植物土传病原菌拮抗细菌的筛选与鉴定

从作物根际土壤中分离到1056株细菌，筛选出7个具有较强拮抗活性的菌株。室内测定对水稻纹枯病菌、辣椒疫病菌、瓜果腐霉、油菜菌核病菌、棉花枯萎病菌、茄根腐病菌、棉花黄萎病菌、番茄青枯病菌、甘蓝黑腐病菌等重要土传病原菌有较强拮抗作用；温室盆栽试验对番茄青枯病表现出较好防效，其中以BOH2和OH11效果较为明显，防效分别为90．9％和86．4％。通过形态观察、生理生化试验和16SrDNA序列分析，确定OH11为产酶溶杆菌。BOH3为荧光假单胞菌，其余5个菌株为不同芽孢杆菌。     

小单孢菌E2 菌株对辣椒疫霉的抑制及抗菌物质的初步测定

利用平板稀释法从辣椒疫病病圃根冠土中分离获得一株放线菌E2,根据形态特征和培养性状初步鉴定为小单孢菌Micromonospora sp..用E2菌株的发酵液和菌丝的提取物进行抑菌试验,结果表明,该菌对多种真菌和细菌具有抑菌作用,其含菌发酵液对辣椒苗期人工接种疫霉的防治效果达75%～85%.捷克八溶剂系统纸层析和抗生素特征化学反应表明,E2菌株产生多烯类抗生素.     

拮抗放线菌JD211的抑菌活性及其鉴定

本实验室从江西庐山珙桐树中分离筛选出的一株拮抗放线菌,编号为JD211。为明确放线菌JD211菌株在植物病害生物防治中的应用潜力,采用菌丝生长速率法测定其抑菌作用,通过多相分类法研究其分类地位。结果表明：JD211的粗提物可显著抑制水稻纹枯病菌、稻瘟病菌、烟草黑胫病菌、根霉、胶孢炭疽病菌、犁头霉、西瓜枯萎病菌等真菌的生长,其中对稻瘟病菌、烟草黑胫病菌等植物病原真菌的相对抑制率高达90％以上。根据形态特征、培养特征、生理生化特征及16SrDNA序列分析结果,将菌株JD211初步鉴定为奈良链霉菌（Streptomyees naraensis）。     

棉花枯萎病拮抗放线菌K5的分离与鉴定

从扬州地区土壤中分离到207株放线菌,以棉花枯萎病菌为目标菌,采用抑菌带法测定,筛选到9株有拮抗活性菌株,占所分离总数的4.3%.其中K5对棉花枯萎病菌的拮抗作用最强,抑茜带宽为8mm,同时对其它病菌也有一定的拮抗作用.盆栽试验结果表明菌株K5对棉花枯萎病防治效果达46.5%,与多菌灵相当.通过对菌株K5的形态观察、培养特征、生理生化反应和16S rDNA测定,将其鉴定为玫瑰产色链霉菌Streptomyces roseochromogenus.     

孜然种子中杀菌活性成分分离及结构鉴定

以小麦白粉病菌Blumer graminis、黄瓜霜霉病菌Pseudoperonispora cubensis、杨树溃疡病菌Dothiorella gregaria、棉花枯萎病菌Fusarium oxysporum f.sp.vasinfectum、白菜黑斑病菌Alternaria oleracea和稻瘟病菌Pyricularia grisea 6种病原菌为指示菌种,对孜然种子中的杀菌活性成分进行了跟踪分离及活性测定。采用柱层析分离技术,从孜然种子乙醇浸膏石油醚萃取液中分离得到两个具有杀菌活性的化合物,其结构经质谱、核磁共振氢谱及碳谱等分析确定其分别为枯茗酸(对异丙基苯甲酸,p-isopropyl benzoic acid)和枯茗醛(对异丙基苯甲醛,p-isopropyl benzaldehyde)。采用菌丝生长速率法测定了两化合物对油菜菌核病菌Sclerotinia sclerotiorum和辣椒疫霉Phytophthora capsici的毒力,其中枯茗醛对两种病原菌的EC50值分别为2.093和 15.40 mg/L,枯茗酸的EC50值分别为7.298和19.66 mg/L。     

拮抗真菌HTC的鉴定及其对辣椒疫病的生物防治潜力

为明确拮抗真菌HTC对辣椒疫霉Phytophthora capsici的拮抗机制,采用平板对峙、形态学鉴定和18S rDNA序列比对分析等方法对菌株HTC进行了鉴定,并研究其发酵液与抗生物质粗提液对辣椒疫霉不同发育阶段的影响。经鉴定,菌株HTC为金色毛壳菌Chaetomium aureum。在平皿对峙试验中,菌株HTC的红色分泌物能抑制辣椒疫霉菌丝的生长,抑制率为59.1%,后期HTC菌丝可缠绕并降解辣椒疫霉菌丝。发酵液与抗生物质粗提液对辣椒疫霉不同发育阶段均有抑制作用,发酵液对辣椒疫霉菌丝生长的抑制率高达97.58%,对辣椒疫病的防治效果均高于70%。浇灌发酵液后,可提高辣椒苗的苯丙氨酸解氨酶、过氧化物酶、多酚氧化酶的活性,并明显促进辣椒苗生长,鲜重和干重增加32.27%和18.09%。研究表明,金色毛壳菌菌株HTC是一株具有开发潜力的生防菌株。     

土壤拮抗放线菌的分离及其抗菌活性筛选

以常见植物病原真菌和细菌为指示菌,从分离的422株放线菌中筛选出28株可代谢抗细菌活性物质的菌株,其中N331、N393菌株发酵液对供试12种真菌有显著的抑制作用。离体条件下,N331、N393菌株发酵液对供试真菌菌丝生长的抑制中浓度(EC50)均小于15 mL/L;对玉米弯孢病菌、棉花枯萎病菌、小麦根腐病菌、黄瓜霜霉病菌孢子萌发的EC₅₀亦小于15 mL/L。盆栽试验结果表明,N331、N393菌株发酵原液对黄瓜霜霉病的保护效果分别为97.4%和85.6%,治疗效果分别为83.3%和59.3%。     

石榴枯萎病菌拮抗放线菌对南方根结线虫的毒力

从云南蒙自万亩石榴园、建水石榴园采集的土样中分离筛选得到对石榴枯萎病菌有较强拮抗活性的放线菌,其中菌株JS1对石榴枯萎病菌的室内抑菌半径为11mm。测定了菌株JS1发酵液对南方根结线虫的抑杀活性,结果表明,50%JS1菌株发酵液处理南方根结线虫卵6d,卵孵化率与对照相比降低了51.67%;50%JS1菌株发酵液处理南方根结线虫2龄幼虫72h,2龄幼虫的校正死亡率增加了88.10%;菌株JS1兼有拮抗石榴枯萎病菌及抑杀南方根结线虫的活性。结合菌株JS1在鉴定培养基上的培养特征、形态学观察及16SrDNA序列分析,将JS1初步鉴定为壮观链霉菌(Streptomyces spectabilis)。     

豆科植物多花木蓝抑菌活性成分研究

活性追踪下,采用柱层析分离多花木蓝籽石油醚提取物中的抑菌活性物质。采用生长速率法和牛津杯法,选取稻瘟病菌、辣椒枯萎病菌、油菜菌核病菌、梨黑星病菌为测试真菌,青枯菌、大肠杆菌、金黄葡萄球菌、绿脓杆菌、枯草芽胞杆菌为测试细菌,测试了一级柱层析各流分的抑菌活性,测试结果显示:A-4和A-5流分对所有测试菌均具有一定的抑菌活性,对梨黑星病菌的抑制作用最好,抑制率分别达到77.66%和74.67%;对青枯菌的抑菌圈最大,分别为1.37cm和1.66cm。分离出2个活性化合物,经13 C、1 H NMR波谱方法鉴定,为BalansenateⅠ、Ⅱ,为首次从该植物中分离得到。生测结果显示2个化合物具有较低的抑菌活性,其对金黄葡萄球菌活性最好,但最小抑菌浓度(MIC)和最小杀菌(MBC)浓度均只为250mg/L和500mg/L。     

生防细菌BMY-1对几种植物病原真菌的抑制作用

采用平板对峙培养法和滤纸片培养法测定细菌BMY-1菌株对番茄灰霉病菌(Botrytis cinerea)、辣椒疫霉病菌(Phytophthora capsici)、玉米小斑病菌(Bipolaria maydis)、玉米弯孢霉叶斑病菌(Curvularia lunata)等11种植物病原真菌生长的抑制作用。结果表明,BMY-1菌株对供试病原菌生长具有很好的抑制效果。平板对峙培养法中,BMY-1菌株对番茄灰霉病菌、玉米弯孢霉叶斑病菌、玉米小斑病菌的抑制效果最好,抑菌宽度分别达到15.0 mm,14.0 mm,13.0mm。滤纸片培养法中,BMY-1菌液对番茄灰霉病菌抑制效果最好,抑菌带宽度为8.0 mm。BMY-1菌液及其无菌发酵液对病原菌孢子萌发也有很强的抑制作用。其中对辣椒疫霉病菌和玉米小斑病菌孢子萌发的抑制效果最好。BMY-1菌液处理后,孢子萌发率分别为0.97%和1.00%,而对照萌发率为分别为96!29%和93.04%。     

Studies on Endophytic Actinomycetes ( I ) <Emphasis Type="Italic">Streptomyces </Emphasis>sp. Isolated from Rhododendron and Its Antifungal Activity

To survey endophytic actinomycetes as potential biocontrol agents against fungal diseases of rhododendron, young plants of rhododendron were surface-sterilized for use as an isolation source. Nine, six and two isolates, with distinguishing characteristics based on the macroscopic appearance of colonies, were obtained from roots, stems and leaves, respectively, suggesting that various species of actinomycetes grow in the respective organs of this plant as symbionts or parasites. On an agar medium, only isolate R-5 commonly formed a clear growth-inhibition zone against two major fungal pathogens of rhododendron, Phytophthora cinnamomi and Pestalotiopsis sydowiana, indicating that this isolate can produce antifungal material(s). Acetone extracts of a liquid culture of R-5 had a broad antimicrobial spectrum against Gram-positive bacteria, yeast and filamentous fungi. Isolate R-5 was identified as a Streptomyces sp. based on morphological, physiological and chemotaxonomical characteristics. The present results indicate that isolate R-5 is a suitable candidate for the biocontrol of diseases of rhododendron. Received 25 March 2000/ Accepted in revised form 18 May 2000     

An Endophytic Actinomycete, Streptomyces sp. AOK-30, Isolated from Mountain Laurel and Its Antifungal Activity

To survey endophytic actinomycetes as potential biocontrol agents against fungal diseases of mountain laurel, young plants of mountain laurel (Kalmia latifolia L.) were used as an isolation source. From a total of 73 actinomycetes isolates obtained from leaves, stems and roots of test plants, Streptomyces sp. AOK-30 was selected, because i) it had a broad and intense antimicrobial spectrum against various yeasts and fungal pathogens of Ericaceae, ii) it grew on the multiplication and rooting media for tissue culture for this plant, and iii) the tissue-cultured seedlings that had been treated with this isolate became resistant to Pestalotia disease without any adverse growth effects such as malformation, dwarfing, discolorization and defoliation. This isolate was identified as Streptomyces sp., based on cultural, physiological and morphological characteristics. The present results indicate that isolate AOK-30 is a potential biocontrol agent against diseases of mountain laurel. Received 10 September 2002/ Accepted in revised form 22 November 2002     

黄河中上游半干旱区典型盐渍土中拮抗性放线菌筛选

对采自陕西、宁夏和内蒙等地典型盐渍土中放线菌组成及拮抗性进行了研究。结果表明:(1)该地区盐渍土中放线菌以链霉菌属为主,非链霉菌仅占9.27%,链霉菌属中以球孢类群,粉红孢类群,灰褐类群,黄色类群为主,占到放线菌总数的55.62%。(2)供试的706株放线菌中有拮抗活性的占到76.44%,但广谱性拮抗菌株较少;对细菌拮抗能力较真菌强;拮抗性放线菌以粉红孢类群、灰褐类群和球孢类群为优势类群;(3)发酵滤液拮抗性试验筛选出4株能产生较强抑菌活性物质的菌株,其中3株都是粉红孢类群。     

Isolation of actinomycetes and endospore-forming bacteria from the cacao pod surface and their antagonistic activity against the witches’ broom and black pod pathogens

In this study, actinomycetes and endospore-forming bacteria were isolated from the surface of cacao pods. The activity of these microorganisms againstCrinipellis perniciosa andPhytophthora palmivora, causal agents of witches’ broom and black pod diseases of cacao, respectively, was investigated. A total of 336 isolates of actinomycetes and endosporeforming bacteria were tested on a detached pod assay againstC. perniciosa. The screening procedure used proved to be fast and inexpensive, allowing the selection of five actinomycetes as the most promising isolates for the biocontrol ofC. perniciosa. Under laboratory conditions the actinomycetes were able to inhibit 100% ofC. perniciosa basidiospore germination. However, under field conditions the selected actinomycetes were unable to protect cacao pods against both pathogens. In these experiments, inhibition ofC. perniciosa ranged from 6% to 21% in relation to the control, whereas there was no inhibition of black pod caused byP. palmivora. Formulations need to be improved in order to enhance the activity of the actinomycetes against cacao pathogens in the field. Molecular identification of the selected isolates showed that they are species of the genusStreptomyces.     

黄瓜内生放线菌的分离、筛选及其活性菌株鉴定

 从植物内生放线菌中筛选拮抗活性菌株和寻找新的农用活性代谢产物,可为植物病害防治提供新的资源。本研究结果发现黄瓜幼苗的根、茎、叶中均有内生放线菌的存在,根组织中的数量、种类明显多于叶片和茎,占总分离株的72.7%,以链霉菌的淡紫灰类群、灰褐类群为主。活性筛选试验结果表明,黄瓜内生放线菌中对12种靶标真菌和6种细菌具有拮抗作用的菌株分别占46.8%和39.0%,主要为分离自根组织的链霉菌淡紫灰类群。分离自黄瓜叶片的gCLA4菌株抑菌谱较广,其发酵滤液对供试12种靶标真菌均具有较好的抑菌效果,但对靶标细菌有选择性抑制作用。形态学、细胞化学和16S rDNA序列分析鉴定该菌株为淡紫灰链霉菌(Streptomyces lavendularectus)。     

Leaf blast disease reduction by rice-phyllosphere actinomycetes producing bioactive compounds

Fungal leaf blast, caused by Pyricularia oryzae, is a devastating disease of rice plants that annually causes severe production losses worldwide and is one of the top 10 fungal diseases that threaten global food security. Thus, a reliable control strategy against this disease is essential. In this study, the antagonistic activity of indigenous phyllosphere actinomycetes was elucidated against P. oryzae in vitro and in planta to develop an efficient, effective and environmental friendly approach to protect rice plants against P. oryzae. Of 75 isolates of actinomycetes isolated from the rice phyllosphere, 18 isolates inhibited P. oryzae by >45%. According to analysis of their 16 S rRNA gene sequences, the majority of the 18 isolates belonged to Streptomyces genera; others were identified as belonging to Saccharothrix, Gordonia, or Lentzea. Isolates that potentially produced a bioactive compound(s) were identified among the 18 isolates: 17 isolates (94.44%) had a domain marker for nonribosomal peptide synthetase (NRPS) gene and 12 (66.67%) had type-I polyketide synthase (PKS) gene in their corresponding genome. Interestingly, isolates JSN1.9, SKB2.14, and SKB2.3 suppressed disease suppression by approximately 88%. To our knowledge, this is the first report on the application of rice-phyllosphere actinomycetes producing bioactive compounds to control leaf blast disease in Indonesia. Thus, these findings have escalated the potential application of phyllosphere actinomycetes as a supreme biocontrol agent against fungal leaf blast disease.     

Predictive factors for the suppression of fusarium wilt of tomato in plant growth media

ABSTRACT Fusarium wilts are economically important diseases for which there are no effective chemical control measures. However, biological control and fertility management are becoming efficient alternatives for controlling this disease. Growth media formulated with composts that are able to suppress Fusarium wilt of tomato provide a control system that integrates both strategies. The aim of this study was to predict Fusarium wilt suppression of growth media using abiotic and biotic variables. Grape marc compost was the most effective medium used to suppress Fusarium wilt. Cork compost was intermediate, and light peat and expanded vermiculite were the most conducive growth media. The growth media evaluated were in a pH range of 6.26 to 7.97. Both composts had high beta-glucosidase activity. When pH and beta-glucosidase activity were taken into account as predictive variables, more than 91% of the variation in severity of Fusarium wilt was explained. This relationship illustrates the effect of nutrient availability and the degree of microbiostasis, two key factors in this pathosystem. Microbial populations involved in suppressiveness were cellulolytic and oligotrophic actinomycetes, fungi, and the ratios cellulolytic actinomycetes/cellulolytic bacteria, oligotrophic bacteria/copiotrophic bacteria, and oligotrophic actinomycetes/oligotrophic bacteria. Based on community level physiological profiles, different community structures were evident among growth media evaluated.     

杨浦链霉菌WLU210的筛选、鉴定及其对苦瓜枯萎病的生防作用研究

本研究从河南黄河湿地采集土壤样品,采用梯度稀释涂布平板法分离和平板对峙培养法筛选获得一株对苦瓜枯萎病菌具有稳定拮抗活性的链霉菌WLU210。在形态学鉴定和生理生化特征测定的基础上,对菌株WLU210的16S rRNA基因序列进行比对分析,将WLU210鉴定为杨浦链霉菌Streptomyces yangpuensis;拮抗谱测定结果表明,杨浦链霉菌WLU210对苦瓜枯萎病菌、苹果轮纹病菌、小麦茎基腐病菌、辣椒疫霉病菌、西瓜炭疽病菌、小麦全蚀病菌和玉米大斑病菌等7种植物病原菌均具有一定的拮抗活性;盆栽试验和田间试验结果表明,杨浦链霉菌WLU210发酵液具有有效防治苦瓜枯萎病和促进苦瓜生长的作用,田间防治效果达68.89%。由此可见,杨浦链霉菌WLU210具有作为防治苦瓜枯萎病生防菌剂研发的潜力。     

不同生境芦苇根茎生长发育与根际微环境的比较研究

以临泽县4种生境(沙丘生境、盐渍生境、盐渍-沙丘过渡生境、沼泽生境)芦苇根茎和芦苇根际土壤为研究对象,测定不同生境下芦苇根茎生长发育指标、根际土壤微生物数量及水分、盐分含量,分析了不同生境下根茎生长发育与根际土壤微生物数量、土壤含水量及含盐量的关系。结果表明:(1)对比4种生境下芦苇根茎发育,从盐渍生境,到盐渍-沙丘过渡生境、沙丘生境,再到沼泽生境,根茎节间距呈缩短趋势,而节直径、根茎长度及不定芽数呈增长趋势,根茎含水量和根茎生物量、株高、基径也呈增加趋势。(2)根际土壤微生物数量在4种生境之间差异显著,盐渍生境下芦苇根际土壤真菌数量最多,细菌、放线菌数量最少;沼泽生境芦苇根际土壤真菌数量最少,细菌、放线菌数量最多。(3)相关性分析表明,芦苇根际土壤微生物数量对芦苇根茎数量特征的驱动作用不同。土壤细菌、放线菌数量、B/F值是芦苇根茎长度、根茎节直径、根茎生物量、根茎含水量、株高、基径的主要影响因子,土壤真菌是芦苇根茎节间距的主要影响因子。土壤细菌、真菌、放线菌、B/F是芦苇根茎不定芽数的主要影响因子。土壤细菌、放线菌、B/F值为正向驱动,真菌为负向驱动。     

鄂尔多斯沙地油蒿根际土壤微生物数量的季节动态

对油蒿根际土壤微生物数量的季节动态、垂直分布及影响微生物分布的环境因子进行了研究。结果表明:(1)微生物总数、细菌数量的季节动态为:春季>夏季>秋季>冬季,放线菌数量的季节分布为秋冬季多于春和夏季,真菌数量的季节动态为夏季达到最高值,其他季节变化不显著。(2)微生物数量的垂直分布总体上表现为随土层加深而逐渐减少,表层土壤(0-10cm)微生物数量是下面3层(10-40cm)的1.05-9.79倍。(3)土壤微生物根际效应明显,且细菌>真菌>放线菌,R/S值介于1.16-3.76之间。微生物根际效应的季节变化为春、夏季高于秋和冬季。(4)微生物分布与水热等环境因子密切相关,细菌数量与水热组合条件共同作用有关,真菌受温度影响较大,放线菌对土壤水分含量变化较敏感。