醚菌酯对番茄灰霉病菌的生物活性及其防病方式研究

离体试验表明,醚菌酯对番茄灰霉病菌菌丝生长、孢子萌发、孢子产量和菌核产量均有一定的抑制作用,其EC50分别为935.670、0.667、0.479和6.975mg/L。其中对菌丝生长的抑制作用较低。但是,用10mg/L和20mg/L的醚菌酯叶片喷雾处理前24h,接种灰霉病菌菌碟,醚菌酯对灰霉病的保护效果分别达到92.3%和100%;处理后24h接种,治疗效果分别达83.3%和100%,说明醚菌酯对番茄灰霉病具有很好的保护和治疗作用。醚菌酯具有优异的内吸活性,能在植物体内向顶传导。     

丙烷脒衍生物的合成及其抑菌活性测定

【目的】寻找更高活性的芳香二脒类抑菌化合物。【方法】以对氰基苯酚、1,3-二溴丙烷和1,4-二溴丁烷为原料,合成了A3、B3、B4、C3、C4、D3、D4和E3 8个丙烷脒衍生物,用1H-NMR和MS鉴定了其结构,用生长速率法和黄瓜真叶法测定了其活性,并与丙烷脒进行了比较。【结果】生长速率法测试表明,8种丙烷脒衍生物对番茄灰霉病菌(Botrytis cinerea)菌丝生长均具有较强的抑制作用,其中N,N-二正丙基丙烷脒(B3)、N,N-二正丙基丁烷脒(B4)、N,N-二异丙基丙烷脒(C3)、N,N-二异丁基丁烷脒(D3)对番茄灰霉病菌生长抑制的毒力均高于丙烷脒,其EC50为0.304 3~3.473 0 mg/L;黄瓜真叶法测定表明,50 mg/L剂量下B3、B4、C3的3 d保护效果均在60%以上,优于丙烷脒处理的57.55%;B3、B4的3 d治疗效果分别为71.70%和68.87%,与丙烷脒防效(64.15%)相当。【结论】B3、B4较丙烷脒抑菌活性有较大提高,有望成为丙烷脒产品的更新品种。     

苍耳愈伤组织提取物对番茄灰霉病菌的抑菌活性

以野外采集苍耳和室外种植苍耳提取物为对照,利用生长速率法、孢子萌发法及显微观察,研究了苍耳愈伤组织提取物对番茄灰霉菌活性的影响。结果表明:苍耳愈伤组织提取物对番茄灰霉菌具有较强的抑制作用,其抑制菌丝生长的EC50为20.27 mg/mL,在30 mg/mL浓度下,对番茄灰霉菌孢子形成和孢子萌发的抑制率均达到70%以上,其对孢子形成和孢子萌发的EC50分别为9.06 mg/mL和15.21 mg/mL,其抑菌活性与野外采集苍耳和室外种植苍耳抑菌提取物相当.     

绿色木霉·烯酰吗啉水分散片防治番茄灰霉病及大鼠急性毒性研究

[目的]为了确切评价绿色木霉.烯酰吗啉水分散片剂对番茄灰霉病的防治效果。[方法]通过对峙培养,菌间交互作用机制观察及拮抗菌培养液对病菌生长影响的测定,探讨绿色木霉菌对番茄灰霉病菌的拮抗作用及其作用机制。[结果]试验表明,绿色木霉.烯酰吗啉水分散片剂300~600倍液,对番茄灰霉病的防效为83%~95%,且对作物安全;其对病原菌的抑制作用是通过代谢产物(酶或有毒物质)的作用表现出来的。以昆明种大鼠为试验动物,观察1次灌胃不同剂量的生物农药绿色木霉.烯酰吗啉水分散片剂大鼠所产生的毒性反应和死亡情况,结果显示LD50>2150 mg/kg。[结论]生物农药绿色木霉.烯酰吗啉水分散片剂在农业生产中具有广阔的发展前景。     

Relationships of aphid and mite infestations to control ofBotrytis cinerea byClonostachys rosea in rose(Rosa hybrida) leaves

Infestations of aphids(Macrosiphum rosae L.) and of twospotted spider mites(Tetranychus urticae Koch) were examined in relation to growth and sporulation ofClonostachys rosea andBotrytis cinerea, and to suppression of the pathogen by the agent, in green rose leaves. Leaves were infested artificially with 10 aphids/leaflet for 3 h, or naturally with 15-30 aphids/leaflet for 7-12 days or with undetermined numbers of mites for 10-12 days. Leaves that had or had not been infested were inoculated withC. rosea, withB. cinerea, or withC. rosea plusB. cinerea. Germination incidence and germ tube growth ofC. rosea andB. cinerea on the phylloplane in most instances were much greater in leaves previously infested with aphids or mites compared with noninfested leaves. After combined inoculation,C. rosea suppressed germination ofB. cinerea from 47% to 19% in noninfested leaves, but in leaves that had been infested the agent was ineffective and germination incidence of the pathogen increased to 75-93%. Previous infestation with naturally introduced aphids or mites, but not brief infestations of artificially introduced aphids, markedly increased sporulation ofC. rosea after the leaves died during an initial 7-15 days of incubation on a paraquat agar medium, regardless of whether or notB. cinerea was present. Sporulation ofB. cinerea was similarly increased when inoculated alone. After 15-20 days, however, conidiophores of the agent or pathogen covered most of the leaf surface in these treatments. In leaves inoculated withC. rosea plusB. cinerea, the agent suppressed sporulation of the pathogen almost completely in both previously infested and noninfested leaves. Thus, aphid and mite infestations did not compromise the ability ofC. rosea to suppress inoculum production byB. cinerea in the leaves. Increased nutrient availability on the phylloplane through exudation or as honeydew or frass is proposed as a basis to explain effects of the pest infestations onC. rosea andB. cinerea.     

Molecular Approaches to Assist the Screening and Monitoring of Postharvest Biocontrol Yeasts

Nineteen yeast isolates obtained from the surface of several fruits and vegetables grown in Southern Italy and Israel were compared by molecular analysis using arbitrarily primed polymerase chain reaction (AP-PCR) and random amplified polymorphic DNA technique (RAPD-PCR). Genetic analysis made it possible to distinguish between closely-related genitically different strains which had the same morphological characteristic, and to discard isolates which were genetically identical. Following PCR characterisation, 6 isolates were selected and tested for their biocontrol activity against major postharvest pathogens (Penicillium digitatum on grapefruit, Botrytis cinerea, Rhizopus stolonifer and Aspergillus niger on table grape and B. cinerea and R. stolonifer on cherry tomato). All the isolates showed a good biocontrol efficacy on both wounded and non-wounded fruits. Furthermore, the preharvest application of the most effective antagonist (LS15) on table grape resulted in a significant reduction in grey mold ranging from 28.3% to 38.2% compared to the untreated control. The RAPD-PCR technique was also useful for identifying and monitoring the survival of the antagonist after field application.     

天然化合物丁香酚对灰葡萄孢菌丝脂质过氧化和膜损伤的影响

研究了天然化合物丁香酚对灰葡萄孢菌丝细胞膜的影响。结果表明,丁香酚可以作用于真菌的膜系统,诱导菌丝膜脂质过氧化,使膜受到损伤,透性改变,丙二醛(MDA)流出细胞。进一步研究发现,丁香酚处理使得菌丝内过氧化氢含量逐渐升高;菌丝内钙离子浓度先升高,30 min后逐渐下降,这可能是由于细胞膜破损导致钙离子外流的缘故。同时,通过DNA梯状条带的检测,初步表明丁香酚不能诱导灰葡萄孢菌丝细胞凋亡。     

Nested PCR‐RFLP is a high‐speed method to detect fungicide‐resistant Botrytis cinerea at an early growth stage of grapes

BACKGROUND: Grey mould caused by the fungus Botrytis cinerea Pers. ex Fr. is one of the major diseases in grapes. The use of fungicides is a simple strategy to protect grapes against B. cinerea disease. However, phenotypes exhibiting resistance to fungicides have been detected in B. cinerea populations. The variation of fungicide‐resistant B. cinerea isolates renders B. cinerea disease control difficult in grapevine fields. RESULTS: The authors have developed a nested polymerase chain reaction–restriction fragment length polymorphism (PCR‐RFLP) method to detect fungicide‐resistant B. cinerea isolates at an early growth stage of grapes in grapevine fields. The nested PCR‐RFLP method was carried out to detect benzimidazole‐, phenylcarbamate‐ and/or dicarboximide‐resistant B. cinerea isolates from grape berries and leaves at Eichorn–Lorenz growth stage 25 to 29. This method successfully detected fungicide‐resistant B. cinerea isolates at an early growth stage of grapes. In addition, only 8 h was required from tissue sampling to phenotyping of fungicide resistance of the isolates. CONCLUSION: It is proposed that the early diagnosis of fungicide‐resistant B. cinerea isolates would contribute to further improvement of integrated pest management against B. cinerea in grapevine fields, and that the nested PCR‐RFLP method is a high‐speed, sensitive and reliable tool for this purpose. Copyright © 2008 Society of Chemical Industry     

Characterisation of QoI‐resistant field isolates of Botrytis cinerea from citrus and strawberry

BACKGROUND: In 2004, field isolates of Botrytis cinerea Pers. ex Fr., resistant to strobilurin fungicides (QoIs), were first found in commercial citrus orchards in Wakayama Prefecture, Japan. Subsequently, QoI‐resistant isolates of this fungus were also detected in plastic strawberry greenhouses in Saga, Ibaraki and Chiba prefectures, Japan. Biological and molecular characterisation of resistant isolates was conducted in this study. RESULTS: QoI‐resistant isolates of B. cinerea grew well on PDA plates containing kresoxim‐methyl or azoxystrobin at 1 mg L^?1, supplemented with 1 mM of n‐propyl gallate, an inhibitor of alternative oxidase, whereas the growth of sensitive isolates was strongly suppressed. Results from this in vitro test were in good agreement with those of fungus inoculation tests in vivo. In resistant isolates, the mutation at amino acid position 143 of the cytochrome b gene, known to be the cause of high QoI resistance in various fungal pathogens, was found, but only occasionally. The heteroplasmy of cytochrome b gene was confirmed, and the wild‐type sequence often present in the majority of resistant isolates, indicating that the proportion of mutated cytochrome b gene was very low. CONCLUSION: The conventional RFLP and sequence analyses of PCR‐amplified cytochrome b gene are insufficient for molecular identification of QoI resistance in B. cinerea. Copyright © 2009 Society of Chemical Industry     

绿色木霉HT-01的生物学特性和抑菌特性

从淮安市城南栽培番茄的菜园土中分离出绿色木霉HT-01,研究不同培养条件对该木霉菌丝生长和孢子产生的影响,以及该木霉对番茄灰霉病的抑制效果.结果显示,25℃、pH为5时最有利于菌丝生长和产孢;葡萄糖和蔗糖为菌丝生长和产孢的最佳碳源;光照处理对菌丝生长影响不明显,但明显促进孢子产生;菌株在玉米粉培养基上生长最快,但在纯糠培养基上产孢量最大.木霉HT-01对番茄灰霉病(Botrytis cinerea Pers)的抑制率为0.452±0.020,用木霉HT-01孢子悬浮液喷布番茄叶面和灌根处理接种灰霉病菌的番茄幼苗,使叶片感病率分别降低38.1%和42.1%,植株感病率分别降低15%和20%.