Effect of inoculum type and timing of application of Coniothyrium minitans on Sclerotinia sclerotiorum: influence on apothecial production

The effects of different inocula of the mycoparasite Coniothyrium minitans on carpogenic germination of sclerotia of Sclerotinia sclerotiorum at different times of year were assessed. A series of three glasshouse box bioassays was used to compare the effect of five spore-suspension inocula of C. minitans , including three different isolates (Conio, IVT1 and Contans), with a standard maizemeal–perlite inoculum. Apothecial production, as well as viability and C. minitans infection of S. sclerotiorum sclerotia buried in treated soil, were assessed. Maizemeal–perlite inoculum at 10⁷ CFU per cm³ soil reduced sclerotial germination and apothecial production in all three box bioassays, decreasing sclerotial recovery and viability in the second bioassay and increasing C. minitans infection of sclerotia in the first bioassay. Spore-suspension inocula applied at a lower concentration (10⁴ CFU per cm³ soil) were inconsistent in their effects on sclerotial germination in the three box bioassays. Temperature was an important factor influencing apothecial production. Sclerotial germination was delayed or inhibited when bioassays were made in the summer. High temperatures also inhibited infection of sclerotia by C. minitans . Coniothyrium minitans survived these high temperatures, however, and infected the sclerotia once the temperature decreased to a lower level. Inoculum level of C. minitans was an important factor in reducing apothecial production by sclerotia. The effects of temperature on both carpogenic germination of sclerotia and parasitism of sclerotia by C. minitans are discussed.     

盾壳霉寄生核盘菌过程中葡聚糖酶、几丁质酶及电导率的变化

为了解盾壳霉寄生核盘菌的生化机理,采用了紫外分光光度法测定了盾壳霉寄生菌核过程中,葡聚糖酶和几丁质酶的变化,并用电导率仪测定了这个过程中电导率的变化。结果表明:从被盾壳霉寄生的活菌核检测到(处理)葡聚糖酶和几丁质酶活性明显高于活菌核和被盾壳霉寄生的死菌核,其中处理的葡聚糖酶活性从15 d以后就明显高于菌核自身也高于盾壳霉本身的酶活性,处理的几丁质酶活性从10 d起就维持在较高水平,处理的电导率从10 d以后明显高于活菌核。在这个过程中这两种酶活性提高,电导率增大,膜透性增强。     

盾壳霉寄生核盘菌过程中胞壁降解酶的变化及对核盘菌的影响

通过测定盾壳霉寄生核盘菌过程中几丁质酶和β-1,3-葡聚糖酶的变化及对核盘菌菌丝和菌组织的影响,结果表明:盾壳霉在寄生核盘菌初期其几丁质酶和β-1,3-葡聚糖酶的活性均会明显的升高,具有明显的诱导酶特征,同时盾壳霉对照在没有核盘菌存在时,也能检测到几丁质酶和β-1,3-葡聚糖酶活性,说明盾壳霉产生的几丁质酶和β-1,3-葡聚糖酶都有组成酶存在。核盘菌菌丝和菌核组织的大量消解都出现在这两种酶活性达到高峰之后,表明两种酶协同作用更有利于核盘菌菌丝和菌核组织的消解腐烂。     

中国腔孢纲的2个新记录种

本文报道了半知菌亚门腔孢纲的一个国内新记录属:Tryblidiopicnis Hohn.和2个新记录种:Coniothyrium ahmadii Sutton,Tryblidiopicnis pinastri Hohn.新记录种有描述和形态线条图。     

Effectiveness of Coniothyrium minitans and Trichoderma atroviride in suppression of sclerotinia blossom blight of alfalfa

The effects of the mycoparasites Coniothyrium minitans and Trichoderma atroviride on the suppression of alfalfa blossom blight caused by Sclerotinia sclerotiorum were evaluated under indoor and field conditions. When T. atroviride (9·0 × 10⁴ conidia/floret) + S. sclerotiorum (6·0 × 10³ ascospores/floret) or C. minitans (9·0 × 10⁴ conidia/floret) + S. sclerotiorum (6·0 × 10³ ascospores/floret) were applied to detached young alfalfa florets, T. atroviride effectively inhibited saprophytic growth of S. sclerotiorum, whereas C. minitans showed no inhibition under the same conditions. When T. atroviride (6·9 × 10⁴ conidia/floret) + S. sclerotiorum (6·0 × 10³ ascospores/floret) or C. minitans (6·9 × 10⁴ conidia/floret) + S. sclerotiorum (6·0 × 10³ ascospores/floret) was applied to young alfalfa petals in vivo just after pollination, the percentage of pod formation was higher for T. atroviride+S. sclerotiorum than that for C. minitans+S. sclerotiorum, and the percentage of pod rot was lower for T. atroviride+S. sclerotiorum than that for C. minitans+S. sclerotiorum. However, when they were applied to senescent petals attached to developing pods of alfalfa at 9·2 × 10⁴ conidia/floret together with S. sclerotiorum at 4·5 × 10³ ascospores/floret at 14 days after pollination, C. minitans was more effective than T. atroviride in suppressing sclerotinia pod rot and seed rot of alfalfa. Field experiments showed that three applications of C. minitans (5·4 × 10⁶ conidia mL⁻¹) or T. atroviride (5·4 × 10⁶ conidia mL⁻¹) at a 7-day interval to blossoms of alfalfa effectively suppressed sclerotinia pod rot in two out of three annual trials. Coniothyrium minitans effectively suppressed sclerotinia seed rot in all three years, whereas T. atroviride was not effective against seed rot in any of the trial years. The efficacy of C. minitans was not significantly different (P > 0·05) from benomyl (250 µg ai mL⁻¹). This study suggests that C. minitans has potential as a biocontrol agent to control blossom blight of alfalfa caused by S. sclerotiorum.     

盾壳霉几丁质酶和葡聚糖酶酶动力学性质初步研究

盾壳霉几丁质酶和葡聚糖酶酶动力学性质研究表明:几丁质酶和葡聚糖酶的酶促反应,在本试验计量范围内,其反应速度与酶量和底物浓度均呈正相关,但两种酶促反应适宜的温度和pH值有较大差。几丁质酶降解几丁质的最适反应温度为50℃,最适pH值为8.0,葡聚糖酶降解葡聚糖的最适反应温度为60℃,最适pH值为3.0。     

盾壳霉抗菌核净菌株的诱导及其特性初步研究

为筛选可与菌核净配合防治作物菌核病的盾壳霉菌株,进行了紫外线辐射和药剂驯化试验,分别得到4个和2个盾壳霉抗菌核净突变菌株,并研究了其生物学特性。结果表明:除菌株D-1外,其余突变菌株生长速率(菌落直径57.00 ~62.75 mm,生物量6.38 ~8.63 mg/mL)均大于亲本菌株(菌落直径54.00 mm,生物量5.91 mg/mL);大部分突变菌株产孢量(对数值7.33 ~8.16)小于亲本菌株(8.40);菌核净对突变菌株的EC50值是亲本菌株的2.23 ~191.84倍;突变菌株D-3、U-6有微弱的几丁质酶活性,且除U-4外的突变菌株葡聚糖酶活性增强(亲本菌株8.96 U,突变菌株9.18 ~17.61 U)。综合分析,U-6菌株既有较高的葡聚糖酶活性又有几丁质酶活性,并能保持较快的生长速率,是值得开发的优良菌株。     

盾壳霉固态发酵基质筛选和发酵条件优化

为了探索盾壳霉固体发酵的条件,提高其生防效果,比较了6种天然培养基对盾壳霉固体发酵的影响,测定了在不同发酵培养基、温度、初始湿度、接种比例条件下盾壳霉的产孢量。结果表明：盾壳霉在麦粒和谷粒培养基上产孢量最大;而在麸皮和棉籽培养基上生长和产孢也较好,且成本低,成为后续试验的发酵材料。在含麦麸、棉籽壳及无机盐的培养料作为基质时,采用温度为20℃,发酵料初始湿度为60%,接种比例为7%,最终孢子含量可达109个/g以上。同时对发酵的条件做了进一步优化,发现麦粒与谷粒的混合比例、光照条件、甘露糖和甘氨酸含量对盾壳霉产孢量的影响较大。     

重寄生真菌盾壳霉与核盘菌对峙培养的差异代谢物分析

运用基于液相质谱联用技术的代谢组分析手段,检测重寄生真菌盾壳霉和宿主植物病原菌核盘菌对峙培养时的代谢物差异。结果发现:盾壳霉与核盘菌对峙接触能显著诱发盾壳霉代谢物的分泌,构建盾壳霉特定代谢物数据库检测到3种代谢物涉及重寄生过程,分别为Macrosphelide A,Benzenediol和5-Aminopentanoate,且对峙接触2d时检测到的代谢物积累量最大。对盾壳霉重寄生过程代谢物变化的分析结果表明,真菌重寄生能诱发代谢物水平的交流和次级代谢物积累。     

生防菌盾壳霉对油菜根际土壤微生物群落结构的影响

为了明确油菜菌核病生防菌盾壳霉施用后对土壤微生物群落结构的影响,为盾壳霉在实际生产中的应用提供理论依据。以喷施盾壳霉为处理组(CM),喷施清水为对照组(CK),调查不同处理下油菜菌核病的发生情况,并采用高通量测序技术分析土壤微生物群落变化。结果表明：对照组油菜菌核病的病株率和病情指数分别高达74.64%和46.13;而处理组菌核病病株率和病情指数分别降低至23.33%和9.37,防治效果达79.70%;此外,喷施盾壳霉还改变了油菜根际土壤微生物群落结构,使得土壤真菌种群的丰富度和多样性程度显著增加;在门水平上,喷施盾壳霉促使土壤中变形菌门、芽单胞菌门、子囊菌门、被孢霉门和担子菌门等的相对丰度增加,导致放线菌门的相对丰度比例降低。在属水平上,施加盾壳霉可增加鞘氨醇单胞菌属、溶杆菌属、独岛菌属、短梗蠕孢属、镰刀菌属、曲霉属、Plectosphaerella等属的丰富度,会显著降低unidentified_Chitinophagaceae、寡养单胞菌、地杆菌属、梭孢壳属和Cutaneotrichosporon等的相对丰度。喷施盾壳霉能有效防治油菜菌核病,增加土壤真菌种群丰富和多样性,改善土壤微生物群落结构,研究结果为盾壳霉的田间大面积推广提供依据。