泰安市“香野”草莓采后主要病原菌的分离鉴定及精油抑菌效果评价

为明确山东省泰安市草莓采后主要致腐病原菌的种类及不同植物精油的抑菌效果,从腐败的"香野"草莓果实上分离、纯化优势菌株,进行形态学和分子生物学鉴定,并通过室内熏蒸毒力试验评价7种植物精油对上述病原菌的抑制效果。结果表明,泰安地区草莓分离得到3种病原菌,分别为灰葡萄孢菌Botrytis cinerea、链格孢菌Alternaria alternata和枝孢菌Cladosporium tenuissimum。7种植物精油对3种病原菌均表现出不同程度的抑制作用,百里香精油对灰葡萄菌和链格孢菌的室内毒力均最高,EC₅₀值分别为39.998和18.456 μL/L;柠檬草精油对枝孢菌的活性最高,EC₅₀值为30.523 μL/L。活体抑菌试验结果表明,百里香精油对草莓果实灰葡萄孢菌和链格孢菌的活体抑菌效果最高,防效分别为86.21%和89.29%;柠檬草精油对枝孢菌的防效最高,为75.86%。综合比较可以发现,百里香精油、玫瑰草精油、柠檬草精油及肉桂精油对草莓采后3种病原菌均具有较好的抑制效果。     

野生药用植物内生真菌的分离及拮抗菌株筛选

为挖掘生防新资源,采用组织分离法从21种野生药用植物不同组织部位分离纯化内生真菌,以6种植物病原菌为靶标菌筛选拮抗菌株,并根据形态学及分子生物学对菌株进行鉴定,在此基础上研究了高效拮抗菌株对靶标菌菌丝生长及孢子萌发的影响。结果表明:从分离纯化的478株内生真菌中筛选出11株高活性拮抗菌株,分属于青霉属Penicillium、平脐蠕孢属Bipolaris、棘壳孢属Pyrenochaeta、镰孢属Fusarium、粒毛盘菌属Lachnum、垫壳孢属Coniella和Neonectria 7个属,其中青霉属占比最高,达36.36%。平皿对峙试验表明, 棘壳孢菌12-R-5对灰葡萄孢表现出高效性和专一性,其对菌丝抑制率达66.67%。含药平皿试验表明,菌株12-R-5发酵滤液10倍稀释液对灰葡萄孢菌丝生长抑制率达100%,与其他处理差异显著。菌株12-R-5 10倍发酵滤液处理灰葡萄孢10 h,孢子不萌发;100倍稀释液处理对孢子萌发抑制率为91.01%。显微镜观察发现,棘壳孢菌使灰葡萄孢菌丝扭曲变形,膨大肿胀及断裂,部分菌丝发生抑缩、消融现象。说明棘壳孢菌12-R-5菌株具有很好的生防潜力及应用前景。     

洛阳市牡丹灰霉病病原菌的鉴定

为明确洛阳市牡丹灰霉病的病原菌种类,于2015年从该地区的4个牡丹Paeonia suffruticosa Andr.种植区采集灰霉病样品,采用组织分离法对病原菌进行分离和纯化,结合形态学和基因序列分析对分离物进行鉴定,采用针刺接种法测定不同分离物对牡丹离体叶片的致病性。结果表明,从灰霉病样品中分离获得40株分离物,这些分离物分别属于Ⅰ、Ⅱ、Ⅲ型菌,I型菌不易产孢但产菌核;Ⅱ型菌易产孢,后期产生少量菌核;Ⅲ型菌易产孢,且产生大量菌核。产孢分离物均形成直立状分生孢子梗,孢子梗分枝顶端聚生葡萄穗状分生孢子,分生孢子卵圆形或长卵圆形。供试菌株的ITS序列和G3PDH基因序列与灰葡萄孢Botrytis cinerea Pers.的同源性达到99%;致病性测定结果表明,各菌群代表菌株对牡丹均有致病性,但不同菌群间致病力有差异。研究表明,引起洛阳市牡丹灰霉病的病原菌为灰葡萄孢,且菌群类型多样。     

新疆葡萄枝孢果腐病病原菌的鉴定及生物学特性

2019年在新疆葡萄上发现一种新病害,主要危害成熟葡萄果实。感病果实病部产生橄榄绿色霉层,导致葡萄产量和品质严重下降。为明确该病害病原菌种类及其生物学特性,从南北疆11个葡萄园采集80个病样,用常规稀释分离法和单孢分离法对病原菌进行分离和纯化,根据采集地点、葡萄品种、菌落生长速度等特征选取21个代表性菌株进行形态学鉴定、分子生物学鉴定和致病性测定以明确病原菌种类;并研究了温度、光照、培养基、pH对病原菌生长及产孢的影响。结果表明,引起新疆葡萄果腐病的病原菌为枝孢菌,有两个种,即枝状枝孢Cladosporium cladosporioides和柠檬形枝孢Cladosporium limoniforme,其中枝状枝孢为优势种。两种病原菌的适宜生长和产孢的温度均为25℃,适宜生长和产孢的培养基是PDA,不同光照条件下都可大量产生分生孢子,最适宜生长和产孢的pH为7。明确了新疆葡萄枝孢果腐病病原菌及其生物学特性,为今后该病害的预测预报、发生规律和防治方法的研究打下基础。     

淡紫紫孢菌防治采后番茄果实灰霉病

本文通过孢子萌发检测、平板生长抑制法和果实病害防治试验,测定了淡紫紫孢菌Purpureocillium lilacinus PL36-1防治储藏期番茄果实灰霉病的效果。结果表明,菌株PL36-1发酵滤液不仅能够抑制灰葡萄孢分生孢子萌发和芽管伸长,而且可以抑制灰葡萄孢菌丝生长。对峙培养的菌株PL36-1显著抑制灰葡萄孢的菌丝生长,菌丝生长抑制率可达到74.45%,菌株PL36-1产生的挥发性物质也对灰葡萄孢菌丝生长有一定的抑制作用。菌株PL36-1发酵液处理樱桃番茄果实后,能有效控制番茄果实灰霉病的发生,含有2×10~8孢子/m L的发酵液10 d的防治效果达70.64%。     

枯草芽胞杆菌NCD-2菌株抑菌蛋白的分离及鉴定

枯草芽胞杆菌NCD-2菌株对灰葡萄孢菌具有较强的抑菌活性,抑菌蛋白是其产生的抑菌物质之一。本研究为明确NCD-2菌株所产抑菌蛋白的作用方式和特性,采用牛津杯法测定抑菌蛋白的抑菌活性及其对病原菌菌丝生长的影响,采用混合培养法测定其对病原菌分生孢子萌发的影响,同时采用阴离子交换层析和非变性聚丙烯酰胺凝胶电泳技术对其进行分离纯化,并利用MALDI-TOF-MS进行鉴定。结果表明,NCD-2菌株产生的粗蛋白能够显著抑制灰葡萄孢菌分生孢子的萌发和菌丝生长,并导致病原菌菌丝分枝增多、局部膨大肿胀。通过分离纯化获得具抑菌活性的蛋白组分D1-3,经鉴定该蛋白的肽指纹图谱与leucyl aminopeptidase[Bacillus subtilis](WP_041057629.1)的氨基酸序列匹配率最高,达到65%,相对分子质量为53.936 k Da,功能分析表明组分D1-3可能是一种新的抑菌蛋白。     

两种蚕豆赤斑病菌对常用杀菌剂的抗药性检测及比较

对采自浙江、湖北和安徽3省的蚕豆赤斑病样品进行了病原菌的分离和鉴定,采用菌丝生长速率法检测了引起赤斑病的2种病原菌——蚕豆葡萄孢Botrytis fabae和灰葡萄孢B.cinerea 的抗药性发生情况,并在离体条件下通过抗药性诱导试验比较了二者的抗药性风险。结果共分离得到153个菌株,其中蚕豆葡萄孢122株(占79.7%),灰葡萄孢31株(占20.3%)。共检测到37株多菌灵高水平抗药性菌株(其中蚕豆葡萄孢9株)和42株异菌脲低水平抗药性菌株(其中蚕豆葡萄孢17株);嘧霉胺对153个菌株的EC50值在0.01~5.13 μg/mL之间,平均EC50值为0.72±0.15 μg /mL;表明蚕豆赤斑病菌对常见杀菌剂已表现出一定的抗药性,且灰葡萄孢的抗药性问题比蚕豆葡萄孢要严重得多。抗药性诱导试验进一步证实,灰葡萄孢的抗药性风险明显高于蚕豆葡萄孢。     

棉花枯萎病拮抗短小芽胞杆菌筛选鉴定及生防研究

为获得对棉花枯萎病有较好生防效果的拮抗细菌，从健康海岛棉植株根围土壤中分离筛选棉花枯萎病拮抗细菌，探索研究拮抗菌株的抑菌作用，为其生物防治提供潜在资源菌。采用平板对峙法，以海岛棉枯萎病尖孢镰刀菌为靶标菌，从土壤中分离到的120株细菌菌株中筛选拮抗菌株。测定拮抗细菌发酵液抑菌活性，通过促芽和盆栽试验筛选生防效果最好的菌株，同时测定该菌株对棉花枯萎病的抑菌效果和耐盐碱性。基于形态特征、生理生化特性和16S rDNA序列分析鉴定菌株分类地位。从120株细菌中筛选到1株对棉花枯萎病拮抗作用很强的菌株，编号为KX-33。促芽分析和盆栽试验结果表明，菌株KX-33能明显缩短出芽时间，促进棉苗生长。对病原菌DD64、DD89、DD11和DD22防效分别为75.32%、72.77%、69.48%和68.81%，均显著高于对照药剂处理（P<0.05）。耐盐碱分析表明，菌株KX-33具一定的耐盐碱性。菌株KX-33镜检为革兰氏阳性菌、呈杆状、有芽胞，16S rDNA和序列与Bacillus pumilus（FJ763643.1）同源性最高。海岛棉根围土壤微生物中含有棉花枯萎病拮抗细菌，经鉴定菌株KX-33为短小芽胞杆菌Bacillus pumilus，促生和抑菌作用显著，在海岛棉枯萎病生物防治中具有潜在的应用价值。     

水稻纹枯病生防细菌筛选及其与病原菌侵染垫形成的关系

为筛选对立枯丝核菌Rhizoctonia solani具有拮抗作用的生防细菌,利用稀释涂布平板法,从吉林省水稻纹枯病样中分离筛选出对立枯丝核菌AG-1A具有高拮抗活性的生防菌株,通过gyrB基因序列分析鉴定其分类地位,并采用离体侵染试验和温室防治试验测定筛选生防菌株对水稻纹枯病的防效,在显微镜下观察生防菌株预处理后接种病原菌的水稻叶片表面的菌丝生长情况,分析其生防机理。结果显示,从水稻纹枯病样中共分离获得35株菌株,其中菌株ND11对立枯丝核菌AG-1A的抑制率和抑菌圈直径最大,分别为74.12%和31.50 mm。基于gyrB基因序列分析最终将菌株 ND11鉴定为短小芽胞杆菌Bacillus pumilus;该菌株能够减缓立枯丝核菌AG-1A的侵染速度,抑制其侵染垫的形成,对水稻纹枯病的温室防效达70.69%以上。表明短小芽胞杆菌菌株ND11能够通过抑制立枯丝核菌侵染垫的形成来防治水稻纹枯病,且防效较好,具有开发为生防菌剂的潜力。     

隐球酵母对长枣果实的生防效果及诱导抗性

隐球酵母（Cryptococcus sp.）C331菌株是从灵武长枣果实表面分离筛选得到的一种拮抗酵母菌。接种优势致病菌黑根霉(Rhizopus sp.)后，研究拮抗菌和致病菌诱导果实自身抗真菌性能的影响。通过离体抑菌法和活体抑菌法，以及PPO、POD和SOD酶活测定，用EXCEL和SAS软件分析数据。结果表明,隐球酵母C331菌株能有效抑制长枣采后病害的发病率和病斑直径，其拮抗效果与拮抗菌的使用浓度成正比；拮抗菌能显著地提高PPO和SOD活性，降低POD活性。研究证明隐球酵母C331菌株能显著抑制长枣采后病害，提高果实自身抗真菌性能。     

Hymenopteran Parasitoids on Fruit-infesting Tephritidae (Diptera) in Latin America and the Southern United States: Diversity, Distribution, Taxonomic Status and their use in Fruit Fly Biological Control

We first discuss the diversity of fruit fly (Diptera: Tephritidae) parasitoids (Hymenoptera) of the Neotropics. Even though the emphasis is on Anastrepha parasitoids, we also review all the information available on parasitoids attacking flies in the genera Ceratitis, Rhagoletis, Rhagoletotrypeta, Toxotrypana and Zonosemata. We center our analysis in parasitoid guilds, parasitoid assemblage size and fly host profiles. We also discuss distribution patterns and the taxonomic status of all known Anastrepha parasitoids. We follow by providing a historical overview of biological control of pestiferous tephritids in Latin American and Florida (U.S.A.) and by analyzing the success or failure of classical and augmentative biological control programs implemented to date in these regions. We also discuss the lack of success of introductions of exotic fruit fly parasitoids in various Latin American countries. We finish by discussing the most pressing needs related to fruit fly biological control (classical, augmentative, and conservation modalities) in areas of the Neotropics where fruit fly populations severely restrict the development of commercial fruit growing. We also address the need for much more intensive research on the bioecology of native fruit fly parasitoids.     

Brief Guide for Authors

正Journal of Arid Land(JAL)is an international journal(ISSN 1674-6767;CN 65-1278/K)for the natural sciences,sponsored by the Xinjiang Institute of Ecology and Geography,Chinese Academy of Sciences and Science Press.It is published by Science Press and Springer-Verlag Berlin Heidelberg bimonthly.JAL publishes original,innovative,and integrative research from arid and semiarid regions,ad     

The parasitoid complex ofLiriomyza cicerina on chickpea (Cicer arietinum)

Liriomyza cicerina (Diptera: Agromyzidae) is an important pest on chickpea in Turkey. The objective of this study was to determine the parasitoids and rates of parasitism ofL. cicerina on chickpea (Cicer arietinum L.) during the 2005 and 2006 seasons in ?anl?urfa province, Turkey. Leaves with mines were sampled weekly and kept in the laboratory to observe and count emerging leafminer and parasitoid adults. Eight parasitoid species were collected: the braconidsOpius monilicornis Fischer andOpius tersus Foerster and the eulophidsDiaulinopsis arenaria (Erdös) andNeochrysocharis formosa (Westwood), which occurred in both the winter and summer seasons;Diglyphus crassinervis Erdös,Neochrysocharis ambitiosa Hansson,Neochrysocharis sericea (Erdös) andPediobius metallicus (Nees), which occurred only in the summer growing areas.Diaulinopsis arenaria was the predominant parasitoid with 4–7.7% parasitism rate whileN. ambitiosa andO. monilicornis were the second and third most predominant species. The results of these trials show that sinceDia. arenaria occurred throughout every season, it could potentially be used for control of the leafminerL. cicerina.     

Plant–herbivore asynchrony necessitates augmentative releases of the exotic beetle,Zygogramma bicolorata,to enhance the biological control of P arthenium hysterophorus

Systematic information on the quantitative impact of Z ygogramma bicolorata on the biology of P arthenium hysterophorus is crucial as the seeds of this weed continue to germinate from the accumulated soil seed bank throughout the year in the form of different germinating flushes, while the activity of the beetle ceases during winter as it enters diapause. Therefore, plant–herbivore interactions need to be explored to develop predictions of the overall impact of the introduced beetle on the weed. The findings revealed that defoliation by Z . bicolorata had a significant impact on the plant height, density and flower production in flushes F ₃, F ₄ and F ₅, but not in F ₁ and F ₂ that exhibited longer periodicity, profuse branching, a longer flowering period and maximum flower production and contributed mostly to the existing seed soil bank. Therefore, total depletion of the existing soil seed bank was not possible. Consequently, the effect of augmentative field releases of laboratory‐reared beetles was explored on F ₁ and F ₂ in February for three consecutive years (2011–2013). Before initiating the trial, random soil samples were taken from the plots that were assigned to the paired treatments (i.e. with the beetle and without the beetle [insecticide‐treated]) and it was found that the seed bank in those samples did not differ. The single release of Z . bicolorata adults at five per plant at the six‐leaf stage significantly reduced the soil seed bank, compared to without the biocontrol agent, irrespective of the flushes at the end of the season.     

Richtlinien der Verbände des ökologischen Landbaus zum Vorratsschutz

Zusammenfassung Verbände des ökologischen Landbaus wie z. B. Bioland, Gäa, Demeter; Naturland, Ernte für das Leben oder Bio Suisse beschränken ihre Mitglieder bei der Wahl von Vorratsschutzmaßnahmen. Vorrang besitzen Maßnahmen zur Vermeidung von Schädlingen gegenüber Bekämpfungsmaßnahmen. Fallen müssen zur Befallsüberwachung eingesetzt werden, um einen Befall durch Vorratsschädlinge frühzeitig zu erkennen. Diese Maßnahmen sollen den weitgehenden Verzicht auf chemisch-synthetische Mittel ermöglichen. In diesem Beitrag werden die Empfehlungen der Verbände mit den derzeit verfügbaren chemischen Mitteln für den Vorratsschutz abgeglichen. Erfahrung in der praktischen Umsetzung von physikalischen und biologischen Verfahren werden diskutiert und Defizite bei der Befallsüberwachung und Bekämpfung beschrieben.     

Development of protocols for detection of Colletotrichum acutatum and monitoring of strawberry anthracnose using real-time PCR

Real-time PCR (TaqMan®) assays were developed for the specific detection and discrimination of Colletotrichum spp., C. acutatum and C. gloeosporioides causing anthracnose in strawberry using the most divergent area of the internal transcribed spacers (ITS1 and ITS2) and 5·8S ribosomal RNA (rRNA) gene region. The specificity of the new assays was tested using DNA from six species of Colletotrichum and nine fungal species commonly found associated with strawberry material, and additionally by comparing the sequences with those from databases using a blast search. The sequences only showed identity with homologous sequences from the desired target organisms. The new assays were 10–100 times more sensitive than conventional PCR methods previously published for the diagnosis of strawberry anthracnose. When real-time PCR was compared with ELISA methods, PCR improved the sensitivity of the identification by obtaining positive results for samples of strawberry plant material that tested negative with ELISA. The development of C. acutatum was monitored using artificially infected strawberry crowns from two strawberry cultivars (Camarosa and Ventana) and a real-time PCR assay specific for this species between January and June 2006. The amount of C. acutatum detected using real-time PCR varied significantly by month ( P  < 0·001), but not by cultivar ( P  = 0·394). The new assays were shown to be useful tools for rapid detection and identification of these pathogens and to allow rapid and accurate assessment of the casual agents of anthracnose in strawberry.     

Effects of the saprophytic leaf mycoflora on growth and productivity of winter wheat

The effects of the saprophytic mycoflora and its interference with cereal aphids on growth and yield of winter and spring wheat was studied in field experiments in 1980, 1981 and 1982.Yields varied between 5000 and 8000 kg dry matter of kernels per ha. The effect of the saprophytic mycoflora on yield was determined in different treatments: A) no control measures against cereal aphids and saprophytic and necrotrophic fungi, B) no control of cereal aphids, control of saprophytic and necrotrophic fungi, C) control of cereal aphids and control of saprophytic and necrotrophic fungi, D) control of cereal aphids and stimulation of saprophytic mycoflora and E) control of cereal aphids, no control of saprophytic and necrotrophic fungi nor stimulation of saprophytic mycoflora.Considerable differences in top densities of saprophytic mycoflora (10 times as large in A and D as in B and C) were determined. The consequences of these differences for the growth and productivity of wheat were minor. A negative effect of saprophytic mycoflora on the yield could not be detected in 1981 and 1982, whereas a small positive significant effect was found in 1980. This stimulation may have been due to competition between necrotrophic fungal pathogens and saprophytic mycoflora. As a result of favourable weather conditions necrotrophic fungal pathogens were very numerous in 1980 and could form an important yield reducing factor. Yield levels may effect the importance of the necrotrophic and saprophytic mycoflora as yield reducing factors. Additionally, in the presence of aphid honeydew captafol was found to be relatively ineffective against saprophytic fungi.     

Relationship Between Production of the Phytotoxin Prehelminthosporol and Virulence in Isolates of the Plant Pathogenic Fungus Bipolaris sorokiniana

A gas chromatographic method was developed to quantify the phytotoxin prehelminthosporol, which is a sesquiterpene metabolite of the plant pathogen Bipolaris sorokiniana. The toxin was extracted from mycelium or culture filtrates, pre-cleaned using solid phase extraction, and analyzed by gas chromatography as a trimethylsilyl-derivative. The detection limit of the method was 5ngl^–1 (signal to noise ratio 4:1) which corresponds to ca. 15ng prehelminthosporol per mg dry weight of mycelium or 15ng prehelminthosporol per ml culture filtrate. The total amount of prehelminthosporol (mycelium plus culture filtrate) increased with cultivation time when examined in six isolates of B. sorokiniana after 6, 9, 12 and 15 days of incubation. The screening experiment of 17 isolates for prehelminthosporol production after 8 days of incubation revealed significant differences in the toxin production between the isolates. The isolates with low toxin production had lower virulence towards barley roots compared to those with higher production of the toxin. However, the virulence did not increase with prehelminthosporol level among the high producing isolates. Prehelminthosporol was also analyzed in a number of related Bipolaris and Drechslera species. In addition to B. sorokiniana, three out of six Bipolaris species (B. setariae, B. zeicola, B. victoriae) produced prehelminthosporol, which indicates that ability to produce prehelminthosporol is conserved among closely-related Bipolaris species.