天山一号冰川产蛋白酶酵母菌的筛选及其系统发育分析

[目的]筛选天山一号冰川底部沉积层耐低温的产蛋白酶菌株,研究其生理生化特性并做系统发育分析.[方法]以天山一号冰川底部沉积层中耐低温产蛋白的酵母菌株为材料,用Yeast Extract Peptone Dextrose Medium(YPD)培养基活化60株酵母菌;再用选择性培养基筛选产蛋白酶的菌株,观察菌落形态;测定产酶菌株最适温度;最后基于26S r DNA基因D1/D2区序列进行系统发育分析.[结果]从60株酵母菌中共筛选出10株产蛋白酶菌株,其中产酶优势菌为BCY-8、BCY-20,产酶酵母细胞平均大小为2.60～5.00μm,最适生长温度为24℃.经系统发育分析可知,10株产酶酵母菌属于隐球菌(Cryptococcus)、红酵母属(Rhodotonla)、南极酵母(Antarcticyeast)、Uncultured fungus、Mazocraeoides gonialosae.[结论]研究可为低温蛋白酶生物技术的研发奠定基础.     

鸡粪中具有产酒精能力酵母的初步筛选

[目的]筛选能发酵产酒精的酵母茵。[方法]在鸡粪样品中经过初筛、复筛，以及生理生化试验和发酵试验，确定所筛选的酵母菌株产酒精的能力，并对所筛选出菌株的适宜生长条件做初步研究。[结果]共筛选出4株可发酵菌株：HY-1、HY-2、HY-3和HY．4，其中HY-1产酒精能力最高。[结论]HY-1菌株的适宜生长条件为：pH值6．5，温度30℃，最适碳源为葡萄糖。该茵株的致死温度为50℃。在最适奈件下进行HY-1的酒精发酵试验，测得发酵液中的酒精度达6．90％。     

长白山区土壤中高毒力苏云金杆菌的分离及生物测定

[目的]分离筛选出对鳞翅目害虫高毒力的新苏云金杆菌.[方法]通过温度筛选法从长白山区土壤中分离出苏云金杆菌,再通过生物测定和毒力测定筛选出高毒力苏云金杆菌.[结果]从150份土壤样品中分离出18株苏云金杆菌,其分离率为12.0％,其中山地分离率为8.5％,农田分离率为16.2％.活性测定结果,18株菌株中对小菜蛾、斜纹夜蛾及甜菜夜蛾致死率超过90％的菌株分别有17株、5株和4株,其中YN1-1菌株对3种害虫都表现出高活性.毒力测定结果,YN1-1菌株对甜菜夜蛾和斜纹夜蛾的速效性最好;对甜菜夜蛾高毒力菌株依次为YN6-2＞YN1-1 ＞YN4-2＞YN2-6;对斜纹夜蛾高毒力菌株依次为YN4-4＞YN1-1 ＞YN6-1 ＞YN4-1＞YN2-1.[结论]经活性测定和毒力测定,最终筛选出广谱性、速效性及高毒力的YN1-1为目标菌株.     

蛹虫草子囊孢子单孢分离与交配型基因鉴定

[目的]明确蛹虫草亲本和子囊孢子单孢菌株交配型基因类型.[方法]以自主分离保存的CM1901菌株新鲜子实体为材料,采用孢子弹射和梯度稀释培养法,获得子囊孢子单孢菌株.采用PCR方法对蛹虫草的亲本及单孢菌株的交配型基因进行鉴定.[结果]获得102个子囊孢子单孢菌株.亲本菌株同时含有MAT1-1和MAT1-2两种交配型;102株单孢分离株中,仅含MAT1-1交配型菌株57株,仅含MAT1-2交配型菌株24株,另外21株为亲本型(同时含有MAT1-1和MAT1-2),MAT1-1∶MAT1-2为78∶45.[结论]分离获得102个蛹虫草子囊孢子单孢菌株,发现这些单孢菌株交配型有3类,且MAT1-1∶MAT1-2比例为78∶45,与理论值1∶1有较大差值,所以蛹虫草不属于典型的二极性异宗配合型子囊真菌.     

高温木质素分解菌的分离、筛选及鉴定

[目的]为获得在高温条件下有较强分解木质素能力的菌株.[方法]以马粪为材料,采用BM培养基,经分离、筛选和纯化获得4株高温木质素分解细菌.[结果]通过对木质素分解菌所产生的漆酶、过氧化物酶和锰过氧化物酶3种酶的活性进行测定,筛选出1株产酶活性较高的菌株M-2.采用CTAB法提取目的菌株DNA,经过PCR扩增、纯化,对16S rDNA测序进行鉴定,确定为假单胞菌属的绿脓杆菌.[结论]该研究为木质素分解菌的选育和菌剂的制备提供菌种资源.     

降解水稻秸秆的好氧性丝状真菌菌株的分离筛选与初步鉴定

[目的]对特定生态环境（腐烂的稻草）中的微生物进行分离筛选,并就这些微生物菌株对水稻秸秆的降解能力进行研究和分析.[方法]利用以微晶纤维素粉等只含纤维素成分的原料作唯一碳源的培养基对从稻草秸秆中分离筛选到的7株可降解稻草秸秆的丝状真菌菌株进行分离和筛选,通过形态学观察和生理生化特性2个层次对其进行鉴定.同时,对这些菌株降解水稻秸秆的能力进行了研究.[结果]试验发现,筛选出的7株菌株中有2株属于多轮青霉组,2株属于烟色曲霉组,1株小孢根霉组,1株梨卵形孢组以及1株总状枝毛霉组.其中1株烟色曲霉组和1株卵形孢组降解秸秆纤维素能力较强,另有1株烟色曲霉组和1株卵形孢组降解秸秆半纤维素能力较强.[结论]研究可为微生物菌种降解水稻秸秆的能力研究提供参考.     

秸秆发酵饲料高产蛋白酵母菌株分离和筛选

为筛选出高产蛋白酵母菌株,推进秸秆发酵饲料在畜牧业中的应用。利用培养基平板划线法从牛瘤胃食糜样本中分离筛选出高产蛋白酵母菌株,通过正交试验优化其培养条件,并依据生长曲线筛选出可作为秸秆发酵饲料用高产蛋白酵母菌。从牛瘤胃食糜样本中共分离获得10株菌落形态和细胞特征较明显的疑似酵母菌株,分别编号为x1~x10。经正交试验L16(44)优化其培养条件,发现在温度32℃、初始p H5.5、接种量12.0%、装液量100ml的条件下,各分离酵母菌的生物产量最高。在10株分离酵母菌株中,生物产量最高的4株分别是x9号、x1号、x10号和x5号。其中,x9号和x1号菌株适的应期较短,在2h后即进入对数生长期,8h后达稳定期;x10号和x5号菌株的适应期较长,4h后进入对数生长期,1 h后进入稳定期。综合酵母菌株的培养条件及生长曲线测定结果,确定假丝酵母属菌株最适宜作为秸秆发酵饲料高产蛋白酵母菌。     

嗜高温灰树花工厂化生产菌株的筛选研究

[目的]筛选适合相对高温环境生产出菇的菌种.[方法]通过比较11个野生灰树花菌株的菌丝生长速率、生长势、拮抗性,并以栽培周期、子实体品质及前期生物学转化率等为指标进行工厂化栽培考察.[结果]菌株"ZDSJ-08"在24 ～32℃生长温度层架式栽培方式下,71 d 2次出菇累计基质生物转化率为88.6％;同时在颜色、风味、叶片厚度、口感及朵形等方面表现优秀.[结论]菌株"ZDSJ-08"适合灰树花高温工厂化栽培.     

筛选产纤维素酶菌株及其产酶条件的优化

[目的]筛选降解菌糠纤维素的菌株,并且优化其发酵条件.[方法]采用纤维素-刚果红培养基进行筛选,筛选出纤维素酶活较高的菌株.通过比较不同氮源、碳氮比例等条件下CMC酶活,研究其最佳发酵条件.[结果]选出一株酶活较高的菌株,命名为N3.其最适有机氮源为豆饼粉,无机氮源是（NH4）2SO4.最适合N3产酶的（NH4）2SO4∶豆饼粉比例为2∶4.最适碳源氮源比例为5∶2.[结论]N3是一株具有研究价值的产纤维素酶的菌株.     

刺激植物响应蛋白基因Epl1的载体构建及酵母转化

[目的]构建刺激植物响应蛋白Epll基因的真核表达载体,筛选多拷贝酵母转化子.[方法]以深绿木霉（Trichoderma atroviride）ACCC30153的cDNA为模板进行PCR获得Epl1基因片段,并将目的片段插入表达载体pPIC9K的相应位置,获得重组表达载体,并将pPIC9K-Epl1转入毕赤酵母（Pichia pastoris）GS115中,并用不同中终浓度的遗传霉素G418筛选多拷贝重组酵母菌株.[结果]对重组载体进行PCR及双酶切检测为阳性;在含有终浓度为2 mg/ml遗传霉素G418的YPD平板上筛选得到7株多拷贝的转化子GS115-Epl1,经PCR鉴定1株转化子呈阳性.[结论]Epl1基因的我体构建及多拷贝酵母转化子筛选为以后大量分离纯化Epl1蛋白并研究其功能奠定基础.     

Web2.0,以个人为中心的互联网时代的到来

本文介绍了Web2．0的知识和背景，对Web2．0出现的基础以及典型技术和应用作了阐述，同时对其作出了前景展望。     

市级综合气象信息分析与管理系统的设计与实现

以抚顺地区3个国家基本站50余年气象资料为依托，采用Microsoft Excel工作表实现由气候和农业气候2部分构成的气象与农业综合信息处理系统的基本思路、方法、内容、用途和使用情况。为了实现气候分析及应用的科学化、客观化、系统化、规范化、程序化及连续化的业务目标，按业务流程设计与实现的，包括资料处理、气候趋势模拟、气候预测、气候鉴定、季节性信息化处理、农作物产量预报、气候生产潜力分析等一系列内容在内的系统建设。关键是解决了大量的计算问题，建立了区域性通用模式。     

Multi-variable grey model (MGM (1,n,q)) based on genetic algorithm and its application in urban water consumption

Urban water consumption has some characteristics of grey because it is influenced by economy, population, standard of living and so on. The multi-variable grey model （MGM（1,n））, as the expansion and complement of GM（1,1） model, reveals the relationship between restriction and stimulation among variables, and the genetic algorithm has the whole optimal and parallel characteristics. In this paper, the parameter q of MGM（1,n） model was optimized, and a multi-variable grey model （MGM（1 ,n,q）） was built by using the genetic algorithm. The model was validated by examining the urban water consumption from 1990 to 2003 in Dalian City. The result indicated that the multi-variable grey model （MGM（1,n,q）） based on genetic algorithm was better than MGM（1,n） model, and the MGM（1,n） model was better than MGM（1,1） model.     

Molecular detection of virulence genes in Campylobacter species isolated from livestock production systems in South Africa

Campylobacter species are a major cause of foodborne bacterial infections in both developed and developing countries worldwide. Campylobacter jejuni is responsible for the majority of infections. This study was conducted to identify virulenceassociated genes in Campylobacter species isolated from livestock production systems in South Africa. A total of 250 fecal samples consisting of cattle(n=50), chickens(n=50), goats(n=50), sheep(n=50) and pigs(n=50) were randomly collected from livestock in Eastern Cape and KwaZulu-Natal provinces of South Africa between April and October 2018. The samples were analyzed for the presence of virulence genes in Campylobacter species using molecular PCR-based methods. It was found that 77 and 23% of Campylobacter jejuni and Campylobacter coli respectively were isolated from all the livestock samples. There were positive significant(P0.05) correlations amongst all the virulence genes that were investigated. Chisquare and Fisher's exact tests were implemented to test for the effect of livestock species on the presence or absence of virulence genes. The study demonstrated that most of livestock species can potentially cause zoonotic infections and food poisoning due to the high prevalence of Campylobacter. The high prevalence of virulence genes highlights the significance of Campylobacter in livestock production systems in South Africa. This requires the implementation of one-health approaches to reduce the impact of foodborne and zoonotic diseases for the welfare of human and animal health.     

不同施肥水平对木薯氮磷钾养分积累、分配及其产量的影响

 【目的】通过比较不同施肥水平下木薯氮磷钾养分积累、分配和产量的差异，探讨粤北坡岗地优质高产木薯氮磷钾养分的最佳用量。【方法】以木薯品种南美119为材料，采用“3414”方案，在粤北翁源进行大田试验。试验共设14个处理。【结果】施肥处理的氮素主要分配到地上部，不施肥处理的氮素则主要分配到根部；不同施肥处理的磷素均主要分配到地上部；不施肥处理和不施钾处理的钾素主要分配到根部，不施氮处理的钾素在根、冠间分配较均衡。在木薯不同生长阶段，植株氮磷钾含量均呈下降趋势，但施氮、磷、钾化学肥料处理植株氮磷钾含量的下降速度小于不施氮、磷、钾化学肥料的处理。处理N2P2K2的氮、磷、钾含量和积累量在各生育期均为最大，不施肥处理（N0P0K0）的氮、磷、钾含量和积累量在各生育期均为最小（P＜0.05）。产量最高的是N2P3K2处理，为22 694.06 kg·hm-2，其次是N2P2K2处理，为21 417.87 kg·hm-2。【结论】木薯产量、氮磷钾养分积累及其在根冠间的分配，对施肥水平高度敏感, 适当比例的氮磷钾肥配合施用既可以显著增加木薯植株的氮磷钾含量和积累量，又可以提高产量。在粤北坡岗地木薯生产中，氮素最重要，在氮素得到满足的条件下，磷素较钾素更重要。在本试验条件下，氮磷钾养分最佳用量为358.80 kgN·hm-2、89.10 kg P2O5·hm-2和187.50 kg K2O·hm-2。     

红苕饲粮中添加铜和/或喹乙醇对猪生产性能及某些血液生化指标的影响

选体重约38公斤的长白(♂)×成华(♀)杂交猪20头,随机分为四组,饲予四种不同的饲粮,研究饲粮中添加铜和/或喹乙醇对猪生产性能和某些血液生化指标的影响。结果表明:基础组、加铜组、加喹乙醇组和两者同时加组全期日增重分别为674、761、796、678克,日采食量分别为2.50、2.65、2.74、2.40公斤,饲料利用率分别为3.73、3.49,3.44、3.54,添加铜和/或喹乙醇对试验第65天血清铜兰蛋白、尿素氨及屠宰率、背膘厚和眼肌面积无显著影响(p>0.05);添加铜使65天血清碱性碱酸酶活性极显著增高(p<0.01),添加喹乙醇有使之升高的趋势,两者同时添加时,碱性酸磷酶活性降低。     

Current progress on genetic interactions of rice with rice blast and sheath blight fungi

Analysis of genetic interactions between rice and its pathogenic fungi Magnaporthe oryzae and Rhizoctonia solani should lead to a better understanding of molecular mechanisms of host resistance, and the improvement of strategies to manage rice blast and sheath blight diseases. Currently, dozens of rice resistance (R) genes against specific races of the blast fungus have been described. Among them, ten were molecularly characterized and some were widely used for breeding for genetic resistance. The Pi-ta gene was one of the best characterized rice R genes. Following the elucidation of its molecular structure, interaction, distribution, and evolution, user friendly DNA markers were developed from portions of the cloned genes to facilitate the incorporations of the Pi-ta mediated resistance into improved rice varieties using marker assisted selection (MAS). However, rice blast is still a major threat for stable rice production because of race change mutations occurring in rice fields, which often overcome added resistance based on single R genes, and these virulent races of M. oryzae pose a continued challenge for blast control. For sheath blight, progress has been made on the exploration of novel sources of resistance from wild rice relatives and indica rice cultivars. A major quantitative trait locus (QTL), named qSB9-2, was recently verified in several mapping populations with different phenotyping methods, including greenhouse methods. The ability to identify qSB9-2 using greenhouse methods should accelerate the efforts on the qSB9-2 fine mapping and positional cloning.     

Virus Location and Lymphocyte Apoptosis in Lymph Nodes of Piglets Infected with PCV-2

The present study has been performed to understand the location of the virus, type of apoptotic cells, and their relation to lymph nodes of piglets infected with porcine circovirus type Ⅱ （PCV-2）. Nine 32-day-old conventional piglets free of infection with PCV-2 were used, and distributed into three groups： control group （n = 3）, piglets inoculated with PCV-2 alone （PCV-2, n = 3）, and PCV-2 inoculated and KLH immunostimulated group （PCV-2 ＋ KLH, n = 3）. Superficial inguinal lymph nodes from all piglets were collected for histological examination after 32 days postinoculation, and immunohistochemistry for PCV-2 detection. Location of apoptotic cells was detected with TdT-mediated dUTP nick end labeling （TUNEL） and cell cycle, and the apoptotic rates were measured by flow cytometry. The characteristic histopathological lesions of the piglets in PCV-2 and PCV-2 ＋ KLH were lymphocyte depletions in the cortex and paracortex of the lymph nodes, epithelioid-like macrophage infiltration, and intracytoplasmic inclusion bodies presented in epithelioid-like macrophages. PCV-2 was mainly found in epithelioid-like macrophages by immunohistochemistry. In the lymph nodes, lymphocytes presented higher apoptotic rates in the cortex by TUNEL, special B-cell areas, and similar apoptotic cells were found in this compartment in the control. The apoptotic rates of the lymph nodes were 0.41, 3.34, and 4.88% in the control, PCV-2, and PCV-2 ＋ KLH groups by flow cytometry, respectively. The apoptotic rates of lymph nodes for PCV-2 and PCV-2 ＋ KLH piglets were significantly higher than those for the control group （P〈0.05 and P〈0.01）. The proliferation index （PI） was 0.17_＋0.01, 0.12_＋0.01 and 0.12_＋0.04 in the control, PCV-2, and PCV-2 ＋ KLH group, the PI of the control group was higher than that of the other groups, but without the statistical difference. PCV-2 can induce lymphocyte depletion in lymph nodes of piglets by blocking cell proliferation and promoting apoptosis. This is one o     

锌对镉·汞复合污染下豌豆幼苗生长的影响

以豌豆为试验材料,采用水培方法研究了锌对镉、汞复合污染下的豌豆种子萌发与生长的影响。结果显示,锌浓度≤100.0 mg/L时促进豌豆种子的萌发,锌浓度为200.0和300.0 mg/L时抑制豌豆种子的萌发。当镉、汞的浓度为3.0 mg/L时,100.0 mg/L锌的解毒效应最明显,表现为能够促进镉、汞胁迫下豌豆种子萌发和幼苗生长,降低豌豆幼苗根尖细胞畸变率,提高豌豆幼苗的叶绿素含量及过氧化物酶活性;但当锌浓度≥200.0 mg/L时,与镉、汞协同迫害豌豆幼苗的生长。     

仙人掌超氧化物歧化酶提取与纯化研究

以仙人掌为试验材料,采用硫酸铵分级沉淀法得到超氧化物歧化酶（Superoxide Dismutase,SOD）粗酶液。然后用Sephadex（葡聚糖凝胶）G-75层析柱层析对粗酶液进行进一步纯化,最后用邻苯三酚法测定酶活力,并对酶活力主要影响因素（包括pH值、温度、抑制剂）进行了研究。结果显示仙人掌SOD酶比活力为751.56U.mg-1。pH值为7.5~8.5时酶活力最大,稳定性也好。温度维持在25~30℃时酶活力最高。由于过氧化氢和氰化钾均能使仙人掌SOD的酶活力显著下降,因而可以初步判断从仙人掌中提取到的SOD是Cu.Zn-SOD。