10种杀菌剂对中华蜜蜂的急性毒性测定及风险评估

明确80%代森锰锌可湿性粉剂、50%啶酰菌胺水分散型粒剂、50%腐霉利可湿性粉剂、50%烯酰吗啉可湿性粉剂、100 g/L氰霜唑悬浮剂、20%叶枯唑可湿性粉剂、2%春雷毒素水剂、12.50%腈菌唑乳油、25%丙环唑乳油和15%咪鲜胺微乳剂等10种杀菌剂对中华蜜蜂Apis cerana cerana工蜂的急性毒性,并评价其对中华蜜蜂工蜂的风险性。在室内采用摄入法和点滴法测定10种杀菌剂对中华蜜蜂工蜂的急性经口和接触毒性。结果表明代森锰锌,啶酰菌胺,腐霉利等7种杀菌剂对中华蜜蜂的急性毒性均为低毒;腈菌唑和丙环唑对中华蜜蜂的急性经口毒性和急性接触毒性均为中毒,咪鲜胺对中华蜜蜂的急性经口毒性为高毒,急性接触毒性为中毒;这3种杀菌剂且均属于中等风险性。建议禁止咪鲜胺在蜜源作物开花授粉期施用,慎重选择中等毒性杀菌剂腈菌唑和丙环唑,尽量选择对蜜蜂低毒的杀菌剂,以保护中华蜜蜂的安全。     

农用杀菌剂对魔芋软腐病菌的抑制效果研究

魔芋软腐病能导致魔芋产量损失严重,甚至绝收,而目前在生产上还未找到防止该病的特效药剂。采用皿内拮抗法测定苯醚甲环唑、甲基硫菌灵、硫酸链霉素、甲霜.噁霉灵、叶枯唑可湿性粉剂、代森锰锌和灰霉1号杀菌剂等七种农用杀菌剂对魔芋软腐病菌的抑制作用。研究结果表明:供试杀菌剂对魔芋软腐病菌均有一定的抑制作用,不同杀菌剂抑菌作用差别较大,80%的甲基硫菌灵和72%的硫酸链霉素对魔芋软腐病病菌有较好的抑制效果。     

苹果套袋果实黑点病菌杀菌剂室内测定研究

为筛选有效抑制套袋苹果黑点病多种病原的杀菌剂,选择25%吡唑醚菌酯悬浮剂、250 g/L嘧菌酯悬浮剂、10%苯醚甲环唑水分散粒剂、430 g/L戊唑醇悬浮剂、30%氟菌唑可湿性粉剂和80%克菌丹水分散粒剂6种杀菌剂,采用菌丝生长速率法,对引起套袋苹果黑点病的6种主要病原菌进行室内抑菌活性测定,获得了对多种黑点病病原具有抑制活性的杀菌剂。嘧菌酯、吡唑醚菌酯、戊唑醇和苯醚甲环唑4种杀菌剂对6种病原具有较好的抑菌活性,EC₅₀均小于20μg/mL,具较好田间应用前景。结果说明甲氧基丙烯酸酯类杀菌剂嘧菌酯、吡唑醚菌酯和三唑类杀菌剂戊唑醇、苯醚甲环唑有望作为交替使用防治套袋苹果黑点病的候选农药。     

干旱胁迫下AMF对云南蓝果树叶片解剖结构的影响

苯菌灵为杀真菌剂,在土壤含水量为32.32％、29.63％、25.86％、19.39％、12.93％和6.46％的条件下,分别添加苯菌灵和不添加苯菌灵,形成“低AMF”和“高AMF”处理。该研究以云南蓝果树幼苗叶片为材料,利用盆栽试验研究了干旱胁迫下丛枝菌根真菌( AMF)对云南蓝果树幼苗叶片解剖结构及抗旱性的影响。结果表明：添加苯菌灵处理显著降低了不同水分处理条件下AMF侵染率,随着干旱胁迫程度加剧,云南蓝果树幼苗根部的AMF侵染率显著降低。轻度胁迫条件下(土壤含水量为29.63％),叶片解剖结构参数未发生显著变化;土壤含水量低于25.86％,云南蓝果树幼苗表现出较高的抗旱性,苯菌灵处理可以显著影响叶片角质层厚度、栅栏组织厚度和上表皮厚度等7个叶片结构指标,证明了高AMF可以增强代表云南蓝果树幼苗叶片抗旱性的结构性状。土壤含水量为25.86％、19.39％和12.93％时苯菌灵处理的效果较土壤含水量为6.46％时更显著,这是因为6.46％的土壤含水量严重抑制AMF的侵染,说明AMF侵染程度会影响云南蓝果树幼苗的抗旱性。进一步用隶属函数值法对10个叶片性状进行综合评价,发现高AMF处理可增强云南蓝果树幼苗的抗旱性。该研究结果为AMF在濒危物种云南蓝果树保护过程中的合理利用提供了理论依据。     

草莓垫壳孢寄主范围测定及室内生物药剂筛选

为明确赤芍根腐病病原菌草莓垫壳孢(Coniella fragariae)的寄主范围并筛选有效的生物杀菌剂,本研究采用叶片刺伤接菌法,对赤芍、樱桃、牡丹等12种植物叶片进行致病性试验,并采用菌丝生长速率法测定了8种生物杀菌剂对C.fragariae的抑菌效果.结果表明,该病原菌可侵染赤芍、樱桃、牡丹、月季、苹果、石榴和核桃叶片,不侵染梨、山楂、竹、小叶黄杨、冬青叶片. 0.3%丁子香酚对C.fragariae的抑制效果最好,EC₅₀仅为0.130 5 mg/L; 3%中生菌素次之,EC₅₀为4.225 6 mg/L; 8%宁南霉素、 6%春雷霉素、 4%嘧啶核苷类抗菌剂、 10%多抗霉素和5.0亿多粘类芽孢杆菌抑菌效果较差,EC₅₀分别为63.586 8 mg/L, 72.167 3 mg/L, 293.900 3 mg/L, 360.950 0 mg/L和405.708 1 mg/L; 24%井冈霉素对赤芍根腐病病原菌的抑制效果最差,抑菌效果不明显.该研究结果为田间赤芍根腐病的诊断及防治药剂的选择提供了理论依据.     

几种常用杀菌剂对胡瓜钝绥螨的影响

采用3种不同的方法测定了5种常用杀菌剂对天敌胡瓜钝绥螨Neoseiulus cucumeris（Oudemans）的影响。结果表明5种杀菌剂中,86．2％铜大师可湿性粉剂8001200倍对胡瓜钝绥螨有较低的影响,喷雾法测定最高浓度800倍校正死亡率为15．00％,而让胡瓜钝绥螨取食经86．2％铜大师可湿性粉剂各浓度药液处理过的山楂叶螨Amphitetranychus viennensis（Zacher）均无二次中毒现象,80％代森锌可湿性粉剂、80％大生M-45可湿性粉剂、50％代森锰锌可湿性粉剂和50％普雅图可湿性粉剂在3种试验方法中各浓度对胡瓜钝绥螨死亡率均与清水对照相当,因此释放捕食螨生防园防治病害可选用对胡瓜钝绥螨影响小的80％代森锌可湿性粉剂800～1000倍、80％大生M-45可湿性粉剂500-600倍、50％代森锰锌可湿性粉剂600-800倍、86．2％铜大师可湿性粉剂12001500倍、50％普雅图可湿性粉剂800-1000倍。     

一种新发生的油桐叶枯病病原真菌

报道了中国广西田林县油桐主产区一种新的叶片真菌病害,定名为油桐叶枯病。该病主要为害叶片,病菌侵染后叶片呈现灰褐色病斑,后期扩至整个叶片,引起过早脱落,影响油桐产量。从采集的带病标本中分离到16个纯培养物,隶属5个分类单元。依据柯赫法则和致病性测定,证明只有菌株VT-04为油桐叶枯病的病原物。在病斑组织中观察不到有性或无性繁殖结构,而该病原菌在诱导培养（含无菌松针的水琼脂+近紫外照射）条件下可产生子实体。比较培养性状、产孢结构特征并结合分析核糖体rDNA基因转录间隔区（ITS）和RNA聚合酶次大亚基基因（RPB2）序列和系统发育关系,将该病原菌鉴定为葡萄座腔菌科Botryosphaeriaceae中的一种无性型菌物,即小新壳梭孢Neofusicoccum parvum。这是首次在油桐叶片上发现由该病菌引起的病害。     

香蕉幼苗三类有机小分子溶质对尖孢镰刀菌侵染的生理响应

为阐明香蕉枯萎病发病机制,研究了尖孢镰刀菌侵染后,香蕉植株中几种对尖孢镰刀菌生长有显著作用的物质(氨基酸、有机羧酸、酚酸)种类和含量的变化。结果表明:(1)病原菌侵染后,伤害逐渐加剧,株高和生物量显著下降。(2)病原菌侵染后,叶片氨基酸总量显著升高,其中丝氨酸、缬氨酸、组氨酸、异亮氨酸和亮氨酸增幅较大,病原菌侵染16 d,其含量分别为侵染前的7.1、6.2、4.4、3.5和2.3倍;而根氨基酸总量开始显著降低,差异逐渐变小。(3)叶片有机羧酸酸含量在病原菌侵染后显著增加,而在根中显著降低。侵染植株叶片中草酸、柠檬酸、苹果酸、琥珀酸和延胡索酸含量分别是未侵染植株叶片的2.6、1.6、1.9、1.8和2.3倍;根中草酸、柠檬酸、苹果酸、琥珀酸和延胡索酸含量分别是未侵染植株的81%、42%、44%、28%和59%。(4)病原菌侵染后,植株叶片和根中酚酸含量都显著升高。叶片中阿魏酸、肉桂酸和水杨酸含量分别是未侵染叶片的2.9、1.7和2.9倍;而根中对羟基苯甲酸和丁香酸含量分别是未侵染根的4.3和1.5倍。研究结果表明,尖孢镰刀菌侵染后,植物与病原菌的相互作用使得植物体内抑菌物质和促菌物质都会相应的增加,植株对病害有一定的抗性,但促菌物质种类和含量较高最终使得感病植株发病。     

几种杀菌剂对黄瓜疫霉菌的毒力测定

采用菌丝生长速率法检测60%嘧菌酯悬浮剂、80%代森锰锌可湿性粉剂、75%百菌清可湿性粉剂、60%唑醚·代森联水分散粒剂、2%蛇床子素微乳剂、64%杀毒矾可湿性粉剂等6种杀菌剂对黄瓜疫霉菌的室内毒力。结果表明,唑醚·代森联和蛇床子素对黄瓜疫霉菌的毒力最高,EC50值分别为3.4143 mg·L-1和10.4641 mg·L-1。将两种原药混配后,表现出增效作用,在两者混配比为1∶2时增效最明显,EC50值为5.8178 mg·L-1。     

秃瓣杜英叶枯病及其病原菌

秃瓣杜英是我国江南地区广泛栽培的园林绿化树种。从2003年开始,在杭州地区秃瓣杜英叶枯病发生严重,发病株率达20%以上。该病害主要在春、秋季为害新梢嫩叶,导致叶片枝条枯死,树势衰退,严重的整株树枯死。通过对侵染秃瓣杜英病原菌的形态学观察、致病性测定及rDNA ITS序列分析,证明侵染秃瓣杜英的病原菌为芒果球座菌Guignardia mangiferae,其无性型为Phyllosticta anacardiacearum。     

Endophytic fungi from leaves of Centella asiatica: occurrence and potential interactions within leaves

Fungal endophytes were isolated from leaves of Centella asiatica (Apiaceae) collected at Mangoro (middle eastern region of Madagascar, 200 km from Antananarivo). Forty- five different taxa were recovered. The overall foliar colonization rate was 78%. The most common endophytes were the non-sporulating species 1 (isolation frequency IF 19.2%) followed by Colletotrichum sp.1 (IF 13.2%), Guignardia sp. (IF 8.5%), Glomerella sp. (IF 7.7%), an unidentified ascomycete (IF 7.2%), the non-sporulating species 2 (IF 3.7%) and Phialophora sp. (IF 3.5%). Using sequences of the ribosomal DNA internal transcribed spacer (ITS) regions, major endophytes (IF > 7%) were identified as xylariaceous taxa or as Colletotrichum higginsianum, Guignardia mangiferae and Glomerella cingulata. Results from in vitro fungal disk experiments showed a strong inhibitory activity of the xylariaceous non-sporulating species 1 against G. mangiferae and C. higginsianum and of C. higginsianum against G. mangiferae. This can be explained by antagonism between dominant taxa.     

Stimulation of asiaticoside accumulation in the whole plant cultures of <Emphasis Type="Italic">Centella asiatica</Emphasis> (L.) Urban by elicitors

The effects of a number of different elicitors on asiaticoside production in whole plant cultures of Centella asiatica were studied, including yeast extract, CdCl₂, CuCl₂ and methyl jasmonate (MJ). Only MJ and yeast extract stimulated asiaticoside production—1.53 and 1.41-fold, respectively. Maximum asiaticoside production was achieved following treatment with 0.1 mM MJ (116.8 mg/l). The highest asiaticoside production (342.72 mg/l) was obtained after 36 days of elicitation in cultures treated with 0.1 mM MJ and 0.025 mg/l 1-phenyl-3-(1,2,3-thidiazol-5-yl)urea (TDZ). Interestingly, MJ not only stimulated the production of asiaticoside but also had an important role in the senescence of C. asiatica. Although asiaticoside content did not change when TDZ was added to medium containing an elicitor, TDZ did increase shoot growth of C. asiatica. We discuss the interactive roles of MJ and TDZ in secondary metabolic production and biomass in whole plants of C. asiatica     

Adriamycin induced myocardial failure in rats: Protective role of Centella asiatica

Generation of reactive oxygen species and mitochondrial dysfunction has been implicated in adriamycin induced cardiotoxicity. Mitochondrial dysfunction is characterized by the accumulation of oxidized lipids, proteins and DNA, leading to disorganization of mitochondrial structure and systolic failure. The present study was aimed to evaluate the efficacy of Centella asiatica on the mitochondrial enzymes; mitochondrial antioxidant status in adriamycin induced myocardial injury. Adriamycin (2.5 mg/kg body wt., i.p.) induced mitochondrial damage in rats was assessed in terms of decreased activities (p< 0.05) of cardiac marker enzymes (lactate dehydrogenase, creatine phosphokinase, amino transferases), TCA cycle enzymes (isocitrate dehydrogenase, α-ketoglutarate dehydrogenase, malate dehydrogenase, respiratory marker enzymes (NADH-dehydrogenase, cytochrome-C-oxidase), mitochondrial antioxidant enzymes (GPx, GSH, SOD,CAT) and increased (p< 0.05) level of lipid peroxidation. Mitochondrial damage was confirmed by transmission electron microscopic examination. Pre-co-treatment with aqueous extract of Centella asiatica (200 mg/kg body wt, oral) effectively counteracted the alterations in mitochondrial enzymes and mitochondrial defense system. In addition, transmission electron microscopy study confirms the restoration of cellular normalcy and accredits the cytoprotective role of Centella asiatica against adriamycin induced myocardial injury. Our results demonstrated elevated oxidative stress and mitochondrial dysfunction in adriamycin treated rats. Moreover, on the basis of our findings it may be concluded that the aqueous extract of C. asiatica not only possesses antioxidant properties but it may also reduce the extent of mitochondrial damage     

Colonization of cucumber plants by the biocontrol fungus Clonostachys rosea f. catenulata

Clonostachys rosea f. catenulata (Gliocladium catenulatum) strain J1446 (formulated as Prestop WP) suppressed Fusarium root and stem rot caused by Fusarium oxysporum f. sp. radicis-cucumerinum (Forc) on cucumber plants grown hydroponically in rockwool medium. Sixty days following application at seeding, the biocontrol agent had proliferated through the rockwool blocks and was present on cucumber roots and the crown region of the stem at populations >1 × 10⁵ CFU/g fresh weight. Scanning electron micrographs showed that C. rosea had rapidly colonized the root surface and was associated with root hairs and epidermal cell junctions. Following transformation of the fungus with Agrobacterium tumefaciens strain AGL-1 containing the hygromycin resistance (hph) and β-glucuronidase (uidA) genes, blue-stained mycelia could be seen growing on the surface and within epidermal and cortical cells of roots, stems and shoots 3 weeks after treatment. Quantification of GUS activity by fluorometric assays showed that fungal biomass was highest in the roots and crown area, while the extent of colonization of upper stems and true leaves was variable. Higher population levels resulted following application to rockwool blocks compared to seed treatment. Application of C. rosea preceding inoculation with Forc significantly reduced pathogen populations on roots compared to plants inoculated with Forc alone. Colonization of infection sites in the root zone reduced pathogen development and disease incidence. Densities of the biocontrol agent appeared to increase in the presence of the pathogen.     

Investigation of HIV-1 Viral Protein R Inhibitory Activities of Twelve Thai Medicinal Plants and Their Commercially Available Major Constituents

Viral protein R (Vpr) is an accessory protein in Human immunodeficiency virus-1 (HIV-1) and has been suggested as an attractive target for HIV disease treatment. Investigations of the ethanolic extracts of twelve Thai herbs revealed that the extracts of the Punica granatum fruits, the Centella asiatica aerials, the Citrus hystrix fruit peels, the Caesalpinia sappan heartwoods, the Piper betel leaves, the Alpinia galangal rhizomes, the Senna tora seeds, the Zingiber cassumunar rhizomes, the Rhinacanthus nasutus leaves, and the Plumbago indica roots exhibited the anti-Vpr activity in HeLa cells harboring the TREx plasmid encoding full-length Vpr (TREx-HeLa-Vpr cells). Moreover, the investigation of the selected main constituents in Punica granatum, Centella asiatica, A. galangal, and Caesalpinia sappan indicated that punicalagin, asiaticoside, ellagic acid, madecassic acid, madecassoside, zingerone, brazilin, and asiatic acid possessed anti-Vpr activities at the 10 μM concentration. Among the tested extracts and compounds, the extracts from Centella asiatica and Citrus hystrix and the compounds, punicalagin and asiaticoside, showed the most potent anti-Vpr activities without any cytotoxicity, respectively.     

西瓜花叶病毒诱导对不同抗性美洲南瓜防卫基因表达的影响

为了明确防卫基因PAL与美洲南瓜抗西瓜花叶病毒(watermelon mosaic virus,WMV)之间的关系,通过室内接种和实时荧光定量PCR技术,测定了WMV侵染对不同抗性美洲南瓜体内防卫基因PAL表达的影响。结果表明:(1)室内测定显示,抗病品种GBRV-8发病率和病情指数(15.6%和14.2)显著低于感病品种‘光板’(91.1%和65.9)。(2)实时荧光定量PCR表明,接种WMV后不同抗感品种不同组织部位PAL基因相对表达量随着接种时间增加,整体呈现出先增加后降低的趋势,而且不同组织部位PAL基因相对表达量总体呈现出叶片较高,叶柄和茎秆次之。(3)接种后5个品种不同组织部位PAL基因相对表达量与对照相比均存在显著差异,且抗病和中抗品种不同组织部位PAL基因相对表达量显著高于感病品种,尤其抗病品种GBRV-8不同组织部位PAL基因相对表达量最高,感病品种光板最低。研究认为,防卫基因PAL表达量与美洲南瓜品种抗病毒病强弱密切相关。     

Isolation and structural characterisation of two antibacterial free fatty acids from the marine diatom, <Emphasis Type="Italic">Phaeodactylum tricornutum</Emphasis>

One solution to the global crisis of antibiotic resistance is the discovery of novel antimicrobial compounds for clinical application. Marine organisms are an attractive and, as yet, relatively untapped resource of new natural products. Cell extracts from the marine diatom, Phaeodactylum tricornutum, have antibacterial activity and the fatty acid, eicosapentaenoic acid (EPA), has been identified as one compound responsible for this activity. During the isolation of EPA, it became apparent that the extracts contained further antibacterial compounds. The present study was undertaken to isolate these additional antibacterial factors using silica column chromatography and reverse-phase high-performance liquid chromatography. Two antibacterial fractions, each containing a pure compound, were isolated and their chemical structures were investigated by mass spectrometry and nuclear magnetic resonance spectroscopy. The antibacterial compounds were identified as the monounsaturated fatty acid (9Z)-hexadecenoic acid (palmitoleic acid; C16:1 n-7) and the relatively unusual polyunsaturated fatty acid (6Z, 9Z, 12Z)-hexadecatrienoic acid (HTA; C16:3 n-4). Both are active against Gram-positive bacteria with HTA further inhibitory to the growth of the Gram-negative marine pathogen, Listonella anguillarum. Palmitoleic acid is active at micro-molar concentrations, kills bacteria rapidly, and is highly active against multidrug-resistant Staphylococcus aureus. These free fatty acids warrant further investigation as a new potential therapy for drug-resistant infections.     

36种中药材体外抗菌活性筛选研究

该文研究36种常用中药材80%乙醇提取物在体外抗临床常见致病菌的抗菌活性。采用药敏纸片法测耐药菌的耐药谱,中药粗粉用80%乙醇浸泡提取,提取液减压浓缩得浸膏,通过琼脂打孔法测定提取物抑菌圈,再通过微量倍比稀释法测定最低抑菌浓度(MIC)和最低杀菌浓度(MBC)。结果表明:36种中药材醇提物中,有15种具有广谱抗菌活性,对实验中各标准菌表现出不同程度的抑制作用,对MRSA抗菌活性也较强。其中,岩陀、卷柏、首乌藤、苏木、乌药、夏枯草6种药材的抗菌活性比较突出,抑菌圈均大于11 mm,细菌对其表现为中高度敏感;它们对7株标准菌的MIC/MBC值除个别为12.5 mg·m L~(-1)以外,均小于1.563 mg·m L~(-1),对16株MRSA的MIC/MBC值均小于1.563 mg·m L~(-1),它们的萃取层活性均小于1 mg·m L~(-1)。所筛选出的15种抗菌活性较强的中药材,可为后续研究其活性单体化合物和作用机制,研发有效的抗多重耐药菌的中药制剂以及解决细菌耐药性问题提供一定的参考。     

车前对铝胁迫生理响应的研究

为探讨车前(Plantago asiatica)对铝胁迫的耐受特性及生理机理,在不同铝浓度及胁迫时间下,对其叶片的渗透调节物质、膜脂过氧化程度和体内保护酶系统进行了研究。结果表明,低浓度铝处理对车前的生理指标无明显影响。随着铝浓度的升高,叶片脯氨酸、可溶性糖、可溶性蛋白质含量呈先升高后下降趋势,细胞质膜透性显著增大、MDA含量显著增加。500 mg L–1的Al3+处理,车前叶片的SOD、CAT、POD活性均明显提高。因此,在铝胁迫下,野生草本植物车前能通过体内的生理保护机制来减少Al胁迫,表现出较强的耐铝特性。