金龟子绿僵菌小孢变种对筛胸梳爪叩甲幼虫致病力的生物测定

从筛胸梳爪叩甲Melanotus cribricollis僵死幼虫体内分离得到1株金龟子绿僵菌小孢变种Metarhizium anisopliae var.anisopliae菌株WP,首次采用拌土法对该菌寄生筛胸梳爪叩甲幼虫的致病力进行测定,并分析外界条件对绿僵菌致病力的影响。在WP菌株1×104~1×107孢子/g干土7个孢子浓度梯度下,随着浓度的提高,筛胸梳爪叩甲的校正死亡率从20%上升至100%。在15、20、25、30℃4个温度下,该菌对筛胸梳爪叩甲的致死率随着温度升高而上升。30℃时,筛胸梳爪叩甲死亡率达96.7%;当降至15℃,筛胸梳爪叩甲不会被寄生。在6%~18%土壤湿度条件下,土壤湿度的升高会提高WP菌株对筛胸梳爪叩甲的致死率,6%的土壤湿度不会导致该虫染病。     

复合微生物菌肥对苹果再植病害调控及对根围土壤真菌群落结构的影响

 为研究复合微生物菌肥对再植苹果幼树的影响,从再植土壤酶活性、真菌群落结构和功能的方面分析,探究复合微生物菌肥的作用机理。本研究设置了KMM和对照CK两个处理,再植苹果树于春秋两次分别施用复合微生物菌肥(KMM)和有机肥(CK)。通过土壤常规农业化学分析法和Illumma MiSeq高通量测序技术,系统研究了两个处理再植苹果树的生长、根围土壤酶活性以及真菌群落结构和功能的变化。试验结果表明,施用复合微生物菌肥显著提高了再植苹果幼树株高、干径、叶绿素、分枝个数和分枝长度;提高再植土壤酶活性(中性磷酸酶、脲酶、蔗糖酶和过氧化氢酶);降低再植土壤真菌群落丰富度,改变真菌群落结构。相对丰度高于1%的优势真菌属中有21个菌属与再植果树的5项生长参数具有相关性,并且在不同分类水平上具有一定的差异,缝壳菌属(Lophiostoma)、织球壳菌属(Plectosphaerella)、赤壳菌属(Nectria)和维希尼克氏属(Vishniacozyma)显著增加,丝孢菌属(Anguillospora)和假丝酵母菌属(Pseudeurotium) 显著降低。通过对测序片段功能水平的分析,在二级功能水平上显著降低了真菌群落氨基酸、核苷和核苷酸的生物合成、电子转移、发酵、呼吸、CTP生物合成嘧啶脱氧核糖核酸功能序列的丰度值,共6种功能类群均与土壤酶活性呈负相关。施用复合微生物菌肥,改变了真菌群落结构,减弱真菌群落相关代谢的功能,提高土壤酶活性,促进再植苹果幼树的生长。     

不同生长状态祁连圆柏针叶内生真菌群落结构与多样性分析

为掌握西北干旱地区祁连圆柏内生真菌群落结构以及多样性,探索内生真菌与祁连圆柏针叶生长状态的联系,通过高通量测序技术对健康和枯黄的圆柏针叶进行了分析。研究结果显示：6组样品测序共得到2,338,753条有效序列,683个操作分类单元(Operational Taxonomic Units, OTUs)。ACE、Chao1、Shannon指数表明发病组样品的物种丰富度与多样性略高于健康组。门分类水平上,两组样品的优势菌群均为子囊菌门,在属分类水平上,枝孢属Cladosporium是健康组样品中的优势属,球腔菌属Mycosphaerella是发病组样品中的优势属。在群落结构组成上,健康组样品中枝孢属Cladosporium、球腔菌属Mycosphaerella、丝状担子菌属Filobasidium相对丰度均高于发病组,组成占比也较为均匀,发病组样品中除Mycosphaerella之外其余属真菌占比均较低,没有明显的层次结构。Beta多样性显示6组样品大致分为两个分布区,各样品组间差异较小,两组样品差异较为明显。属分类水平上大多数属间呈正相关,负相关主要集中球腔菌属Mycosphaerella...     

氯虫苯甲酰胺与高效氯氟氰菊酯复配防治沟金针虫配方筛选与田间应用效果

为筛选对地下害虫沟金针虫Pleonomus canaliculatus具有增效作用的氯虫苯甲酰胺与高效氯氟氰菊酯最佳复配组合,并评价其田间应用效果。采用浸渍法测定氯虫苯甲酰胺与高效氯氟氰菊酯不同配比下对沟金针虫3龄幼虫的共毒因子和共毒系数,并选择最佳配比加工成制剂进行室内安全性试验和田间药效试验,评价该复配剂对玉米田沟金针虫的防治效果。结果显示,氯虫苯甲酰胺与高效氯氟氰菊酯以配比2∶5、4∶5和2∶1复配时的共毒因子分别为41.16、23.88和36.07,经细化配比后,以配比1∶3、2∶5、4∶5、1∶1、2∶1和3∶1复配时共毒系数分别为121.77、228.03、284.41、175.05、150.98和135.80,其中以4∶5复配时的增效作用最显著;按照此比例配制的15%氯虫苯甲酰胺·高效氯氟氰菊酯悬浮种衣剂,以该种衣剂0.5、1、1.5、2、2.5 g (a.i.)/kg剂量处理后在玉米出苗后24 d的防治效果分别为54.76%、59.52%、66.67%、71.43%和78.57%。氯虫苯甲酰胺与高效氯氟氰菊酯复配防治地下害虫金针虫幼虫增效作用明显,15%氯虫苯甲酰胺·高...     

不同寄主植物对美国白蛾肠道细菌群落的影响

为明确美国白蛾Hyphantria cunea在取食不同寄主植物后其肠道细菌群落的变化情况,采用高通量测序技术分析其肠道细菌16S rDNA的V3～V4区域,分析肠道细菌的多样性、物种组成和菌群结构差异,并对其生物学功能进行预测。结果显示,美国白蛾4龄幼虫在取食不同寄主植物后肠道细菌Shannon多样性指数出现明显差异,由高到低依次为取食寄主法国梧桐Platanus orientalis、柿Diospyros kaki、胡桃Juglans regia、杨树Populus×canadensis和杏Armeniaca vulgaris的群体。美国白蛾肠道优势菌门主要为变形菌门和厚壁菌门,而属分类水平上的物种在不同处理之间存在较大差异,在取食法国梧桐和柿树的美国白蛾肠道中Burkholderia-Caballeronia-Paraburkholderia的相对丰度较高,分别为63.16%和34.50%;取食杏树和杨树的美国白蛾肠道优势菌属分别是肠球菌属Enterococcus和肠杆菌属Enterobacter,相对丰度分别为87.58%和37.93%。不同寄主也导致美国白蛾肠道细菌群的组成结...     

云南省不同海拔区草地贪夜蛾幼虫肠道与粪便细菌多样性

为探明草地贪夜蛾Spodoptera frugiperda不同海拔种群的幼虫肠道及粪便细菌组成,采用分离培养法、形态特征及16S rDNA测序分析法对云南省5个海拔22个草地贪夜蛾种群的幼虫肠道及粪便细菌进行分离培养、鉴定。结果表明,从不同海拔种群的幼虫肠道和粪便中分别获得可培养细菌4门15科22属45种和4门13科18属36种,优势菌门均为变形菌门和厚壁菌门。不同海拔种群的幼虫肠道和粪便细菌组成均不同,500 m以下、1 001～1 500 m、1 501～2 000 m和2 000 m以上4个海拔种群的幼虫肠道细菌优势属和优势种分别为肠球菌属Enterococcus和铅黄肠球菌Enterococcus casseliflavus,500～1 000 m海拔种群的优势属和优势种分别为克雷伯氏菌属Klebsiella和变栖克雷伯氏菌Klebsiella variicola;500 m以下、500～1 000 m、1 001 m以上海拔种群的幼虫粪便细菌优势属分别为假单胞菌属Pseudomonas、克雷伯氏菌属和肠球菌属,优势种分别为韩国假单胞菌Pseudomonas koreensis...     

基于高通量测序技术检测草地贪夜蛾病毒多样性

为探索草地贪夜蛾Spodoptera frugiperda病毒的多样性,通过宏转录组和小RNA测序技术手段分析鉴定了自云南省采集的草地贪夜蛾田间自然种群携带的病毒类型。结果显示,宏转录组测序数据分析可比对得到14个病毒科,包括5个昆虫相关的病毒科,5个植物病毒科和4个细菌病毒科。昆虫相关的病毒分别属于杆状病毒科Baculoviridae、呼肠孤病毒科Reoviridae、分体RNA病毒科Partitiviridae、传染性软腐病病毒科Iflaviridae和弹状病毒科Rhabdoviridae,其中传染性软腐病病毒和分体RNA病毒为首次在草地贪夜蛾中报道。同时利用小RNA深度测序技术从草地贪夜蛾样品中检测到鳞翅目杆状病毒、分体RNA病毒和传染性软腐病病毒的序列,进一步验证了宏转录组的分析结果。表明利用高通量测序技术可初步揭示存在于草地贪夜蛾自然种群的昆虫病毒多样性。     

Changes in diversity,composition and assembly processes of soil microbial communities during Robinia pseudoacacia L. restoration on the Loess Plateau,China

Robinia pseudoacacia L. (RP) restoration has increased vegetation cover in semi-arid regions on the Loess Plateau of China, but ecological problems have also occurred due to RP restoration, such as reduced soil moisture. Further, it is still uncertain how microbial diversity, composition and assembly processes change with RP restoration in semi-arid regions. Therefore, amplicon sequencing of small subunit ribosomal ribonucleic acid (16S rRNA) and internal transcribed spacer (ITS) genes was performed to study soil bacterial and fungal diversity, composition and assembly processes at four study sites with different stand ages of RP plantations (Y10, RP plantation with stand ages less than 10 a; Y15, RP plantation with stand ages approximately 15 a; Y25, RP plantation with stand ages approximately 25 a; and Y40, RP plantation with stand ages approximately 40 a) along a 40-a chronosequence on the Loess Plateau. The diversity of soil bacteria and fungi increased significantly during the restoration period from 10 to 15 a (P<0.05). However, compared with Y15, bacterial diversity was lower at Y25 and Y40, and fungal diversity remained stable during the restoration period between 25 and 40 a. The relative abundances of Proteobacteria and Ascomycota increased during the restoration period from 10 to 15 a. Conversely, after 15 a of restoration, they both decreased, whereas the relative abundances of Actinomycetes, Acidobacteria and Basidiomycota gradually increased. The variations in soil bacterial communities were mainly related to changes in soil total nitrogen, nitrate nitrogen and moisture contents, while soil fungal communities were mainly shaped by soil organic carbon and nitrate nitrogen contents. Bacterial communities were structured by the heterogeneous selection and stochastic process, while fungal communities were structured primarily by the stochastic process. The RP restoration induced an increase in the relative importance of heterogeneous selection on bacterial communities. Overall, this study reveals the changes in microbial diversity, community composition and assembly processes with RP restoration on the Loess Plateau and provides a new perspective on the effects of vegetation restoration on soil microbial communities in semi-arid regions.     

Decomposition of different crop straws and variation in straw-associated microbial communities in a peach orchard,China

Crop residue is a major source of soil organic matter;therefore,application of crop straw to soil contributes to the sustainable development of organic agriculture.To better understand the transformation of crop straw in orchard soils,we investigated the relationship between the characteristics of straw decomposition and functional diversity of associated microbial communities in a long-term peach orchard,China.Mesh bags,each containing 30 g of corn or bean straw,were buried at a soil depth of 20 cm in a 12-year-old peach orchard for 360 d(October 2011–October 2012).Three treatments were applied,i.e.,fresh corn straw,fresh corn straw with nitrogen fertilizer(urea,10.34 g/kg),and fresh bean straw.Changes in straw residual rate,straw water content and soil conditions were monitored after treatment.The functional diversity of straw-associated microbial communities was analyzed by the Biolog-Eco microplate assay.During the decomposition process,straw residual rates did not vary considerably from 10 d(30.4%–45.4%)to 360 d(19.0%–30.3%).Irrespective of nitrogen addition,corn straw decomposed faster than bean straw.Corn straw with nitrogen fertilizer yielded the highest average well color development(AWCD)values(1.11–1.67),followed by corn straw(1.14–1.68)and bean straw(1.18–1.62).Although the AWCD values did not differ significantly among the three treatments,substantial differences occurred across various time periods of the decomposition process(P<0.01).In terms of carbon source utilization,the dominant microbial groups fed mainly on saccharides.Hard-to-decompose substances gradually accumulated in the middle and late stages of straw decomposition.Of the six categories of carbon sources tested,the utilization rate of aromatics was the lowest with corn straw,whereas that of polymers was the lowest with bean straw.Among different treatments,straw residual rate was negatively correlated to soil available phosphorous,soil available potassium and soil temperature(P<0.05),but not to soil water content.In some cases(corn straw with or without nitrogen fertilizer),straw residual rate was negatively correlated to straw water content,amino acid utilization and carboxylic acid utilization,and positively correlated with microbial species richness and evenness(P<0.05).Microbial community associated with corn and bean straw decomposition in soil was respectively dominated by aromatic-and polymer-metabolizing groups during the middle and late stages of this process,which could reduce the stability of microbial community structure and decrease the rate of straw decomposition in the fruit tree orchard.     

棉隆对辣椒疫霉病的防效及对土壤微生物群落的影响

为明确不同剂量棉隆对辣椒疫霉的防效及对土壤微生物群落的影响,在温室大棚条件下采用密封熏蒸法测定了不同剂量(300、450、600和750 kg/hm2)棉隆土壤熏蒸对土壤疫霉病菌的抑制率及田间防效,并采用Biolog法研究了其对土壤微生物功能多样性的影响。结果表明,不同剂量棉隆在熏蒸后揭膜当天(0)、80、140 d对辣椒疫霉病菌的抑制率分别为86.84%~100%、75.26%~96.37%和73.24%~95.46%;且80、140 d时对辣椒疫霉病的防效分别为77.19%~96.49%和70.00%~93.33%;在棉隆熏蒸揭膜当天各剂量处理下微生物对碳源的利用(用平均每孔颜色变化率表示)以及微生物多样性指数中的丰富度指数、均匀度指数及Mc Intosh指数均显著低于对照,而Simpson指数显著高于对照,随着试验时间的延长,各剂量对其影响逐渐减小,但高剂量尤其是750kg/hm2处理在140 d时仍与对照差异显著。表明棉隆剂量越高对辣椒疫霉病的防效越显著,对微生物活性影响越大,高剂量处理能明显抑制土壤微生物的活性,降低土壤微生物的多样性。     

Molecular characterization of Erwinia amylovora strains from different host plants through RFLP analysis and sequencing of hrpN and dspA/E genes

A total of 73 Erwinia amylovora strains obtained from 13 Maloideae host species and from Rubus spp., and isolated from different geographic areas, were assessed using RFLP and DNA sequencing analysis of the 3' hrp N gene and/or of a fragment of 1341 bp of the dsp A/E region. An Erwinia pyrifoliae strain, used as outgroup, was checked in the same way. For the three strains isolated from Rubus spp. and for one strain from Amelanchier sp., RFLP analysis of the hrp N gene using the Rsa I enzyme yielded a PCR product 60 bp smaller than that of all the other strains. Sequence analysis of the gene revealed this was due to the absence of a 60 bp fragment in the noncoding region downstream of the gene. The strain PD 2915, isolated from Amelanchier sp. grown in Canada, showed five same-sense substitutions and one missense substitution at position 868 of the hrp N gene, converting aspartic acid into asparagine. Also, restriction analysis of a fragment of 613 bp of the dsp A/E region with Cfo I revealed an RFLP pattern suitable for differentiating the E. amylovora strains isolated from Rubus spp. and Amelanchier sp. from all the others. In the dsp A/E coding region, the four strains showed 13–14 missense point mutations, in some cases yielding drastic amino acid substitutions. In addition, partial sequencing of the dsp A/E region of PD 2915 from Amelanchier sp. indicated a higher similarity to E. amylovora strains isolated from Rubus spp. than towards strains from other Maloideae hosts. The E. pyrifoliae strain showed 23 single nucleotide substitutions along the hrp N gene and 88% of nucleotide identity with E. amylovora strains in the portion of dsp A/E region. Artificial inoculations on immature pear fruits and young shoots of Maloideae and Ruboideae showed a restricted pathogenicity for the strains from Rubus and Amelanchier , with the latter inciting blight symptoms only on Amelanchier .     

基于高通量测序检测甜菜生尾孢中真菌病毒多样性

为明确甜菜生尾孢Cercospora beticola中真菌病毒的多样性,2020年自新疆维吾尔族自治区、内蒙古自治区、黑龙江省和北京市采集感染褐斑病甜菜病叶,鉴定获得78株甜菜生尾孢菌株,运用高通量的宏转录组测序技术检测甜菜生尾孢所携带的真菌病毒种类;将分离到的甜菜生尾孢按照菌落生长表型分为4组,进行宏转录组测序,将得到的序列经过拼接、聚类和基因功能注释,筛选出真菌病毒相关同源序列。序列分析结果显示,4组样品共比对到6个真菌病毒科,包括双分体病毒科Partitiviridae、双核糖核酸病毒科Birnaviridae、低毒病毒科Hypoviridae、裸露RNA病毒科Narnaviridae、泛欧尔密病毒科Botourmiaviridae和转座病毒科Metaviridae,以及未分科的负义单链RNA病毒。研究结果丰富了甜菜生尾孢真菌病毒资源库。