发生和未发生连作芦蒿枯萎病土壤的尖孢镰刀菌数量、产毒能力和致病力

尖孢镰刀菌引起的枯萎病是芦蒿连作障碍主要病害.对芦蒿不同种植年限发病和未发病土壤中尖孢镰刀菌数量、产毒素(镰刀菌酸)能力、致病力等开展研究.对采集的11份土壤样品分析结果显示:相较于不发病土壤,发病率≥40%的土壤尖孢镰刀菌数量显著增加;不发病和发病率≤25%的土壤尖孢镰刀菌数量没有显著差异(P>0.05).发病率为85%和40%的土壤尖孢镰刀菌高产毒素(镰刀菌酸产量>500μg L-1)的菌株比例显著(P<0.05)高于发病率为11%和不发病土壤.致病力测定显示发病率为85%和40%土壤中高致病力菌株比例显著(P<0.05)高于发病率为11%的土壤和不发病土壤,不致病菌株比例显著(P<0.05)低于发病率为11%的土壤和不发病土壤.土壤尖孢镰刀菌数量与连作年限没有显著相关性(P>0.05).发病土壤中,发病率、尖孢镰刀菌高产毒素菌株比例和高致病力菌株(病情指数>2)比例与连作年限显著(P<0.05)相关;未发病土壤中,尖孢镰刀菌高产毒素菌株比例和高致病力菌株比例与连作年限没有显著相关性(P>0.05).随着连作芦蒿发病率的增加,土壤中尖孢镰刀菌菌株数量、高产毒素菌株比例、致病菌株比例均显著增加(P<0.05),未发病连作土壤尖孢镰刀菌菌株数量、高产毒素菌株比例、致病菌株比例均没有显著变化.本研究结果可为连作芦蒿枯萎病发病机理的研究及其防治提供理论基础.     

一株草莓根腐尖孢镰刀菌拮抗内生细菌的分离鉴定及抑菌特性

为筛选对草莓根腐尖孢镰刀菌(Fusarium oxysporum)有较好抑菌活性的草莓内生细菌,从草莓的根、茎、叶和果实中分离内生细菌,采用平板对峙法筛选具有抑菌活性的菌株,经形态特征观察、生理生化特性及分子鉴定后,进行拮抗菌抑菌特性分析.结果筛选到1株对草莓根腐尖孢镰刀菌具有较高抑菌活性(抑菌带达9.8 mm)、稳定性较好的菌,标记为SY-4.此外,该菌对桃褐腐病菌(Monilinia fruticola)、棉花红腐病菌(Fusarium moniliforme)和草莓根腐石楠拟盘多毛孢(Pestalotiopsis photiniae)等也有较好的拮抗作用.SY-4经鉴定确定为枯草芽胞杆菌.SY-4发酵液在培养时间48 h、培养温度37℃、pH值为7时,抗菌效果最好,但发酵产物不耐高温、不耐酸、较耐碱性,对蛋白酶敏感,初步说明存在蛋白类抑菌活性物质.电镜观察发现SY-4发酵液处理草莓根腐尖孢镰刀菌后,菌丝形态变得不规则.该拮抗内生细菌是潜在的植物病原真菌生防菌资源,具有一定的开发与应用价值.     

尖孢镰刀菌致病相关因子及其分子生物学研究进展

由致病性尖孢镰刀菌(Fusarium oxysporum)侵染引起的植物枯萎病是一种世界性的土传真菌病害,对农业生产造成了巨大经济损失.研究表明,尖孢镰刀菌的致病与多种致病因子相关,包括信号传导系统(环腺苷酸单磷酸-蛋白激酶A和促分裂素原活化蛋白激酶信号途径)、细胞壁降解酶(木聚糖酶、果胶酶、果胶裂解酶、细胞毒素、纤维素酶、半纤维素酶和木质素降解酶等)、克服植物防御响应系统(降解抗真菌化合物和破坏植物细胞壁的化学修饰)以及其它(细胞毒素和过氧化物酶)等.此外对今后的研究工作提出展望.参26     

天祝高寒草地三线镰孢的分离鉴定及β-tubulin基因序列分析

按30 m的梯度将海拔2 880～3 360 m分为16个采样点,在同一海拔高度按照"Z"字形取样,共取5个点,对16个采样点的土样混匀后以根残体分离法在镰刀菌选择性培养基上进行分离,对分离得到的镰刀菌菌落进行纯化和单孢分离后,以形态学为基础,参照Nelson分类系统进行鉴定.结果表明:在分离到的212个镰刀菌菌株中,有5个菌株为三线镰孢(Fusarium tricinctum).随机选取2株三线镰孢菌株进行β-tubulin基因序列分析,将PCR产物回收测序后在Genbank上比对,菌株TZh-4-3-7和TZh-11-5-2与GenBank上登记的6个新西兰菌株的亲缘关系最近,同源性达96%.利用DNAStar软件绘制其系统发育树状图,结果显示,菌株TZh-4-3-7和TZh-11-5-2与以上6个F.tricinctum菌株均位于系统发育树的同一分支,聚为一类.本研究采用的β-tubulin基因序列分析结果验证了利用形态学鉴定镰刀菌的准确性;将β-tubulin基因序列分析应用于三线镰孢的鉴定中,拓宽了鉴定途径,提高了鉴定的准确性;并了解了三线镰孢在天祝高寒草原不同海拔高度的分布,丰富了我国草原镰刀菌资源库,为进一步探讨镰刀菌与草原退化的关系奠定了基础.     

木霉菌辅助合成纳米银对甜瓜的防病促生作用研究

尖孢镰刀菌(Fusarium oxysporum)侵染甜瓜引起的枯萎病造成甜瓜减产、品质下降,生产上多采用化学农药防治枯萎病,但频繁使用农药容易造成镰刀菌抗药性增强和对生态环境造成影响,甚至危及人类健康。因此,新型生防制剂的研制是解决甜瓜枯萎病绿色防控的急需措施。纳米银(Silver Nanoparticles,Ag NPs)由于其强杀菌性和不易使病原菌产生抗药性的特点,在农业病虫害防治中越来越受到关注。利用不同木霉菌(T.atroviride、T.crassum、T.longbranchunes、T.spirale、T.virens、T.afroharzianum、T.koningiopsis、T.hamatum、T.citrinoviride和T.velutinum)发酵液与Ag NO_3生物合成Ag NPs,通过Ag NPs(25、50、100、200 mg·L~(-1))对尖孢镰刀菌的抑制作用筛选优势Ag NPs并对其进行表征分析,最后使用不同浓度的优势Ag NPs对甜瓜种子萌发及甜瓜防病进行探究。结果表明,木霉菌合成的Ag NPs对镰刀菌均有抑制作用,其中Ag NPsT.longbranchunes的抑菌作用显著高于其他Ag NPs。UV-Vis显示Ag NPs-T.longbranchunes表面等离子体共振发生在417 nm处有最大吸收峰;XRD证明了Ag NPs-T.longbranchunes高度结晶;FTIR研究证实Ag NPs-T.longbranchunes包含稳定的封闭剂;TEM观察到大多数Ag NPs-T.longbranchunes粒子是分散的,直径大小为5—15 nm。低浓度Ag NPs-T.longbranchunes(25 mg·L~(-1))可促进甜瓜种子萌发及生长,而高浓度(大于25 mg·L~(-1))则有抑制作用。Ag NPs-T.longbranchunes可有效降低甜瓜枯萎病发病率和病情指数。木霉菌生物合成Ag NPs为甜瓜枯萎病的防治提供了新材料,为新型生防制剂的研制奠定了基础。     

田间连续应用生物育苗基质育苗增加设施黄瓜产量

为评估田间连续移栽由根际促生菌与普通育苗基质联合研制成的生物育苗基质所育种苗的增产效果,通过田间试验,研究连续5季,移栽含解淀粉芽胞杆菌SQR9生物育苗基质所育种苗对设施黄瓜产量、生物量、土壤理化及尖孢镰刀菌数量的影响.结果显示,连续5季移栽生物育苗基质所育黄瓜种苗处理(OFNS)的黄瓜产量,每一季均高于移栽普通育苗基质所育黄瓜种苗对照(OF)的产量,在第1、2、3、4和5季,分别提高了2.74%、4.35%、5.95%、15.14%和11.07%,且第4和5季产量间具有显著性差异.相比于OF处理,OFNS处理连续5季均提高了黄瓜的株高和茎粗,有效提高了土壤中速效钾、速效磷、电导率、全氮和总钾的含量,同时降低了土体土壤和根际土壤中尖孢镰刀菌的数量.本研究表明,连续5季移栽生物育苗基质所育种苗能够有效促进黄瓜生长和提高黄瓜产量,结果可为根际有益微生物新产品的开发提供新思路以及为设施蔬菜高产栽培提供指导.     

粮食与饲料中七种单端孢霉烯族真菌毒素的测定

本文提出了一种同时检测粮食和饲料中七种主要单端孢霉烯族真菌毒素的电子捕获气相色谱分析方法.粮食样品经醋酸乙酯、氯仿、乙醇提取后,减压浓缩.浓缩液用活性炭-中性氧化铝(2:1,W/W)柱净化,84％乙腈水溶液洗脱.洗脱液减压浓缩至于,残渣用N-七氟正丁酰咪唑衍生化.电子捕获气相色谱的分析结果表明,活性炭-中性氧化铝柱能有效地除去粮食中的干扰物,有很好的分离效果.T-2毒素、HT-2毒素、二乙酰藨草镰刀菌烯醇、新茄病镰刀菌烯醇、脱氧雪腐镰刀菌烯醇、雪腐镰刀菌烯醇、镰刀菌烯酮-X等七种毒素在小麦、大麦、玉米及大米中的回收率在浓度范围为50ng/g—500ng/g的范围内均为80％以上.     

轮作联用生物有机肥促进香蕉生长

为促进连作蕉园恢复香蕉高效栽培,采集连作蕉园土壤,利用盆栽实验研究连作蕉园土壤轮作菠萝与玉米联合生物有机肥施用,对连作土壤理化性质、可培养微生物和总细菌、总真菌数量及下茬香蕉生长的影响.结果显示:较之香蕉连作对照,连作土壤轮作菠萝或玉米后再植香蕉,其叶片数、株高、茎粗、叶面积、地下部鲜重和地上部鲜重均有不同程度提高,其中株高和叶面积显著高于连作对照处理,轮作菠萝处理促生效果优于轮作玉米处理;联合生物有机肥施用后,进一步促进了香蕉的生长;菠萝和玉米轮作后均对土壤pH有一定影响,菠萝-香蕉处理和玉米-香蕉处理土壤pH值高于对照,且菠萝-香蕉处理效果优于玉米-香蕉处理;菠萝-香蕉和玉米-香蕉处理土壤可培养镰刀菌属数量均减少,根际可培养芽孢杆菌和假单胞菌的数量均增加,且菠萝-香蕉处理提高了土壤总细菌拷贝数.本研究表明菠萝和玉米轮作均能显著改善连作香蕉土壤生态环境,促进下茬香蕉生长,缓解连作生物障碍,并且联合生物有机肥施用后效果更优.     

鱼类镰刀菌的研究Ⅲ：几种药物对鱼类镰刀菌的抑制作用

研究发现制霉素（14．4～16．0mg·L－1）、K制剂（4．0mp·L－1）和孔雀绿（3，0～40mp·L-1）对镰刀菌Fu2和尖F1抑制效果好，而对Fu1-2抑制效果差；优氯净（2．0mg·L－1）能抑制Fu1-2，而不抑制Fu2和尖F1；龙胆紫（1．0mg·L-1）可抑制尖F1，而不抑制Fu2和Fu1-2制霉素对鱼毒性最小，而孔雀绿毒性最大．     

八种镰刀菌毒素的电子捕获气相色谱分析

通过对各种反应条件的选择,建立了八种主要镰刀菌毒素同时检测的电子捕获气相色谱分析方法(GC-ECD法).在SE-54涂布的弹性石英毛细管交链柱上,八种镰刀菌毒素达到完全分离。T-2毒素,HT-2毒素、新茄病镰刀菌烯醇、二乙酰蘸草镰刀菌烯醇,玉米赤霉烯酮等的最低检测限为5×10~(-11)g,脱氧雪腐镰刀菌烯醇、雪腐镰刀菌烯醇、镰刀菌烯酮-X的最低检测限为3×10~(-12)g.     

High performance liquid chromatographic determination of pesticides in soluble phase and suspended phase in river water

A high performance liquid chromatographic method has been developed to determine pesticides in river water as both dissolved phase and suspended phase. The target pesticides were eight herbicides, asulam, diuron, flazasulfuron, linuron, MCPB, mecoprop, pyrazosulfuron‐ethyl and siduron, and two fungicides, oxine‐copper and thiram. The pesticides in filtered river water were extracted with styrene‐divinylbenzene copolymer and were eluted with acetonitrile. The pesticides on suspended solids were extracted ultrasonically with acetonitrile. Each eluate was concentrated and analyzed by HPLC with multiwavelength detector. Recoveries of the pesticides in the overall procedure of this method were 78–114% for filtered river water and 75–107% for suspended solids. The limits of detection in water and suspended solids ranged from 0.1 to 0.9 μg/L and 1 to 7 μg/g, respectively. Pesticide distribution between soluble phase and suspended phase in river water was measured by this method.     

The effects of compost prepared from waste material of banana plants on the nutrient contents of banana leaves

In this study, the possible utilization of removed shoots and plant parts of banana as compost after fruit harvest were investigated. Three doses (15-30-45 kg plan(-1)) of the compost prepared from the clone of Dwarf Cavendish banana were compared with Farmyard manure (50 kg plant(-1), Mineral fertilizers (180 g N + 150 g P + 335 g K plant(-1)) and Farmyard manure + Mineral fertilizers (25 kg FM + 180 g N + 150 g P + 335 g K plant(-1)) which determined positive effects on the nutrient contents of banana leaves. The banana plants were grown under a heated glasshouse and in a soil with physical and chemical properties suitable for banana growing. The contents of N, P, K and Mg in compost and in farmyard manure were found to be similar. Nitrogen, phosphorus and potassium contents of leaves in all applications except control, and Ca, Mg, Fe, Zn, Mn, Cu contents in all applications were determined between optimum levels of reference values. There were positive correlations among some nutrient contents of leaves, growth, yield and fruit quality characteristics. Farmyard manure, Farmyard manure + Mineral fertilizers and 45 kg plant(-1) of compost increased the nutrient contents of banana leaves. According to obtained results, 45 kg plant(-1) of compost was determined more suitable in terms of economical production and organic farming than the other fertiliser types.     

The effects of compost prepared from waste material of banana on the growth, yield and quality properties of banana plants

Three doses (15-30-45 kg plant(-1)) of the compost prepared from the clone of Dwarf cavendish banana were compared with farmyard manure (50 kg plant(-1)), mineral fertilizers (180 g N+ 150 g P +335 g K plant(-1)) and farmyard manure+ mineral fertilizers (25 kg FM+ 180 g N+ 150 g P +335 g K plant(-1)) which determined positive effects on the growth, yield and fruit quality characteristics of banana plants. The banana plants were grown under a heated glasshouse and in a soil with physical and chemical properties are suitable for banana growing. The contents of N, P, K and Mg in compost and in farmyard manure were found to be similar. Farmyard manure, farmyard manure + mineral fertilizers and 45 kg plant(-1) of compost increased growth, yield and fruit quality characteristics. According to obtained results, 45 kg plant(-1) of compost was determined more suitable in terms of economical production and organic farming than the other fertilizer types.     

福美双对斑马鱼胚胎Ⅲ型脱碘酶基因表达的影响

福美双是一种二硫代甲氨基甲酸盐类农药,在我国应用较为广泛,但其残留毒性引起广泛重视。试验采用斑马鱼胚胎作为动物模型,探讨了福美双对斑马鱼胚胎的毒性。结果表明福美双导致斑马鱼胚胎孵化率下降,在受精后72 h(72 hpf),对照组的孵化率是100%,1×10-7mol·L-1福美双染毒组的孵化率降至46%,而1×10-6mol·L-1福美双染毒致使孵化率降到0%。III型脱碘酶与斑马鱼的发育和变态息息相关,以上2种浓度的福美双在24 hpf分别增高了III型脱碘酶基因的表达6.71和14.84倍,结果表明福美双具有一定的内分泌搅乱作用,也表明斑马鱼胚胎作为农药安全评价模型的理论可行性。     

蕉园土壤因子与香蕉枯萎病发病的相关性研究

香蕉枯萎病（Fusarium oysporum f.cubense）是当前严重威胁我国华南地区香蕉生产的重要土传病害,目前尚未找到有效的防治方法。于2010年采集了3个典型香蕉枯萎病患病样地（尖峰、冲坡、十月田）和2个健康样地（临高、广州）的根际与非根际土壤样品共46份,采用常规分析方法测定了19项土壤理化指标含量。旨在从农艺措施的角度考虑香蕉枯萎病的防治提供理论依据。结果表明：健康蕉园的大多理化性状和养分含量高于患病蕉园。尤以土壤粘粒、有机质、CEC、全N、全P、有效P、有效Cu、有效Fe、有效B和交换性Ca含量表现更为明显,其在健康蕉园中的含量均为患病蕉园的1.5倍以上。同一患病蕉园根际土壤中的pH值随植株感病级别增加而上升,而有效P和有效B含量随植株感病级别增加而降低。由此可以推论,在蕉园适当增施P肥和B肥有助于抑制香蕉枯萎病发生。     

Pesticides alter ion transport across frog (Pelophylax kl. esculentus) skin

We have measured, in the edible frog (Pelophylax kl. esculentus), the effect of two fungicides (8-hydroxyquinoline and captan), and four herbicides (DCMU, glyphosate, paraquat, and propachlor) on the short-circuit current, whose value gives an estimate of the net ion transport taking place across isolated skin. Glyphosate and paraquat treatment produced a modest increase in short-circuit current, corresponding to 2.6±0.7 and 4.6±0.8 μA·cm^?2, whereas the other substances had a more sustained effect, ranging from 9.1±0.6 (propachlor) to 14.8±0.9 μA·cm^?2 (captan), which is mainly attributable to an increase in the Na⁺ absorption, and, to a lesser extent, Cl^? secretion. The increase in short-circuit current after pesticide treatment, was partially abolished by AF12198, indomethacin, SC58125, SQ 22536, and W7; these results suggest that pesticides, independently from their chemical structure, induce the release of interleukin-1, which triggers the activity of cyclooxygenase-2, whose products, via a concentration in intracellular cAMP and Ca²⁺ concentration, increase Na⁺ absorption. The resulting Na⁺ disequilibrium must be compensated for by other epithelia, with the only consequence being the dissipation of energy. However, our results are important because they indicate that pesticides interact with the basic cellular machinery, which is responsible for the myriad of biological functions of different cell types.     

COSMOS,a spatially explicit model to simulate the epidemiology of Cosmopolites sordidus in banana fields

A stochastic individual-based model called COSMOS was developed to simulate the epidemiology of banana weevil Cosmopolites sordidus, a major pest of banana fields. The model is based on simple rules of local movement of adults, egg laying of females, development and mortality, and infestation of larvae inside the banana plants. The biological parameters were estimated from the literature, and the model was validated at the small-plot scale. Simulated and observed distributions of attacks were similar except for five plots out of 18, using a Kolmogorov–Smirnov test. These exceptions may be explained by variation in predation of eggs and measurement error. An exhaustive sensitivity analysis using the Morris method showed that predation rate of eggs, demographic parameters of adults and mortality rate of larvae were the most influential parameters. COSMOS was therefore used to test different spatial arrangements of banana plants on the epidemiology of C. sordidus. Planting bananas in groups increased the time required to colonise plots but also the percentage of banana plants with severe attacks. Spatial heterogeneity of banana stages had no effect on time required to colonise plots but increased the mean level of attacks. Our model helps explain key factors of population dynamics and the epidemiology of this tropical pest.     

Influence of manufacturing process on the residues of certain fungicides used on tea

Green tea shoots which were sprayed with fungicides such as hexaconazole, propiconazole, carbendazim, and tridemorph were collected from tea bushes after 24 h of fungicide application to study the effect of manufacturing process on the residues. Significant loss of residues was noted at each stage of the manufacturing process. The total loss in residues upon manufacturing of these fungicides ranged from 12.20% to 57.14%. Among the various influential factors that determine the loss of residues, vapor pressure of the fungicides was most prominent in the contribution to residue loss. A high vapor pressure of the fungicide tridemorph amounted to a residue loss of 57.14%, while the low vapor pressure of the other three fungicides caused a loss of less than 22.22%.     

Genome-wide identification and hormone expression patterns of the MaPIF family of banana genes北大核心CSCD

植物光敏色素作用因子(phytochrome interacting factor,PIF)是广泛分布于植物体内的一种转录因子,在植物的生长发育方面有着重要的作用.基于香蕉基因组数据,对香蕉MaPIF基因家族进行基因组鉴定,采用生物信息学分析方法对其进行命名,分析理化性质、蛋白质结构、基因结构、启动子顺势作用元件以及构建系统进化树;分析PIF家族在不同激素处理下的表达情况.结果显示,香蕉MaPIF家族有7个成员,均含有高度保守的bHLH结构域;编码区长度在1 116-2001bp之间,至少包含5个内含子,且大部分位于细胞外;进化树结果可以发现与拟南芥、水稻以及玉米PIF的亲缘关系较近;顺式作用元件预测结果显示,MaPIF上存在多种与激素和光相关的响应元件.qRT-PCR结果显示,MaPIF3-1、MaPIF4、MaPIF4-1在生长素(IAA)、赤霉素(GA)、生长素抑制剂(NPA)处理中均有显著表达,除此之外,所有成员在脱落酸(ABA)处理下均有明显表达.本研究表明MaPIF在香蕉生长发育中激素调控有重要作用.(图7表3参45)     

常用杀菌剂对啶虫脒中华蜜蜂毒性潜在增效作用研究

啶虫脒对蜜蜂急性毒性较低,且允许在作物花期施用,而杀菌剂也是蜜源植物花期常用药剂。本文采用点滴法和摄入法测定了10种常用杀菌剂对啶虫脒中华蜜蜂毒性的潜在增效作用。结果表明,点滴田间实际暴露剂量的杀菌剂使啶虫脒对中华蜜蜂的毒性不同程度的增加。点滴杀菌剂和啶虫脒24 h、48 h后,咪鲜胺、丙环唑、腈菌唑、戊唑醇、苯醚甲环唑、嘧菌酯、己唑醇、吡唑醚菌酯、三唑酮、氟硅唑使啶虫脒毒性分别增加14.02、10.74、8.50、7.92、4.77、4.28、4.19、4.07、2.74、2.67倍和8.01、10.14、4.80、7.09、2.66、2.56、3.62、2.69、2.24、1.49倍。摄入田间实际暴露剂量的杀菌剂和啶虫脒混剂24 h、48 h后,啶虫脒对中华蜜蜂的毒性增加。其中,丙环唑、氟硅唑、苯醚甲环唑、嘧菌酯、咪鲜胺使啶虫脒的毒性分别增加3.62、2.46、2.10、1.98、1.56倍和2.07、2.81、2.20、2.58、2.23倍。因此蜜源植物花期应避免丙环唑、咪鲜胺、戊唑醇、腈菌唑与啶虫脒先后喷施或混合喷施,慎用己唑醇等其他6种杀菌剂,以防啶虫脒残留导致中华蜜蜂采集蜂中毒。