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.     

Production,survival and evaluation of solid-substrate inocula ofConiothyrium minitans againstSclerotinia sclerotiorum

Coniothyrium minitans grew on all ten solid-substrates (barley, barley-rye-sunflower, bran-vermiculite, bran-sand, maizemed-perlite, millet, oats, peat-bran, rice and wheat) tested, producing high numbers of germinable pycnidiospores (1.9–9.3×10⁸ g^–1 air dry inocula). All solid substrate inocula survived better in the laboratory at 5 and 15 °C than at 30 °C for at least 64 weeks.In pot bioassays carried out in the glasshouse and field, soil incorporations of each inoculum almost completely inhibited carpogenic germination ofS. sclerotiorum. In the field bioassay, no sclerotia were recovered after 38 weeks fromC. minitans-treated pots compared to 56% from control pots. In the glasshouse bioassay, 9–30% of sclerotia were recovered after 20 weeks fromC. minitans-treated pots, but 88–100% of these were infected by the antagonist. The antagonist also spread to infect sclerotia in control pots.In larger scale glasshouse trials, single preplanting soil-incorporations of five inocula (barley-ryesunflower, maizemeal-perlite, peat-bran, rice and wheat) controlled Sclerotinia disease in a sequence of lettuce crops, with only small differences between the types of inocula tested. At harvest,C. minitans reduced sclerotial populations on the soil surface and over 74% of sclerotia recovered fromC. minitans-treated plots were infected by the antagonist.C. minitans survived in soil in all solid-substrate inocula-treated plots for at least 39 weeks at levels of 10⁴–10⁵ colony forming units cm^–3 soil and spread to infect over 36% of sclerotia recovered from control plots.     

小麦纹枯病菌菌核形成条件研究

研究了小麦纹枯病菌在不同光照、温度、培养基条件下菌核数量形成的差异.结果表明,小麦纹枯病菌在不同的光照条件下都可以形成菌核,但是形成菌核数量有差异.小麦纹枯病菌在24 h光照(即G24)条件下产生的菌核数量最多,其次是光照12h,而在全黑暗(即G0)的条件下产生的菌核数量较少.小麦纹枯病菌在15～35℃条件下都可以形成菌核,且在15～30℃的条件下,随着温度的升高,菌核数量也在增加,而在高于30C的条件下形成的菌核数量在减少;小麦纹枯病菌在25～30℃条件下最有利于形成菌核且数量较多.最适合小麦纹枯病菌形成菌核的培养基是PDA培养基.     

培养条件对春油菜菌核病产生菌核影响的研究

本文对春油菜菌核病菌（Sclerotinia sclerotiorum(Lib.)Derbay)产生菌核的影响因子进行初步研究。结果表明：菌丝产生菌核的最适温度15－20℃；最适PH3－8；最适碳源为葡萄糖;适氮源为胰蛋白胨；最适培养基为玉米粉琼脂培养基。     

不同施氮量对油菜“浙油50”菌核病及产量的影响

以浙江省油菜主栽品种"浙油50"为对象进行不同施氮量试验,研究不同施氮量对油菜菌核病发生程度及产量的影响。研究结果表明:浙油50每667 m2适宜施N量为15～20 kg,在该施氮水平下,油菜菌核病发生较轻,且产量水平高。     

蔬菜菌核病菌对氟吡菌酰胺的敏感性及防病应用潜力评估

【目的】评价蔬菜菌核病菌（Sclerotinia sclerotiorum）对新型琥珀酸脱氢酶抑制剂类杀菌剂氟吡菌酰胺的敏感性,明确药剂对不同发育阶段菌核病菌的毒力效应以及氟吡菌酰胺防治蔬菜菌核病的作用方式和田间应用效果,以指导氟吡菌酰胺的科学使用。【方法】从山东省昌乐、寿光、青州、临朐和泰安等地的蔬菜产区采集173株来自黄瓜、番茄、茄子、西葫芦、芸豆、辣椒等6种作物上的田间菌核病发病组织,室内分离纯化后采用菌丝生长速率法测定其对氟吡菌酰胺的敏感性;测定氟吡菌酰胺对病菌菌核产量、菌核形态和菌核菌丝型萌发的影响;采用离体茄子叶片接种试验确定氟吡菌酰胺防治菌核病的作用方式;最后通过两年田间药效试验进一步验证其实际应用效果。【结果】氟吡菌酰胺对菌核病菌的菌丝生长具有较强的抑制活性,且对来自不同作物的173株菌核病菌的抑制中浓度（EC₅₀）差异不大,分布在0.02-0.30 μg·mL^-1,表明这些菌株可被用来分析蔬菜菌核病菌对氟吡菌酰胺的敏感性水平。EC₅₀频率分布图呈单峰偏正态曲线分布,变异系数较小,表明该地区的蔬菜菌核病菌对氟吡菌酰胺均表现敏感。氟吡菌酰胺对菌核病菌的菌核产量、菌核形态以及菌核菌丝型萌发具有较高的抑制活性。在氟吡菌酰胺1.6 μg·mL^-1处理浓度下,该药剂对3株来自不同作物的菌核病菌表现出相同的抑制趋势,其菌核的数量和干重均明显降低,形态明显变小,表明该药剂能够有效地减少菌核病菌的初侵染源数量,并降低其侵染活性;而经过连续3 d的观察,5 μg·mL^-1氟吡菌酰胺对3株病菌的菌核菌丝型萌发的抑制率均在95%以上,表明该药剂有能力抑制菌核病菌的这一侵染方式,从而保护作物茎基部免受菌丝侵染。离体叶片接种防治试验表明,氟吡菌酰胺具有保护作用与治疗作用,40 μg·mL^-1的保护效果为100.00%,治疗效果为88.81%,显著高于对照药剂多菌灵和菌核净的防治效果,但该药剂对菌核病的保护作用明显优于治疗作用,表明该药剂在田间使用时应在病害发病前或发病初期使用,从而获得较好的防治效果。2016和2017两年的田间药效试验中,氟吡菌酰胺200 g a.i./hm²处理对茄子菌核病的防治效果分别为90.30%和87.60%,显著高于氟吡菌酰胺其他施用剂量以及对照药剂菌核净600 g a.i./hm²、多菌灵1 150 g a.i./hm²的防治效果。【结论】新型琥珀酸脱氢酶抑制剂氟吡菌酰胺对菌核病菌的菌丝生长、菌核形成及萌发均具有较高的抑制活性,且在田间能够有效地控制菌核病发生,因此该药剂是防治菌核病的高效药剂,可作为现有防治药剂的补充。     

水稻纹枯病菌核的浮沉特性与超微结构的关系研究

研究水稻纹枯病菌核的浮沉现象与超微结构的关系及不同生态条件下的菌核结构的差异性,结果表明,菌核具有浮水性,沉水性和转变性等特点,电镜扫描显示菌核的浮沉特性与菌核内外层组织解剖结构有密切关系。外层空细胞腔的厚薄决定菌核的浮沉。浮核外层较厚。沉核外层较薄,用化学药剂处理可使菌核浮或沉,甚至多次反复转变,此外,不同生态条件下形成的菌核其形成结构存在明显差异。     

Saccharides of ectomycorrhizal fungal sclerotia as sources of forest soil polysaccharides

ABSTRACT

The neutral monosaccharide composition of forest soils differs from that of non-forest soils suggesting there is an accumulation of microbial saccharides. Ectomycorrhizal (ECM) fungi can be responsible as the fungi are typical in forest soils. We investigated neutral saccharides of ECM fungal sclerotia to determine what part it might play in the origin of forest soil polysaccarides. Sclerotial grain (SG) was collected from the O, A1 and A2 horizons of a soil of subalpine forest of Mt. Ontake, central Japan. Neutral saccharides in soil and SG were analyzed by two step hydrolysis with sulfuric acid and gas-chromatography of alditol acetate derivatives. Saccharides accounted for 6.0?16% of the SG by carbon content. The SG contained predominantly easily hydrolysable (EH)-glucose, which accounted for 75–85% of the composition depending on grain size and the soil horizon, followed by mannose (7.7?15%), galactose (2.2?4.8%) and non-easily hydrolysable (NEH)-glucose (1.7?6.1%). The SG contained all of these sugars irrespective of its size. The SG collected from the A1 and A2 horizons contained all sugar components found in that from the O horizon, except for fucose in that from A2 horizon. The monosaccharide composition of SG indicates that accumulation of ECM fungal sclerotial polysaccharides might have been responsible for enlarging the molar ratios of (galactose + mannose) /(arabinose + xylose) and EH-glucose/NEH-glucose of forest soils. The proportions of SG saccharides relative to soil saccharides were 3.6, 1.2, and 0.83% for the O, A1 and A2 horizons, respectively. These levels of the proportion are considerable as ECM fugal sclerotia are the products of a limited species among hundreds and thousands of microbial species inhabiting forest soils. The sclerotia forming ECM fungal species such as Cenococcum geophilum may be key sources of forest soil polysaccharides.     

油菜菌核病菌秋季与春季萌发菌株生物学特性的比较

通过室内和田间试验,观察并比较采自湖北省咸宁市核盘菌秋季萌发菌株与春季萌发菌株的生物学特性。结果表明:9个秋季萌发菌株(aSs-1至aSs-9)和9个春季萌发菌株(sSs-1至sSs-9)在菌丝生长速率、菌核产量、菌核大小和对油菜的致病力等方面均无显著差异(P0.05);室内和田间菌核的萌发均受菌核形成温度和诱导菌核萌发温度的影响,高温形成的菌核易于萌发;核盘菌秋季萌发菌株和春季萌发菌株在菌核萌发特性方面存在明显差异。     

链孢粘帚霉寄生核盘菌菌核差异表达基因的筛选及分析

为克隆和研究链孢粘帚霉Gliocladium catenulatum寄生核盘菌菌核的相关基因,应用抑制消减杂交技术构建了cDNA消减文库并进行了筛选。通过PCR技术从文库中共筛选到1315个阳性克隆,克隆中插入片段大小主要集中于300～600bp之间。随机挑取120个克隆,经测序和同源性分析,获得60条有效序列,其中部分序列所编码的血红素加氧酶、核糖体蛋白L11、细胞色素P450及热激蛋白等均参与机体对胁迫条件的应答反应。11条序列在NCBI数据库中未找到显著匹配的序列,可能为新基因片段。分别将寄生于核盘菌菌核上的粘帚霉cDNA和粘帚霉与核盘菌纯培养的cDNA混合物经RasⅠ酶切后进行标记作为探针,利用反向Northern杂交技术验证了所选取的25条序列全部为差异表达基因片段。