草莓病害生物防治初探

用平板稀释法从２０份根围土样中分离出６７株芽孢杆菌,用异步培养法筛选出对草莓灰霉菌和立枯丝菌核有强拮抗作用的菌株２２个,其中对２种病原菌有一强一弱拮抗作用的菌株９个,对２种病原菌均有强拮抗作用的菌株５个。朱１０,１２,３４和６３还对梨轮纹病菌和玉米大斑病有拮抗作用,表现为广谱抗菌丝,其中菌株１０和１２对４种病原真菌均有强拮抗作用。     

Significance of ABC transporters in fungicide sensitivity and resistance

ATP-binding cassette (ABC) transporters are members of a protein superfamily which can be responsible for efflux of drugs from cells of target organisms. In this way, the transporters may provide a mechanism of protection against cytotoxic drugs. In laboratory-generated mutants of fungi, overproduction of ABC transporters can cause multi-drug resistance to azoles and other non-related toxicants. The impact of this mechanism of resistance in field populations with decreased sensitivity to azoles remains to be established. Inhibitors of ABC transporter activity may synergize activity of azoles to populations of both sensitive and azole-resistant pathogens. The natural function of ABC transporters in plant pathogenic fungi may relate to transport of plant-defence compounds or fungal pathogenicity factors. Therefore, inhibitors of ABC transporter activity may act as disease control agents with an indirect mode of action. ©1997 SCI     

3种植物免疫诱抗剂对番茄灰霉病菌的抑制作用

为明确植物免疫诱抗剂对番茄灰霉菌的抑菌效果,选用3%氨基寡糖素水剂、0.5%几丁聚糖水剂和1%香菇多糖水剂3种常规植物免疫诱抗剂,分别配制高、中、低3个浓度,采用含药培养基的方法开展室内抑菌试验,明确3种植物免疫诱抗剂相应最佳抑菌浓度后,在番茄生育期开展田间诱导抗灰霉病试验。室内抑菌试验结果表明：3种植物免疫诱抗剂均可抑制番茄灰霉菌的生长,其中,1%香菇多糖水剂对番茄灰霉病菌具有较好的速效性和持效性,1%香菇多糖水剂20倍液接菌1d后的抑菌率为36.1%,4d后抑菌率为67.43%;3%氨基寡糖素水剂抑菌效果次之,3%氨基寡糖素水剂100倍液接菌1d后抑菌率为22.1%;0.5%几丁聚糖水剂抑菌效果相对较低,抑菌率维持在0.0%～14.3%。田间诱导抗病试验结果表明：0.5%几丁聚糖水剂200倍液对番茄灰霉病具有较好的诱抗效果,连续施用4次后,番茄灰霉病防效达61.83%。     

解淀粉芽孢杆菌菌株P-r21抑菌广谱性及对番茄灰霉病的防治

为明确内生解淀粉芽孢杆菌(Bacillus amyloliquefaciens)菌株P-r21的抑菌广谱性及对番茄灰霉病的防治效果,本研究优化了菌株P-r21发酵条件,利用平板对峙法测定了该菌株对6种病原菌的抑菌活性及对番茄灰霉病菌(Botrytis cinerea)孢子萌发的抑制作用,并通过离体接种试验研究了对番茄灰霉病的防效。结果表明：菌株P-r21最佳发酵条件为在PYJ中培养初始pH 6.0,培养时间为12 h。菌株P-r21对供试的6种植物病原菌(Botrytis cinerea、Fusarium proliferatum、Rhizoctonia cerealis、Gaeumannomyces graminis、Fusarium graminearum、Colletotrichum karstii)的生长均表现出抑制作用;对C. karstii和B.cinerea抑制作用最强,抑菌率达到88.0%和84.5%。发酵滤液对B. cinerea孢子萌发抑制率达80.6%,对叶片和果实灰霉病的防效达到100%和80.0%;发酵液10倍稀释后防效也可达到67.1%和60.7%。本研究明...     

拟南芥转录因子IDD4不同突变体对灰葡萄孢菌的抗性分析

为明确IDD家族IDD4基因在拟南芥Arabidopsis thaliana抵抗灰葡萄孢菌Botrytis cinerea侵染过程中的作用,通过统计病情指数检测拟南芥野生型（wild type,WT）植株、过表达植株IDD4-OE和缺失突变体idd4植株感染灰葡萄孢菌情况,利用组织染色检测叶片细胞死亡和H₂O₂的积累情况,采用实时荧光定量PCR（real-time quantitative-PT-PCR,qRT-PCR）技术分析灰葡萄孢菌肌动蛋白基因Bc. ACTIN在3种植株叶片中的表达情况,并施加0.1 mmol/L外源水杨酸（salicylic acid,SA）后测定IDD4-OE植株的病情指数。结果显示,不同株系对灰葡萄孢菌的抗性由高到低依次为idd4＞WT＞IDD4-OE,IDD4-OE植株中病原菌感染部位的寄主细胞死亡程度比idd4植株严重。染色结果表明,病原菌侵染拟南芥后4 h,接种部位已有H₂O₂积累。qRT-PCR反应结果显示,Bc. ACTIN在IDD4-OE中比在idd4植株中的表达水平更高,表明灰葡萄孢菌在IDD4-OE植株中的繁殖速率更快。对IDD4-OE植株外源施加SA后,其病情指数、Bc. ACTIN表达量与WT植株间均无显著差异,说明SA能将感病植株的抗性提高至WT植株的水平,表明IDD4作为负调控因子参与了拟南芥对灰葡萄孢菌的抗性调控,SA在其中发挥着重要作用。     

Fusaricidins in Paenibacillus polymyxa A21 and their antagonistic activity against Botrytis cinerea on tomato

Four kinds of antifungal compounds from an extract of Paenibacillus polymyxa A21 with molecular masses of 883.56, 897.59, 947.55, and 961.58 Da were characterized as the members of fusaricidin-type of antibiotics according to LC-MS analysis. Fusaricidins isolated from culture filtrate displayed high antagonistic activity against several plant fungal pathogens, especially Botrytis cinerea, the causal agent of gray mold. The fusaricidins biosynthetic gene cluster (BGC) from A21 was cloned by PCR and comparative cluster analysis revealed that gene fusTE, the 3′ boundary of the fusaricidin BGC in strain PKB1, was not present in fusaricidin BGC of A21, indicating that fusTE is not necessary for fusaricidin synthesis. Fusaricidin extract from A21 significantly reduced gray mold disease incidence and severity on tomato. The mRNA levels for three patho-genesis-related proteins (PRs) revealed that treatment of tomato leaves with fusaricidin extract induced the expression of PR genes to different levels, suggesting that one reason for the reduction of gray mold infection by fusaricidin is induction of PR proteins, which lead to increased resistance to pathogens. This is the first report of the application of fusaricidins to control tomato gray mold and the comparative cluster analysis provides important molecular basis for research on fusaricidin biosynthesis.     

Long‐lasting β‐aminobutyric acid‐induced resistance protects tomato fruit against Botrytis cinerea

Minimizing losses to pests and diseases is essential for producing sufficient food to feed the world's rapidly growing population. The necrotrophic fungus Botrytis cinerea triggers devastating pre‐ and post‐harvest yield losses in tomato (Solanum lycopersicum). Current control methods are based on the pre‐harvest use of fungicides, which are limited by strict legislation. This investigation tested whether induction of resistance by β‐aminobutyric acid (BABA) at different developmental stages provides an alternative strategy to protect post‐harvest tomato fruit against B. cinerea. Soil‐drenching plants with BABA once fruit had already formed had no impact on tomato susceptibility to B. cinerea. However, BABA application to seedlings significantly reduced post‐harvest infection of fruit. This resistance response was not associated with a yield reduction; however, there was a delay in fruit ripening. Untargeted metabolomics revealed differences between fruit from water‐ and BABA‐treated plants, demonstrating that BABA triggered a defence‐associated metabolomics profile that was long lasting. Targeted analysis of defence hormones suggested a role of abscisic acid (ABA) in the resistance phenotype. Post‐harvest application of ABA to the fruit of water‐treated plants induced susceptibility to B. cinerea. This phenotype was absent from the ABA‐exposed fruit of BABA‐treated plants, suggesting a complex role of ABA in BABA‐induced resistance. A final targeted metabolomic analysis detected trace residues of BABA accumulated in the red fruit. Overall, it was demonstrated that BABA induces post‐harvest resistance in tomato fruit against B. cinerea with no penalties in yield.     

Triggering the biocontrol of Botrytis cinerea by Trichoderma harzianum through inhibition of pathogenicity and virulence related proteins

This study reports a strain of Trichoderma harzianum CCTCC-SBW0162 with potential to enhance biocontrol activity against gray mold pathogen, Botrytis cinerea, and with a pivotal role in tomato (Solanum esculentum) plant growth enhancement. A total of 254 Trichoderma isolates were screened by in vitro antagonistic assay. Of these, 10 were selected for greenhouse experiments based on their greater inhibition of B. cinerea. The in vitro antagonistic assay and greenhouse experiments indicated that T. harzianum CCTCC-SBW0162 gave the highest inhibition rate (90.6%) and disease reduction (80.7%). Also, to study the possible mechanism associated with antifungal activity of CCTCC-SBW0162 against B. cinerea, molecular docking was used to assess the interactions between CCTCC-SBW0162-derived metabolites, and pathogencity and virulence related proteins of B. cinerea. The molecular docking results indicated that the combination of harzianopyridone, harzianolide and anthraquinone C derived from CCTCC-SBW0162 could synergistically improve antifungal activity against B. cinerea through the inhibition/modification of pathogenicity and virulence related proteins. However, this computerized modeling work emphasized the need for further study in the laboratory to confirm the effect T. harzianum-derived metabolites against the proteins of B. cinerea and their interactions.     

2-噻唑酰氨基环己烷基磺酰胺的合成与杀菌活性

为进一步研究环烷基磺酰胺类化合物的杀菌活性与构效关系,在前期工作基础上,合成了11个未见文献报道的2-噻唑酰氨基环己烷基磺酰胺类化合物 ( 7a ~ 7k ),其结构均经1H NMR、13C NMR、质谱和元素分析确证。分别采用菌丝生长速率法、黄瓜活体叶片法、孢子萌发法和番茄活体盆栽法对目标化合物进行了生物活性测定。结果表明:目标化合物对番茄灰霉病菌Botrytis cinerea表现出较好的抑制活性,其中化合物 7a 和 7c 在10 mg/L下对番茄灰霉病菌孢子萌发的抑制率分别为90%和67%;在200 mg/L 施药剂量下,对活体黄瓜叶片、番茄叶片和番茄花上灰霉病的防治效果,化合物 7a 分别为75%、78%和30%,化合物 7c 分别为78%、62%和44%,均优于对照药剂腐霉利,有进一步研究的价值。     

海洋生境贝莱斯芽孢杆菌TCS001的鉴定及抑真菌活性

为研究海洋生境芽孢杆菌TCS001的分类地位和抑菌活性,通过形态和生理生化特征观察,并结合gyrA序列同源性分析对菌株进行了鉴定;通过平板对峙培养法测定了菌株TCS001对多种植物病原真菌的抑菌谱;采用菌丝生长速率法和凹玻片法,测定了不同浓度TCS001菌株发酵滤液对靶标菌黄瓜灰霉病菌Botrytis cinerea菌丝生长和孢子萌发的影响。结果显示:该菌株为贝莱斯芽孢杆菌Bacillus velezensis,其对6种供试病原菌均有一定的抑制效果,其中对黄瓜灰霉病菌的抑制率最高,达87.66%。不同稀释倍数下,TCS001发酵滤液对黄瓜灰霉病菌均有一定的抑制作用,其中,稀释5倍时对菌丝生长和孢子萌发的抑制率最高,分别为96.24%和98.05%,稀释20倍时抑制率也均达90%以上。形态学观察发现,TCS001发酵滤液可导致黄瓜灰霉病菌孢子萌发芽管中间或顶端膨大畸形。研究表明,海洋生境贝莱斯芽孢杆菌TCS001极具开发为微生物农药的潜能。