冬凤兰非共生萌发和低温离体保存（英文）

[Objective] The aim of the research was to establish asymbiotic germination and low-temperature in vitro conservation technique system of Cymbidium dayanum by using plant tissue culture technique to realize its rapid propagation and long-term conservation in vitro.[Method] With mature seeds of C.dayanum as explants,different media were selected to establish asymbiotic germination technique system.With protocorms as materials,conservation,resumptive proliferation and plant regeneration conditions were selected to establish low-temperature in vitro conservation technique system preliminarily.[Result] Mature seeds of C.dayanum could germinate after cultured 90 days on MS media as well as "Hyponex 1" media.The germination rate reached more than 98%.Protocorms inoculated in "Hyponex 1" media could be conserved continuously at 5 ℃ in dark for more than 18 months and the survival rate could reach 90%.Conserved protocorms could realize resumptive proliferation culture both on 1/2 MS and "Hyponex 1" media.The seedling-strengthening and rooting media were 1/2 MS media.[Conclusion] This research provided practical basis for in vitro conservation and rapid propagation of C.dayanum germplasm resource.     

木质素高效降解菌血红密孔菌产酶条件研究（英文）

[Objective] The purpose was to study the optimum conditions of Pycnoporus sanguineus for producing ligninase.[Method] A strain of lignin-degrading white-rot fungus was selected from 5 strains of collected fungi and its ligninase production and the optimum conditions for producing ligninolytic enzyme were measured.[Result]It could produce two kinds of ligninase with good thermal stability. Different temperatures, carbon sources, nitrogen sources,acidities, as well as the additions of surfactant had distinct influence on the development of lignin-degrading enzymes of the fungus. The optimum condition was drawn out: 38 ℃, pH=4.5, 10.0 g/L glucose, 1.0 g/L tartaric acid ammonium.[Conclusion] The aim of research was to provide a basis for lignin degradation in practical production.     

寒地早粳花培培养基中铁的效应(英文)

In this study, through vitro culturing anthers of 7 F1 progenies of early Japonica rice in cold region on medium with different Fe2+ contents, it was found that Fe2+ content generated greater impacts on the induction rate and green plantlet differentiation. The result demonstrated that if Fe2+ increased from 32 to 40 mg/kg, the induction rate of early Japonica rice anther culture in N6 culture media was more then 1.4 times higher than that in N6 culture media containing 5.6 mg/kg Fe2+. In this concentration range, the induction rate increased with the increase of Fe2+ content, while if the concentration was over this concentration range, the induction rate decreased with the increase of Fe2+, showing single peak distribution. When the Fe2+ was 40 mg/kg in differentiation medium, the differentiation rate decreased dramatically. The green plantlet differentiations of callus which were induced on culture media containing 32-40 mg/kg Fe2+ were different, when they were cultured on MS culture media, and 85.7% materials could increase green plantlet productivity to about 7.8%. Therefore, increasing Fe2+in induction media properly could increase anther culture efficiency of early Japonica rice in cold region.     

“胰岛素-转铁蛋白-硒钠”对山羊卵母细胞体外成熟及胚胎发育的影响(英文)

[Objective] The research aimed to enhance culture efficiencies of oocyte and embryo of goat in vitro and to explore serum-free culture system in vitro.[Method] At present,the conventional solutions of oocyte maturation and embryo development in vitro were always added into 1% ITS(Insulin-transferrin-selenium) or using 1% ITS to replace FBS in 2 kinds culture solutions for conducting in vitro cultures of goat oocyte and parthenogenetic embryo.The influences of ITS on their developments were detected.[Result] ITS in maturation liquid of oocytes could not increase oocytes maturation rate but significantly increased blastocyst rate (58.06% vs. 48.19%)of parthenogenetic embryo.If FBS in maturation liquid of oocytes was replaced by ITS, the maturation rate, cleavage rate and blastocyst rate were basically unchanged.Adding ITS into embryo medium could increase blastocyst rate (68.30% vs. 56.82%)of parthenogenetic embryo of goat.If FBS in embryo medium was replaced by ITS,the cleavage rate didn’t change basically,while the blastocyst rate in ITS was obviously lower than that in FBS group(42.33% vs.56.82%).[Conclusion] ITS could promote maturation of oocyte in vitro and early embryonic development, in addition,ITS could replace serum in maturation medium of oocyte as serum-free culture system for conducting relevant researches.     

金龟子绿僵菌诱发UV-B抗性导致分生孢子产孢和孢子逆境耐受性的转化

The conidial tolerance of Metarhizium anisopliae var.anisopliae isolate ARSEF 2575 to UV-B irradiation is greatly influenced by growth-environment alterations. In this review, we report high variability in conidial UV-B tolerance in response to altered culture conditions.Conidia produced on insect cadavers[Zophobas morio(Coleoptera) or Galleria mellonella(Lepidoptera)] had low tolerance to UV-B radiation; and conidia produced on potato dextrose agar supplemented with yeast extract (PDAY) had medium UV-B tolerance; whereas conidia produced on a minimal medium without any carbon source (MM), on MM with a non-preferred carbon source such as lactose (=MML), on PDAY plus 1 M NaCl or KCl, or PDBY with high alkalinity had the highest UV-B tolerances. All of the above conditions that induced high UV-B tolerance, however, also greatly reduced conidial production. Comparisons between stress tolerance and conidial production, particularly with conidia produced under osmotic and nutritive stress, point out that the benefits of producing very tolerant conidia have the enormous cost of low conidial production. Growth under visible light also greatly improved conidial UV-B tolerance, but light did not negatively influence conidial production. Therefore, culture on rich media under light is proposed as the most promising approach to producing conidia with improved UV-B tolerance for biological control of pest insects in agriculture.     

Preparation and Amplification of Colony of Goat Transgenic Fetal Fibroblast and Mammary Gland Epithelial Cell with Human Lactoferrin Gene

[ Objective] The aim was to explore technical system of making single transgenic positive cells become colony cells by amplification culture. [ Method] Fetal fibroblasts and mammary gland epithelial cells of single goat fetus of pBLM-C1 which specifically expressed human lactoferrin were cloned. Single cell colony of single transfection cell was prepared with 3 concentrations of 0%, 50% and 100% conditioned culture media. Transfection cell and non-transfection cell were carried out amplification culture by con-culture, neo gene was as screened gene, genome DNA of transfection cell was detected by PCR method. Chromosome karyotype analysis of single colony cell was tested. [ Result] Compared with non-conditioned culture medium, 100% conditioned culture medium could greatly increase survived rate of single colony cells （ FF： 53.33% vs. 10.00%; MGE： 33.33% vs. 6.67%）. Compared with control, con-culture of transfection cell and non-transfection cell could greatly increase rate of transfection cell single colony after amplification culture （ FF： 53.33% vs. 10.00% ; MGE ： 33.33% vs. 6.67% ）, confluence time of amplification culture was significantly decreased （20 -30 d）. The result of PCR showed that the colony cell obtained by above method contained hLF target gene. The result of karyotype analysis showed that most cloned cell chromosomes were normal. [ Conclusion] The study provides a reliable method for separating transgenic cell, inserting and diagnosing ideal vector, and can save expense and time for transgenic animal production.     

Current Status and Possibilities for the Development of the Cattle Breeding in the Primorsko-Goranska County,Croatia

At the beginning of 2007 the milk producers from the Primorsko-Goranska County, Croatia were surveyed through the questionnaire in order to determine the current status and to highlight the obstacles and the possibilities of development and improvement of the modern cattle breeding in the observed county. The questionnaire spanned 2001-2006 period and all together 260 families were surveyed. The survey aimed to get as much as possible useful information＇s that would indicate the problems concerning the cattle breeding in this area, and based on these results to give certain guidelines for the improvement of current status. Older age of milk producers, significant number of family farms with small number of cattle and difficult implementation of selective measures due to the small number of cows that were included in milk recording were determined as the main obstacles for cattle breeding development in the surveyed county. It is emphasized that development of cattle breeding in this area should be based upon the increased number of family farms with larger herds. To achieve this it is necessary to retain existing cattle fund, and then gradually increase the number of cattle in the mentioned county through the import of breeding heifers of dairy and combined breeds from neighboring European countries with highly developed cattle breeding. Besides that, it is also necessary to create conditions for the production of organic （ecological） products, which represents the future of agriculture, livestock production, bearing in mind that demanding European market has recognized the value of organic agricultural products that were produced through environment friendly production, which Primorsko-Goranska County, due to its significant natural resources, could easily assure.     

Detection of Staphylococcus aureus in Dairy Products by Polymerase Chain Reaction Assay

A polymerase chain reaction （PCR） assay was employed for direct detection of Staphylococcus aureus without enrichment in dairy products. A solvent extraction procedure was successfully modified for the extraction of Staphylococcus aureus DNA from artificially contaminated whole milk, skim milk, and cheese. A primer targeting the thermostable nuclease gene （nuc） was used in the PCR. A DNA fragment of 279 bp was amplified. The PCR product was confirmed by DNA sequencing. In this study, the PCR, GB-4789.10-94, Perifilm RSA.Count Plate, and Baird-Parker ＋ RPF Agar were compared. The sensitivity of the PCR was 10 CFU mL^-1 of whole milk, skim milk, and 55 CFU g^-1 of cheese. The developed methodology allowed for detection of Staphylococcus aureus in dairy products in less than 6 h. The time taken for the development of this PCR assay was 12-24 h, less than the time taken by the general PCR assay using the enrichment method, and the coincidence rate of this developed PCR was 94.3%, the sensitivity was 100%. It was a rapid, sensitive, and effective method for PCR to detect Staphylococcus aureus in milk and milk products.     

Effects of Different Compound Trace Element Premixed Materials on Beef Cattle Production Performance and Anti-oxidation Capacity

In this study, the effects of two types of premixed materials with different combinations of trace elements on the production performance and antioxidant capacity of simmental beef cattle were examined.Fifteen healthy simmental beef cattle of similar weight(approximately 330 kg), the same age(12 months), without castration, and a good physique were divided into three groups, with five beef cattle in each group.Food of GroupⅠ beef cattle was supplemented with a commercially marketed 5% compound trace element premixture for fine beef cattle.Food of GroupⅡbeef cattle was supplemented with a 5% compound microelement premixture for beef cattle that was designed to address local nutrient deficiencies and surpluses.In the blank control group, the beef cattle were not fed a premixture.The pretest period was 15 days, and the test period was divided into prefattening(45 days) and postfattening(45 days) stages.Body weight and body size indices were recorded at 1, 2 and 3 months, and blood samples were collected regularly.In GroupⅠ, the daily weight gain increased significantly by 15.7% compared with that of the control group.The largest daily weight gain was in Group Ⅱ, which increased by 31.6% compared with that in the control.During the test period of 90 days, the body size indices of the three different groups increased in different months, with significant increases in the indices for both test groups compared with those of the control.In GroupⅠ, the activity of CP, the total activity of SOD and Cu-Zn-SOD increased significantly(p0.05) compared with those in the control group, with a highly significant increase observed in GSH-PX activity(p0.01).In GroupⅡ, the increases in the activity of CP and the total activity of SOD and Cu-Zn-SOD were highly significant compared with those in the control group(p0.01).In addition, a significant increase was observed in GSH-PX activity(p0.05).Based on pretest results, the concentrations of Cu, Zn, Mo, Mn, Se and Co in the blood of experimental beef cattle were lower than those of the normal range.After feeding for 90 days, the concentrations of Cu, Zn, Mo, Mn, Se and Co in the blood of GroupsⅠ andⅡ were significantly higher than those in the control group(p0.05).The concentrations of elements in the blood of Group II were close to the appropriate levels.Thus, the effects of the specifically designed compound microelement premixture on the prevention of nutrient imbalances and control of beef cattle nutrition metabolism and the production of fattened beef cattle were significant.     

Research on Rapid Propagation of Gongshui Pomelo by Tissue Culture

[Objective] This study aimed to investigate the optimal medium and hor-mone combinations for efficient rapid propagation of Gongshui pomelo and analyze key technical measures in the tissue culture process. [Method] Stem tips and stem segments with buds were col ected from four varieties of pomelo adult trees as explants, to investigate the main effect and key regulatory factors of vegetative organs and tissue culture explants and to propose a series of measures to prevent and control microbial contamination. Final y, an efficient rapid propagation technology system of Gongshui pomelo was established. [Result] Spring shoot explants contained large amounts of auxin, cytokinins, gibberel ins and other growth regulators, which could be used for tissue culture with high bud generation rate and rapid growth. Different conditions led to various culture results. Specifical y, mature pomelo seeds should be generated on semisolid 1/2MS medium and transferred to solid MS medium for incubation. The propagation coefficient of stem segments with axillary buds was greater than that of stem tips, exhibiting significant differences. In ad-dition, the optimal hormone combination was 6-BA 0.5 mg/L ＋ NAA 0.5 mg/L, which significantly promoted the induction and differentiation of adventitious buds. [Conclusion] This study provided basis for basic research, production and application of pomelo germplasm resources.     

培养条件对奶牛金葡菌5型荚膜多糖产量的影响

[目的]研究不同培养条件对牛源金葡菌5型荚膜多糖产量的影响,以便于CP5的生产制备,从而为开展奶牛金葡菌新型多糖疫苗的研究奠定基础。[方法]从患隐性乳腺炎奶牛乳样中分离金葡菌,鉴定荚膜多糖血清型,对5型荚膜多糖菌株,采用BHI,哥伦比亚固体培养基,mod110三种培养基,每种培养基分别采用固体培养,碳源为葡萄糖的液体培养和碳源为乳糖的液体培养三种方式,共组成9种不同培养条件进行培养,研究培养条件对金葡菌荚膜多糖产量的影响。[结果]不同培养结果表明,与常用哥伦比亚培养基比较,BHI培养基能降低荚膜多糖产量,而mod110培养基能提高荚膜多糖产量;同种培养基,固体培养方式比液体培养方式有更高的荚膜多糖产量,同时乳糖作为碳源可提高荚膜多糖产量。     

培养条件对奶牛金葡菌5型荚膜多糖产量的影响

奶牛乳腺炎是奶牛生产上的重要疾病,不仅影响奶牛的产奶量、降低牛奶品质,而且严重威胁消费者健康,已成为制约奶牛业发展的主要原因之一。据国内外相关资料显示,50％以上的奶牛乳腺炎是由金葡菌（Staphylococcus aureus,S.aureus）所引起的。研究发现,94％～100％的从患乳腺炎奶牛乳汁中分离到的金葡菌菌株表面带有多糖物质构成的保护性荚膜荚膜具有11种血清型,     

金黄色葡萄球菌血清5型生长曲线的测定

[目的]为了测定金黄色葡萄球菌血清5型的生长曲线。[方法]在不同培养基和不同培养方式下,用分光光度计定时测定菌液吸收值,绘制了金黄色葡萄球菌的生长曲线。[结果]在哥伦比亚培养基中,液体培养基在振荡条件下比固体培养基更有利于金黄色葡萄球菌的生长,在静止条件下最不利于金黄色葡萄球菌的生长;在Mod110培养基中,金黄色葡萄球菌的生长速度为固体液体(振荡)液体(静止)。在相同培养方式下,Mod110固体培养基比哥伦比亚固体培养基更有利于金黄色葡萄球菌的生长。[结论]该生长曲线反映了细菌的生长趋势,说明细菌可以进行较长时间的研究。     

金黄色葡萄菌5型荚膜多糖偶联鞭毛蛋白疫苗对小鼠乳腺炎模型的保护作用

[目的]金黄色葡萄菌5型荚膜多糖(CP5)分别与鼠伤寒沙门氏菌鞭毛蛋白Flagellin、牛血清白蛋白(BSA)进行了偶联,并在小鼠模型上比较了这2种偶联疫苗所引起的体液免疫和粘膜免疫反应。[方法]以鼠伤寒沙门氏菌鞭毛蛋白为载体蛋白,与CP5进行化学偶联,并在小鼠乳腺部位免疫,然后通过检测小鼠乳汁中的抗体分析其对小鼠的免疫保护作用。[结果]乳腺局部免疫后flagellin-CP5组在乳腺部位引起了sIgA为代表的粘膜免疫反应和IgG为主的体液免疫反应。CP5和鞭毛蛋白的偶联物能够诱导更高的体液免疫以及粘膜免疫反应。[结论]鞭毛蛋白具有较强的免疫佐剂作用。     

一株粘性多糖产生菌LV-1的发酵条件研究

1材料与方法 1．1粘性多糖产生菌的分离与培养 1．1．1菌株分离。将2g土样加入100ml含有3g异麦芽酮糖和0．15g酵母浸粉的培养基中,28℃摇床（150r／min）培养2d,然后将菌悬液用生理盐水梯度稀释法涂平板,于28℃培养箱内培养过夜。     

胞外表达L-天冬酰胺酶发酵条件的研究

[目的]优化胞外表达L-天冬酰胺酶工程菌的发酵条件。[方法]用正交试验研究培养基的组成,用单因素试验研究诱导条件以及通气量对酶产量的影响。[结果]发酵培养基采用8.0%玉米浆和1.0%甘油组成;诱导剂采用终浓度为0.5g/L的乳糖,扩大培养后10h加入,诱导时间为12h,通气量为0.83VVM。[结论]该研究为胞外表达L-天冬酰胺酶的工业化生产提供了参考。     

金黄色葡萄球菌荚膜多糖的提取及其多糖含量的测定

为提取纯化金黄色葡萄球菌荚膜多糖并测定多糖含量.采用超声波破碎菌液,用CTAB结合其中的多糖、CaCl2解离、乙醇沉淀等化学方法提取纯化金黄色葡萄球菌荚膜多糖,利用苯酚-硫酸法测定其含量.结果显示,得到荚膜多糖0.31 g,含量为65.06%.结果表明,成功提取金黄色葡萄球菌荚膜多糖,并对其含量进行了测定,初步确定了粗提金黄色葡萄球茵荚膜多糖的方法.     

里氏木霉RutC-30产纤维素酶条件优化研究

[目的]优化里氏木霉RutC-30产纤维素酶的液体发酵条件。[方法]以里氏木霉RutC-30为出发菌株,通过单因素试验研究培养基不同氮源(硫酸铵、尿素、蛋白胨)及浓度、不同碳源(纤维素、乳糖、甘油、葡萄糖)及浓度和不同的初始pH(3.0、4.0、5.0、6.0、7.0)对产酶的影响,在此基础上选取氮源、碳源和pH为影响因子采用正交试验探讨里氏木霉RutC-30产纤维素酶的优化条件。[结果]正交试验分析表明,各因素对产酶影响顺序依次为碳源>氮源>pH,里氏木霉RutC-30产纤维素酶的最佳条件是:以1%纤维素为碳源、以0.5%蛋白胨为氮源,初始pH值为4.0,在30℃产酶发酵培养5 d,纤维素酶活力高达7.303 U。[结论]里氏木霉RutC-30经优化培养后,产酶能力可得到大幅度提高,具有潜在的工业应用价值。     

魔芋细胞悬浮培养生产葡甘露聚糖的研究

[目的]对影响魔芋细胞悬浮培养及其次生代谢物葡甘露聚糖产生的主要影响因子进行研究。[方法]利用3,5-二硝基水杨酸测定葡甘露聚糖含量。[结果]在魔芋细胞悬浮液体培养葡甘露聚糖的过程中,采用MS培养基为基本培养基,pH值为6.0时有利于葡甘露聚糖的合成,25g/L乳糖可作为最适碳源;2,4-D在1.0mg/L时葡甘聚糖的积累效果明显;细胞分裂素6-BA使葡甘露聚糖积累明显高于KT,且1.5mg/L时葡甘露聚糖积累量较高,培养15d左右葡甘露聚糖含量达到最大。[结论]MS＋2,4-D1.0mg/L＋6-BA1.5mg/L＋乳糖25g/L,pH6.0,为魔芋细胞悬浮培养生产葡甘聚糖的较优培养基。     

奶牛子宫内膜炎病原菌的分离·鉴定及药敏试验

[目的]了解奶牛子宫内膜炎的病原菌感染情况与耐药现状.[方法]随机采集库尔勒地区某牛场患子宫内膜炎奶牛的子宫黏液20份,进行实验室细菌学的分离、生化鉴定及药敏试验.[结果]共分离出89株菌,其中链球菌45株,占50.56％;葡萄球菌24株,占26.97％;大肠杆菌13株,占14.60％;蜡样芽胞杆菌7株,占7.87％.分离出的单一菌株的检出率为80％ （16/20）.引起该地区奶牛子宫内膜炎的主要病原菌是链球菌、葡萄球菌.对链球菌、葡萄球菌、大肠杆菌、蜡样芽胞杆菌这4种菌抑菌效果最好的药物依次分别为奥复星、氟苯尼考、甲氧苄啶、氟苯尼考.[结论]该研究可为指导子宫内膜炎的临床用药和预防提供理论依据与技术支持.