生防枯草芽孢杆菌L1特性的初步研究

通过对峙培养,测定出枯草芽孢杆菌L1的抑菌谱较宽,特别是对水稻纹枯病菌、大豆菌核病菌、禾谷镰孢菌、辣椒灰霉病菌、玉米小斑病菌抑菌效果明显;枯草芽孢杆菌L1不同发酵时间经湿热灭菌处理后,5 d发酵液中抑菌活性物质含量最高,对水稻纹枯病菌菌丝生长抑制率达83.23%,发酵液随时间延长抑菌效果不再增加;枯草芽孢杆菌L1抑菌活性物质对温度不敏感;枯草芽孢杆菌L1发酵液用硫酸铵梯度沉淀法提取粗蛋白在硫酸铵饱和度达60%～70%(不含60%)沉淀的活性物质抑菌效果最好,对水稻纹枯病菌平均抑菌半径达1.15;枯草芽孢杆菌L1对玉米、大豆、小麦、番茄、菜豆、黄瓜、水稻无致病性,而且还有保鲜和促生作用。     

培养条件对枯草芽孢杆菌G8抗菌蛋白含量的影响及蛋白液对黄瓜菌核病的生防效果

为确定不同培养条件对枯草芽孢杆菌G8抗菌蛋白含量的影响,用硫酸铵沉淀法提取了不同pH,以及不同温度、装液量和培养时间下获得的发酵液中的抗菌粗蛋白,测定了其浓度和对黄瓜菌核病菌Sclerotinia sclerotiorum (Lib.) de Bary的抑制活性。结果表明,在Luria Bertani培养基条件下,适宜发酵产生抗菌蛋白的pH值、温度、装液量、培养时间和摇床转速分别为7.0、37℃、75 mL/250 mL(250 mL容器中装75 mL液体)、36 h和180 r/min。在此优化条件下,1 L发酵液经硫酸铵沉淀后的蛋白量达 48.4 mg。对在上述条件下获得的G8抗菌粗蛋白进行了温室及田间试验。结果表明,浓度为310.5 μg/mL的G8对黄瓜菌核病的保护效果和防治效果分别为 76.14%和 69.30%;G8抗菌粗蛋白与高岭土1∶ 2(V ∶ W)制成的膏剂对田间黄瓜茎部菌核病斑的治愈率为65.07%。     

枯草芽胞杆菌GB519抗菌蛋白的理化性质及生防效果

枯草芽胞杆菌GB519是从土壤中分离对稻瘟病菌具有显著抑菌活性的菌株。为了明确菌株GB519抗菌粗蛋白的理化性质，分析了光照、紫外线、温度、pH和酶（蛋白酶K、胰蛋白酶和胃蛋白酶）对抑菌活性的影响。结果表明，抗菌蛋白对光照、紫外线和3种酶作用稳定；40℃~100℃处理1 h抑菌活性稳定，121℃处理30 min活性仅降低了7.4%，具有较好的耐高温特性；在pH 2.0~10.0活性稳定，但pH 11.0~12.0活性有所下降，分别降低了14.9%和18.1%。抗菌蛋白对多种植物病原菌具有抑菌作用，其中对茄子菌核病菌的抑菌活性为72.7%。在温室条件下对水稻叶瘟的防效达到80.1%。因此，菌株GB519可作为防控稻瘟病生防潜力菌进行开发。     

玉米内生菌YY1菌株抗菌蛋白的稳定性和活性组分分析

为探究枯草芽胞杆菌Bacillus subtilis菌株YY1抗菌蛋白的基本特性,采用硫酸铵分级沉淀法对蛋白粗提液进行提取,测定温度、酸碱性、蛋白酶和金属离子对抗菌蛋白稳定性的影响,利用阴离子交换柱层析和葡聚糖凝胶柱层析对其组分进行分离,通过MALDI-TOF质谱对其活性组分进行初步鉴定。结果表明,当硫酸铵饱和度为50%时,菌株YY1产生的粗蛋白对玉米大斑病菌Setosphaeria turcica的抑制效果最好,抑菌圈半径达2.20 cm。该粗蛋白具有较高的热稳定性,90℃处理120 min其抑菌活性未受到明显影响,121℃处理120 min其抑菌活性仍可保持原有活性的85.0%。该粗蛋白具有较好的耐碱性,pH 5～11条件下其抑菌活性稳定,pH 3～4条件下其抑菌活性显著下降。该粗蛋白对蛋白酶K和胰蛋白酶不敏感,两者处理后的抑菌活性分别为对照的97.5%和98.9%;不同浓度金属离子对其抑菌活性的影响存在差异,Cr³⁺可使该粗蛋白完全失活,Zn²⁺、Li⁺、K⁺、Na⁺、M...     

内生解淀粉芽孢杆菌LP-5抗菌蛋白的分离纯化及特性

解淀粉芽孢杆菌Bacillus amyloliquefaciens LP-5是一株对梨黑斑病菌Alternaria alternata具有抑制作用的内生细菌,其产生的抗菌物质主要是抗菌蛋白.为明确该抗菌蛋白的理化性质,采用盐酸沉淀、DEAE-52层析和Sephadex G-100层析进行纯化,测定了所得抗菌蛋白对热、蛋白酶、紫外辐射及酸、碱的稳定性.结果表明:纯化的抗菌蛋白分子量为20.3 kD,等电点为6.33.该蛋白对高温敏感,100℃处理30 min,抑菌活性仅为对照的18.4%;对蛋白酶K和胰蛋白酶稳定,酶处理后的抑菌活性分别为对照的92.5%和88.9%;对紫外辐射稳定,照射12 h后抑菌活性仍达对照的92.5%;对酸稳定、对碱敏感,pH 13处理后抑菌活性下降为原有活性的74.1%.抑菌活性测定结果显示,该蛋白对梨黑斑病菌的孢子萌发和菌丝生长有明显抑制作用,可造成孢子萌发端泡囊化、菌体破裂、原生质外渗.     

芽孢杆菌B11拮抗蛋白性质及其对西瓜枯萎病菌的作用机理

芽孢杆菌(Bacillus sp.)B11分离自土壤,其拮抗蛋白粗提液在温度60、80℃各处理30 min后仍能保持抗菌活性,对紫外线、蛋白酶K、RNase处理稳定,对胃蛋白酶、胰蛋白酶、氯仿部分敏感,作用活性pH(pH4～10)范围较宽.B11拮抗蛋白对西瓜枯萎病菌具有较强的拮抗作用,其抑菌机制主要使病菌菌丝畸形,细胞质浓缩,孢子不萌发或萌发异常.     

短小芽孢杆菌BSH-4抗菌物质的提取及其特性

为明确短小芽孢杆菌BSH-4所产生抗菌物质的理化性质,以黄瓜菌核病菌为指示菌,采用平板打孔法研究了温度、pH变化、紫外照射、有机溶剂、金属离子及蛋白酶等对其稳定性的影响.结果表明,该抗菌物质粗提液具有良好的热稳定性,100℃处理1 h后仍具有对照活性的90%左右;对酸碱较敏感,在pH值6～8时活性较高;对紫外线稳定,在紫外光下照射10 h以上时,其活性基本不受影响;经乙醚和丙酮处理后仍具有对照活性的80%以上;对重金属离子敏感,Ag+、Cu2+、Zn2+能够降低对照活性的40%左右;对蛋白酶较稳定.此外,该抗菌粗提物同时对黄瓜蔓枯病菌、番茄早疫病菌和黄瓜立枯病菌有较高的抑制活性,具有分解羧甲基纤维素的能力,但不具有分解几丁质和β-1,3-葡聚糖的能力,并且其抗菌活性与分解羧甲基纤维素的能力无关.     

芽孢杆菌B11拮抗蛋白性质及其对西瓜枯萎病菌的作用机理

芽孢杆菌(Bacillus sp．)B11分离自土壤，其拮抗蛋白粗提液在温度60、80℃各处理30min后仍能保持抗菌活性，对紫外线、蛋白酶K、RNase处理稳定，对胃蛋白酶、胰蛋白酶、氯仿部分敏感，作用活性pH(pH4～10)范围较宽。B11拮抗蛋白对西瓜枯萎病菌具有较强的拮抗作用，其抑菌机制主要使病菌菌丝畸形，细胞质浓缩，孢子不萌发或萌发异常。     

枯草芽孢杆菌G3菌株的抗菌物质及其特性

 产几丁质酶枯草芽孢杆菌G3菌株的固体培养物在黄瓜灰霉病菌和番茄叶霉病菌抑菌试验中证实,抑菌活性物质存在于过滤上清液中,它们是从酸沉淀物中提取出的伊枯草菌素、生物表面活性素和存在于盐析粗蛋白中的几丁质酶。在叶霉孢子萌发试验中,伊枯草菌素微弱地抑制孢子萌发但强烈破坏芽管和新生菌丝;生物表面活性素和几丁质酶则强烈抑制孢子萌发并长久性地抑制芽管伸长。在PDA平板上的灰霉菌丝抑菌试验中,伊枯草菌素抑制菌丝生长,引发菌丝顶端膨大,形成泡囊,泡囊破裂后原生质外泄;几丁质酶抑制菌丝生长,引发产生不规则的菌丝团;生物表面活性素在平皿上对菌丝则不显示出抑菌活性。     

植物内生枯草芽孢杆菌E1R-j脂肽类化合物的分离鉴定及抑菌作用

植物内生枯草芽孢杆菌Bacillus subtilis菌株E1R-j分离自小麦根部。以小麦全蚀病菌作为靶标菌,采用酸沉淀、快速蛋白液相色谱(FPLC)和基质辅助激光解吸附飞行时间质谱(MALDITOF/TOF-M S)技术,对该菌发酵液中的抗真菌活性物质进行分离鉴定和抑菌活性研究。结果表明:菌株E1R-j产生的抗菌活性物质由P1、P2、P3和P4 4类组分组成,其中P3起主要抑菌作用。M ALDI-TOF/TOF-M S分析结果显示,P3为fengycins。经试验发现,P3对温度稳定,121℃下放置30 min仍可保持抑菌活性。电子显微境观察发现,P3对小麦全蚀病菌菌丝生长有强烈的抑制作用,同时还可造成菌丝细胞畸形、细胞壁溃解、细胞质外渗等。     

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.