somaclones to Pythium violae, causal agent of cavity spot

Cavity spot is a major disease of carrots, causing cavities on the surface of the root. Available commercial varieties show a range of susceptibility but no significant resistance. Seed progeny from 46 tissue culture-derived carrot ( Daucus carota ) somaclones were screened for viability, then 19 selected somaclone families were sown under glasshouse conditions, along with commercial cultivars (Bertan, Nandor, Bolero and Vita Longa) as controls. Mature roots were exposed to Pythium violae in a cavity spot bioassay to determine their response as measured by disease incidence and severity. Some somaclones formed fewer lesions than the least susceptible control cultivar, Vita Longa. Seven somaclonal families that showed a range of susceptibility were sown under field conditions and the assessment was repeated. Although there was little relationship between glasshouse and field-trial results, under field conditions one of the somaclones had a mean incidence of disease, as estimated by transformed data, of 1·9 compared with 37·9 for the most susceptible somaclone and 3·5 for Bolero, the most resistant commercial cultivar. The results indicated that significant genetic variation in susceptibility to cavity spot disease was present in the somaclones.