努利虫疠霉感染桃蚜过程的扫描电镜观察

利用扫描电镜观察了努利虫疠霉Pandora nouryi (Remaudière & Hennebert) Humber初生分生孢子接种桃蚜Myzus persicae (Sulzer)后孢子萌发、入侵以及菌体突破虫体的整个侵染过程。结果表明：附着于虫体表面的初生分生孢子在3～5 h后即有60%以上的萌发率,萌发的孢子形成芽管或产生球形或棍棒状的附着胞;12 h后大部分孢子均已萌发,并成功入侵寄主虫体;接种60 h后,呈掌状分枝的假根首先从桃蚜胸部的腹面突破虫体长出体外,明显区别于新蚜虫疠霉Pandora neoaphidis (Remaudière & Hennebert) Humber假根突破虫体的位置;假囊状体不多见,且仅分布于蚜虫身体两侧,这可在一定程度上解释努利虫疠霉产孢对湿度条件要求较高的生物学现象。     

蓝光影响新蚜虫疠霉分生孢子弹射潜力的室内评估

新蚜虫疠霉Pandora neoaphidis是虫霉菌亚门Entomophthoromycotina虫霉目Entomophthorales的代表菌,是世界上流行最广的蚜科专化性病原真菌。其主动弹射分生孢子的能力是引发高强度流行病控制蚜虫种群的关键原因。本研究以新蚜虫疠霉菌丝制备的弹孢胶囊（模拟感病蚜尸）为研究对象,在室内对其在蓝光照射和黑暗条件下的产孢量和分生孢子弹射距离进行了测量;同时利用高速显微摄像技术对弹孢胶囊的分生孢子弹射进行了拍摄观察和软件视频分析,利用MATLAB软件分析图像视频并进行了模型构建。结果表明：新蚜虫疠霉菌丝胶囊经蓝光照射处理后弹射出来的分生孢子数量和孢子弹射速度均显著高于黑暗条件下的处理（P<0.05）;分析高速显微拍摄视频计算所得的孢子弹射初始速度从114mm/s到2.07m/s不等,速度在弹射后期会受到空气阻力的影响而急速下降;采用Stokes模型进行预测的结果显示其分生孢子的弹射路径与实际路径拟合度较高（R²=0.8941）。综上,本研究结果为深入理解新蚜虫疠霉弹射相关的流行扩散模型和生物学适态机制提供了基础数据。     

福建厦门地区蚜虫病原真菌调查及其发生规律

从2014年至2015年对厦门地区蚜虫虫霉发生和流行情况进行初步调查,共鉴定出6种虫霉目病原真菌:新蚜虫疠霉Pandora neoaphidis、努利虫疠霉Pandora nouryi、普朗肯虫霉Entomophthora planchoniana、暗孢耳霉Conidiobolus obscurus、弗雷生新接霉Neozygites fresenii、根虫瘟霉Zoophthora radicans。新蚜虫疠霉和普朗肯虫霉是春季蔬菜蚜虫种群中发生、流行的最重要病原真菌,夏季未观察到虫霉的发生。     

飞虱虫疠霉杀蚜效果及开发应用前景

通过比较飞虱虫疠霉 (Pandoradelphacis)与其它几种常见蚜虫专化性病原真菌新蚜虫疠霉(P .neoaphidis)、安徽虫瘟霉 (Zoothphoraanhuiensis)和根虫瘟霉 (Z .radicans)对桃蚜 (Myzuspersicae)毒力 ,证实了该菌的杀蚜应用潜力。时间 -剂量 -死亡率模型分析表明 ,飞虱虫疠霉杀蚜的各项毒力指标与其它几种虫霉相似或略高 ,但杀蚜速率更快。同时对培养性状、产孢特性和生长适温等的比较研究发现 ,该菌具有易发酵生产、产孢能力强和温度生长范围宽等特点 ,是一种较为理想的有望研制开发成杀蚜真菌制剂的病原菌。此外 ,根据虫霉孢子主动弹射的生物学特性 ,提出了该菌的适用剂型 ,并对它们的流行学特性及其在设栽培中的应用前景进行了讨论。     

温湿度对玫烟色棒束孢IF‐1106菌株孢子萌发及对烟粉虱致病力的影响(英文)

B型烟粉虱Bemisia tabaci Gennadius是一种重要的世界性农业害虫。玫烟色棒束孢Isaria fumosorosea是防治烟粉虱的一种重要的虫生真菌。对不同温度(20、23、26、29、32℃)、不同湿度(53%、65%、75%、85%、95%)下玫烟色棒束孢IF‐1106菌株分生孢子萌发及其对烟粉虱的致病力进行了测定。结果表明:温度对孢子的萌发有显著影响,26℃时孢子萌发率最高。当相对湿度低于75%时,孢子不萌发或萌发率较低;当相对湿度为85%–95%时,孢子萌发率显著升高。26℃时,烟粉虱2龄若虫的累计死亡率最高。随着相对湿度增大,病菌的致病力增强。当相对湿度为53%–95%时,烟粉虱2龄若虫累计死亡率从54.55%增加到88.89%。玫烟色棒束孢的致病力与孢子萌发率呈正相关,但温度比相对湿度对其影响更明显。结果表明,玫烟色棒束孢IF‐1106菌株侵染烟粉虱的最佳条件是温度26℃和相对湿度大于85%。     

诺氏虫疠霉生物学特性的研究

通过对诺氏虫疠霉生物学特性的研究,明确了该菌孢子萌发,生长的适温范围为15℃-25℃,萌发最适湿度为100％,最适pH为9－10。紫外线对诺氏虫疠霉分生孢子有强烈杀伤作用,照射10分钟,萌发率降低41．9％,不同光照处理间菌落生长差异显,全光照可完全抑制菌落生长,明暗交替最地菌落生长和产孢。     

安徽虫瘟霉的黍米培养及其对桃蚜的侵染力

安徽虫瘟霉Zoophthora anhuiensis (Li) Humber是较难人工培养的蚜虫专化性病原真菌。将灭菌并适度熟化的黍米Panicum miliaceum L.作为固体基质与挑碎的安徽虫瘟霉平板菌落混合,在20℃和12L∶12D的温光条件下静止固体培养,获得了产孢潜能大、杀蚜活性强的米粒培养物。培养7天的黍米的产孢量达13.0×10⁴个孢子/粒,产孢持续时间长达6天。用此黍米培养物弹射的孢子对桃蚜Myzus persicae (Sulzer) 若蚜进行7.9～134.9个孢子/mm²共9个剂量的孢子浴接种,所获数据很好拟合时间 剂量 死亡率模型。接种后第5～7天各天的LC₅₀依次为59.8, 39.5和33.5个孢子/mm²,LC₉₀依次为354,234和198个孢子/mm²。在57.7～134.9个孢子/mm²的接种剂量范围内,致死中时LT₅₀从5.1天下降到4.3天。由此表明,安徽虫温霉的黍米培养不仅简单易行,而且菌种的产孢和侵染生物学特性在培养物中被充分体现,每颗米粒如同自然罹病而死的蚜尸,值得进一步研究开发和利用。     

飞虱虫疠霉分生孢子在不同温湿度组合条件下的存活率

用荧光染色法研究了不同温度（10－30℃）和湿度（51－100％RH）组合条件下飞虱虫疠霉Pandora delphacis分生孢子的存活率。在无放回抽样观察中,孢子弹射后第24h的存活率在51％、74％、85％、90％、95％和100％RH与不同温度的组合中分别为42－81％、69－89％、70－95％、67－100％、76－100％和56－100％。存放4个月后,不同温度下的孢子存活率在51％、74％、85％和90％RH下分别为55－74％、52％－87％、38－73％和1－65％。而在≥95％RH下孢子存活率24h后锐减,至第7d几乎全部失活。以上结果表明,弹射后的飞虱虫疠霉孢子在头24h的存活易受低湿影响,但存活下来的孢子能继续在低湿下存活较久;饱和或接近饱和的高湿度最不利孢子长久存活。     

球孢白僵菌对温室蚜虫的侵染生物学

研究了球孢白僵菌在不同浓度、施菌方式、温度和相对湿度下对温室蚜虫的侵染力以及接种后对蚜虫的侵染速率。结果表明,高(109个孢子.mL-1)、中(108个孢子.mL-1)、低(107个孢子.mL-1)3个浓度剂量对蚜虫都有较强的致病力,且浓度越大,蚜虫的死亡率越高,死亡时间越提前。用浸渍法和孢子浴法接种蚜虫,6 d的累积死亡率分别为100%和31.1%。在测试的3个温度(22、26、30℃)中,26℃时蚜虫侵染力最强,第3天出现死亡高峰,第5天时累积死亡率就达到100%,明显高于22、30℃的处理。相对湿度越大,球孢白僵菌的致病力越强,蚜虫死亡速度越快。在温湿度组合中相对湿度为95%时,温度对白僵菌的侵染力几乎无影响,但影响发病速度,相对湿度低于95%时,26℃的侵染力始终高于22和30℃时的侵染力。通过接种后不同时间段用0.2%百菌清处理蚜虫测定该菌株的侵染速率,结果表明接种后24 h是该球孢白僵菌有效侵染蚜虫的关键时期。     

哈茨木霉的培养及其对烟草疫霉生长的抑制研究

哈茨木霉是一类重要的植病生防因子。哈茨木霉TH-1分别在PDA培养基、麦芽糖培养基、查氏培养基和琼脂培养基上培养均能产孢,其中PDA培养基为最适培养基。PDA培养基上,菌丝生长适宜温度27．5℃～35℃,最适温度32．5℃,产孢最适温度27．5℃。菌丝生长适宜pH值为3～7,产孢适宜pH值为5-9,生长与产孢最适pH值为5。光照对菌丝生长影响不大但明显影响菌株的产孢数量,光照时间越长产孢量越大。对峙培养试验表明TH-1明显抑制疫霉菌的生长速率,其无菌滤液明显抑制烟草疫霉菌游动孢子的萌发,并抑制游动孢子芽管的伸长,TH-1对游动孢子萌发的相对抑制率为12．7％,对芽管生长长度的相对抑制率为63．1％。水解酶平板活性测定显示,TH-1产生β-1,3葡聚糖酶与纤维素酶,从而使烟草疫霉菌细胞壁的消解,产生非挥发性抗生素抑制烟草疫霉菌孢子萌发,但对菌丝生长影响不大。     

Factors affecting sporulation and germination of Pandora nouryi (Entomophthorales: Entomophthoraecae), a pathogen of Myzus persicae (Homoptera: Aphididae)

Pandora nouryi discharged large numbers of primary conidia between 8 and 25°C from cadavers on the surface of water-agar. At 8°C conidial discharge lasted for 120 h, but most conidia were produced within 48 h when temperature was >15°C. Saturated humidity alone was not enough to allow for sporulation to occur freely and where RH?<?95%, no conidia were discharged. Light did not affect the pattern of conidial production nor the total number of conidia. Germination percentages of conidia on the surface of water-agar were 40 and 66% at 8 and 30°C, respectively, and were significantly lower than that at 15–25°C where germination was >95%. Conidia on leaves germinated well when RH?>?74%, while no germination occurred when RH?<?100% on cover slips. All eight insecticides tested entirely inhibited conidial germination at recommended doses (R), in particular, both the organophosphorus pesticides Lorsben (chlorpyrifos) and the organochlorine pesticides Thiodan (endosulfan) completely inhibited conidial germination even at 0.2R dose.     

Effects of temperature and relative humidity on sporulation of Metarhizium anisopliae var. acridum in mycosed cadavers of Schistocerca gregaria.

The effects of relative humidity (RH) and temperature on the sporulation of Metarhizium anisopliae var. acridum on mycosed cadavers of desert locust, Schistocerca gregaria, were assessed in the laboratory. Quantitative assessments of conidial production over 10 days under constant conditions showed that sporulation was optimized at RH > 96% and at temperatures between 20 and 30 degrees C. Under both these conditions >10(9) conidia/cadaver were produced. At 25 degrees C, conidial yield was maximized under conditions in which cadavers remained in contact with damp substrate. Relatively little sporulation occurred at 15 degrees C (< 3 x 10(7) conidia/cadaver) and 40 degrees C (< 4 x 10(6) conidia/cadaver) and no sporulation occurred at 10 or 45 degrees C. Following incubation, conidial yield was closely related to the water content of locust cadavers. In separate tests, locust cadavers were incubated for 10 days under diurnally fluctuating temperature and RH that comprised favorable (25 degrees C/100% RH) alternating with unfavorable (40 degrees C/80% RH) conditions for sporulation. In this case, fewer conidia were produced compared with cadavers that were incubated under the favorable conditions for an equal period cumulatively but were not periodically exposed to unfavorable conditions. However, this reduced sporulation observed with the fluctuating condition was not observed when cadavers were similarly incubated under favorable/unfavorable conditions of temperature but were not periodically exposed to the low RH condition. This result implies that sporulation is a dynamic process, dependent not only on periodic exposure to favorable RH but also on the interrelation of this with low RH. Associated tests and the monitoring of changes in cadaver weights imply that the mechanism driving the reduced sporulation under fluctuating RH is the net water balance of cadavers, i.e. the cumulative ability of the fungus/cadaver to adsorb water necessary for sporulation at high RH is restricted by water loss associated with intermittent exposure to a low RH. The duration of daily exposure to high humidity appears to be a crucial constraint to the recycling ability of M. anisopliae var. acridum.     

The effects of culture media, solid substrates, and relative humidity on growth, sporulation and conidial discharge of Valdensinia heterodoxa

Valdensinia heterodoxa (Sclerotiniacae) is a potential fungal bioherbicide for control of salal (Gaultheria shallon). The effect of culture media, substrates and relative humidity (RH) on growth, sporulation and conidial discharge of V. heterodoxa was determined for two isolates PFC2761 and PFC3027 in vitro. Culture media significantly affected the growth, sporulation, and conidial discharge of V. heterodoxa. Of eight agar media used, colony radial growth was optimal on salal oatmeal agar and salal potato dextrose agar for isolates PFC2761 and PFC3027, respectively; whereas sporulation was at an optimum on salal oatmeal agar for both isolates. Of the eight liquid media tested, mycelial production was highest on wheat bran–salal–potato dextrose broth. Growth on solid substrates greatly stimulated sporulation and conidial discharge of V. heterodoxa. Of the 12 solid substrates used, the greatest numbers of discharged conidia were observed from wheat bran and wheat bran–salal within 14 d of sporulation. Sporulation on solid substrates continued for 42 d. RH significantly affected the sporulation and conidial discharge for both isolates across all solid substrates tested. No conidia were produced or discharged below 93 % RH on wheat bran–salal and millet. With an increase of the RH from 93 to 97 %, sporulation and the number of discharged conidia increased significantly for both isolates on wheat bran–salal, but not on millet.     

Effects of Fluctuating Moisture and Temperature Regimes on Sporulation of Beauveria bassiana on Cadavers of Rhodnius prolixus

The effects of both moisture and temperature on the sporulation of an isolate of Beauveria bassiana , highly virulent to the Chagas' disease vector, Rhodnius prolixus , were studied in the laboratory under fluctuating regimes. Quantitative assays for conidial production from funguskilled cadavers from 10-day-old tests showed that recycling was highly dependent on the duration of exposure to high relative humidity (RH). On first-instar larval cadavers, sporulation reached more than 10 5 conidia/nymph under most regimes with a favourable phase combining high humidity (97% RH) and temperatures of 20 or 25 C for at least 12 h/day. Sporulation on third- and fifth-instar nymphs required a longer period (16 h/day or more) of favourable conditions (97% RH and 25 C) per day. The intensity of the conidial production from Rhodnius cadavers was enhanced when diurnal temperatures were high (28 and 35 C). From a vector control standpoint, daily high humidity requirements appear to be a crucial constraint. The recycling ability of B. bassiana on fungus-killed R. prolixus is most likely to contribute to the regulation of this triatomine vector if applications are made during the rainy seasons and in the most favourable habitats.     

Effect of relative humidity,temperature and fungicide on germination of conidia of Cercospora canescens caused the Cercospora leaf spot disease in mungbean

The aim of the present investigation was to determine the impact of relative humidity (RH) and temperature on conidial germination, nuclear position and effect of important fungicides on growth and conidial germination of Cercospora canescens. Germination of conidia was observed at RH range 92–100% at 5–35°C. Significant interaction between temperature and RH indicated that higher humidity and high temperature promoted quick germination both in the presence and absence of free moisture. Although in absence of free moisture at 92–95% RH higher temperatures 25–35°C promoted quick evaporation of moisture and no conidial germination. Number of germtube was increased significantly at the optimum temperature 25–30°C and higher humidity (98–100%). But higher temperature 25–35°C with lower RH did not support the conidial germination. This finding is very important for disease forecasting using meteorological data. The spray of Carbendazim as contact fungicide may not be useful since it is not effective against the conidia of C. canescens. Triadimefon did not inhibit the conidia germination but completely inhibited mycelium development at 50 μg/ml. Propriconazole inhibited both conidia germination and mycelial development. Therefore, Propiconazole may be taken as protective as well as curative spray. In non-systemic fungicide, Copper oxychloride gave anticipated result by inhibiting both conidial germination and mycelium development. Therefore, copper oxychloride can be used as protectant fungicides for Cercospora leaf spot caused by C. canescens.     

Effects of environmental factors on virulence of the entomopathogenic fungus, Nomuraea rileyi, against the corn earworm, Helicoverpa armigera (Lep., Noctuidae)

The effects of environmental factors on infection of the entomopathogenic fungus, Nomuraea rileyi , isolated from the corn earworm, Helicoverpa armigera , in Taiwan, to its host insect were studied in the laboratory. The fungus caused higher larval mortality at 20°C than at 30°C when 5 × 10⁶ conidia/ml were sprayed on the fourth instar. However, mortality of the fifth instar injected with 1 × 10³ conidia/larva was not significantly different when the inoculated larvae were incubated from 15 to 30°C. The fungal development in inoculated larvae was best at 20 and 25°C after shifting from 20°C to either lower or higher temperatures. The germination rate was higher at 20 and 25°C than at 30 or 35°C. Conidial germination was better on the wash-off of insect cuticle than on Sabouraud maltose agar with yeast extract. Sporulation on chill-dried cadavers was maximal at 95 or 100% relative humidity than at lower levels of relative humidity. The time required for sporulation was 2 days less at 100% than at 95% relative humidity. Although photoperiod did not affect fifth instar mortality caused by N. rileyi , the median lethal time (LT₅₀) values were shorter upon incubating under light than in darkness. Incubation of infected cadavers under 12 or 24 h light resulted in 20-fold more conidial production than under full darkness. Therefore, illumination is necessary for development of this isolate on insect cadavers.     

Effect of exposure method to Beauveria bassiana and conidia concentration on mortality, mycosis, and sporulation in cadavers of Chilo partellus (Lepidoptera: Pyralidae)

The effects of exposure methods, conidial concentrations, and temperature on mortality, mycosis and sporulation in second instar Chilo partellus cadavers infected by Beauveria bassiana was investigated in laboratory studies. Larvae directly sprayed with conidia, exposed to conidia-treated leaves, and dipped into conidial suspension resulted in high mortality (98-100%). The longest LT50 (3.5 days) and days to mortality (2.6 days) were observed in the treated-leaves exposure method. The shortest LT50 (1 day) and days to mortality (1 day) were recorded for the dipping method. With increasing conidial concentrations, there were decreasing LT50 and days to mortality. Larvae exposed to treated leaves and larvae directly sprayed with conidial suspensions produced high mycoses in cadavers. Exposure of larvae to treated-leaves resulted in high sporulation. At lower concentrations of conidia, both mycosis and sporulation in cadavers were high. The optimum temperature for mycosis was 20 and 15 degrees C for sporulation.     

Impact of sixteen chemical pesticides on conidial germination of two entomophthoralean fungi: Conidiobolus thromboides and Pandora nouryi

Effects of sixteen chemical pesticides on conidial germination of C. thromboides and P. nouryi were investigated at two concentrations (R and 0.2R, where R=the lowest manufacture's recommended concentration for field use). Of the fungicides tested, propamocarb hydrochloride did not significantly affect conidial germination of C. thromboides, but at R it completely inhibited conidial germination of P. nouryi. At 0.2R of procymidone, iprodione, haloxyfop-methyl and fenoxaprop-p-ethyl, conidia of C. thromboides had some germination after 24 h, although they entirely restricted conidial germination of the fungus at the concentration of R. Tribenuron-methyl, one herbicide, had no adverse effect on conidial germination of C. thromboides and P. nouryi after 6 h at the two concentrations, whereas the other seven herbicides were strongly antagonistic to P. nouryi, except for carfentrazone-ethyl at 0.2R.     

飞虱虫疠霉分生孢子在不同温湿度组合条件下的存活率*

用荧光染色法研究了不同温度（１０～３０℃）和湿度（５１～１００％ＲＨ）组合条件下飞虱虫疠 霉 Ｐａｎｄｏｒａ ｄｅｌｐｈａｃｉｓ分生孢子的存活率。在无放回抽样观察中,孢子弹射后第２４ ｈ的存活 率在５１％、７４％、８５％、９０％、９５％和１００％ＲＨ与不同温度的组合中分别为４２～８１％、６９～８９％、７０～ ９５％、 ６７～１００％、 ７６～１００％和 ５６～１００％。存放４个月后,不同温度下的孢子存活率在 ５１％、 ７４％、 ８５％和 ９０％ＲＨ下分别为 ５５～７４％、 ５２～８７％、 ３８～７３％和 １～６５％。而在≥９５％ ＲＨ下孢子存活率 ２４ｈ 后锐减,至第７ ｄ几乎全部失活。以上结果表明,弹射后的飞虱虫疠霉孢子在头２４ ｈ的存活 易受低湿影响,但存活下来的孢子能继续在低湿下存活较久;饱和或接近饱和的高湿度最不 利孢子长久存活。