Resistance of peas to Sclerotinia sclerotiorum in the Pisum core collection

In this study, 497 pea accessions from the Pisum core collection located at the USDA-ARS, Western Regional Plant Introduction Station (WRPIS), Pullman, WA and seven woody-stem pea lines from a private collection in the UK, were screened for resistance to Sclerotinia sclerotiorum , the cause of white mould. All of the Pisum genotypes screened were susceptible to infection, and 237 of the 504 genotypes were highly susceptible since these did not survive 2 weeks post-inoculation. However, 22 pea accessions and one woody-stem line were identified with quantitative partial resistance to white mould. Pea accessions 103709, 166084, 169603, 240515 and 270536 from the core collection demonstrated the greatest quantitative partial resistance to S. sclerotiorum based on nodal resistance and plant survival in replicated greenhouse and laboratory tests. Only five of the 504 genotypes screened had a mean lesion length of between 0 and 1 cm when assessed 3 days post-inoculation. Pea stem diameter was significantly ( P  ≤ 0·03) negatively correlated with stem lesion length in replicated greenhouse and laboratory experiments, and was determined to be the best predictor of quantitative partial resistance to S. sclerotiorum based on lesion length.     

Spatial characteristics of white mould epidemics and the development of sequential sampling plans in Australian bean fields

To improve sampling efficiency and precision in the assessment of white mould (caused by Sclerotinia sclerotiorum) disease incidence on bean (Phaseolus vulgaris), the spatial characteristics of epidemics were characterized in 54 linear transects in 18 bean fields during 2008–2010 in northern Tasmania, Australia. The incidence of diseased pods and plants was assessed prior to harvest. Distributional and correlation‐based analyses indicated the incidence of diseased pods was characterized by a largely random pattern at the individual plant scale, with some patches of similar disease levels on pods occurring at a scale of 1·5 m or greater. Collectively, these results suggested epidemics may be dominated by localized sources of inoculum. Sequential sampling approaches were developed to estimate or classify disease incidence above or below provisional thresholds of 3, 5 and 15% incidence on pods near harvest. Achieving prespecified levels of precision by sequential estimation was possible only when disease incidence on pods was greater than approximately 4% and sampling was relatively intense (i.e. 10 pods evaluated on each of at least 64 plants). Using sequential classification, correct decisions on disease status were made in at least 95% of independent validation datasets after assessment of only 10·1–15 plants, depending on classification threshold and error rates. Outcomes of this research provide the basis for implementing more efficient sampling and management strategies for this disease in Australian fields.     

向日葵菌核病拮抗木霉菌筛选及防病效果

采用平板对峙、离体叶片检测、盆栽和田间试验,筛选出3株对向日葵菌核病有较强拮抗能力的木霉菌,分别为深绿木霉Trichoderma atroviride N862、长枝木霉T. longibrachiatum N246和哈茨木霉 T. harzianum X102。3株木霉对向日葵菌核病菌的生长抑制率均超85%,离体叶片试验显示,N862、N246和X102对病斑扩展的抑制率分别为90.6%、96.5%和63.8%,表现出良好的防病效果。分别采用木霉孢子拌土、灌根、拌土+灌根3种处理方式对向日葵菌核病进行盆栽防治试验,结果显示拌土+灌根处理防病效果最佳,3株木霉的防效均超过73%。采用拌土+灌根处理进行防治向日葵菌核病田间试验,N862、N246和X102对病害的田间防效分别为67.54%、49.96%和49.23%,其中以N862的防效最高,3株木霉对向日葵菌核病均表现出良好的防病效果,有重要的开发应用潜力。     

绿色木霉发酵配方与防治油菜菌核病的研究

以绿色木霉Tv36为农抗产生菌,比较了多种液体培养基对Tv36产生抗生代谢物能力差异。根据几种较优配方的组分与用量,经正交试验得到产抗生代谢物的最优配方OTF。Tv36用OTF发酵并过滤后,对抗生代谢物进行了防治油菜菌核病的生物测定和田间试验。结果表明,Tv360－OTF发酵滤液对核盘菌菌丝生长有明显的抑制效果;Tv36－OTF滤液10、20、50倍3种不同的稀释倍数在油菜离体叶试验中均有明显的抑制病斑扩展的效果;Tv36－OTF农抗液剂20、50倍对油菜核病的田间防治效果分别达80．5％、68．6％,与菌核净防效相当。     

Evaluation of Microbial Antagonists for Biological Control of Botrytis cinerea Stem Infection in Cucumber and Tomato

Of fifteen isolates of yeasts, filamentous fungi and bacteria and a commercial product, tested in a bioassay with stem segments, eleven isolates consistently reduced incidence of disease and sporulation of Botrytis cinerea Pers; Fr in tomato and seven isolates in cucumber. Several isolates reduced disease by more than 75% in all experiments. Six antagonists that performed well in the bioassays and that were fairly easy to produce in vitro, were selected for further testing in two glasshouse experiments with cucumbers. After application of spores of B. cinerea and the antagonists or the fungicide tolylfluanid to pruning wounds, disease incidence was reduced by 50–100% by all antagonists in both experiments and only in one experiment by tolylfluanid.For Trichoderma harzianum T39, Aureobasidium pullulans and Cryptococcus albidus, biological control efficacy in bioassays with cucumber stem segments was not strongly influenced by temperatures in the range between 18 and 30°C, but at 24°C the efficacy of the three antagonists strongly decreased at relative humidities of 90% and 80% (vapour pressure deficits 0.299 and 0.598kPa, respectively) compared to 100.     

影响盾壳霉寄生核盘菌菌核的几个生态因子的分析

 在室内测定了盾壳霉(Coniothyrium minitans)对不同寄主上的核盘菌(Sclerotinia sclerotiorum)菌核的寄生致腐作用,研究了温度、含水量和土壤类型等生态因子对核盘菌菌核的寄生致腐作用的影响,通过检测土壤的呼吸速率探讨了它在土壤中定殖与核盘菌菌核的关系。结果表明:盾壳霉能寄生致腐核盘菌属所有供试菌株的菌核;寄生致腐菌核的最适温度是20℃,最适相对含水量为50%~60%;盾壳霉在供试的8种土壤中均能寄生致腐菌核,对它们的pH值要求不严格,但土壤类型影响其寄生致腐速度;在土壤中添加菌核和菌核提取液都可不同程度地刺激它的生长。     

哈茨木霉防治茉莉白绢病效果试验

1977年从水稻叶面分离获得木霉82(T—82)经鉴定为哈茨木霉Trichoderma harzianum Rifai。用这种木霉的分生孢子浓度0.5％和0.7％,防治茉莉白绢病Sclerotium rolfsii,效果分别为97.62％和100％。本文也讨论了防治机理,以及对其它病原真菌的拮抗作用。     

棘孢木霉TCS007菌株固体发酵产孢条件优化及其抑菌活性

棘孢木霉 (Trichoderma asperellum) TCS007菌株分离自海洋极端环境并进行了驯化培养和诱变稳定,该菌株及其代谢产物对植物病原真菌具有广谱的抑制活性,且能诱导植物促生、抗逆等,具有较好的开发前景。为探索TCS007菌株固体发酵产孢量的最佳发酵条件,明确其代谢产物的抑菌活性,本研究通过单因素试验,结合响应曲面法,对TCS007菌株固体发酵培养基配方和发酵培养条件进行优化,采用菌丝生长速率法测定其发酵滤液和粗提物对6种植物病原真菌的抑制活性。结果表明：影响TCS007菌株固体发酵产孢量的3个主要因素为培养时间、加水量和光照周期时间。优化后的发酵培养基配方为麦麸和黄豆粉按质量比5 : 1混合,并添加1%硫酸锰、1%硫酸镁;优化后的发酵条件为：培养时间15 d、加水量39.7 mL、光照周期时间21 h、培养温度28 ℃、发酵接种量2 mL (1 × 10⁷个/mL)、初始pH值5。在此条件下,按优化后的发酵培养基配方,TCS007菌株的孢子产量为7.14 × 10⁹个/g,与响应曲面法预测的孢子产量 (6.85 × 10⁹个/g) 拟合,比基础发酵条件孢子产量提高了6倍。抑菌活性测定结果表明,TCS007菌株固体发酵滤液(稀释10倍)和粗提物 (10 mg/L) 对油菜菌核病菌(Sclerotinia sclerotiorum)均具有较好的抑制效果,抑制率分别为92.4%和95.8%。该结果可为后续产品发酵工艺研究及其木霉菌剂的研制提供理论依据。     

哈茨木霉菌(Trichoderma harzianum)和终极腐霉菌(Pythium ultimum)对玉米蛋白质组的影响(I)

 哈茨木霉(Trichoderma harzianum) T22菌株已普遍用于防治包括由终极腐霉(Pythium ultimum)引起的苗病或根腐病在内的各种病害。玉米自交系Mo17种子经T22处理后播种在接种腐霉或未接种的田间土壤内,5 d后取幼苗的根系或幼茎提取蛋白。结果表明:在接种腐霉菌的土壤内,未进行T22处理的5 d龄幼苗长势明显比对照差,而经T22处理的幼苗长势明显比对照好。T22和腐霉菌复合处理及T22单独处理对幼苗生长影响基本相同。本研究建立了蛋白质提取和双向电泳分离技术。通过双向电泳及相应的分析软件(PDQuest^TM 2-D softw are)可将不同处理的幼苗自交系蛋白进行分离。T22菌株处理的根系产生104种上游调控蛋白和164种下游调控蛋白,T22与腐霉菌复合处理可产生97种上游调控蛋白和150种下游调控蛋白,而用腐霉菌单一处理诱导的上游或下游蛋白的数量明显少于上述2个处理。T22或腐霉菌单一或复合处理的根系蛋白质组图谱与空白对照相比差异显著,它们与对照的蛋白质组图谱相似系数分别为0.72、0.51和0.49;T22与腐霉菌分别处理的蛋白质组图谱间也相差明显,两者的相似系数仅为0.65。进一步研究发现,T22菌体蛋白质组图谱与上述各种处理的蛋白质组图谱相似系数均很低,说明各种处理诱导后的幼苗根系蛋白质组组分主要来自植物本身,其变化主要因T22或腐霉菌的诱导所致。腐霉菌的侵染对寄主根系蛋白组图谱影响明显高于T22的作用。蛋白质组中各种蛋白质的质谱分析(M ALDI-TOF)与鉴定将另文发表。     

核盘菌菌核萌发多样性的研究

在15、20、25和28℃条件下培养核盘菌菌核,再在20℃下诱导菌核萌发,结果将不同来源的50个菌株分成5类:①菌核易进行菌丝型萌发;②4种温度下形成的菌核均易进行子囊盘型萌发;③高温(25和28℃)下形成的菌核进行子囊盘萌发,而低温(15或20℃)下形成的菌核则不易萌发;④15℃下形成的菌核易产生子囊盘,而其它温度下形成的菌核都不能萌发;⑤4种温度下形成的菌核都不能萌发。5类菌株各占2％、6％、22％、6％和64％。对第2、3类菌株而言,形成菌核时温度越高,菌核越易萌发。进一步分析说明菌核萌发多样性和菌株来源有一定的关系。对其中3个菌株的21个单子囊孢子后代菌核萌发特性的研究结果表明,菌核萌发特性具有遗传稳定性。这说明核盘菌菌株间菌核萌发习性存在着明显的多样性,且具有遗传稳定性,可用于研究这一病菌的群体结构。     

来源于佳木斯茄子上的核盘菌菌株多样性的研究

 从我国黑龙江省佳木斯市同一茄子田采集的菌核样品中分离得到21个核盘菌(Sclerotinia sclerotiorum)单菌核分离物。通过比较其菌丝生长速度、菌落形态、菌核产量及菌核在PDA平板上的萌发特性等将其分成3种类型,记为A、B、C,各包含5、7、9个菌株。A型和B型菌株为正常菌株,而C型菌株为异常菌株,生长慢且呈扇形扩展。对代表性菌株转代培养物及菌核后代的培养特性比较结果说明它们的分化特性是稳定的。致病性的测定结果表明参比菌株Cor-6的致病性最强,A型菌株Ep-1PB和B型菌株Ep-1次之,C型菌株Ep-1PD最弱。3类菌株菌核的可溶性蛋白和酯酶同工酶电泳谱带没有明显的差异。     

控病栽培技术对油菜菌核病发生效应的研究

大小行栽培对油菜菌核病的控病研究结果表明,油菜大小行组合以行距70cm和32.5cm为宜,其发病期比常规等行栽培区的见病期延迟5～7d,自然控病效果达41.6%～47.4%,相当于常规等行栽培1次用药的防治效果;增产效果达17.9%.起到了减轻发病程度,减少用药次数,降低污染的作用.     

粉红粘帚霉67-1菌株寄生核盘菌研究

 用诱捕法从海南乐东县菜园土壤中获得一株对核盘菌菌核具有强寄生能力的粉红粘帚霉(Gliocladium roseum)菌株67-1,寄生频率为100%。该菌的PDA平板回接核盘菌菌核,一周后寄生率可达100%。对峙培养发现其对核盘菌有明显的抑菌带。保湿条件下,该菌孢子在24h内成功侵入核盘菌菌核。切片显微观察证明该菌能高效侵染菌核,造成组织溃解。寄生过程中菌核内蛋白组成发生明显变化。这一菌株在22~35℃均能很好生长,菌丝及产孢最适温度为24℃,产孢量大。认为这一菌株具有良好的菌核病生防应用潜力。     

油菜菌核病菌对多菌灵和乙霉威的抗药性机理

生物测定结果表明,油菜菌核病菌田间菌株对多菌灵(MBC)敏感性表型呈多样性,即存在MBCS、MBCLRLR、MBCHRHR和MBCVHR表型.而对乙霉威(DIE)则只检测到DIES和DIEHR表型.MBCS、MBCLR和MBCHR菌株中除JD2-3菌株为DIES外,其余菌株均为DIEHR,MBCVHR菌株对DIE表现为DIES.MBC和DIE之间存在典型的负相关交互抗性.序列分析结果表明,表型为MBCVHRDIES菌株的β-微管蛋白基因,第198位氨基酸由Glu(GAG)突变为Ala(GCG);表型为MBCHRDIEHR菌株的β-微管蛋白基因的突变位点在第200位,由Phe(TTC)突变为Tyr(TAC);而表型为MBCSDIES和MBCLRDIEHR的菌株在所扩增的β-微管蛋白基因片段中未发生突变.初步表明,β-微管蛋白基因198和200位氨基酸的突变是引起油菜菌核病菌对多菌灵抗药性呈多样性的分子机理.     

木霉对土传病原真菌的拮抗作用

 分别在体外及温室测定了筛选菌株哈茨木霉Trichoderma harzianum(T₈₂)和Tricho-derma sp.(NF9)对土传病原真菌的拮抗作用。体外测定表明,木霉菌株T₈₂和NF₉对白绢病菌Sclerotium rolfsii,立枯丝核菌Rhizoctonia solani,瓜果腐霉Pythium aphanidermatum刺腐霉P.spinosum和尖镰孢Fusarium oxysporum在对崎培养中的拮抗系数分别为2或2~3和2。温室测定表明,用0、6%(W/W)T₈₂麸皮培养物(10⁷cfu/g)处理土壤。在人工接种白绢病菌,立枯丝核菌及瓜果腐霉20天后,黄瓜发病率分别比未用木霉处理的对照减少46、5%,28.4%和81。2%;用T₈₂和NF₉木霉孢子悬浮液(10⁸cfu/ml)处理黄瓜种子,人工接种白绢病菌11天后,黄瓜成苗率分别比未用木霉处理的对照增加14%的20%。分别在光学显微镜和扫描电镜下观察到木霉T₈₂对白绢病菌菌丝和菌核的重寄生以及木霉T₈₂和NF₉对立枯丝核菌菌丝的缠绕。穿入及寄生。作者认为重寄生可能是试验木霉菌株T₈₂和NF₉对白绢病菌和立枯丝核菌的主要拮抗机制。     

生物拮抗菌Trichoderma harzianum处理玉米种子诱导实生苗的抗病性分析

 本文研究了生物拮抗菌Trichoderma harzianum(T22)处理对玉米(Mo17)实生苗生长的影响,以及对病原菌Colletrichonum graminicola的抗病性。试验结果表明:用拮抗菌T.harzianum处理后能明显地提高玉米苗根系发育和叶片生长,显著的诱导玉米叶片和根系中防御相关的酶活性和蛋白含量。用病原菌C.graminicola接种玉米叶片后观测发现,处理的实生苗叶片上病斑直径明显小于不处理的对照。尽管拮抗菌T.harzianum诱导植物抗病性的作用已被证实,但该研究更进一步揭示了T.harzianum处理玉米种子后植株的抗性应答与促进实生苗根系发育和叶片生长,以及诱导抗性相关的酶。     

基于转录组研究哈茨木霉WF2诱导的烟草抗病分子机制

为了深入探究哈茨木霉WF2诱导烟草抗病性的分子机制,利用转录组测序技术与荧光定量PCR技术,将经过哈茨木霉WF2处理的烟苗与对照的转录组进行对比与分析。结果发现,WF2处理共诱导了377个差异表达基因(DEGs),其中341个DEGs上调表达,36个DEGs下调表达。通过对差异表达基因进行GO和KEGG富集分析,发现WF2主要诱导乙烯(ET)信号通路和MAPK信号通路相关基因的上调表达,其中包括大量植物防御抗性相关基因,如编码ERF、WRKY、TGA等转录因子的基因。此外,哈茨木霉WF2还诱导了非生物胁迫相关的信号调节因子如DREB、CRK、PYL等的上调表达。     

壳聚糖对水稻恶苗病菌和油菜菌核病菌的作用

用平板含毒介质法测定了壳聚糖对水稻恶苗病菌(Fusarium moniliforme)和油菜菌核病菌(Sclerotinia sclerotiorum)的影响。结果表明,壳聚糖对F.moniliforme抑制作用相对较强,EC_(50)低于1000mg/L,而对S.sclerotiorum的作用较弱,EC_(50)高于3000mg/L。壳聚糖抑制F.moniliforme菌丝生长,导致菌丝畸形肿胀,产生不正常分支,并且抑制其分生孢子的萌发和芽管伸长。壳聚糖处理后F.moniliforme菌丝细胞膜的透性增加,导致蛋白质渗漏,但对其细胞膜的主要结构成分麦角甾醇的含量影响不明显。在离体培养时,壳聚糖对S.sclerotiorum产草酸毒素无明显影响,但预先经壳聚糖处理后,再接种S.sclerotiorum的油菜叶片中草酸含量显著减少。