大别山的两种刺束梗孢

采自大别山的两种刺束梗孢,其中长孢刺束梗孢（Akanthomyceslongisporus）为新种,有拉丁文描述和图版。本种孢梗束多,密集,棕褐色,棒状。产孢结构和纤细刺束梗孢（A.gracilis相似,瓶梗以密集的子实层状被覆孢梗束。分生孢子多单细胞,少数双细胞,纺锤形至柱形,8.8～14.8×2．0～3.0μm。新几内亚刺束梗孢的标本同时并存刺状和棒状的孢梗束,这支持Samson将虫束梗孢属视为刺束梗孢属异名的观点。标本保存于安徽农业大学虫生真菌室（RCEF）     

莲状虫草及其刺束梗孢无性型

报道采自安徽省万佛山自然保护区的一种寄生在蜘蛛上的虫草标本,经鉴定为莲状虫草Cordycepsnelumboides。经多批次利用子囊孢子分离得到其刺束梗孢无性型Akanthomyces,属首次报道,并在人工固体培养基上得到了与野生标本一致的子实体,从丽确证了有性型与元性型之间的对应关系。经研究发现,KOBAYASI和SHIMIZU对莲状虫草次生子囊孢子的原始描述可能有误。     

青蛤Kazal型丝氨酸蛋白酶抑制剂基因的克隆及其在组织间的表达分析

利用构建的SMART-cDNA文库及高通量测序方法,获得了青蛤Kazal家族丝氨酸蛋白酶抑制剂基因(Serine Protease Inhibitor,SPI)的全长序列,采用荧光定量PCR方法分析了SPI在青蛤各组织的表达情况,并在鳗弧菌胁迫下分析了SPI在青蛤血液中的时序表达关系。结果表明,青蛤SPI基因全长587 bp,CDS为67~520 bp,编码151个氨基酸,具有19个氨基酸的信号肽序列;荧光定量PCR结果显示,该基因在血液、肝脏、外套膜、鳃和闭壳肌等组织中表达差异明显,其中血液中表达水平最高,在闭壳肌中表达水平较低,其他组织表达量极少;在鳗弧菌刺激后3 h和48 h青蛤血液中SPI的表达量出现明显上调的趋势且与对照组有极显著性差异(P0.01),说明SPI基因在青蛤的免疫反应中具有重要的作用。     

寄主为蚧的层束梗孢一新记录种

在贵州茂兰国家自然保护区发现寄生蚧壳虫的层束梗孢属真菌gzuifr-hym201307jk。主要特征为孢梗束灰白色单生,14¹7 mm×0.9 mm,产孢细胞柱状向上收缩变尖,顶部具有117 mm×0.9 mm,产孢细胞柱状向上收缩变尖,顶部具有1³个叉状的产孢小颈,大小10.83个叉状的产孢小颈,大小10.8¹4.0×4.314.0×4.3⁵.4μm;分生孢子梭形,7.65.4μm;分生孢子梭形,7.6¹0.8×4.310.8×4.3⁵.4μm。经鉴定该真菌为中国新记录种——Hymenostilbe furcata。     

对桃蚜高毒力的蜡蚧菌菌株筛选

【目的】桃蚜Myzus persicae是一种世界性的重要农作物害虫,近年来已对多种化学杀虫剂产生抗性。本研究旨在筛选对桃蚜具有高毒力且培养性状优良的蜡蚧菌菌株。【方法】选用蜡蚧菌属Lecanicillium 2个种的7个菌株,即刀孢蜡蚧菌L.psalliotae 3个菌株(HFLP006, HFLP021和HFLP025)和渐狭蜡蚧菌L.attenuatum 4个菌株(HFLA032, HFLA041, HFLA064和HFLA066),接种在SDAY平板上培养并观察菌落生长与产孢性状;以2.0×10⁷孢子/mL悬浮液用浸渍法测定对桃蚜无翅成蚜的致死率和致死中时(LT₅₀);进而用系列浓度(1.0×10⁴-1.0×10⁸孢子/mL) HFLP006和HFLA032菌株孢子悬浮液测定其对桃蚜无翅成蚜的致死中浓度(LC₅₀);利用体视显微镜逐日观察1.0×10⁸孢子/mL HFLP006菌株孢子悬浮液侵染后无翅成蚜的染病症状变化。【结果】在SDAY平板上7个...     

蜡蚧轮枝菌和球孢白僵菌的共发酵

【目的】球孢白僵菌(Beauveria bassiana)和蜡蚧轮枝菌(Verticillium lecanii)是国内外目前研究应用最广泛的杀虫生防真菌,欲扩大其防治范围、增强防治效果、降低生防成本。【方法】采用共发酵技术,通过组合菌株产孢能力和杀虫毒力比较试验,确定蜡蚧轮枝菌和球孢白僵菌共发酵的可行性。【结果】蜡蚧轮枝菌L-31和球孢白僵菌Q-55共发酵的最佳配比为1:1时,按10%总量接种于发酵培养液中(培养液按酵母膏5.0 g/L、葡萄糖20.0 g/L、麦芽糖提取物5.0 g/L、KH2PO43.0 g/L、黄小米200.0 g/L,pH 6.5配制),23.0°C±0.1°C恒温静置发酵12 d,共发酵液的含孢量可达1×109CFU/mL以上,杀虫毒力比较强,其对温室白粉虱和菜青虫可同时显效,处理9 d后的致死中浓度LC50分别为2.09×104±0.12 CFU/mL和3.17×105±0.11 CFU/mL,发酵液浓度为1×108CFU/mL时的致死中时间LT50分别为2.11±0.14 d和4.27±0.43 d,温室小区试验校正防效在80%以上,与其各单一菌株发酵液的防效之间存在显著性差异。【结论】通过两株生防真菌的共发酵研究,为杀虫真菌的扩谱增效以及植物害虫的有效防治提供科学依据和有效途径。     

叉角厉蝽丝氨酸蛋白酶及抑制剂基因的克隆与表达分析

为明确叉角厉蝽Eocanthecona furcellata (Wolff)丝氨酸蛋白酶基因EfSP1及抑制剂基因EfSPI20的基因序列特征和时空转录特征,为其生理功能研究奠定基础。利用PCR克隆技术获得叉角厉蝽唾液腺EfSPI20和EfSP1的完整开放阅读框(Open reading frame, ORF)序列,使用生物信息学软件进行序列分析以及系统进化分析,采用实时荧光定量PCR (Real time quantitativate PCR,RT-qPCR)分析两个基因分别在叉角厉蝽不同发育时期和组织中的表达特征。结果表明,EfSPI20与EfSP1基因完整开放阅读框长度分别为378 bp和921 bp,分别编码125个氨基酸和306个氨基酸,预测均为亲水蛋白质,理论分子量分别为13.48 kDa和33.82 kDa,等电点分别为6.68和5.80,分别有30个和23个氨基酸残基的信号肽序列,EfSPI20有跨膜结构域,EfSP1无跨膜结构域。序列比对显示叉角厉蝽EfSPI20与茶翅蝽Halyomorpha halys PPI同源性最高,氨基酸序列一致性达58%;EfSP1与稻绿蝽Nezara viridula SP同源性最高,氨基酸序列一致性达66%;系统发育树显示叉角厉蝽与同为蝽科的茶翅蝽和稻绿蝽物种亲缘关系近。EfSPI20基因在雌雄成虫和唾液腺中高表达,推测EfSPI20可能具有抑制胰蛋白酶活性的功能和与叉角厉蝽的捕食消化相关;EfSP1基因在卵期、卵巢和肠道中高表达,推测EfSP1可能与叉角厉蝽的生殖功能和蛋白消化相关。     

蝉棒束孢菌孢梗束培植过程中主要成分含量的变化规律及其复方对小鼠睡眠的影响

分析检测蝉棒束孢菌株BAI0101在培植过程中不同时间点孢梗束的转化率和孢梗束中多糖、虫草酸、腺苷和N ⁶-(2-羟乙基)腺苷含量的变化规律,对配伍的孢梗束复方辅助改善睡眠功能进行了初步评价。结果表明,从培植18d起,孢梗束的转化率呈现上升趋势,在培植30d左右,有保持稳定趋势;孢梗束中的多糖、虫草酸和腺苷含量均随着培植时间的延长呈现增高趋势,均在22-24d达到最高值,其后开始呈现降低趋势,而孢梗束中的N ⁶-(2-羟乙基)腺苷含量随着培植时间的延长呈现降低趋势,在24-26d左右达到最低值,其后呈现上升趋势,在培植至36d左右达到含量的最高值并保持稳定;对利用该菌株培植的孢梗束,经中药配伍组成的保健品小复方进行了辅助改善睡眠保健功能的初步评价,结果表明该复方有明显促进阈下剂量戊巴比妥钠诱导的小鼠睡眠作用,能明显降低巴比妥钠诱导小鼠睡眠潜伏期,且无直接睡眠作用,不影响戊巴比妥致小鼠睡眠时间,根据《保健食品检验与评价技术规范》,判定该复方具有辅助改善睡眠的保健功能。     

昆虫病原真菌长孢蜡蚧菌TF-2菌株对豌豆蚜的侵染过程和致病力

【目的】豌豆蚜Acyrthosiphon pisum是世界性的重要农业害虫,给豆类作物造成巨大的经济损失。本研究在前期筛选已获得豌豆蚜致病菌长孢蜡蚧菌Lecanicillium longisporum TF-2菌株的基础上,检测了该致病菌分生孢子对豌豆蚜的毒力效用及侵染方式,以期为利用昆虫病原真菌防治豌豆蚜提供理论基础。【方法】采用离体叶片饲养法检测在不同浓度长孢蜡蚧菌TF-2菌株孢子悬浮液(1.0×10³～1.0×10⁷孢子/mL)中浸渍后对豌豆蚜成虫的致病力;应用扫描电镜和体视显微镜观察豌豆蚜成虫接种TF-2菌株(1.0×10⁷孢子/mL)后长孢蜡蚧菌TF-2菌株侵染症状和侵染过程。【结果】不同浓度长孢蜡蚧菌TF-2菌株分生孢子浸染豌豆蚜成虫致病力随分生孢子浓度升高而逐渐增强,侵染后6 d对豌豆蚜成虫的半致死浓度(LC₅₀)为3.26×10⁴孢子/mL,最高浓度分生孢子(1.0×10⁷孢子/mL)对豌豆蚜成虫的半致死时间(LT₅₀...     

影响蜡蚧轮枝菌发酵产孢量和孢子活力的基本因素分析

不同接种浓度、发酵时间、菌株和基质对蜡蚧轮枝菌固体发酵的产孢量都有显著影响,而对孢子活力的影响不明显,萌发率均在90%以上.在5至7天内,随发酵时间的延长产孢量有明显增长;7至10天则变化不大;15天时产孢量和孢子活力均显著降低.随接种孢子浓度每增加1倍,菌株(VLFNL95-01)发酵的产孢量平均提高15.6%,浓度加大至32倍则提高71.9%.3个菌株比较,VLFNL95-01产孢量明显高得多.不同固体基质发酵的结果有显著差异,产孢量最高的(正交3号配方)比其次的(黄豆饼粉+玉米碎粒+磷酸盐)就提高了95.1%.     

A morphological study on autumnal morphs of Aphis glycines (Hemiptera: Aphididae)

The soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), is one of the most important pests of soybean. The complex life cycle of A. glycines is characterized as heteroecious and holocyclic, and has seasonal polymorphisms occurring during its life cycle. In the autumn, A. glycines occur as winged and wingless virginoparae in soybeans, as gynoparae and males that migrate from soybeans to Rhamnus spp. and as gynoparae, males and oviparae on Rhamnus spp. In this study, wingless virginoparae, gynoparae, males and oviparae of A. glycines were successfully induced in the laboratory and morphological parameters of these morphs were selected for quantitation. To aid in identification of these A. glycines autumnal morphs, these aphids were imaged by microscope and significant differences in morphological characteristics were found: distal parts of hind leg femurs of gynoparae were grayish black, which were darker than those of winged virginoparae; the 4th instars of gynoparae, males and winged virginoparae differ greatly in dorsal abdomen coloration and covering. Our results provide an important guide for distinguishing adults of gynoparae and winged virginoparae, and for identifying nymphs of gynoparae, males and winged virginoparae.     

Leaf temperature of soybean grown under elevated CO2 increases Aphis glycines (Hemiptera: Aphididae) population growth

Abstract Plants grown under elevated carbon dioxide (CO₂) experience physiological changes that influence their suitability as food for insects. To determine the effects of living on soybean (Glycine max Linnaeus) grown under elevated CO₂, population growth of the soybean aphid (Aphis glycines Matsumura) was determined at the SoyFACE research site at the University of Illinois, Urbana‐Champaign, Illinois, USA, grown under elevated (550 μL/L) and ambient (370 μL/L) levels of CO₂. Growth of aphid populations under elevated CO₂ was significantly greater after 1 week, with populations attaining twice the size of those on plants grown under ambient levels of CO₂. Soybean leaves grown under elevated levels of CO₂ were previously demonstrated at SoyFACE to have increased leaf temperature caused by reduced stomatal conductance. To separate the increased leaf temperature from other effects of elevated CO₂, air temperature was lowered while the CO₂ level was increased, which lowered overall leaf temperatures to those measured for leaves grown under ambient levels of CO₂. Aphid population growth on plants grown under elevated CO₂ and reduced air temperature was not significantly greater than on plants grown under ambient levels of CO₂. By increasing Glycine max leaf temperature, elevated CO₂ may increase populations of Aphis glycines and their impact on crop productivity.     

Identification and expression profiles analysis of odorant‐binding proteins in soybean aphid,Aphis glycines (Hemiptera: Aphididae)

The soybean aphid, Aphis glycines, is an extreme specialist and an important invasive pest that relies on olfaction for behaviors such as feeding, mating, and foraging. Odorant‐binding proteins (OBPs) play a vital role in olfaction by binding to volatile compounds and by regulating insect sensing of the environment. In this work we used rapid amplification of complementary DNA ends technology to identify and characterize 10 genes encoding A. glycines OBPs (AglyOBPs) belonging to 3 subfamilies, including 4 classic OBPs, 5 Plus‐C OBPs, and one Minus‐C OBP. Quantitative real‐time polymerase chain reaction demonstrated variable specific expression patterns for the 10 genes based on developmental stage and aphid tissue sampled. Expression levels of 7 AglyOBPs (2, 3, 4, 5, 7, 9, and 10) were highest in the 4th instar, indicating that the 4th nymphal instar is an important developmental period during which soybean aphids regulate feeding and search for host plants. Tissue‐specific expression results demonstrated that AglyOBP2, 7, and 9 exhibited significantly higher expression levels in antennae. Meanwhile, ligand‐binding analysis of 5 OBPs demonstrated binding of AglyOBP2 and AglyOBP3 to a broad spectrum of volatiles released by green leaf plants, with bias toward 6‐ to 8‐carbon chain volatiles and strong binding of AglyOBP7 to trans‐β‐farnesene. Taken together, our findings build a foundation of knowledge for use in the study of molecular olfaction mechanisms and provide insights to guide future soybean aphid research.     

有翅型和无翅型大豆蚜卵巢和胚胎发育的差异

【目的】比较大豆蚜Aphis glycines有翅型和无翅型个体的卵巢发育,为从生理生化角度研究蚜虫翅型分化发生的繁殖适应性提供依据。【方法】在卵巢形态显微镜观察基础上,比较有翅型和无翅型大豆蚜卵巢小管数量、胚胎总数量、成熟胚胎数量以及卵巢小管中最大胚胎体积及其发育速率。【结果】有翅型和无翅型大豆蚜的3龄和4龄若蚜以及成蚜均有2个卵巢/雌,大多数蚜虫每个卵巢包含4条卵巢小管,可见一个卵巢包含5条卵巢小管的现象。有翅型和无翅型个体卵巢中胚胎总数量和成熟胚胎数量以及最大胚胎体积随发育时间增加均显著增大,但有翅型个体胚胎总数量和成熟胚胎数量以及最大胚胎体积均显著小于同一发育阶段无翅型个体。有翅型和无翅型的3龄和4龄若蚜以及成蚜卵巢中胚胎总数量与后足胫节长度均呈显著正相关关系。有翅型和无翅型大豆蚜卵巢小管中最大胚胎体积随发育时间均呈指数增长,但有翅型个体最大胚胎体积显著小于无翅型个体。【结论】有翅型大豆蚜相比于无翅型个体存在卵巢发育延迟现象,卵巢发育程度的差异可能与翅的发育相关。     

Inhibition of kallikrein-related peptidases by the serine protease inhibitor of Kazal-type 6

Kallikrein-related peptidases (KLKs) are a group of serine proteases, expressed in several tissues. Their activity is regulated by inhibitors including members of the serine protease of Kazal-type (SPINK) family. Recently, we discovered that SPINK6 is expressed in human skin and inhibits KLK5, KLK7, KLK14 but not KLK8. In this study we tested whether SPINK6 inhibits other members of the KLK family and caspase-14. Using chromogenic substrates, SPINK6 exhibited inhibitory activity against KLK12 and KLK13 with K_i around 1 nM, KLK4 with K_i = 27.3 nM, KLK6 with K_i = 140 nM, caspase-14 with a K_i approximating 1 μM and no activity against KLK1, KLK3 and KLK11. Taken together, SPINK6 is a potent inhibitor of distinct KLKs members.     

热应激和吡虫啉对大豆蚜hsp70和hsc70基因mRNA表达的影响

【目的】昆虫在高温或农药的胁迫下,通过高效表达热休克蛋白(HSP)等建立应激自我保护机制。本研究为从转录组水平上认识大豆蚜Aphis glycines在热应激和吡虫啉胁迫下hsp70和hsc70 mRNA表达分子机制,进而寻找自我保护应激反应中的薄弱环节,为大豆蚜的生物防治提供理论基础。【方法】采用同源克隆、RACE技术和实时荧光定量PCR等方法研究不同热激时间和热激后不同恢复时间及不同吡虫啉浓度对大豆蚜4龄若虫hsp70和hsc70的表达影响。【结果】37℃热激后,大豆蚜4龄若虫中hsp70表达量先上调,1 h时升至对照组的10.36倍（P<0.05）,然后逐渐下降。同样热激后恢复时间的长短对大豆蚜若蚜中hsp70的表达具有显著影响。热激处理后,大豆蚜若蚜中hsp70立即大量表达,表达量为对照组的8.78倍（P<0.05）,随后表达量下降至对照组水平,而hsc70的表达量并没有显著变化（P>0.05）。大豆蚜若蚜受吡虫啉的胁迫时,其hsp70和hsc70的表达量受吡虫啉的浓度及胁迫的时间的影响,呈现先升高后下降的趋势,具有明显的短期效应。【结论】吡虫啉诱导大豆蚜hsp70和hsc70表达量的上调;而热胁迫对hsp70和hsc70 mRNA具有不同的表达模式,高温可以诱导hsp70的表达,但对hsc70没有明显的诱导作用。     

基于MaxEnt模型的大豆蚜全球潜在地理分布分析

【目的】为预测和分析大豆蚜Aphis glycines的全球潜在地理分布,研究大豆蚜分布与环境变量之间的联系。【方法】利用最大熵法生态位模型(maximum entropy niche-based modeling, MaxEnt)和地理信息系统软件ArcGIS,根据收集的大豆蚜已知分布点和环境变量,预测大豆蚜的全球潜在地理分布区,推测环境变量对大豆蚜分布的影响。【结果】结果表明,大豆蚜适生区主要分布在低海拔地区,高 度适生区集中在25°~50°N的中国、日本、韩国、朝鲜、加拿大、美国、意大利和格鲁吉亚。决定大豆蚜分布地点的关键环境变量为最暖季度降水量、最暖季度平均温度、最湿季度平均温度、最干月降水量、月平均昼夜温差和温度季节性变化标准差。【结论】大豆蚜潜在地理分布区域广泛,应在各国大豆农产品贸易时做好检验检疫工作,以防止大豆蚜的扩散。     

大豆蚜寄生蜂豆柄瘤蚜茧蜂室内生物学特性观察

豆柄瘤蚜茧蜂Lysiphlebus fabarum( Marshall)是大豆蚜Aphis glycines Matsumura的一种重要寄生蜂.在实验室条件下,对豆柄瘤蚜茧蜂的发育历期、繁殖力、寿命和性比等指标,以及成蜂的羽化、取食、产卵行为等生物学特性进行了观察描述.结果表明:在19 ～31℃范围内,发育历期随着温...     

室内人工诱导大豆蚜产生有性世代的研究

【目的】大豆蚜Aphis glycines(Matsumura)是大豆上最重要的害虫之一,短暂的有性阶段是蚜虫种群基因交流的关键环节。通过探索影响大豆蚜产生有性世代的室内人工诱导条件,对研究种群遗传和生态适应机制具有重要意义。【方法】采用豆苗罩笼培养法,在实验室条件下用一定的光温组合处理,测定了处理1代和连续处理3代的大豆蚜各蚜型产生的比率,并且分析了各相关因素的影响。【结果】本试验成功诱导大豆蚜产生无翅孤雌蚜、有翅孤雌蚜、有翅产雄性母和有翅雄性蚜。结果表明,人工诱导雄性母产生的最适条件为15℃,10L︰14D处理3代,雄性蚜产生的最适条件为12℃,10L︰14D处理3代,而15℃,8L︰16D条件下大豆蚜翅型分化率最高。对各相关因子进行方差分析结果表明,温度与光周期互作对有性世代的产生影响最为显著,其次是温度、诱导日龄的单独作用,诱导世代只有和光温条件互作才有显著影响。【结论】在特定的光周期内,温度是大豆蚜有性世代产生的重要因素,同时光温条件组合对此有一定的累积效应,表现为光、温、诱导世代三者交互对有性世代的产生影响显著。