华重楼根际土芽胞杆菌多样性研究

为了解华重楼根际芽胞杆菌种群多样性信息，本研究采用稀释平板法分离福建省光泽县华重楼品种根际的芽胞杆菌，并对其进行16S rRNA序列分析。结果表明：从华重楼根际共分离到17个形态差异的芽胞杆菌菌株；16S rRNA序列分析表明，17个菌株鉴定为8个种，归属于4个属，芽胞杆菌属(Bacillus)、赖氨酸芽胞杆菌属(Lysinibacillus)、类芽孢杆菌属(Paenibacillus)和绿芽胞杆菌属(Viridibacillus)，其中芽胞杆菌属的种类和数量最多。经抑菌试验分析，芽孢杆菌属的特基拉芽孢杆菌FJAT-43012对尖孢镰刀菌具有抑制作用。本研究结果表明，华重楼根际芽胞杆菌种群多样性较为丰富。本研究对芽胞杆菌资源的开发和利用提供良好的实验材料和理论基础。     

14株芽胞杆菌菌株的分子鉴定及其促生机理分析

以前期筛选的14株植物促生细菌为实验材料,运用16S rRNA基因序列同源性对菌株进行分子鉴定,通过生物测定评价菌株对番茄的促生效果,并对其促生机理进行初步分析。实验结果表明,促生菌株来自芽胞杆菌科的2个属、9个种。对番茄种子促生特性研究表明,14株芽胞杆菌均能提高种子发芽率,增长率为0.74%~11.85%;其中12株芽胞杆菌能促进胚根生长,增长率为4.03%~65.33%;而8株芽胞杆菌促进胚芽生长,增长率为1.24%~18.17%。对不同芽胞杆菌的促生因子相关分析结果表明,芽胞杆菌的6个促生因子（解无机磷、解有机磷、固氮效能、吲哚-3-乙酸含量、植酸酶活性、吡咯喹啉醌浓度）之间无相关关系（P>0.05）,与活菌数无相关关系（P>0.05）,与种子发芽率、胚根长、胚芽长也无显著相关关系（P>0.05）。种子发芽率与胚根长、胚芽长无相关关系（P>0.05）,而胚根和胚芽长之间具有显著性相关（P<0.01）。室内番茄盆栽实验表明,试验的6株芽胞杆菌对番茄幼苗均有促生作用,不同芽胞杆菌对番茄促生特性存在差异,如图瓦永芽胞杆菌FJAT-47648能显著增加根长,提升地上部与地下部干重,而暹罗芽胞杆菌FJAT-47226的促生效果则主要表现在增加株高和地上部生物量上。     

芽胞杆菌JK05的鉴定及其对香蕉、玉米的促生和生防潜能研究

解淀粉芽胞杆菌、枯草芽胞杆菌和苏云金芽胞杆菌等是重要的有益微生物,被广泛应用于植物病虫的生物防治。前期分离获得1株芽胞杆菌JK05,对其形态、生理生化特征、16S rRNA和gyrA基因序列进行分析,将其鉴定为解淀粉芽胞杆菌植生亚种Bacillus amyloliquefaciens subsp. plantarum。对峙培养实验结果显示,JK05菌株对多种植物病原镰刀菌具有拮抗作用。植物生长促进实验表明,JK05菌株对香蕉和玉米生长具有明显促进作用。盆栽实验结果显示,JK05菌株对香蕉枯萎病具有良好的防治效果。采用特异引物对JK05菌株基因组DNA进行PCR扩增,其可扩增到表面活性素（surfactin）、丰原素（fengycin）、伊枯草菌素A（iturin A）等抗生素合成基因。综上,JK05菌株具有良好的生防潜力,有望应用于生物农药和微生物肥料。     

白僵菌与苏云金芽胞杆菌水悬浮剂研制及田间防治玉米螟研究

通过室内生物测定,研究球孢白僵菌和苏云金芽胞杆菌联合防治玉米螟的效果。研究表明,确定白僵菌与苏云金芽胞杆菌浓度分别为1.0×106孢子/mL、1.0×109孢子/mL条件下,杀虫效果达到最佳,LT90为6.64(±0.07)d,较1.0×109的Bb LT90缩短26.06%。通过助剂悬浮率、乳化效果以及助剂对微生物活性影响等确定1个白僵菌与苏云金芽孢杆菌水悬浮剂剂型配方。田间防治玉米螟结果表明,两种生防菌均在防治过程中起作用,田间玉米受害株数、活虫数均明显低于未施药对照,防治效果优于单一使用白僵菌或苏云金芽胞杆菌。     

酸碱条件对枯草芽胞杆菌FJAT-14254代谢物产生的影响

为考察酸碱条件对枯草芽胞杆菌FJAT-14254代谢物产生的影响,采用气相色谱-质谱联用分析枯草芽胞杆菌FJAT-14254菌株的胞内成分。结果表明：通过谱库扫描得到32种高匹配率代谢物的初步鉴定结果,主要包括氨基酸类、酸类、烷烃类等,其中含量相对较高的有7种;枯草芽胞杆菌FJAT-14254代谢物的种类和含量与其培养环境的酸碱度变化呈一定的相关性,强酸性(pH＝3)条件下培养的胞内代谢物具有特异性,而接近中性(pH＝5、7)条件及碱性(pH＝9、11)条件下培养胞内代谢物则相似。由此推断,不同pH条件影     

莺歌海盐场30%盐度晒盐池底泥中可培养芽胞杆菌资源遗传多样性研究

采用80℃水浴处理与稀释涂布法从莺歌海盐场30%盐度晒盐池底泥中分离获得215株芽胞杆菌。采用16S rDNA PCR-RFLP与16S rDNA序列分析技术,对分离的菌株进行遗传多样性分析。16S rDNA PCR-RFLP酶切图谱UPGMA聚类分析结果表明,在100%的相似性水平上,分离菌株归属到9种酶切类型。16S rDNA测序结果显示,这些菌株主要分布在Pontibacillus、Halobacillus、Oceanobacillus和Bacillus等4个属,其中Pontibacillus为优势属。有7株芽胞杆菌的16S rDNA序列与数据库中相应模式菌的最大相似性在97.4%~99.0%之间,有可能为芽胞杆菌新种资源。     

芽胞杆菌对玉米幼苗生长及土壤酶活性和养分含量的影响

以解淀粉芽胞杆菌NECC10723(Bacillus amyloliquefaciens NECC10723,Bam723)和阿耶波多氏芽胞杆菌NECC10295(Bacillus aryabhattai NECC10295,Bar295)为试验材料,通过盆栽试验,探究接种不同芽胞杆菌对于玉米幼苗的生长指标、土壤酶活性和土壤养分含量的影响。结果表明,盐碱胁迫下接种不同芽胞杆菌均对玉米幼苗起到不同程度的促生作用,Bam723和Bar295均在1×10⁷cfu/mL浓度下促生效果最为显著,提高盐碱胁迫下玉米幼苗的株高、茎粗、地上部及地下部干重和根冠比。土壤中施加Bam723可显著提升盐碱胁迫下玉米幼苗种植土壤中的酶活和土壤养分含量,与对照处理相比,土壤中脲酶、碱性磷酸酶、蔗糖酶活性分别提高19.31%、31.77%和45.78%;土壤全氮、速效磷、速效钾含量分别提高137.54%、295.49%和3.19%,增加幅度均大于Bar295处理。     

香蕉枯萎病生防微生物产品的功效限制因子

由尖孢镰刀菌古巴专化型引起的香蕉枯萎病，是目前香蕉产业发展中的限制因子，采用生物防治的方法是目前非常有潜力的防控手段。生防微生物的效果主要与土壤水分、作为活菌制剂的微生物种类和性质、生防微生物在土壤中短期定殖影响因素、生防微生物长期定殖影响因素有关。采用茎叶滴注的方法进行香蕉内生芽胞杆菌接种或辅以适当的载体，和保水微生物一起混合接种到香蕉根围，可以有效防控香蕉枯萎病。     

巨大芽胞杆菌(A6)在红黄壤中对油菜的促生作用

在有效磷缺乏的鄂东南红黄壤稻田土壤条件下，接种巨大芽胞杆菌A6，研究其对油菜生长发育和盆栽产量的影响。试验结果表明，A6能促进油菜生长，表现不同程度地增加植株的叶面积、干重、全磷含量和盆栽产量。这一结果揭示了利用油菜促生根际细菌A6的可能性。     

短短芽胞杆菌FJAT-0809-GLX超氧化物歧化酶基因的克隆及原核表达

短短芽胞杆菌FJAT-0809-GLX具有抑菌、抗褐变和保鲜功能。以短短芽胞杆菌菌株FJAT-0809-GLX总DNA为模板,采用PCR扩增超氧化物歧化酶(Supseroxide dismutase,SOD)基因,将该片段纯化回收后与p MD18-T连接并转入大肠杆菌(Escherichia coli)DH5α,进行序列测定,结果显示SOD基因序列长度为609 bp(Gen Bank登录号:KM255665),编码202个氨基酸残基。将SOD基因片段与同样酶切的表达载体p ET-28a连接,构建重组表达载体p ET-28a-SOD,转入大肠杆菌BL-21,采用异丙基-β-D-硫代吡喃半乳糖苷(Isopropyl-β-D-thiogalactopyranoside,IPTG)进行诱导表达。SDS-PAGE结果表明,在菌体中存在约25 ku的蛋白表达产物,与该基因ORF预期大小接近。以上结果为进一步研究该酶的生理生化特性奠定了基础。     

白叶枯病菌拮抗菌筛选及水稻叶围微生物互作研究初报

 1998年7月～10月, 于田间汕优63水稻叶片上分离采集微生物, 并进行叶围微生物鉴定,以筛选白叶枯病拮抗菌。结果表明:除极少量分布的真菌和细菌外, 主要种群包括真菌类群Alternaria和 Fusarium属; 细菌类群Flavobacterium, Xanthomonas, Pseudomonas和Bacillus属。对白叶枯病菌（Xanthomonas oryzae pv. oryzae）具有明显拮抗作用的微生物为Bacillus属,而且Bacillus属细菌对其余大部分叶围微生物具有程度不同的拮抗作用,可以初步断定其产生的拮抗物质具有一定的广谱性。     

An Evaluation of the Microflora Associated with Fermented African Oil Bean (Pentaclethra Macrophylla Bentham) Seeds During Ugba Production

The microorganisms associated with fermented African oil bean (Pentaclethra macrophylla Bentham) seed during ugba production was studied. Only bacteria were isolated from the ugba samples used. Although the bacteria included Bacillus spp., Lactobacillus spp., Staphylococcus spp., Micrococcus spp. and members of the family Enterobacteriaceae, only the Bacillus spp. were found to ferment African oil bean seeds to ugba. Bacillus spp. were the predominant microorganisms present, constituting over 95% of the total microbial population density. An increase in the number of Bacillus cells of about 2 log units daily, which attained a maximum density of log₁₀ 9.00 - log₁₀ 11.90 cfu/g after 3 days was observed. Contrarily, the Lactobacillus spp. increased minimally and attained a maximum value of log₁₀ 4.20 - log₁₀ 6.35 cfu/g within the same period. The Staphylococcus spp., Micrococcus spp. and the members of the family Enterobacteriaceae remained fairly steady in number for 24h, increased slightly till the 3rd day followed by exponential increases which attained maximum values of between log₁₀ 9.20 - log₁₀ 11.00, about the 7th day. Bacillus spp. cells also had the highest protease activities which were significantly (p < 0.05) higher than the values for the other bacterial isolates. The Bacillus spp. responsible for the fermentation of African oil bean seeds to ugba were identified as Bacillus coagulans, B. macerans, B. megaterium, B. pumilis and B. subtilis.     

Chemical Diversity and Biological Properties of Secondary Metabolites from Sea Hares of Aplysia Genus

The marine environment is an important source of structurally-diverse and biologically-active secondary metabolites. During the last two decades, thousands of compounds were discovered in marine organisms, several of them having inspired the development of new classes of therapeutic agents. Marine mollusks constitute a successful phyla in the discovery of new marine natural products (MNPs). Over a 50-year period from 1963, 116 genera of mollusks contributed innumerous compounds, Aplysia being the most studied genus by MNP chemists. This genus includes 36 valid species and should be distinguished from all mollusks as it yielded numerous new natural products. Aplysia sea hares are herbivorous mollusks, which have been proven to be a rich source of secondary metabolites, mostly of dietary origin. The majority of secondary metabolites isolated from sea hares of the genus Aplysia are halogenated terpenes; however, these animals are also a source of compounds from other chemical classes, such as macrolides, sterols and alkaloids, often exhibiting cytotoxic, antibacterial, antifungal, antiviral and/or antifeedant activities. This review focuses on the diverse structural classes of secondary metabolites found in Aplysia spp., including several compounds with pronounced biological properties.     

Secondary Metabolite Production Potential of Mangrove-Derived Streptomyces olivaceus

Mangroves are intertidal extreme environments with rich microbial communities. Actinobacteria are well known for producing antibiotics. The search for biosynthetic potential of Actinobacteria from mangrove environments could provide more possibilities for useful secondary metabolites. In this study, whole genome sequencing and MS/MS analysis were used to explore the secondary metabolite production potential of one actinobacterial strain of Streptomyces olivaceus sp., isolated from a mangrove in Macau, China. The results showed that a total of 105 gene clusters were found in the genome of S. olivaceus sp., and 53 known secondary metabolites, including bioactive compounds, peptides, and other products, were predicted by genome mining. There were 28 secondary metabolites classified as antibiotics, which were not previously known from S. olivaceus. ISP medium 2 was then used to ferment the S. olivaceus sp. to determine which predicted secondary metabolite could be truly produced. The chemical analysis revealed that ectoine, melanin, and the antibiotic of validamycin A could be observed in the fermentation broth. This was the first observation that these three compounds can be produced by a strain of S. olivaceus. Therefore, it can be concluded that Actinobacteria isolated from the mangrove environment have unknown potential to produce bioactive secondary metabolites.     

芝麻种子带菌检测及药剂消毒处理效果

采用吸水纸检测法对我国6个芝麻主产区的13个广泛栽培的芝麻品种进行了带菌检测,并测定了5种杀菌剂对芝麻种子的消毒效果。结果表明,芝麻种子外部携带真菌的主要类群包括镰孢属（Fusarium spp.）、棒孢属（Corynespora spp.）、长蠕孢属（Helminthosporium spp.）、壳球孢属（Macrophomina spp.）、链格孢属（Alternaria spp.）、曲霉属（Aspergillus spp.）和青霉属（Penicillium spp.）,其中引起芝麻茎点枯病的壳球孢属分离频率和带菌率最高。种子内部携带的真菌类群主要包括镰孢属、棒孢属、壳球孢属和曲霉属等,引起芝麻叶枯病的棒孢属真菌分离频率和带菌率最高。供试的5种杀菌剂对芝麻种子均有一定的消毒效果,其中50 %扑海因和80%多菌灵处理对芝麻种子消毒效果最显著。     

海南省甘蔗寄生线虫调查与鉴定

2011～2014年,调查了海南省6个县市125份甘蔗根际土样的植物线虫发生情况。结果显示,海南甘蔗地每百克土壤中平均含有植物寄生线虫57条,其中沙性土壤中线虫平均含量达到143条,高于粘性土壤的42条。所分离出的线虫分布于13个属,包括矮化线虫属(Tylenchorhynchus spp.)、根结线虫属(Meloidogyne spp.)、短体线虫属(Pratylenchus spp.)、剑线虫属(Xiphenema spp.)、长针线虫属(Longidorus spp.)、毛刺线虫属(Trichodorus spp.)、拟毛刺线虫属(Paratrichodorus spp.)、拟长针线虫属(Paralongidorus spp.)、螺旋线虫属(Helicotylenchus spp.)、肾形线虫属(Rotylenchulus spp.)、垫刃线虫属(Tylenchus spp.)、滑刃线虫属(Aphelenchus spp.)和小环线虫属(Criconemella spp.)等。根据形态学特征及测量值,初步鉴定饰环矮化线虫(Tylenchorhynchus annulatus)及玉米短体线虫(Pratylenchus zeae)为海南甘蔗上的优势种类。     

Relative abundance and damage by target and non-target insects on Bollgard and BollgardII cotton cultivars

Bollgard^® and BollgardII^® cotton cultivars were evaluated for their efficacy for control of bollworms and their effects on sucking insects and the abundance of natural enemies under bollworm insecticide protected and non-insecticide protected field conditions during the 2004 and 2005 cropping seasons. Bollgard cultivars are genetic transforms that produce Cry1Ac toxins from Bacillus thuringiensis (Bt) to control American bollworm, Helicoverpa armigera. Second generation BollgardII cultivars contain 2 toxins, Cry1Ac and Cry2Ab, and are more effective in controlling a broader range of caterpillar species. Bollgard cultivars were free of H. armigera damage until harvest under insecticide protected as well as non-protected conditions. Both BollgardII and Bollgard cultivars were infested with larvae of spotted bollworm, Earias vitella, and spiny bollworm, Earias insulana, at later crop growth stages. Neither BollgardII nor Bollgard cultivars were free of square (fruiting body), open boll and loculi damage. Bollworm damage did not reach economic threshold levels up to harvest. Densities of sucking insects (Amrasca biguttula biguttula, Bemisia tabaci, Aphis gossypi, and Thrips tabaci), of the foliage feeder Myllocerus undecimpustulatus and of predators (Chrysoperla spp., Orius spp., Coccinella spp., Brumus spp., Vespa spp., Lycosa spp., and Aranews spp.) were similar on Bollgard, BollgardII and conventional cultivars. The time of the first appearance of bollworms, sucking insects and predators on Bt cotton did not vary from conventional cotton varieties. Insecticidal protection based on economic threshold levels resulted in significant reductions in bollworm damage on conventional cultivars.     

木麻黄小枝内含物及其对木毒蛾抗性的关系

为探讨木麻黄小枝内含物与其对木毒蛾抗性的关系,明确其抗性机理,对抗虫品系湛江3、惠76、惠83、广东A8-2,中抗品系平潭2、惠1、龙7-18,感虫品系东山2号、广东501、抗风10个木麻黄品系的黄酮、单宁、总酚、蛋白质、可溶性糖、多糖6种内含物含量进行了测定和分析比较。结果表明:总体上木麻黄小枝内次生物质黄酮、单宁、总酚在抗性品系中含量较高,在感虫品系中含量较低;而蛋白质和可溶性糖却在感虫品系中较高,在抗虫品系中较低;多糖含量则无明显规律性。灰色关联度分析结果显示,抗虫等级、平均化蛹率和平均蛹重均与蛋白质含量和可溶性糖含量关联系数较大,均大于或接近0.500 0。     

海洋青霉菌抗真菌活性物质高产菌株的诱变选育

采用紫外线(UV)、激光(Laser)、亚硝基胍(NTG)复合诱变HN89青霉菌后，再用HN89菌株的次生代谢产物(HN89A)对复合诱变菌株进行了自我抗性的选育．结果表明，与出发菌相比，菌株经诱变后相对效价显提高．自我抗性诱变结果表明：在67株经复合诱变的抗性菌株中，其抗真菌活性物质相对效价高于出发菌株的正变株有60株，正变率达到90％。其中相对效价提高50％以上的菌株为2l株，占所筛选菌株总数的31％；效价提高100％以上的菌株为9株，占13％，其中突变株HN89-68活性物质的相对效价可提高到286％。活性物质效价提高幅度与其自身产量提高幅度存在一定的正相关。诱变高产菌株遗传稳定性好．     

Recent Advances in Drug Discovery from South African Marine Invertebrates

Recent developments in marine drug discovery from three South African marine invertebrates, the tube worm Cephalodiscus gilchristi, the ascidian Lissoclinum sp. and the sponge Topsentia pachastrelloides, are presented. Recent reports of the bioactivity and synthesis of the anti-cancer secondary metabolites cephalostatin and mandelalides (from C. gilchristi and Lissoclinum sp., respectively) and various analogues are presented. The threat of drug-resistant pathogens, e.g., methicillin-resistant Staphylococcus aureus (MRSA), is assuming greater global significance, and medicinal chemistry strategies to exploit the potent MRSA PK inhibition, first revealed by two marine secondary metabolites, cis-3,4-dihydrohamacanthin B and bromodeoxytopsentin from T. pachastrelloides, are compared.