5种杀螨剂对不同螨态荔枝叶螨的毒力评价和田间防效

为了综合评价常用杀螨剂对荔枝叶螨Oligonychus litchii的室内毒力,在室内分别采用Potter喷雾法、浸叶碟法和浸叶法测定了5种常用杀螨剂对荔枝叶螨成螨、卵和若螨的毒力,并测定了5种杀螨剂对荔枝叶螨的田间防效。室内毒力结果表明:哒螨灵、阿维菌素对荔枝叶螨成螨和若螨有较高的毒力,联苯肼酯对成螨和若螨的毒力较低,而螺螨酯和乙螨唑对成螨基本无活性,对若螨有一定的毒力;5种杀螨剂对卵的毒力大小依次为乙螨唑螺螨酯哒螨灵阿维菌素联苯肼酯。田间防效试验中,15%哒螨灵乳油和1.8%阿维菌素乳油对荔枝叶螨的总体防效较好,药后1 d防效高于70%,药后10 d防效达100%,药后20 d防效仍在90%以上,速效性和持效性均较好;240 g/L螺螨酯悬浮剂和110 g/L乙螨唑悬浮剂对荔枝叶螨药后3 d防效均低于70%,而药后20 d的防效高于90%,说明两药剂速效性较差但持效性较好;43%联苯肼酯悬浮剂药后1、3 d防效低于阿维菌素和哒螨灵,高于螺螨酯和乙螨唑,药后15 d防效开始下降。在田间防治荔枝叶螨时,若荔枝叶螨发生早期、成螨较少时建议施用螺螨酯或乙螨唑,而荔枝叶螨种群中成螨、若螨较多时推荐施用阿维菌素、哒螨灵或联苯肼酯。     

开口箭甾体皂甙元的分离鉴定及其抗荔枝霜疫霉菌活性

在以荔枝霜疫霉菌为指示菌的生物活性测定导向跟踪下,运用TLC、硅胶柱层析、反相硅胶柱层析和凝胶柱层析等分离纯化方法,从开口箭(Tupistra chinensis)甲醇提取物的乙酸乙酯分部萃取物中分离纯化得到2个抗菌化合物。经现代光谱(MS、IR、1D NMR和2D NMR)分析,确定化合物1为1β,2β,3β,4β,5β,7α-六羟基螺甾-25(27)-烯-6-酮,化合物2为螺甾-25(27)-烯-1β,2β,3β,4β,5β,6β,7α-七醇。离体抗菌活性测定化合物1和化合物2抑制荔枝霜疫霉菌(Peronophythora litchii)孢子囊萌发的EC50分别为100.28 mg.mL-1和124.37 mg.mL-1。用质量浓度为500 mg.mL-1的两个化合物分别处理后接种霜疫霉菌,供试荔枝果实的发病率分别为16.7%和18.9%。     

液氮保存疫霉属菌种存活期检测

对Phytophthora和Peronophythora所属12个种29株菌在液氮中保存5年零8个月至6年零3个月后检测证明有68.9％的菌种存活下来。但有些种未能存活,这些种有Phytophthora colocasiae(XH30),P.drechsleri(ATCC15428),P.erythroseptica(ATCC36302)与Peronophythora litchii(ATCC 34595)。即使存活的菌种也不一定每个保存的菌株与菌块都存活。说明在液氮中也有存活力下降的现象。除菌种本身耐深冻贮藏特性不同外,贮藏前菌种培养的旺盛程度明显影响存活。对一些菌的致病性测定证明长期保存后致病力维持不变。液氮保藏不失为保持菌种长期不变的良好方法,只是要严格按要求掌握,并对菌的耐冻力先行了解。     

绿僵菌SC0924含氮杂环类代谢产物及其抗荔枝霜疫霉活性

从绿僵菌(Metarhizium sp.SC0924)固体发酵物中分离得到12个含氮杂环类化合物,通过波谱分析,分别鉴定为环(丙氨酸-脯氨酸)(1)、环(脯氨酸-酪氨酸)(2)、环(缬氨酸-亮氨酸)(3)、环(亮氨酸-异亮氨酸)(4)、腺嘌呤(5)、腺嘌呤核苷(6)、尿嘧啶(7)、胸腺嘧啶(8)、胸腺嘧啶脱氧核苷(9)、4-乙酰氨基咪唑(10)、焦谷氨酸正丁酯(11)和光敏素(12).滤纸片琼脂扩散法试验表明化合物2和3具有较弱的抗荔枝霜疫霉活性.     

绿僵菌SC0924酚酸类代谢产物及其抗荔枝霜疫霉活性

从绿僵菌SC0924固体发酵物中分离得到8个酚酸类化合物,通过波谱分析,分别鉴定为香草酸(1)、丁香酸(2)、邻氨基苯甲酸(3)、苯乙酸(4)、阿魏酸(5)、二氢阿魏酸(6)、2-羟基-3-苯丙酸(7)和2-羟基-3-对羟基苯丙酸丁酯(8).以滤纸片琼脂扩散法对以上化合物进行抗荔枝霜疫霉活性试验,结果表明除化合物2和6外,其余化合物均有抑菌活性.     

Fungicide Effectiveness during the Various Developmental Stages of Peronophythora litchii In Vitro

Litchi downy blight caused by Peronophythora litchii is one of the most destructive diseases suffered by litchi in China. This study has evaluated the activities of the fungicides dimethomorph (DMM), azoxystrobin (AZB), famoxadone (FMD), metalaxyl (MTL), cymoxanil (CYX) and mancozeb (MCB) on the mycelial growth, sporulation, zoospores release, and germination of sporangia as well as of cystospores of P. litchii . Dimethomorph and MTL inhibited mycelial growth more effectively than the other fungicides tested. Mycelial growth was affected less by MCB, and only weakly by AZB, FMD and CYX. Sporangia production was more strongly reduced by DMM and MTL than by AZB, FMD, CYX or MCB. Zoospore release from the sporangia was most sensitive to AZB and FMD, less sensitive to MCB and insensitive to DMM, MTL and CYX. Direct germination of sporangia of P. litchii was the most strongly inhibited by AZB and FMD, followed by DMM and MCB, whilst the sensitivity of this life stage to MTL and CYX was relatively low. Germination of encysted zoospores of P. litchii was the most sensitive to AZB and FMD, followed by DMM. Mancozeb was moderately active, while MTL and CYX did not inhibit this development stage. This is the first report on the in vitro response of the litchi pathogen, P. litchii, to fungicides. These findings can be valuable tools in setting up efficient disease management programmes to control litchi downy blight.     

疫霉属菌种的液氮超低温保藏

对疫霉属的15个种68株菌和霜疫霉2株菌进行了液氮超低温的保藏试验并得到成功。比较了冷冻速度、保护剂和解冻速度对菌存活的影响。严格控制每分钟降温摄氏1度直到-40℃后再放入液氮罐中,对疫霉和霜疫霉来说都是非常必要的。这种降温程序可通过简单设备人工操作达到。而直接由室温降到-150℃以下会损伤菌种以致死亡。在所用的保护剂中,不论10％甘油还是5-15％二甲基亚砜都能起到保护作用。尚看不出对那种保护剂有特别的要求,似可任意选用。至于解冻条件,由液氮中取出放置在38℃水浴中快速融化与在20℃水浴中中等速度融化效果相等,对菌的成活都没有太大影响。我们的试验肯定了在有保护剂存在下,用慢速冷冻可以在液氮中保存疫霉和霜疫霉。     

荔枝霜疫霉的研究

对华南的荔枝霜疫霉(Peronophythora litchii Chen ex Ko et al)的形态和营养特性进行了研究,并和新模式种进行了比较。发现此菌孢囊梗的生长是一种有限-无限生长类型,或称之为多级有限生长。即孢囊梗上的小分枝大多数是有限生长的,在其顶端同时形成孢子囊。但有时在同一孢囊梗上有的小分枝会继续生长,形成二级、三级甚至四级孢囊梗。在营养要求上与疫霉无大差别,能在天然和合成培养基上旺盛生长,需要硫胺素,ca~(++)和有机二元酸,能利用NH_4~+ 或No_3~-为其氮源,并能利用淀粉为其碳源,菌体匀浆中测出淀粉酶活性。根据孢囊梗的独特生长方式,我们认为完全有理由承认这菌是一个新属,并可成为新科,霜疫霉科。本文中已将霜疫霉科作了修改描述。孢囊梗已被修改为:孢囊梗多级有限生长。无疑这菌是腐霉科和霜霉科的中间类型。在营养类型与有性器官上和疫霉相近,而其孢子囊的形成和霜霉相似。但其孢囊梗的多级有限生长方式则和这两科都不相同。     

An RXLR effector PlAvh142 from Peronophythora litchii triggers plant cell death and contributes to virulence

Litchi downy blight, caused by the phytopathogenic oomycete Peronophythora litchii, results in tremendous economic loss in litchi production every year. To successfully colonize the host cell, Phytophthora species secret hundreds of RXLR effectors that interfere with plant immunity and facilitate the infection process. Previous work has already predicted 245 candidate RXLR effector-encoding genes in P. litchii, 212 of which have been cloned and tested for plant cell death-inducing activity in this study. We found three such RXLR effectors could trigger plant cell death through transient expression in Nicotiana benthamiana. Further experiments demonstrated that PlAvh142 could induce cell death and immune responses in several plants. We also found that PlAvh142 localized in both the cytoplasm and nucleus of plant cells. The cytoplasmic localization was critical for its cell death-inducing activity. Moreover, deletion either of the two internal repeats in PlAvh142 abolished the cell death-inducing activity. Virus-induced gene silencing assays showed that cell death triggered by PlAvh142 was dependent on the plant transduction components RAR1 (require for Mla12 resistance), SGT1 (suppressor of the G2 allele of skp1) and HSP90 (heat shock protein 90). Finally, knockout of PlAvh142 resulted in significantly attenuated P. litchii virulence on litchi plants, whereas the PlAvh142-overexpressed mutants were more aggressive. These data indicated that PlAvh142 could be recognized in plant cytoplasm and is an important virulence RXLR effector of P. litchii.     

霜疫霉菌侵染对荔枝氧化作用的影响

淮枝、桂味和糯米糍 3种荔枝经接种霜疫霉菌后 ,果皮超氧化物歧化酶 (SOD)和过氧化氢酶 (CAT)活性下降 ;而过氧化氢 (H2 O2 )含量升高 ,超氧阴离子自由基 (O2 .)产生速率增加 ,丙二醛 (MDA)积累增多 ,与果实感病指数增加相一致 ,表明霜疫霉菌侵染加速荔枝氧化作用的进程。实验还表明不同荔枝品种对霜疫霉菌抵抗力与其自由基清除能力有关