拮抗球胞链霉菌AM6代谢产物防治烟草赤星病的田间效果测定

 拮抗球胞链霉菌AM6代谢产物田间试验结果表明,以含有AM6拮抗物质的液剂喷雾烟草叶片,可有效防治烟草赤星病。其防效随液剂中拮抗物质的含量的增加而升高,以100倍稀释防效最好,田间防效可达80.3%;施用次数2次为佳,平均防效达64.9%,病害发生严重时,可增施1次。同时,试验过程中发现含有AM6拮抗物质的液剂对烟草无明显药害。     

黄麻链霉菌NF0919菌株发酵培养基的优化

以黄麻链霉菌(Streptomyces corchorusii)NF0919菌株发酵液对草莓炭疽病菌(Glomerella cingulata)的抑菌率为效价指标。通过单因素试验筛选出NF0919菌株发酵用最佳的碳源、无机盐和氮源,在此基础之上,再通过正交试验和二次通用旋转组合设计试验,对NF0919菌株发酵培养基进行优化。结果表明:与发酵初始培养基相比,优化后得到的最佳培养基发酵液对草莓炭疽病菌的相对毒力效价提高了13.77倍。筛选得到的最佳发酵培养基组成为:10.2%马铃薯淀粉、2.6%棉籽蛋白、0.1%CaCO3、0.05%K2HPO4、0.05%MgSO4。     

广谱拮抗放线菌C28的鉴定及其对甜瓜蔓枯病的防治效果

【目的】研究放线菌C28对12种植物病原真菌的抑菌作用及对盆栽甜瓜蔓枯病的防治效果,并对该放线菌进行分类鉴定。【方法】采用琼脂块法、生长速率法研究放线菌C28对靶标病原真菌的抑菌作用;通过盆栽试验研究C28对甜瓜蔓枯病的防治效果;根据形态特征、生理生化特性及16S rDNA序列分析确定C28的分类地位。【结果】①放线菌C28对供试病原真菌具有一定的广谱拮抗性,拮抗圈直径为10.0～22.5 mm;其无菌发酵滤液对病原真菌菌丝生长具有明显的抑制作用,无菌发酵滤液原液对木贼镰刀菌（Fusarium equiseti）、层出镰刀菌（F.proliferatum）及葡萄座腔菌（Botryosphaeria dothidea）的抑菌率分别高达95.0%,90.0%及93.1%。②用放线菌C28无菌发酵滤液稀释10⁴倍处理甜瓜种子后,甜瓜胚轴、胚根长度较对照（无菌水处理）增加37.1%和12.5%;种子简明活力指数高达141.7,与对照差异显著（P<0.05）。③用C28无菌发酵滤液原液喷施处理后,甜瓜蔓枯病病情指数较对照显著下降（P<0.05）,预防和治疗效果分别为42.4%和40.0%。④根据形态特征、生理生化特性及16S rDNA序列分析,将放线菌C28鉴定为球孢链霉菌球孢亚种（Streptomyces globisporus subsp.globisporus）。【结论】球孢链霉菌球孢亚种C28对多种病原真菌具有广谱拮抗性,对甜瓜蔓枯病具有良好的抑菌和生防效果,对瓜类种子萌发和生长具有一定的促进作用。     

大丽轮枝菌菌体对链霉菌胞外蛋白酶活性及抑菌效果的影响

 为研究拮抗链霉菌对棉花黄萎病原大丽轮枝菌（Verticillium dahliae Kleb.）的抑菌机理,以大丽轮枝菌菌体为唯一碳氮能源,采用液体培养法及生长速率法研究供试病原菌菌体对4株拮抗链霉菌胞外蛋白酶合成的诱导作用及抑菌活性物质产生的影响。结果表明：大丽轮枝菌菌体可以诱导供试链霉菌合成蛋白酶;诱导粗酶液对病原真菌菌丝有溶解作用;当菌体添加量为10 g·L^-1,28℃培养7 d时菌株Z13蛋白酶活性高达4.24 U;以大丽轮枝菌菌体为碳氮能源时供试链霉菌能产生活性较强的抑菌物质;当菌体添加量为20 g·L^-1,发酵液稀释5倍时菌株B49所产抑菌活性物质的最大抑菌率达95.7%。     

Solid-Phase Extraction Embedded Dialysis (SPEED), an Innovative Procedure for the Investigation of Microbial Specialized Metabolites

Solid-phase extraction embedded dialysis (SPEED technology) is an innovative procedure developed to physically separate in-situ, during the cultivation, the mycelium of filament forming microorganisms, such as actinomycetes and fungi, and the XAD-16 resin used to trap the secreted specialized metabolites. SPEED consists of an external nylon cloth and an internal dialysis tube containing the XAD resin. The dialysis barrier selects the molecular weight of the trapped compounds, and prevents the aggregation of biomass or macromolecules on the XAD beads. The external nylon promotes the formation of a microbial biofilm, making SPEED a biofilm supported cultivation process. SPEED technology was applied to the marine Streptomyces albidoflavus 19-S21, isolated from a core of a submerged Kopara sampled at 20 m from the border of a saltwater pond. The chemical space of this strain was investigated effectively using a dereplication strategy based on molecular networking and in-depth chemical analysis. The results highlight the impact of culture support on the molecular profile of Streptomyces albidoflavus 19-S21 secondary metabolites.     

苹果树腐烂病生防链霉菌A144的鉴定及其代谢产物的抑菌活性

旨在明确菌株A144的分类地位及其代谢产物的抑菌活性。基于形态学观察、生理生化特征以及16S rDNA序列分析将菌株A144鉴定为娄彻氏链霉菌(Streptomyces rochei)。采用含药平板法测定菌株A144的抑菌谱,结果表明其发酵滤液对苹果树腐烂病菌(Valsamali var.mali)、西瓜枯萎病菌(Fusarium oxysporum)、水稻恶苗病菌(Fusarium moniliforme)以及番茄早疫病菌(Alternaria solani)均有抑菌活性,抑菌率分别为68.15%±2.62%、28.90%±1.79%、33.77%±2.59%和11.43%±0.81%。以苹果树腐烂病菌为指示菌,从12种不同培养基中筛选出ISP2培养基为菌株A144的最佳发酵培养基,其发酵滤液的抑菌率达85.06%±0.86%。平板涂抹法和平板对扣法试验结果表明,其脂肽粗提物、蛋白粗提物和挥发性物质对苹果树腐烂病菌均有抑菌活性,抑菌率分别为81.44%±0.92%、47.92%±2.25%和22.87%±4.40%。可见,菌株A144在苹果树腐烂病的生物防治中具有良好的开发潜力。     

防病促生功能性微生物的筛选及应用研究

从温室番茄根际分离得到具有抗病和促生功能的链霉菌 (Streptomyces sp)S5 0 6菌株。盆栽法测定 ,S5 0 6对番茄苗期立枯病Rhizoctonia sp和枯萎病Fusarium sp的防效分别达到了 86 1 %和 89 4% ,与化学药剂的防效无显著差别 ;与常规处理比较 ,对番茄株高、整株鲜重、地上部干重、叶面积、主根长、毛细根数量和根系干重的促进效果均达极显著水平 (P <0 0 1 )。在连茬 4年冬春茬番茄生产上应用 ,S5 0 6有效预防了苗期立枯病的发生 ,对毛粉 40 2和宝冠都表现出显著的促生长效果 ,单株产量分别比对照提高了 1 5 9%和1 7 6%。     

链霉菌702生物活性物质抑菌的致死浓度和致死时间的测定

通过对链霉菌702所产生物活性物质和纳他霉素及Nisin分别对黑曲霉、青霉、绿色木霉、枯草芽孢杆菌、蜡质芽孢杆菌、苏云金芽孢杆菌、产朊假丝酵母、啤酒酵母的抑制作用研究,试验结果得出链霉菌702所产生物活性物质对各种指示菌的半致死浓度和半致死时间,证明链霉菌702所产生物活性物质抑制效果比纳他霉素及Nisin强。试验结果为链霉菌702所产活性物质应用提供了有益的试验数据。     

将透明颤菌血红蛋白基因(vgb)转移到链霉菌染色体整合载体的构建

链霉菌抗生素的生物合成对培养环境中氧的供应相当敏感,为了改善这一溶氧问题,本研究通过将透明颤菌血红蛋白基因(vgb)克隆到含ФC31int,attP基因的诱导型表达载体pU8600上,构建了可接合转移到链霉菌的整合型表达vgb基因的质粒pHZ1276。pHZ1276通过接合转移导入变铅青链霉菌(Streptomyces lividans),所得重组子经Southern杂交验证后,进行诱导表达,菌体经破碎后所得蛋白粗提物经Western杂交和CO结合试验表明vgb基因在该菌中表达出了有活性的VHb蛋白。     

Cliona varians-Derived Actinomycetes as Bioresources of Photoprotection-Related Bioactive End-Products

Sunscreen and sunblock are crucial skincare products to prevent photoaging and photocarcinogenesis through the addition of chemical filters to absorb or block ultraviolet (UV) radiation. However, several sunscreen and sunblock ingredients, mostly UV filters, have been associated with human and environmental safety concerns. Therefore, the exploration and discovery of promising novel sources of efficient and safer compounds with photoprotection-related activities are currently required. Marine invertebrates, particularly their associated microbiota, are promising providers of specialized metabolites with valuable biotechnological applications. Nevertheless, despite Actinobacteria members being a well-known source of bioactive metabolites, their photoprotective potential has been poorly explored so far. Hence, a set of methanolic extracts obtained from Cliona varians-derived actinomycetes was screened regarding their antioxidant and UV-absorbing capacities (i.e., photoprotection-related activities). The active extract-producing strains were identified and classified within genera Streptomyces, Micrococcus, Gordonia, and Promicromonospora. This is the first report of the isolation of these microorganisms from C. varians (an ecologically important Caribbean coral reef-boring sponge). The in vitro cytotoxicity on dermal fibroblasts of oxybenzone and the selected active extracts revealed that oxybenzone exerted a cytotoxic effect, whereas no cytotoxic effect of test extracts was observed. Accordingly, the most active (SPFi > 5, radical scavenging > 50%) and nontoxic (cell viability > 75%) extracts were obtained from Streptomyces strains. Finally, LC-MS-based characterization suggested a broad chemical space within the test strains and agreed with the reported streptomycetes’ chemodiversity. The respective metabolite profiling exposed a strain-specific metabolite occurrence, leading to the recognition of potential hits. These findings suggest that marine Streptomyces produce photoprotectants ought to be further explored in skincare applications.