Dehydro-alpha-lapachone isolated from Catalpa ovata stems: activity against plant pathogenic fungi

The methanol extract of stems of Catalpa ovata G Don exhibits potent in vivo antifungal activity against Magnaporthe grisea (Hebert) Barr (rice blast) on rice plants, Botrytis cinerea Pers ex Fr (tomato grey mould) and Phytophthora infestans (Mont) de Bary (tomato late blight) on tomato plants, Puccinia recondita Rob ex Desm (wheat leaf rust) on wheat plants and Blumeria graminis (DC) Speer f. sp. hordei Marchal (barley powdery mildew) on barley plants. An antifungal substance was isolated and identified as dehydro-alpha-lapachone from mass and nuclear magnetic resonance spectral data. It completely inhibited the mycelial growth of B. cinerea, Colletotrichum acutatum Simmonds, Colletotrichum gloeosporioides Simmonds, M. grisea and Pythium ultimum Trow over a range of 0.4-33.3 mg litre(-1). It also controlled the development of rice blast, tomato late blight, wheat leaf rust, barley powdery mildew and red pepper anthracnose (Colletotrichum coccodes (Wallr) S Hughes). The chemical was particularly effective in suppressing red pepper anthracnose by 95% at a concentration of 125 mg litre(-1).     

Control efficacy against rice sheath blight of Platycladus orientalis extract and its antifungal active compounds

The control efficacy of Platycladus orientalis extract against Rhizoctonia sonali Kühn, the causal agent of rice sheath blight, was evaluated by pot experiments under greenhouse conditions, and the antifungal compounds were isolated and identified through antifungal bioassay-guided fractionation using R. sonali as a tested fungus. The results indicate that the extracts from P. orientalis exhibited a significant reduction in the severity of rice sheath blight. The petroleum ether fraction partitioned from the ethanolic crude extract, showing the highest antifungal activity, was further separated, and two diterpenoid compounds with antifungal property, totarol and sclareol, were isolated and identified from the active subfractions. Totarol and sclareol possessed antifungal activity against most of the tested fungal pathogens of cereal crops such as R. solani, R. cerealis and Fusarium graminearum, indicating a similar broad antifungal spectrum. These findings suggest that the P. orientalis extract and its derived active compounds may be promising candidate agents for controlling plant fungal diseases like rice sheath blight.     

对甲氧基肉桂醛的抗真菌活性及田间药效分析

通过对植物香料产物对甲氧基肉桂醛的抗真菌活性研究,发现该化合物对鞭毛菌亚门卵菌纲、子囊菌亚门和半知菌亚门的8种植物病原真菌,番茄晚疫病菌(Phytophthora infestans)、水稻纹枯病菌(Rhizoctonia solani AG-1IA)、油菜菌核病菌(Sclerotinia sclerotiorum)、西瓜枯萎病菌(Fusarium oxysporum)、番茄灰霉病菌(Botrytis cinerea)、草莓褐色轮斑病菌(Phomopsis obscurans)、草莓炭疽病菌(Colletotrichum gloeosporioides)和烟草赤星病菌(Alternaria alternata)具有高效、广谱的特性;田间药效试验表明其对黄瓜霜霉病和水稻纹枯病具有较高的防治效果。这为其在植物病害防治中的应用以及进一步开发提供了依据。     

一些植物提取物的抗菌活性筛选

 Methanol-extracts from nine plant species were screened for their bioactivity against Peronophythora litchii, Xanthomonas oryzae pv. oryzae and Ralstonia solanacearum. The plant extracts were applied on detached litchi young leaves before inoculation with sporangial suspension of P. litchii. The results showed that methanol extracts (at conc. of 100mg dry methanol extract weight per mL) of Hamamelis mollis and Derris scabricanlis possessed the highest antifungal activity against litchi downy blight with the inhibitory effectiveness of 88.89%, followed by Phryma leptostachya (77.78%), Clematis armcmdii (66.67%), and Daphniphyllum macropodum (61.11%). At the dose of lmg dry methanol extract weight, none of the tested plant methanol extracts showed inhibitory activities against X. oryzae pv. oryzae, whereas, H. mollis had the strongest antibacterial activity against R. solanacearum with the inhibitory zone diameter of 21 mm, followed by Aristolochia mollissima, D. macropodum, Pachyrhizus erosus, Vicia unijuga, and C. armcmdii with the inhibitory zone diameter between 9 to 11 mm.     

过表达嗜热毛壳菌Cu/Zn-SOD基因提高转基因水稻对3种病害的抗性

超氧化物歧化酶(Superoxide dismutase,SOD,EC1.15.1.1)是生物体中清除氧自由基、保护细胞免受氧化损害的关键保护酶之一,SOD活性的提高可以增加植物对各种不良环境的适应和耐受能力。本研究将一个源自嗜热毛壳菌(Chaetomium thermophilum,Ct)的Cu/Zn-SOD基因Ct SOD通过农杆菌介导的遗传转化方法转入水稻,转基因植株的SOD活性均得到显著提高。对转基因植株分别接种水稻白叶枯病、细菌性条斑病以及纹枯病的病原,发现过量表达Ct SOD能够显著提高转基因植株对3种病害的抗性。这种对多种病害具有抗性的单个基因在植物抗病育种中具有重要应用价值。     

In vitro and in vivo antifungal activities of decursin and decursinol angelate isolated from Angelica gigas against Magnaporthe oryzae, the causal agent of rice blast

Blast is considered the most important fungal disease of rice due to its wide distribution and destructiveness under favorable conditions. Development of new effective and environmentally benign agents against the causal pathogen, Magnaporthe oryzae, is of great interest. In the course of a search for natural antifungal compounds in medicinal plants, we found that the methanol extract of Angelica gigas roots showed a potent control efficacy against rice blast caused by M. oryzae. We isolated antifungal coumarins from the extract, and they were identified as decursin and decursinol angelate. Antifungal activities of these compounds, along with kasugamycin, were tested on M. oryzae in vivo and in vitro. In an in vivo assay, the three compounds effectively suppressed the development of rice blast at concentrations more than 100 μg/mL. Coumarins showed relatively weak inhibitory effect on fungal mycelial growth when compared to kasugamycin. However, they strongly inhibited M. oryzae spore germination, which was not observed in kasugamycin treatments. This is the first report demonstrating that decursinol angelate can provide control against rice blast and that the two coumarins inhibit M. oryzae spore germination. In addition, the wettable powder formulation of the crude extract of A. gigas prohibited the development of blast symptoms on rice plants more effectively than liquid concentrate formulation of kasugamin, a commercial fungicide. Based on our study, we propose that coumarin compounds as well as the A. gigas root crude extract can be used as natural, benign fungicides for controlling rice blast.     

Screening extracts of Achyranthes japonica and Rumex crispus for activity against various plant pathogenic fungi and control of powdery mildew

Methanol extracts of fresh materials of 183 plants were screened for in vivo antifungal activity against Magnaporthe grisea, Corticium sasaki, Botrytis cinerea, Phytophthora infestans, Puccinia recondita and Erysiphe graminis f sp hordei. Among them, 33 plant extracts showed disease-control efficacy of more than 90% against at least one of six plant diseases. The methanol extracts of Achyranthes japonica (whole plant) and Rumex crispus (roots) at concentrations greater than 11 g fresh weight of plant tissue per litre of aqueous Tween 20 solution effectively controlled the development of barley powdery mildew caused by E graminis f sp hordei in an in vivo assay using plant seedlings. At a concentration of 300 g fresh weight of plant tissue per litre of Tween 20 solution, the two extracts were as efficient as the fungicide fenarimol (30 mg litre(-1)) and more active than the fungicide polyoxin B (100 and 33 mg litre(-1)) against Sphaerotheca fuliginea on cucumber plants in glasshouse trials.     

2014年上高县早稻主要病虫为害损失试验报告

本文对上高县2014年早稻纹枯病、稻瘟病、二化螟、稻纵卷叶螟和稻飞虱这几类水稻主要病虫害在防治与不防治条件下进行了田间试验对比,对不同处理病虫情况及产量情况进行调查,对防治效益进行了评估。结果表明,完全不防治产量损失最大,单一病虫害中纹枯病损失最大,其次是稻纵卷叶螟、稻飞虱、二化螟和稻瘟病;完全不防治的效益损失最大,单一病虫害中纹枯病损失效益最大,其次是稻纵卷叶螟、稻飞虱、二化螟和稻瘟病。2014年上半年雨水多,湿度大,造成纹枯病重发生,稻纵卷叶螟和稻飞虱中等发生,二化螟和稻瘟病轻发生。     

水稻细菌性条斑病和纹枯病生防细菌的复筛、室内防效及应用评价

采用活体生测法对前期工作中离体初筛出的5株水稻细菌性条斑病生防菌和5株水稻纹枯病生防菌分别进行复筛、温室防效测定和应用评价。结果表明,菌株GN233、GN222和GN223的菌悬液对水稻苗期细菌性条斑病有一定的防治效果,其中菌株GN233对细菌性条斑病的温室防效最好,达到28.90%,而药剂"稻丰TM多·福"可湿性粉剂的温室防效为34.20%;5株生防菌对水稻纹枯病均有不同防治效果,其中菌株GN222和GN211菌悬液的防治效果较好,分别达到44.4%和27.8%,而药剂"稻丰"可湿性粉剂的防效为33.3%;此外,筛选出的生防菌株GN222、GN233和GN211均对水稻生长具有促生作用。本研究结果可为水稻细菌性条斑病和水稻纹枯病的生物防治奠定基础。     

枯草芽孢杆菌Bs-916的抑菌活性及其抑菌物质初探

为了揭示从土壤中分离出的拮抗菌—枯草芽孢杆菌Bs-916的抑菌作用和抑菌机理,采用管蝶法、平板涂抹法及孢子盟发法测定了Bs-916及其代谢液对水稻纹枯病菌、蚕豆枯萎病菌等8种病原菌的抑菌活性,结果表明其对供试的大多数病原菌均具有较强的抑菌活性,如Bs-916及其代谢液对水稻纹枯病菌的抑制率分别为95.4％和70.7％,对水稻稻曲病菌的抑制率均为100%。为了探索Bs-916胞外物质的种类及特性,分别采用丙酮沉淀、PEG沉淀、等电点沉淀和超微浓缩等方法从Bs-916代谢液中获得了沉淀物或截留物,它们均具有抑菌活性。沉淀物经蛋白酶K 处理后,活性减弱,说明Bs-916胞外抗菌物质具有蛋白质的性质。     

Control of sheath blight disease in rice by thermostable secondary metabolites of <Emphasis Type="Italic">Trichothecium roseum</Emphasis> MML003

The aim of the study is to investigate the biocontrol mechanisms of Trichothecium roseum MML003 against the rice sheath blight (ShB) pathogen, Rhizoctonia solani as the former exhibited strong antagonistic activity against the latter. It has been found that T. roseum MML003 did not show any hyperparasitic interaction against R. solani. Further, it did not produce siderophores and hydrogen cyanide. However, the culture filtrate of T. roseum MML003 strongly inhibited the mycelial growth and sclerotial formation, its germination and viability, which proved that the biocontrol activity is antibiosis-mediated. The extracellular crude antifungal metabolites of T. roseum MML003 were thermo and photo-stable. Potted plant experiment showed that the crude metabolites of T. roseum MML003 effectively reduced the ShB disease in rice up to 47.7%. Thus, this study assumes significance as it provided further scope for the identification of antifungal metabolites from T. roseum MML003 and their possible use against sheath blight disease of rice.     

烯丙苯噻唑和氟唑环菌胺复配颗粒剂研究及其对水稻苗期稻瘟病和纹枯病的防效

水稻苗期病害的发生及防治备受关注。为明确诱抗剂烯丙苯噻唑与杀菌剂氟唑环菌胺复配颗粒剂对水稻幼苗的安全性和对苗期病害的防治效果,研制了16%烯丙苯噻唑·氟唑环菌胺复配颗粒剂配方并检测了理化性质,采用高效液相色谱法检测了有效成分含量及其缓释效果,并测定了温室条件下烯丙苯噻唑对水稻幼苗的安全性,以及颗粒剂对水稻稻瘟病和纹枯病的防治效果。结果表明,所制备颗粒剂的各项指标均符合国家相关标准要求。释放动态结果显示:杀菌剂氟唑环菌胺在V甲醇:V水=30:70的混合溶液中释放4 h后,累积释放率(质量分数)即达到峰值33%,且随后趋于稳定;而诱抗剂烯丙苯噻唑则在7 d后累积释放率方达到最高峰值;将两种有效成分复配可实现药剂对作物苗期病害的速效性与持效性协同作用。安全性试验显示,16%烯丙苯噻唑颗粒剂制剂用量不高于150 g/m2时对水稻幼苗安全。人工接菌条件下,试验剂量的烯丙苯噻唑可诱导水稻幼苗产生稳定的抗稻瘟病和纹枯病效果。16%烯丙苯噻唑·氟唑环菌胺颗粒剂对稻瘟病和纹枯病的防治效果在制剂用量150 g/m2剂量下最佳,分别为64.79%和68.32%,实现了1次用药同时防治两种病害的效果。研究结果可为诱抗剂与杀菌剂复配制剂的研发提供理论依据,同时可为缓释颗粒剂的田间科学施用提供参考。     

不同时期嫁接感染甘薯病毒病害SPVD对甘薯产量的影响

以苹果腐烂病菌、苹果轮纹病菌、棉花枯萎病菌、柑橘炭疽病菌与白菜黑斑病菌为供试菌,采用生长速率法测定25种中药材甲醇粗提物的抑菌活性。结果表明,在浓度为5．0 mg／mL 时,大部分中药材的甲醇粗提物对至少一种植物病原真菌都表现出不同程度的抑制活性,其中知母、青蒿、细辛与白鲜皮对3种供试植物病原菌的抑制率大于75％,而茜草与姜黄对4种供试植物病原菌的抑制率均在80％以上,具有广泛的抑菌谱。进一步采用系统溶剂法得到的茜草不同溶剂萃取物中,水饱和正丁醇相抑菌活性明显优于石油醚及乙酸乙酯相,表明茜草抑菌活性成分主要为中等及大极性化合物。     

A fragment of the Xanthomonas oryzae pv. oryzicola harpin HpaG Xooc reduces disease and increases yield of rice in extensive grower plantings

Harpins of phytopathogenic bacteria stimulate defense and plant growth in many types of plants, conferring disease resistance and enhanced yield. In a previous study, we characterized nine fragments of the harpin protein HpaG(Xooc) from Xanthomonas oryzae pv. oryzicola for plant defense elicitation and plant growth stimulation activity relative to the intact protein. In plants grown under controlled conditions, the fragment HpaG10-42 was more active in both regards than HpaG(Xooc). Here, we demonstrate that the activity of HpaG10-42 in rice under field conditions significantly exceeds that of HpaG(Xooc), stimulating resistance to three important diseases and increasing grain yield. We carried out tests in 672 experimental plots with nine cultivars of rice planted at three locations. Application protocols were optimized by testing variations in application rate, frequency, and timing with respect to rice growth stage. Of the concentrations (24, 24, 12, and 6 microg/ml), and number and timing of applications (at one to four different stages of growth) tested, HpaG10-42 at 6 microg/ml applied to plants once at nursery seedling stage and three times in the field was most effective. Bacterial blight, rice blast, and sheath blight were reduced 61.6 and 56.4, 93.6 and 76.0, and 93.2 and 55.0% in indica and japonica cultivars, respectively, relative to controls. Grain yields were 22 to 27% greater. These results are similar to results obtained with typical local management practices, including use of chemicals, to decrease disease severities and increase yield in rice. Our results demonstrate that the HpaG10-42 protein fragment can be used effectively to control diseases and increase yield of this staple food crop.     

茜草等25种中药材对5种植物病原真菌的抑制活性

以苹果腐烂病菌、苹果轮纹病菌、棉花枯萎病菌、柑橘炭疽病菌与白菜黑斑病菌为供试菌,采用生长速率法测定25种中药材甲醇粗提物的抑菌活性。结果表明,在浓度为5.0mg/mL时,大部分中药材的甲醇粗提物对至少一种植物病原真菌都表现出不同程度的抑制活性,其中知母、青蒿、细辛与白鲜皮对3种供试植物病原菌的抑制率大于75%,而茜草与姜黄对4种供试植物病原菌的抑制率均在80%以上,具有广泛的抑菌谱。进一步采用系统溶剂法得到的茜草不同溶剂萃取物中,水饱和正丁醇相抑菌活性明显优于石油醚及乙酸乙酯相,表明茜草抑菌活性成分主要为中等及大极性化合物。     

5种甾醇生物合成抑制剂类(SBIs)抗真菌药物对9种植物病原真菌的活性筛选

为了寻找高效、低毒以及环境友好型的农药先导化合物,通过菌丝生长速率法测定了5种甾醇生物合成抑制剂类(SBIs)抗真菌药物(益康唑、氟康唑、伏立康唑、酮康唑和咪康唑)对7种植物病原菌的抑制效果,选择其中活性较高的药物进行了防治小麦白粉病和水稻纹枯病的盆栽试验及防治小麦条锈病和水稻纹枯病的田间药效试验。室内毒力测定结果表明:伏立康唑对供试的7种植物病原真菌的杀菌活性最高,其EC50值均低于0.349 mg/L,咪康唑对小麦赤霉病菌、梨黑斑病菌、西瓜枯萎病菌和香樟炭疽病菌,益康唑对梨黑斑病菌和西瓜枯萎病菌,以及酮康唑对水稻稻瘟病菌均表现出较强的杀菌活性,且均高于对照药剂苯醚甲环唑。盆栽试验结果显示:在药剂质量浓度为37.5 mg/L时,伏立康唑和氟康唑对小麦白粉病的防治效果分别为98.26%和89.11%,明显高于商品化杀菌剂三唑醇;在质量浓度为150 mg/L时,益康唑对水稻纹枯病的防治效果最好,为86.14%。田间试验结果表明:在有效剂量为240 g/hm^2时,氟康唑对小麦条锈病的防效为98.42%,益康唑对水稻纹枯病的防效为75.21%。研究结果表明,临床上的抗真菌药物氟康唑、伏立康唑和益康唑对植物病原真菌也具有很高的活性,可望作为农用杀菌剂的先导化合物进一步研究。     

丙环唑对水稻纹枯病菌的抑制作用及对纹枯病的防治效果

为明确丙环唑对水稻纹枯病的防治作用,通过离体抑菌试验,温室盆钵试验和田间试验研究了丙环唑对水稻纹枯病菌的抑制作用和对纹枯病的防治效果.离体抑菌试验表明,丙环唑能够强烈抑制水稻纹枯病菌(Rhizoctonia solani)的菌丝生长,EC50为0.045 μg/mL,并能够强烈抑制菌核产生,但不能抑制菌核萌发.温室试验表明,纹枯病菌经丙环唑处理后,致病力随药剂浓度提高而显著减弱;丙环唑可以很好地被水稻叶片和根系吸收,并输送到水稻的茎部和叶鞘;丙环唑对水稻纹枯病具有保护和治疗作用,400 μg/mL丙环唑处理水稻,抑制病斑长度分别达到82.16％和76.61％,并随药剂处理浓度的降低而降低;丙环唑对立枯丝核菌有较长的持效期,800 μg/mL处理水稻11 d后再接种的防效为40.10％.田间药效试验表明,25％的丙环唑乳油在有效剂量150 g/hm2下3次喷药后14 d的防效达到89.15％,丙环唑在本试验剂量范围内对水稻安全.表明丙环唑对水稻纹枯病有很好的防治作用,可为生产上防治水稻纹枯病和科学用药提供依据.     

Management of bakanae and bacterial seedling blight diseases in nurseries by irradiating rice seeds with atmospheric plasma

The control of seedborne rice seedling diseases in the seed beds is important to avoid epidemics in rice nurseries and paddies, which may result in severe yield loss. Recently, irradiation with plasma containing electrons, creating positive or negative ions and neutral species, has been shown to have an antimicrobial effect, probably via generation of reactive oxygen species. This study examines whether two seedborne rice seedling diseases, bakanae disease caused by the fungal pathogen Fusarium fujikuroi, and bacterial seedling blight caused by Burkholderia plantarii, are suppressed by irradiation of infected rice seeds with atmospheric plasma. Seed germination and seedling growth were not inhibited in plasma‐treated healthy seeds. When F. fujikuroi‐infected rice seeds were irradiated with plasma after being immersed in sterile distilled water, bakanae disease severity index and the percentage of plants with symptoms were reduced to 18.1% and 7.8% of non‐irradiated control, respectively, depending on the duration of plasma irradiation. The bacterial seedling blight disease index was also reduced by plasma irradiation in vacuum‐inoculated seeds to 38.6% of the non‐irradiated control, and in infected seeds harvested from spray‐inoculated heads of rice plants to 40.1% of the control. Therefore, plasma irradiation seems to be effective in controlling two independent seedborne rice seedling diseases.     

大蒜与黄瓜轮作控制黄瓜疫病及其化感作用初探

传统农业种植中常将目标作物与化感作物轮作控制土传病害,但对化感作物控病机制研究较少。本文研究了温室大蒜与黄瓜轮作对黄瓜疫病的控制效果,并进一步研究了大蒜组织挥发物和浸提液对甜瓜疫霉Phytophthora melonis及黄瓜种子萌发的影响,以期为利用大蒜与黄瓜轮作控制黄瓜疫病提供理论指导。结果表明,大蒜与黄瓜轮作可以有效降低黄瓜疫病的发生;大蒜不同组织挥发物和浸提液对P.melonis的各生长阶段都表现出显著的抑菌活性,其中蒜瓣的抑菌活性最强,挥发物(紫皮蒜/白皮蒜)在含量0.5 g/皿时对菌丝生长的抑制率均为100%;蒜瓣浸提液(紫皮蒜/白皮蒜)在浓度1.67 mg/mL时对菌丝生长的抑制率分别为32%和45%;在浓度0.25 mg/mL时对游动孢子萌发的抑制率分别为98%和53%。另外,大蒜挥发物和浸提液在低浓度或含量对黄瓜种子的萌发并无显著抑制效果,但在中高浓度或含量会表现出延缓生长的现象。综上所述,大蒜不同组织产生和释放的化合物对P.melonis具有抑制活性,在生产上可以利用大蒜或大蒜秸秆与黄瓜轮作控制黄瓜疫病的发生。     

PGPR and entomopathogenic fungus bioformulation for the synchronous management of leaffolder pest and sheath blight disease of rice

BACKGROUND: The biological control of plant pests and diseases using a single organism has been reported to give inconsistent and poor performance. To improve the efficacy, bioformulations were developed possessing mixtures of bioagents. RESULTS: Bioformulations combining Pseudomonas fluorescens Migula strains Pf1 and AH1 and Beauveria bassiana (Balsamo) Vuill. isolate B2 were developed and tested for their efficacy against leaffolder pest and sheath blight disease on rice under glasshouse and field conditions. The combination of Pf1, AH1 and B2 effectively reduced the incidence of leaffolder insect and sheath blight disease on rice compared with other treatments. An in vitro assay of leaffolder preference to rice leaf tissues treated with Pf1 + AH1 + B2 biformulation showed variation from normal growth and development of leaffolder larvae. Plants treated with the Pf1 + AH1 + B2 combination showed a greater accumulation of enzymes, lipoxygenase and chitinase activity against leaffolder insect compared with other treatments. Similarly, the plants showed a higher accumulation of defence enzymes, peroxidase and polyphenol oxidase activity against sheath blight pathogen in Pf1 + AH1 + B2 treatment compared with the untreated control. The bioformulation mixture attracted the natural enemy population of leaffolder under field conditions. In addition, a significant increase in rice grain yield was observed in Pf1 + AH1 + B2 treatment compared with the untreated control. CONCLUSION: The combination of P. fluorescens strains and B. bassiana isolate effectively reduced the incidence of leaffolder insect and sheath blight disease on rice plants and showed the possibility of controlling both pest and disease using a single bioformulation. Copyright © 2010 Society of Chemical Industry