Endomelanconiopsis microspora发酵产物乙酸乙酯提取物对人参核盘菌的抑制机理

【背景】人参菌核病是人参的主要病害之一,严重影响人参的产量。【目的】探索白花蒲公英内生菌(Endomelanconiopsis microspora)发酵产物乙酸乙酯提取物对人参核盘菌的抑制机理。【方法】采用人参核盘菌菌丝生长和孢子萌发试验测定抑制效果;采用显微镜观察菌丝形态变化,通过电导率和核酸含量的变化测定细胞膜通透性,通过丙二醛(malondialdehyde,MDA)含量和超氧化物歧化酶(superoxide dismutase,SOD)、过氧化物酶(peroxidase,POD)和过氧化氢酶(catalase,CAT)活力的变化测定膜脂过氧化程度。【结果】内生菌E. microspora发酵产物乙酸乙酯提取物能显著抑制人参核盘菌菌丝生长,最小抑菌浓度为3.75 mg/mL,培养6 d后抑制率为76.22%。该提取物能显著抑制人参核盘菌孢子萌发,15.00 mg/mL时抑制效果最好,抑制率达90.69%。提取物影响菌丝形态,增加人参核盘菌细胞膜通透性,造成菌丝内含物外渗,7.50 mg/mL处理10 h后电导率和核酸含量分别比对照组增加30.11%和62.85%。同时提取物显著增加人参核盘菌MDA含量和SOD、POD、CAT活力,7.50 mg/mL处理组呈现先上升后下降的变化趋势,并在12 h时达到最高值。【结论】内生菌E. microspora发酵产物乙酸乙酯提取物通过改变人参核盘菌细胞膜通透性,加剧膜脂过氧化,破坏细胞膜完整性,导致细胞内含物流失,显著抑制孢子萌发和菌丝生长。     

黄瓜枯萎病生防菌HD-087产抗菌物质条件的 优化及抑菌作用初探

【目的】优化比基尼链霉菌HD-087摇瓶发酵条件,提高菌株发酵液对黄瓜枯萎病菌HU-M的抑制率,并通过拮抗试验初步评价发酵液的抑菌作用效果。【方法】采用单因素筛选及正交试验对HD-087的发酵培养基和发酵条件进行优化。经发酵液处理后,光学显微镜观察HU-M菌丝形态和孢子萌发抑制率,测定HU-M菌丝电导率。【结果】改进的发酵培养基配方为:淀粉1.00%、黄豆粉0.80%、酵母粉0.12%、CaCO30.40%。对发酵条件的研究表明:pH为6.8,180 r/min、28°C条件下,250 mL三角瓶装液量为40 mL,接种1 mL种龄为2 d的种子,发酵5 d为最佳培养条件。抑菌结果表明,HD-087产抗菌物质能造成病原菌HU-M菌丝细胞质渗漏,菌丝畸形,分生孢子萌发受抑制,5倍发酵稀释液孢子抑制率达72.1%;除此之外还能引起菌丝电解质渗漏,造成菌丝细胞膜受损。【结论】优化后的摇瓶发酵条件能提高生防菌HD-087发酵液抑菌效果,并且发酵液可破坏细胞膜明显抑制病原菌HU-M生长,具有较大开发应用潜力。     

假单胞菌YL11对扩展青霉的抑制作用及其机理初探

【背景】苹果青霉病是由扩展青霉引起的一种重要的果实采后病害,影响果实品质导致苹果腐烂从而造成经济损失。【目的】研究假单胞菌YL11对扩展青霉的抑制作用和苹果采后青霉病的防治效果,并对抑菌机理进行初步探讨。【方法】以扩展青霉为供试菌株,研究不同浓度的假单胞菌YL11无菌发酵液对扩展青霉菌落直径、孢子萌发率、菌丝体干重、苹果损伤接种病斑直径扩展的影响,利用对电导率、核酸及蛋白释放量、AKP含量、SDH活性、ATP酶活性和ATP含量的影响对抑菌机理进行探究。【结果】假单胞菌YL11无菌发酵液能有效抑制扩展青霉生长,抑菌圈直径为22.33±0.27 mm,抑菌效价为71.67 mm/mL;能有效抑制孢子萌发,100%无菌发酵液对孢子萌发抑制率达到80.2%;对扩展青霉的生物量也有一定抑制作用,体积分数为100%时,菌丝体干重为4.7mg/mL,抑制率达到39.74%;无菌发酵液处理能有效抑制苹果青霉病病斑的扩展,3d时对病斑扩展的抑制率最大,达到47.1%;无菌发酵液处理均能引起电导率升高、胞内核酸和蛋白释放量增大、胞外AKP含量升高、SDH活性降低、ATP酶活性和ATP含量均降低,且随着发酵液浓度的增加效果越明显。【结论】假单胞菌YL11能显著抑制扩展青霉的生长,破坏细胞膜结构、降低能量代谢酶活性,从而扰乱扩展青霉的正常生长,对苹果青霉病有较好的生防效果,具有潜在的开发价值。     

甘蔗内生解淀粉芽孢杆菌CGB15的分离、鉴定及生防活性

真菌病害占作物病害种类的一半以上,病原真菌是目前已知种类最多的作物病原菌。从作物根际与/或体内分离筛选具有生防活性的微生物,并应用于病害的防控,是除作物品种改良与化学防治外的另一种高效的病害防控策略。【目的】本研究拟筛选并分离鉴定对重要作物病原真菌具有拮抗作用的甘蔗内生细菌,为开发生物防治作物真菌病害新策略提供理论依据。【方法】采用平板对峙法初步筛选对病原真菌具有拮抗能力的甘蔗叶片内生细菌,通过16SrRNA基因测序鉴定其种属;进一步检测候选拮抗内生细菌对甘蔗鞭孢堆黑粉菌(Sporisorium scitamineum)致病发育过程关键步骤：有性配合/菌丝生长、冬孢子萌发的抑制率,田间试验检测其对甘蔗鞭黑穗病的防治效果;检测候选拮抗内生细菌对稻梨孢菌(Pyricularia oryzae)附着胞形成、离体叶片及盆栽条件下叶片病斑形成的抑制作用。【结果】分离自甘蔗叶片的细菌菌株,编号为CGB15,经分子鉴定为解淀粉芽孢杆菌(Bacillus amyloliquefaciens)。CGB15菌株能有效抑制甘蔗鞭孢堆黑粉菌有性配合/菌丝生长,对峙培养条件下使真菌菌落呈现光滑;抑制冬孢子萌发,...     

香芹酚和丁香酚对腐皮镰刀菌的抑菌活性及抑菌机理

【背景】腐皮镰刀菌(Fusarium solani)是世界上最具破坏性的土传病原菌之一,严重影响作物的产量及品质。因此寻找并开发广谱的可持续生物防治药剂迫在眉睫,而植物次级代谢物为自然界筛选生物源天然杀菌剂提供了丰富的原料。【目的】研究香芹酚和丁香酚对腐皮镰刀菌的抑菌活性,探究其可能涉及的抑菌机理。【方法】采用菌丝生长速率法、十字交叉法和孢子萌发法分析香芹酚和丁香酚对菌丝和孢子的抑制活性,通过扫描电镜(scanning electron microscope,SEM)观察菌丝微观形态的变化,利用荧光染料碘化丙啶(propidium iodide,PI)观察细胞膜的损伤情况,并测定其对腐皮镰刀菌细胞膜胞外电导率、蛋白质含量和麦角固醇合成的影响。【结果】香芹酚和丁香酚对腐皮镰刀菌菌丝生长和孢子萌发均有显著的抑制效果,并呈现剂量依赖效应,EC50值分别为92.39μL/L和263.00μL/L。SEM结果表明,2种精油处理腐皮镰刀菌后,其细胞壁和细胞膜均遭到破坏从而不能维持菌丝正常的线性形态,表现出不同程度的弯曲、褶皱和凹陷。PI染色结果发现,2种精油处理严重破坏了腐皮镰刀菌细胞膜的完整性和...     

月腺大戟根总黄酮对尖孢镰刀菌抑制作用的研究

采用生长速率法和孢子萌发法分别测定月腺大戟根总黄酮对尖孢镰刀菌菌丝生长和分生孢子萌发的抑制率,显微观察总黄酮处理尖孢镰刀菌后菌丝体形态结构的变化,并测定菌丝体相对电导率,菌丝体内超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性。结果表明:月腺大戟根总黄酮对尖孢镰刀菌菌丝生长和分生孢子萌发均有显著的抑制作用,抑制率随总黄酮浓度增加而增高,20 mg/L时抑制率达100%。总黄酮处理后的尖孢镰刀菌菌丝较细,分支减少,透明度差,液泡数量增多且形成较大的液泡;菌丝体细胞膜透性增加,SOD、CAT活性呈先上升后下降的趋势。以上实验结果可为植物病害生物防治及开发植物源农药方面提供理论依据和实验技术。     

独蒜兰提取物对植物病原真菌的抑菌活性

【目的】研究独蒜兰假鳞茎乙醇提取物对植物病原真菌的抑菌作用,为植物源杀菌剂的开发提供依据。【方法】采用菌丝生长速率法,研究独蒜兰假鳞茎乙醇提取物对15种植物病原菌的抑制活性;以西瓜尖孢镰刀菌作为供试菌,进一步研究该提取物对病原真菌的菌丝干重、细胞膜、过氧化氢酶（CAT）、过氧化物酶（POD）和超氧化物歧化酶（SOD）等的影响。【结果】独蒜兰假鳞茎提取物对辣椒疫霉病菌、西瓜尖孢镰刀菌、番茄灰霉病菌和非洲隐地疫霉菌的抑菌效果明显,其EC₅₀值分别为0.849、0.782、0.813和1.161 mg·mL^-1;经独蒜兰提取物处理后的西瓜尖孢镰刀菌菌丝干重随着药剂浓度的增加而减少;细胞膜丙二醛含量和相对电导率增加;菌丝体细胞内CAT、POD和SOD 3种保护酶活性增加。【结论】独蒜兰假鳞茎提取物对植物病原真菌具有较好的抑菌活性,其抑菌作用可能与其干扰菌丝生长、使菌丝细胞膜正常功能受损等有关。     

鹅膏菌抑真菌活性及其活性物质的提取工艺优化(英文)

通过鹅膏菌抑菌活性物质对杨树烂皮病病原菌金黄壳囊孢菌的菌丝生长及其孢子抑制效果的研究,分析鹅膏菌的抗真菌活性。将培养15 d的鹅膏菌液体发酵物分成培养液和菌丝体两部分,然后分别将培养液、菌丝体提取物和发酵液本身对金黄壳囊孢菌菌丝生长和孢子萌发抑制效果进行检测。结果表明:培养液对金黄壳囊孢菌菌丝生长的抑制率最高,为68.90%;三者对孢子萌发都有明显的抑制效果,即在处理后10 d孢子都没有萌发。菌丝体在超声波、加热和浸泡的条件下,利用乙醇,丙酮、乙酸乙酯和正丁醇进行提取,其提取物用于病原菌的抑制效果检测,结果表明:用乙醇和丙酮提取时,超声波、浸泡及加热处理,对金黄壳囊孢菌的抑制效果都与对照有显著差异(a=0.01),而正丁醇的提取物的抑制效果较差,与对照没有显著差异。通过不同提取物对金黄壳囊孢菌菌丝生长和孢子萌发的抑制效果进行分析,并对抑菌活性物质提取工艺进行筛选,得到抑菌活性物质提取的最佳方法为摇床培养,乙醇是最适提取溶剂,处理方式(超声波、浸泡或者加热)相互间差异不显著。     

一株表面活性剂产生菌的分离及抑菌活性

【目的】研究分离得到的表面活性剂产生菌的产表面活性剂能力、分类地位和抑菌活性。【方法】采用血平板、油平板进行表面活性剂产生菌的分离,以排油圈法进行表面活性的测定;通过生理生化特性和16S rDNA序列相似性分析对BS1菌株进行初步鉴定;利用对峙培养法和菌丝生长、孢子囊形成、孢子萌发的抑制率测定研究其抑菌活性。【结果】从石油污染土壤中分离到的BS1菌株可产生表面活性剂,在分类学地位上属于假单胞菌属(Pseudomonas sp.)。BS1菌体、发酵上清液、挥发性物质对12种供试病原真菌均表现出一定的抑制作用。BS1菌体、发酵上清液对大豆疫霉菌(Phytophthora sojae)的抑制率最大,分别达到65.31%和95.93%。发酵上清液通过影响大豆疫霉菌菌丝生长、孢子囊形成、孢子萌发等方式抑制病原菌的正常生长,稀释20倍的发酵上清液依然具有明显的抑制作用。BS1菌株产生的挥发性物质对大豆菌核菌(Sclerotinia sclerotiorum)的抑菌效果最好,抑制率达到84.25%。【结论】BS1菌株在产生表面活性剂的同时,还具有生物防治作用潜力。     

关于茭白黑粉菌的正名问题

茭白黑粉菌在文献中出现Yenia esculenta(P.Henn.)Liou及Ustilago esculenta P.Henn.两个名字。现经形态发生学的研究以明确新名字是否有效。此菌冬孢子没有休眠期,孢子成熟后在适宜的环境中即可萌发。萌发的最适pH值为6;最适温度为25％;不需要外界光的诱导,在黑暗条件下也能正常萌发;培养基中的营养成份对孢子萌发的速率有一定影响。冬孢子在不同的环境条件下,其萌发形态及方式都是稳定的:孢子萌发时产生有隔的担子或先菌丝(promycelium)。该先菌丝起初可以是无隔的,但生长到一定程度,便产生分隔,分隔多为2—4个。初生的先菌丝和小孢子是单核的,但成熟的先菌丝、小孢子及短菌丝细胞内均是双核的。可以认为,该菌的双核期在整个生活史中占有相当长的时间,它只有短暂的单核期。没有发现先菌丝、小孢子彼此之间或相互间融合的现象;未见到双核的融合及减数分裂过程。茭白黑粉菌在人工组合培养基上生成厚壁的孢子,较自然界中的冬孢子为大,但萌发方式则相同。最后作者等认为茭白黑粉菌仍应保留在Ustilago属中,而Yenia esculenta(P.Henn.)Liou的名字是无效的。     

Strain Identification and Quantitative Analysis in Microbial Communities

Microbiology has long studied the ways in which subtle genetic differences between closely related microbial strains can have profound impacts on their phenotypes and those of their surrounding environments and communities. Despite the growth in high-throughput microbial community profiling, however, such strain-level differences remain challenging to detect. Once detected, few quantitative approaches have been well-validated for associating strain variants from microbial communities with phenotypes of interest, such as medication usage, treatment efficacy, host environment, or health. First, the term “strain” itself is not used consistently when defining a highly-resolved taxonomic or genomic unit from within a microbial community. Second, computational methods for identifying such strains directly from shotgun metagenomics are difficult, with several possible reference- and assembly-based approaches available, each with different sensitivity/specificity tradeoffs. Finally, statistical challenges exist in using any of the resulting strain profiles for downstream analyses, which can include strain tracking, phylogenetic analysis, or genetic association studies. We provide an in depth discussion of recently available computational tools that can be applied for this task, as well as statistical models and gaps in performing and interpreting any of these three main types of studies using strain-resolved shotgun metagenomic profiling of microbial communities.     

Perceptual strain index for heat strain assessment in an experimental study: An application to construction workers

Although the physiological strain index (PhSI) is universal and comprehensive, its restrictions are recognized in terms of invasive on-site measurements and the requirement of accurate instruments. The perceptual strain index (PeSI) has been proposed as a user-friendly and practical indicator for heat strain. However, the application of this index in assessing the heat strain of construction workers has yet to be examined and documented. This study aims to ascertain the reliability and applicability of PeSI in an experimental setting that simulates a stressful working environment (i.e., environment, work uniform, and work pace) experienced by construction workers. Ten males and two females performed intermittent exercise on a treadmill while wearing a summer work uniform at 34.5 °C and 75% relative humidity in a climatic chamber. Physiological parameters (core temperature, heart rate) and perceptual variables (thermal sensation, perceived exertion) were collated synchronously at 3 min intervals. The results of two-way repeated measures analysis of variance (clothing×time) revealed that the PeSI was useful in differentiating the heat strain levels between different work uniforms. Not only did the PeSI change in the same general manner with the PhSI, but it was also powerful in reflecting different levels of physiological strain. Thus, the PeSI offers considerable promise for heat strain assessment under simulated working conditions.     

低温下水稻幼苗形态生理应变研究

低温下水稻幼苗形态生理应变研究李太贵王磊（中国水稻研究所，杭州３１０００６）ＳｔｕｄｙｏｆＭｏｒｐｈｏＰｈｙｓｉｏｌｏｇｉｃａｌＳｔｒａｉｎｏｆＲｉｃｅＳｅｅｄｌｉｎｇＵｎｄｅｒＬｏｗＴｅｍｐｅｒａｔｕｒｅ．ＬｉＴａｉｇｕｉ，ＷａｎｇＬｅｉ（Ｃｈｉ...     

Construction of an inducible, pfkA and pfkB deficient strain of Escherichia coli for the expression and purification of phosphofructokinase from bacterial sources

As the key obligatory step in the glycolytic pathway, the regulation of phosphofructokinase (PFK-1) has been the focus of study of several laboratories. While standard cloning procedures have opened the door to study PFK from a vast array of sources, a good pfk knockout Escherichia coli strain has not previously been developed. Many laboratories rely on DF1020 or similar derivatives for PFK expression. Unfortunately, DF1020 grows poorly and does not have an inherent means for controlling expression of genes from plasmids. More importantly, however, DF1020 has a tendency to grow on minimal media when glucose is used as the sole carbon source. In this study, a new E. coli PFK expression strain lacking both PFK-1 and PFK-2 has been engineered using lambda-red mediated chromosomal deletion. The resulting strain has been designated RL257. In addition to having both pfkA and pfkB deleted, RL257 contains the lacI(q) allele, which allows for inducible expression when coupled with an expression vector containing either the lac or tac promoter.     

Cadmium sequestration in cells of two stains of Alcaligenes eutrophus

Abstract Alicaligenes eutrophus CH34 and its plasmid-free derivative were cultivated in the presence of cadmium. The wild type strain A. eutrophus CH34 harboured two plasmids and was cadmium-resistant, while the plasmid-free mutant AE104 was cadmium-sensitive. The Cd-resistant strain immobilized more cadmium than the Cd-sensitive one. Cadmium was mainly located in the cell envelopes of the Cd-resistant strain. Several cellular modifications were observed: thickening of the cell envelopes and changes int he peptidoglycan, i.e. its proportion in the envelopes, the ratio between glutamic acid and diaminopimelic acid, the presence of aspartic acid. These modifications should contribute to increase the cadmium sequestration capacity of the envelopes.     

竹红菌素产生菌的筛选与鉴定

通过多点采集、反复筛选,从野生竹黄子实体中筛选出1株产北醌类光敏色素的真菌。通过对菌株的个体形态特征、菌落形态特征进行观察,结合其显微结构特征最终鉴定为竹黄菌（Shiraia bambuscola)。     

Immersion anaesthesia with tricaine methanesulphonate or propofol on different sizes and strains of silver catfish Rhamdia quelen

The efficacy of immersion anaesthesia with tricaine methanesulphonate (MS222) or propofol on albino and grey silver catfish Rhamdia quelen was assessed through induction and recovery times and observation of mortality. Besides reporting a novel, efficient and practical use of propofol as an immersion anaesthetic, the study shows that it is essential to consider size and strain when anaesthetizing R. quelen with MS222 or propofol bath solution in order to minimize physiological impact.     

提高Lovastatin产生菌生物合成能力的研究

用微波等离子体(N^ 20w，4win)诱变M．ruber，筛选到一株高产菌株，再经过发酵工艺优化，最终Lovastaitin的发酵产率提高21．8％。     

磷细菌PB12菌株的选育及培养条件的研究

以PB-1P为出发菌株,经紫外线诱变,获得1株解磷高产菌株PB12。在以磷酸三钙为惟一磷源的培养基中接种PB12菌株,28℃,振荡培养5d,培养基上清液中磷含量达61.21mg/L,是出发菌株的2.03倍。同时研究了各种理化因素对PB12菌株生长的影响,确定了最佳培养条件,即玉米粉1.0%,豆饼粉2.0%,pH7.0,装液量为100mL(500mL三角瓶),36℃培养42h,有效活菌数可达10.2×108cfu/mL。     

Genetic diversity of Neisseria meningitidis strains isolated in Rio de Janeiro, Brazil, evaluated by multilocus enzyme electrophoresis

AIMS: To analyse Neisseria meningitidis isolates from meningococcal meningitis cases in Rio de Janeiro (Brazil) from 1990 to 1993 and 1999-2002, to determine the genetic and relatedness with hypervirulent and epidemic strains. METHODS AND RESULTS: The isolates were analysed by multilocus enzyme electrophoresis (MEE) clustering into 83 electrophoretic types (ET). All isolates from 1999 to 2002, formed a cluster which included one strain of the ET-5 complex worldwide associated with epidemics. CONCLUSIONS: The overall results suggested a panmictic structure probably because of recombination events. The observation of a separated cluster including isolates from 1999 to 2002 and an ET-5 complex strain, also suggested the introduction of strains genetically related with this hypervirulent complex in the State of Rio de Janeiro (Brazil) over the last 5 years. SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of strains related to the ET-5 complex in several states of Brazil was already described elsewhere, but this is the first time it was reported in the State of Rio de Janeiro. Our findings reinforce the necessity to genetically determine the clones which should be considered to produce a national vaccine against meningococcal meningitis.