生物技术专业外文翻译-壳聚糖对红掌组培污染菌的抑制作用

1、外文原文CHITOSAN ON ANDRAEANUM TISSUEINHIBITION OF BACTERIACONTAMINATIONAbstract: The fungi was isolated and purified from polluted anthurium subculture medium, ide ntified to beDeuteromycot in aHyphpmycetesHyphomycetales Dematiaceae and trichoderma. The inhibition effect of the three kinds of chitosa n

2、 on the fungi wasc on ducted.The results showed that the chitosan had inhibition effect on the fungi. The higher concentrations of chitosan had a betteinhibition effect than the lower ones.The inhibition effect of 50000 and 250000 molecular weight chitosa n is better than that of the3000 molecular w

3、eight. It is prelim in arily proved that chitosa n had good in hibiti on effect on the fungi from polluted tissue culture , andwill have good application value in the fields of pollution prevention in flowers tissue culture.Key words: tissue culture; contamination; Chitosan; inhibition effectPlant t

4、issue culture, not only in plant breeding, rapid propagation of seedli ngs and the product ion of useful sec on dary metabolites, such as the use of a wide range of pla nt gen etic engin eeri ng is and Pla nt Molecular Biology,1 one of the important foundation for the study . Pollution, browning, gl

5、ass and tissue culture are the main problems, including pollution of the most2 com mon . Therefore take effective preve nti on and con trol measures to reduce the chance of contamination occurred, tissue culture is an important guara ntee for success. Chitosa n (chitosa n) is the chit in in stro ng

6、alkali ne con diti ons from BAfter the formatio n of acyl an importa nt derivative is determ ined by a nu mber of N- acetyl glucosamine through W) glycosidic bond linking the nature of it more lively. Has natural antibacterial properties of chitosan, and a broad spectrum antimicrobial in recent year

7、s, chitosan in various fields such as agriculture , medicine ,manufacture ,food 5, etc. The use of great importanee. Tissue culture of the chitosan in the application of inhibition has not been reported. This experiment to be from the contaminated medium subculture andraeanum extraction, purificatio

8、n and identification of bacteria contamination, with different molecular weight chitosan and different concentrations of their inhibition, and explore the best inhibitory effect of chitosan molecular weight and the best inhibitory concentration, in order to explore the chitosa n as a new type of an

9、timicrobial age nt to provide specific in formati on.1 Materials and Methods1.1 Test materials1.1.1 IsolatesStreptomyces bacteria pollution (code-named WCHPA), separated from the laboratory con tam in ati on an draea num subculture medium.1.1.2 Test for DrugsChitosa n, a Zhejia ng Xi ng Biotech no l

10、ogy Australia Limited, a molecular weight of 3kd (degree of deacetylati on 80%), 50kd (degree of deacetylati on 90%), 25kd (degree of deacetylatio n 90%).1.1.3 Medium PDA mediumPotatoes 200g, glucose 20g, agar 20g, distilled water 1000ml, pH value of the n atural.1.2 Test Method1.2.1 Isolation and p

11、urificationClea n out the work in Taich ung an draea num subculture medium polluti on fun gi, from PDA medium. Repeat several times un til the colony characteristics and morphology cha nges are no Ion ger, we can ide ntify bacteria have bee n purified.1.2.2 Identification of bacteria contaminationCo

12、ntinuous observation of colony morphology after purification. Film production equipment to further observe the shape strain, mycelium, spores, spore cell spore cell morphology and coni diatio n of the ways and forms and so on, tak ing pictures with a digital microscope.1.2.3 Bacteriostatic TestThat

13、the different molecular weight chitosan with 1M solution of acetic acid dissolved mixed with PDA medium to produce a final concentration of chitosan (0.5,1.0,1.5,2.0,2.5,3.0)mg/ml of culture medium, 1M solution of sodium hydroxide by adjusting pH value of 6.0, in addition to the control without chit

14、osa n, the other with the additi on of chitosa n in the same medium To eliminate all high-pressure cooker sterilization 30min under 121 C and pour plate. Mycelium white to green after 24h and then into a brown, hyphae developed, velvet-like, 3d training around the middle of the brown and white flocc

15、ule nt mycelium of fimbriae produced. Hyphae are separated from a long stalk conidia to produce lateral branches whorled branches meristematic conidiation short bottles. Single-cell conidia oval, smooth.2 Results and An alysis2.1 An draea num symptoms subculture medium polluti onAndraeanum subcultur

16、e in growth medium brown Fimbriae, medium surface was black, brow n and white mycelium on the flocculatio n betwee n the hyphae.2.2 fun gal pure culture and ide ntificati on of the resultsPDA medium in the upper colony diameter, edge neatly, spore germ in atio n of the PDA medium by add ing chitosa

17、n, 4d clearly observed after the colony is surroun ded by a black halo, black halo and colony growth as to the relocation of . The better the general inhibitory effect of the more obvious81black halo . Colony has no control this phenomenon. Chitosan molecular weight and concen tratio n of the bacter

18、iostatic effect.2.3. The same three concentrations of the bacteriostatic effect of chitosan molecular weight comparedConcentration of 0.5mg/ml 3 species of molecular weight chitosan WCHPA colony inhibitoryeffect of six concentrations of three kinds of molecular weight of chitosan on the inhibition r

19、ate of colony WCHPA results0.5mg/ml concen trati on of three kinds of molecular weight chitosa n there is no clear law, the maximum inhibitory rate of 50,000 in molecular weight of chitosan for the culture medium for 20.45%. Inhibitory rate and the time and no close ties. The results show that the c

20、oncentration of 0.5mg/ml of the three kinds of molecular weight of the an tibacterial effect of chitosa n has not bee n satisfactory .5d prior to the first three kinds of chitosan molecular weight the greater the inhibitory rate of the higher molecular weight, and are extended over time to reduce th

21、e rate of inhibition.In paragraph 5d, 3000 molecularweight inhibitory rate of more than two other inhibitory rate of molecular weight, molecular weight and in hibiti on rate of 3000 also reached the highest value. At its bacteriostatic effect after 7d and 5d is similar to before, the greater the in

22、hibitory rate of the higher molecular weight.3 Discussi on and con clusi ons3.1 Chitosan as a potential new antibacterial agentsPollution plant tissue can be broadly divided into two types of operations may produce air pollution mainly mold contamination; explants is itself brought about by bacteria

23、 primarily by fun gal and bacterial con tam in atio n 10. High inhibitory rate was as high as 79.55%. Antibiotics in tissue culture is now the most com mon molecular weight of chitosa n and 3.2 have an importa nt impact on an timicrobial properties of the experime ntal results from the 50,000 and 25

24、0,000 the bacteriostatic effect of chitosa n molecular weight tha n that of 3000; the highest in hibitoryrate in the 250,000 molecular weightconcentration of 3.0mg/ml and 2.0mg/ml of the two. But on thewhole, takinginto account the factors of price and its operation to the conclusion that the experi

25、mental molecular weight of 50,000 as the best concentration of 1.5mg/ml.3.2 Concentrationon the antimicrobial properties of chitosan have asig ni fica nt impactQiu-ping 11, The results show that the antimicrobial effect of chitosan with its concen trati on in creases, the experime ntal results suppo

26、rt the results of the study. A wide range of applicati ons of chit in, chitosa n in the medical, food, en vir onmen tai protect ion, in areas such as chemical products have wide-ranging and important applications. Chitosan flowers will be used for tissue culture would be very good prospects.Referenc

27、es1 MK.Razdan. Zun-Xiao-an, ZHU Yang translation. Introduction to Plant Tissue Culture M. Beijing: Chemical Industry Press ,2006:1-2.2 Hu,ZHANGLi-jun,baixuemei,et al. Plant Tissue Culture PollutionAnalysis and the disinfection of explants J. Anhui Agricultural Scienee, 2007,35 :680-681.3 Yi-Ti ngWan

28、 g,wa nglia nping , Mou Hero, et al. In dustrial producti on oftissue culture studies of pollution and its control J. Anhui Agricultural Scienee, 2005, 33 (12): 2357 - 2358.4 Chia ng big. Chit in M. Beiji ng: Chi na En viro nmental Scie nce Press, 1996.5 Fu Tian Xia, LIU Xiao-yu, Liu Zhiheng. Chitos

29、ans antibacterial propertiesof the progress of applied research J. Liaoning Chemical, 2005,1234 (12) :541-544.6 Yu Han shou, Wu Zhang, Yang Bing. Chitosa n in hibit research progress ofplant diseasesJ.Natural products research and development,1999,12(3) :94-97.7 LiFang,shishaowei. Chitosan Preparati

30、on and application in the foodindustry Mechanism J. Food and Drug, 2006 (03A) :19-22.8 Wu, Xia water. Non-biologicalfactors on the antibacterial activity ofchitosan J. Food Science, 2004,25 :53-55.9 wuxiaoyong,Qing-Xiao Zeng, MOshaofang. Several of the antibacterialproperties of chitosan J. Food and

31、 Fermentation Industry, 2005,206 (2) :18-21.10 Hu, ZHANG Li-jun,baixuemei, et al. Plant Tissue Culture Pollution An alysis and the disi nfectio n of expla nts J. An hui Agricultural Scien ce, 2007,35 :680-681.11 Qiu-ping,Chitosan on mango Colletotrichum gloeosporioides, Diplodiaantagonism bacteria J

32、. Food and machinery, 2005,21, (1) :25-27.中文译文壳聚糖对红掌组培污染菌的抑制作用摘要：在污染的红掌继代培养基中提取污染菌并进行了分离及纯化， 初步鉴定为为半知菌亚门(Deuteromycot in a),丝抱纲(Hyphpmycetes),丝抱目 (Hyphomycetales ),淡色抱科 (Dematiaceae ),木霉属(trichoderma )。研究了壳聚糖对该菌的抑制作用，探索了 3种分子量及 其6种浓度的壳聚糖对该菌的抑制效果。结果表明：壳聚糖对该污染菌有抑制作用，5万和25万分子量的抑菌效果高于3000分子量，浓度越高,抑菌效果越好

33、。初步证实了壳聚糖对组培污染菌类有很好的抑制作用，在 花卉组织培养污染防治方面具有应用价值。关键词：组织培养；污染；壳聚糖；抑制作用植物组织培养不仅在植物育种、苗木快速繁殖和有用次生代谢产物生产等方面应用广泛，也是植物基因工程和植物分子生物学研究的重要基础 之一。污染、褐化、玻璃化等是组织培养存在的主要问题，其中污染最 普遍2。因此采取有效的防控措施，降低污染发生的机率，是组织培养成 功的重要保障。壳聚糖(chitosan )是甲壳素在强碱性条件下进行脱乙酰 基作用后形成的一种重要的衍生物，是由多个 N-乙酰氨基葡萄糖通过B-(1-4)糖苷键连接起来，但它的性质较为活泼。壳聚糖具有天然抑菌性能

34、，且抑菌谱广，近年来壳聚糖在各个领域如农业、医学、制造4、食品等方面中的运用受到很大的重视。有关壳聚糖在组织培养中的抑菌应用尚 未见报道。本实验拟从污染的红掌继代培养基中提取、纯化污染菌并进行 鉴定， 用不同分子量及其不同浓度的壳聚糖对其进行抑制实验，摸索出抑菌效果最好的壳聚糖分子量及最佳抑菌浓度，为探索壳聚糖成为新型的 抑菌剂提供具体信息。1材料与方法1.1 试验材料1.1.1 供试菌株霉菌污染菌(代号 WCHPA，分离于实验室污染的红掌继代培养基中。1.1.2 供试药品壳聚糖，有浙江澳兴生物科技有限公司提供，分子量为3kd （脱乙酰度80% ，50kd （脱乙酰度90%），25kd （脱乙

35、酰度 90%。1.1.3 培养基 PDA培养基土豆200g，葡萄糖20g，琼脂20g，蒸馏水1000ml，pH值自然。1.2 试验方法1.2.1 污染菌的分离及纯化在超净工作台中挑出红掌继代培养基中的污染真菌，接到PDA培养基中。重复若干次，直至菌落特征及其显微形态不再有变化，可确定菌种已 纯化。1.2.2 污染菌的鉴定连续观察纯化后的菌落形态。制作装片，进一步形观察菌株、菌丝、 抱子、产抱细胞的形态及产抱细胞的产抱方式和形态等，用数码显微镜进 行拍照。1.2.3 抑菌试验称取不同分子量壳聚糖用1M的乙酸溶液溶解后与 PDA培养基混合，制成壳聚糖终浓度分别为（0.5、1.0、1.5、2.0、2

36、.5、3.0 ） mg/ml的培养基，用1M的氢氧化钠溶液调节 pH值为6.0，对照除不加壳聚糖外， 其他与加有壳聚糖的培养基相同。高压灭均锅 121 T下灭菌30min后倒平皿。产生白色菌丝24h后变为绿色继而变为褐色，菌丝发达，绒状，培养3d左右，在褐色菌丝中间有白色絮状的菌毛产生。菌丝有隔，分生抱子 梗长，产生侧向分枝，分生枝上轮生短的产抱瓶体。分生抱子单胞卵圆形、 光滑。2结果与分析2.1红掌继代培养基污染症状在红掌继代培养基中长出褐色的菌毛，培养基表面呈黑色，褐色的菌 丝上间有白色絮状的菌丝。2.2真菌纯培养的形态特征及鉴定结果菌落在PDA培养基上等径生长，边缘整齐，抱子萌发加入壳聚

37、糖的 PDA培养基，4d后能明显观察到菌落周围有黑色晕圈，且黑色晕圈随着菌落的生长而向外移。一般抑菌效果越好黑色晕圈越明显8对照菌落始终没有出现这种现象。壳聚糖的分子量和浓度影响抑菌效果。2.3相同浓度的三种分子量壳聚糖抑菌效果的比较浓度为0.5mg/ml 3种分子量壳聚糖对 WCHP/菌落的抑制效果 6个浓 度3种分子量壳聚糖对 WCHP菌落的抑菌率结果0.5mg/ml浓度的3种分子量壳聚糖之间没有明显的规律，最高抑菌率出现在分子量为5万的壳聚糖培养基中为20.45%。抑菌率和时间上也没有紧密的联系。结果表明在0.5mg/ml浓度下3种分子量的壳聚糖的抑菌效果都不理想。第5d之前3种分子量壳聚糖分子量越大抑菌率越高，而且都是随着时间的延长抑菌 率降低。在第 5d，3000分子量的抑菌率超过其它两种分子量的抑菌率， 而3000分子量的抑菌率也达到最高值。在第7d后抑菌效果和第 5d之前的相似，分子量越大抑菌率越高。3讨论与结论3.1壳聚糖成为新型抑菌剂的可能植物组培的污染大致可分为两种，一种是操作中产生得污染，主要是 霉菌的污染；一种是外植体自身所带杂菌主要是