低温环境中T-2毒素降解菌的分离鉴定及特性研究

离筛选低温环境中T-2毒素的降解菌并探明其生化特性,探究T-2毒素降解微生物生化特性异同点,为T-2毒素降解微生物的检验提供生化判断参考。针对暴露于－20 ℃低温环境中的低浓度T-2毒素标准品,采用LC-MS/MS定量分析T-2毒素残留量,利用营养琼脂培养基（NA）和马铃薯葡萄糖琼脂培养基（PDA）共分离出5株降解菌,16S rDNA结合生化鉴定结果为死谷芽胞杆菌（Bacillus vallismortis）、蜡状芽胞杆菌（Bacillus cereus）、阴沟肠杆菌（Enterobacter cloacae）、弯曲假单胞菌（Pseudomonas geniculata）和尼泊尔葡萄球菌（Staphylococcus nepalensis）。这些分离株在－20 ℃条件下对低浓度T-2毒素均有不同程度的降解能力,其中蜡状芽胞杆菌（Bacillus cereus）降解能力最强,降解率为91%。它们的理化特性具有很多相似性,主要表现在5株菌均不能利用D 塔格糖(dTAG）,对ELLMAN都表现出阴性;Bacillus vallismortis Bp1234-7、Pseudomonas geniculata Bp24-4、〖WTBX〗Staphylococcus nepalensis Bp124-5和Enterobacter cloacae Bp123-7都能分解D-葡萄糖,而Bacillus cereus Bp1234-8不能。     

嗜盐单胞菌ST-1的分离及其石油烃类降解特性的研究

从油田石油污染物中分离出了一株菌株ST-1,通过形态和16S r DNA序列的分析,初步鉴定属于Halomonas的一个种。对该菌株降解石油烃的能力进行了实验,结果表明,菌株生长的指数期石油烃降解速率较大,最大降解率达到80%以上,降解的最适温度为27~34℃,最适p H为6.8~8.3。提高接种量可以加快反应速度。     

一株异养型细菌对无机硫化物的降解特性和培养条件优化

畜禽屠宰加工、鱼粉饲料加工等一些食品工业生产过程中会释放出大量的硫化氢恶臭气体,导致周边环境的严重污染。为实现以培养异养型细菌脱除硫化氢气体的目的,取分离到的异养脱硫细菌XJ-2,通过诱变筛选得到一株高效脱硫菌株ZJNB-B3,其脱硫率达97%。基于形态学研究、API 50 CHB生理生化鉴定及16S r RNA基因测序,鉴定该菌为蜡状芽胞杆菌Bacillus cereus ZJNB-B3。该菌Gen Bank登录号为MF679650。降解特性研究表明,ZJNB-B3菌株对有毒的硫化物有较高耐受性,耐受上限高达300 mg/L。采用响应面法优化环境因素对菌株降解硫化物效率的影响,得到在最适培养温度30℃下,初始S~2–浓度为211.8 mg/L、初始p H值6.72、接种量为5.04%时,菌株氧化脱硫效果最显著,经过实测在48 h产生的硫酸盐浓度为63.8 mg/L,脱硫率达97.3%。菌株在氧化硫化物时不会产生硫酸抑制菌株的生长,可以在p H值温和的环境条件下脱硫,因此,该菌有较高的工业应用价值。本研究为异养型细菌应用于工业反应器脱除硫化氢恶臭气体提供了小试研究基础。     

微囊藻毒素降解菌的筛选、鉴定及其胞内粗酶液对藻毒素MCLR的降解

【目的】从巢湖底泥中分离筛选高效的藻毒素降解菌,并初步研究其胞内粗酶液降解藻毒素-LR(MC-LR)的特性,为水体中藻毒素污染的微生物治理提供有效的菌源与理论依据。【方法】利用富集驯化培养技术,以MC-LR为唯一碳源,分离筛选MC-LR降解菌,通过形态观察、生理生化实验及16S rRNA序列分析鉴定菌株,并考察其胞内粗酶液在不同条件下对MC-LR的降解特性。【结果】分离得到1株能高效降解MC-LR的菌株M6。分子鉴定结果表明,该菌株为蜡状芽胞杆菌(Bacillus cereus)。其降解MC-LR的活性物质为胞内酶,而且至少有3种酶参与了MC-LR的降解,它们是菌体本身的组织酶而非诱导酶。当反应体系pH值为8.0,胞内粗酶液浓度为404.9 mg/L,MC-LR的初始浓度为10 mg/L时降解率最高,16 h可达98.7%。【结论】分离出的MC-LR降解菌为蜡状芽胞杆菌,该菌株对MC-LR有较高的降解能力,并且酶促反应受到反应体系的pH值、胞内粗酶液浓度以及藻毒素初始浓度等因素的影响。     

类似S-层蛋白的苏云金芽胞杆菌伴胞晶体蛋白基因的克隆

芽胞杆菌CTC菌被鉴定为苏云金芽胞杆菌,鞭毛血清型H2,幕虫亚种;产生卵圆形伴胞晶体,伴胞晶体蛋白为100kD;测定了该蛋白 N－末端序列,该序列与炭疽芽胞杆菌的细胞表面S－层蛋白具92－93％相似性,根据Southern杂交制作了该晶体蛋白基因ctc所在位置的限制性酶切图谱,分别克隆了该基因5’和3’端所在2．9kb XbaI片段和3．1kb Cla I DNA片段,彼此间具0．6kb重叠,通过拼接获得含完整ctc基因的克隆,含该基因的大肠杆菌与表达S－层蛋白的大肠杆菌具相似生长特征,初步表明CTC菌株的伴胞晶体由细胞表面S－层蛋白组成,苏云金芽胞杆菌区别于蜡状芽胞杆菌和炭疽芽胞杆菌的唯一标准是能形成伴胞晶体,由于S－层是细胞表面的结构成分,本文对CTC菌株鉴定为苏云金芽胞杆菌以及伴胞晶体作为苏云金芽胞杆菌鉴别的唯一标准提出了质疑。     

聚丁二酸丁二醇酯降解菌株的筛选及其降解性能研究

从石油污染土壤中筛选到1株具有聚丁二酸丁二醇酯降解能力的菌株PBS1302,经菌体形态特征、菌落培养特征、生理生化鉴定和16S r DNA基因序列分析,初步鉴定为铜绿假单胞菌(Pseudomonas aeruginosa)。该菌株在培养温度37℃,培养基起始p H 6.8的条件下经6 d的培养,对聚丁二酸丁二醇酯薄膜的降解率可达36.9%。经电子显微镜观察,与降解前相比,聚丁二酸丁二醇酯薄膜表面变得粗糙,出现明显的蚀刻痕迹。     

1株产琼脂酶细菌的分离鉴定及其胞外酶活性测定

从四川省成都市青城山采集土壤,以琼脂作为唯一碳源,筛选到产琼脂酶细菌CMCK136;通过形态观察、生化鉴定、16S r DNA测序及序列分析鉴定其种属;随后测定了菌株CMCK136的胞外酶活性。菌株CMCK136被鉴定为芽胞杆菌属细菌,命名为Bacillus sp.CMCK136。菌株CMCK136的胞外琼脂酶的最适酸碱度为p H 7.0,最适温度为35℃。菌株CMCK136是产琼脂酶细菌家族的新成员,该菌株的发现进一步提示芽胞杆菌属很可能蕴含有尚待开发的琼脂酶资源。     

海洋短小枯草芽胞杆菌的鉴定及抑菌活性分析

对从连云港东西连岛海泥样品中分离得到的菌株Bacillus pumilus HX2-2的分类地位、生长条件和抑菌活性进行了研究。经过形态特征、生理生化性质及16S r DNA序列分析鉴定,该菌属于短小芽胞杆菌。不同温度、盐度、pH培养条件下测定菌液吸光度OD600值,表明该菌是一株轻度嗜盐菌,最适温度、盐度、pH分别为30℃、3%、7.0~8.0。在不同病原真菌的平板抑菌活性试验中,该菌对草莓尖胞镰刀菌、马铃薯炭疽病菌和水稻立枯丝核菌表现出显著的抑菌作用。菌株B.pumilus HX2-2是一株短小芽胞杆菌,具有广谱抑菌活性,具有进一步研究的价值。     

短小芽胞杆菌HLK8-1的分离鉴定及抑菌特性分析

从海南翁田镇凡纳滨对虾(Litopenaeusvannamei)养殖场筛选获得1株能抑制副溶血弧菌(Vibrio parahaemolyticus)等多种病原菌的芽胞杆菌HLK8-1。经形态特征分析、16S r DNA序列分析及Biolog微生物自动鉴定系统鉴定为短小芽胞杆菌(Bacillus pumilus);菌株生长及抑菌性能研究表明该菌株抑菌物质主要产生在生长对数期及平台期;相对于盐度及初始p H等因素,该菌株的抑菌性能受温度影响较大;菌株在p H值范围为7.0~8.5,盐度范围为0~0.2%,温度为30℃等条件下表现出较高的生长水平及抑菌性能;共培养试验表明该菌能将水体中的副溶血弧菌数量控制在较低水平,培养24 h时实验组105cfu/m L,而对照组108cfu/m L。该菌抑制多种水产病原菌,对水体环境耐受能力强,具有防治水产养殖病原菌的应用潜力,且易于培养,抑菌物质分泌到胞外便于分离提取,亦具有发酵工程应用潜力。     

土壤中PHA高产菌株的分离及鉴定

从成都某炼油厂油罐和排污口附近土壤中分离到1株PHA(聚羟基烷酸)的高产菌株,最高含量为19.25%。根据形态特征观察、生理生化特征鉴定及16S rDNA序列比对,结果表明该菌株为蜡状芽胞杆菌。并对其发酵条件进行初步探索,确定培养的最适温度为37℃,最适pH值为7.0,为下一步构建基因工程菌和优化发酵条件提供了理论基础。     

Nystatin and killer toxin sensitivity of free and immobilizedSaccharomyces cerevisiae

The inhibitory effect of nystatin and killer toxin on the growth of free and covalently-immobilizedSaccharomyces cerevisiae cells was studied. The resistance of immobilized cells to both agents was accompanied by increased amounts of phospholipids and sterols. The possible relationship between these changes in the membrane composition and the transduction of a signal across the cytoplasmic membrane is discussed.     

Ovule and seed anatomy ofCistaceae and relatedMalvanae

Ovule and especially seed anatomy of eight species ofCochlospermaceae, Bixaceae, Cistaceae, Monotoideae, Pakaraimaeoideae (two subfamilies ofDipterocarpaceae), andSarcolaenaceae were investigated. All representatives show a bixoid chalazal region in the seed as probable exclusive synapomorphy among angiosperms. The palisade layer of the exotegmen is curved inwards at its proximal end and forms a dome-shaped structure. A plug of hypostase tissue with an annulus/core structure fits into this dome. Moreover, two additional tissue types in the hypostase can be found in some representatives of the group. These and other micromorphological, wood anatomical, and floral morphological characters, indicate that the taxa form a monophyletic group close toMalvales s. str. The form of the starch grains in the endosperm is compared and is described for the first time forPakaraimaea (Dipterocarpaceae) andLeptolaena (Sarcolaenaceae). The position ofDiegodendron close toBixa and the presumably more distant positions ofMuntingia andNeuradaceae are discussed.     

The hernandez and fernandez genes of Drosophila specify eye and antenna

The formation of different structures in Drosophila depends on the combined activities of selector genes and signaling pathways. For instance, the antenna requires the selector gene homothorax, which distinguishes between the leg and the antenna and can specify distal antenna if expressed ectopically. Similarly, the eye is formed by a group of "eye-specifying" genes, among them eyeless, which can direct eye development ectopically. We report here the characterization of the hernandez and fernandez genes, expressed in the antennal and eye primordia of the eye-antenna imaginal disc. The predicted proteins encoded by these two genes have 27% common amino acids and include a Pipsqueak domain. Reduced expression of either hernandez or fernandez mildly affects antenna and eye development, while the inactivation of both genes partially transforms distal antenna into leg. Ectopic expression of either of the two genes results in two different phenotypes: it can form distal antenna, activating genes like homothorax, spineless, and spalt, and it can promote eye development and activates eyeless. Reciprocally, eyeless can induce hernandez and fernandez expression, and homothorax and spineless can activate both hernandez and fernandez when ectopically expressed. The formation of eye by these genes seems to require Notch signaling, since the induction of ectopic eyes and the activation of eyeless by the hernandez gene are suppressed when the Notch function is compromised. Our results show that the hernandez and fernandez genes are required for antennal and eye development and are also able to specify eye or antenna ectopically.     

Development and function of Pisolithus and Scleroderma ectomycorrhizas formed in vivo with Allocasuarina,Casuarina and Eucalyptus

The effect of inoculating seedlings of Eucalyptus grandis, Allocasuarina littoralis and Casuarina equisetifolia with two isolates of Pisolithus and two isolates of Scleroderma from under eucalypts was examined in a glasshouse trial. Ectomycorrhizas formed extensively on Eucalyptus (23–46% fine roots ectomycorrhizal) and Allocasuarina (18–51% fine roots ectomycorrhizal). On Casuarina, the fungi were either unable to colonize the rhizosphere (one isolate of Pisolithus), or sheathed roots, resembling ectomycorrhizas, formed on 1–2% of the fine roots. Colonization of roots by one isolate of Scleroderma resulted in the death of Casuarina seedlings. Inoculation with fungi increased shoot dry weight by up to a factor of 32 (Eucalyptus), 4 (Allocasuarina) and 3 (Casuarina). Ectomycorrhizas formed in associations with Eucalyptus and Allocasuarina had fully differentiated mantles and Hartig nets in which the host and fungal cells were linked by an extensive fibrillar matrix. Sheathed roots in Casuarina lacked a Hartig net, and the epidermis showed a hypersensitive reaction resulting in wall thickening and cell death. The sheaths are described as mantles since the density and arrangement of the hyphae in the sheaths was similar to that in mantles of the eucalypt ectomycorrhizas. The intercellular carbohydrate matrix was not produced in the Casuarina mantle in association with Pisolithus, hence the mantle was not cemented to the root. These structures differ from poorly compatible associations described previously for Pisolithus and Eucalyptus. The anatomical data indicate that ectomycorrhizal assessment based on surface morphological features may be misleading in ecological studies because compatible and incompatible associations may not be distinguishable.     

Y-chromosome Data and Tribal Affiliations of Allenopithecus and Miopithecus

The phylogeny of Old World monkeys has remained unresolved in part because of a lack of resolution in the Cercopithecinae. Competing morphological hypotheses have had Allen's swamp monkey (Allenopithecus nigroviridis) and the talapoins (Miopithecus spp.) as basal branches of either the tribe Cercopithecini or the tribe Papionini. Previous molecular analyses have not adequately addressed the issue. To better understand the evolutionary history of these primates, we sequenced and subjected to phylogenetic analysis 3.1 kb of 2 loci (TSPY and SRY) from the non-recombining portion of the Y-chromosome. Individuals from the genera Allenopithecus, Miopithecus, Erythrocebus, Chlorocebus, and Cercopithecus were surveyed and their sequences compared with those previously published for the Papionini and Colobinae. The results suggest Allenopithecus and Miopithecus are more closely related to the Cercopithecini than Papionini. Our data also support the hypothesis that within the Cercopithecini, Erythrocebus and Chlorocebus share a close evolutionary relationship, distinct from the other members of the tribe.     

Amino acids and sugars in nectar,and their putative evolutionary significance

Individual amino acids and sugars from flower nectar of 32 plant species with different pollination systems were quantified and compared. Data show that there is no correlation between sugar and amino acid concentration. Furthermore there is no correlation between composition and concentration of amino acids and evolutionary advancement, nor any direct relation with pollination systems. However, higher sugar concentrations are often linked with more advanced morphological characters. Nectars from pierced or damaged flowers or nectars contaminated with pollen exhibit modifications and increases in amino acid composition. The presence of proline probably indicates such pollen contamination. Most pollinating animals depend on flower nectar in their energetic requirements, yet innumerable alternative amino acid and protein sources exist. Future research has to consider the relationship between nutritional requirements of pollinating animals and dependence on flower nectars.Dedicated to Prof. Dr.L. van der Pijl, Den Haag, in honour of his 80th birthday.     

Cyclooxygenase inhibitory and antioxidant cyanidin glycosides in cherries and berries

Anthocyanins from tart cherries, Prunus cerasus L. (Rosaceae) cv. Balaton and Montmorency; sweet cherries, Prunus avium L. (Rosaceae); bilberries, Vaccinum myrtillus L. (Ericaceae); blackberries, Rubus sp. (Rosaceae); blueberries var. Jersey, Vaccinium corymbosum L. (Ericaceae); cranberries var. Early Black, Vaccinium macrocarpon Ait. (Ericaceae); elderberries, Sambucus canadensis (Caprifoliaceae); raspberries, Rubus idaeus (Rosaceae); and strawberries var. Honeoye, Fragaria x ananassa Duch. (Rosaceae), were investigated for cyclooxygenase inhibitory and antioxidant activities. The presence and levels of cyanidin-3-glucosylrutinoside 1 and cyanidin-3-rutinoside 2 were determined in the fruits using HPLC. The antioxidant activity of anthocyanins from cherries was comparable to the commercial antioxidants, tert-butylhydroquinone, butylated hydroxytoluene and butylated hydroxyanisole, and superior to vitamin E, at a test concentration of 125 microg/ml. Anthocyanins from raspberries and sweet cherries demonstrated 45% and 47% cyclooxygenase-I and cyclooxygenase-II inhibitory activities, respectively, when assayed at 125 microg/ml. The cyclooxygenase inhibitory activities of anthocyanins from these fruits were comparable to those of ibuprofen and naproxen at 10 microM concentrations. Anthocyanins 1 and 2 are present in both cherries and raspberry. The yields of pure anthocyanins 1 and 2 in 100 g Balaton and Montmorency tart cherries, sweet cherries and raspberries were 21, 16.5; 11, 5; 4.95, 21; and 4.65, 13.5 mg, respectively. Fresh blackberries and strawberries contained only anthocyanin 2 in yields of 24 and 22.5 mg/100 g, respectively. Anthocyanins 1 and 2 were not found in bilberries, blueberries, cranberries or elderberries.     

Pollen morphology ofSonchus and related genera,and a general discussion

The pollen morphology of the taxa belonging to the generaAetheorhiza Cass.,Launaea Cass.,Reichardia Roth andSonchus L. in the Iberian Peninsula has been studied with light and electron microscopy. The pollen is 3(-4)-zonocolporate and echinolophate (without polar lacunae, but in general with prelacunae), with equatorial ridges and 15–20 lacunae: 3–4 poral, 6–8 abporal and 6–8 paraporal. Small to medium size, P × E = 19–36 × 23–42 µm; sometimes two different sizes have been found. Exine 3–9 µm thick and ornamentation microreticulate and echinate. The results clearly show the relationships between genera. Moreno-Socías, E., Mejías, J. A., Díez, M. J., 1994: Morfología polínica deLactuceae (Asteraceae) en la Península Ibérica, I.Lactuca y géneros relacionados. — Acta Bot. Malacitana.19: 103–113.     

Molecular identification and differentiation of Brevibacterium species and strains

Amplified ribosomal DNA restriction enzyme analysis (ARDRA), pulsed field gel electrophoresis (PFGE) and ribotyping were used to differentiate among 24 strains of Brevibacterium linens, Brevibacterium casei and Brevibacterium epidermidis obtained from type culture collections or isolated from various smear ripened cheeses. ARDRA was applied to the 16S rDNA. B. linens was shown to be a quite heterogenic group with 2 to at least 4 copies of rrn operons per strain with aberrant nucleotide sequences. AccI gave genus specific restriction patterns and was used to separate Brevibacterium from Corynebacterium species. The expected species specificity of TaqI applied to B. linens type culture strains, but not to all strains isolated from cheese. By AvaI restriction, B. casei and B. linens were differentiated from B. epidermidis and the orange pigmented Arthrobacter casei, a new species of coryneform bacteria; by XmnI restriction, B. linens and B. epidermidis were differentiated from B. casei. One of 4 B. linens genotypes could not be distinguished from B. casei by this method. Here, the typical orange B. linens pigments were used for classification, which was confirmed by partial sequencing of the 16S rDNA.     

Utilized and potentially utilizable seaweeds on the Namibian coast: biogeography and accessibility

Of the 1500 km coast of Namibia, only 80 km is predominantly rocky, the remaining 1420 km being sandy with only minor rocky outcrops. At present two species are utilized, Gracilaria verrucosa for agar and Laminaria schinzii for human consumption. Other potentially utilizable seaweeds occurring on the coast are Ecklonia maxima for alginates, Suhria vittata for agar, Gigartina radula, G. stiriata and Aeodes orbitosa for carrageenans and Porphyra capensis for human consumption. Laminaria schinzii also can be used for alginate production. Due to the diamond-mining areas around the 80 km rocky area at Luderitz (26° 39 S), only 20 km are accessible; hence, at present the seaweeds are being exploited almost to their full potential. The species currently not being utilized could be used, however, to support the industry that already exists at Luderitz. To increase production, access to the diamond areas would have to be gained or a cultivation program initiated.