实验用封闭群金黄地鼠mtDNA D-环核苷酸组成与变异分析

对兰州、上海、北京和武汉4个封闭群共98只金黄地鼠线粒体DNA(mtDNA)D-环进行分析.结果表明:实验用封闭群金黄地鼠mtDNA D-环序列中,T、C、A、G碱基含量分别为30.9%2、5.2%、29.5%、14.4%,其中A+T为60.4%、G+C为39.6%,G+C含量明显低于A+T;发现的167个多态位点中,113个转换、52个颠换、2个位点既有转换也有颠换,分别占多态位点数的67.7%、31.1%1、.2%,没有插入/缺失同时出现的位点,表明我国实验用封闭群金黄地鼠个体间多态性丰富,群体间无明显变异.     

绵羊脂肪型脂肪酸结合蛋白基因(A-FABP)的变异研究

【目的】明确绵羊A-FABP(adipocyte fatty-acid binding protein,A-FABP)基因的变异和单倍型特征.【方法】基于Ensemble数据库中绵羊A-FABP基因全序列,对7个绵羊品种共20个样本的A-FABP基因全序列进行测序分析.【结果】PCR扩增获得绵羊A-FABP基因全长6474bp,A、T、G、C 4种碱基的比例分别为31.48%、33.02%、17.21%和18.29%,A+T平均含量为64.50%,G+C平均含量为35.50%;共发现48处单碱基变异位点和2处微卫星位点M1((TG)n)和M2((TA)n),单一变异位点、2核苷酸变异位点和3核苷酸变异位点比例分别为28.85%、40.4%和0.02%;共发现转换、颠换、插入和缺失4种变异类型,其占变异位点总数的比例分别为45.83%、31.25%、2.08%及20.83%;共发现19种单倍型,单倍型多样度为0.995.【结论】A-FABP基因19个单倍型序列的NJ树分化为2个聚类簇,表明A-FABP基因单倍型最初由2个主要单倍型衍化形成.     

基于Cyt b基因序列的大叶蝉亚科部分种类分子系统学分析

为探讨吸汁类害虫大叶蝉亚科种类间的系统发生关系,选用线粒体Cyt b基因432 bp片段进行扩增、测序,分析大叶蝉亚科9属21种昆虫的系统发育关系.结果表明:Cyt b基因的多序列中,有227个核甘酸变异位点,变异率为52.5％,简约信息位点206个,占总位点数的47.7％;T、C、A、G碱基平均含量分别为38.8％、17.4％、33.2％、10.6％,A+T平均含量为72.0％,明显高于G+C含量(28.0％).T、C转换大于A、G,T、A颠换占主体.密码子第1、3位点转换已达到一定饱和.以横脊叶蝉亚科(Evacanthinae)的2种类(黑条脊额叶蝉、褐带横脊叶蝉)以及杆叶蝉(Hylicinae)作为外群,基于432 bp编码的144个氨基酸序列构建了系统树,MP树和NJ树结构基本一致,均支持大叶蝉各属的单系性,构建的系统树和雄性外生殖器一些特征演化吻合.故Cyt b基因是大叶蝉亚科属种系统发育十分有效的基因.     

中华豆蟹与太平大眼蟹ITS区比较研究

[目的]为蟹类的分子进化研究积累资料。[方法]采用酚-氯仿抽提法从中华豆蟹和太平大眼蟹的肌肉组织中提取总基因组DNA,对其ITS基因片段进行PCR扩增和测序,同时对其序列同源性、碱基组成和变异情况进行初步研究。[结果]中华豆蟹与太平大眼蟹ITS序列片段分别为739和769 bp,存在明显长度差异。中华豆蟹的AT含量和GC含量分别为49%和51%,而太平大眼蟹分别为40.79%和59.3%。4种碱基(A、T、G、C)在2种蟹中ITS序列的平均含量基本相当。中华豆蟹与太平大眼蟹ITS序列片段共有777位点,其中变异位点346个,碱基转换与颠换比约为0.8。颠换数大于转换数,表现为较高的颠换偏岐。[结论]中华豆蟹和太平大眼蟹间的平均遗传距离为0.603。     

基于柿雌雄花芽转录组测序的SSR和SNP多态性分析

为深入开发柿SSR和SNP分子标记,进而推动柿品种鉴定和遗传多样性等研究,以‘禅寺丸’柿的雌雄花芽转录组序列为基础,对SSR和SNP位点进行发掘。结果表明:转录组测序获得了154 741条unigene,其中有38.49%unigene在数据库中得到注释。利用MISA软件进行SSR位点的搜索,共得到44 304个SSR,包含83种重复基元,其中以A/T类型为主的单核苷酸重复所占的比例最高(20 006个,占47.63%),其次是以AG/CT类型为主的二核苷酸重复(16 055个,占38.23%),再次是以AAG/CTT类型为主的三核苷酸重复(5 402个,占12.87%)和以AAAG/CTTT类型为主的四核苷酸重复(500个,占1.19%)。在转录组得到的unigene中共发现SNP 405 685个,发生频率为1/253bp。6种单核苷酸变异中,转换类型发生频率显著高于颠换类型,转换类型中的C/T(31.51%)和A/G(31.41%)发生频率最高,在颠换类型中A/T发生频率最高。柿SSR和SNP的位点是非常丰富的,可为柿遗传图谱构建、遗传多样性和亲缘关系研究提供丰富的基础数据信息。     

基于454测序的碧桃花瓣组织转录组核苷酸变异1）

通过对碧桃花瓣组织进行转录组测序,获得1556684条序列,平均读长446 bp,共计695．34 Mb数据,其中1492289条高质量序列经拼接后得到22762个重叠群。以拼接后的重叠群作为参考序列,利用SNP／InDel分析软件检测碧桃花瓣组织转录组中的核苷酸变异位点,共得到9836个SNP和1550个InDel位点。在SNP位点中,转换占62．55％、颠换占37．45％,A／G的变异最为丰富,C／G的变异最少。在InDel位点中,有34．71％的位点为单核苷酸插入、缺失突变。     

基于黄地老虎转录组测序的 SSR 和 SNP 特征分析

【目的】基于黄地老虎转录组测序结果，对其SSR及SNP位点信息进行分析。【方法】对黄地老虎 总RNA进行提取，使用Illumina平台对转录组进行测序，利用MISA和GATK软件分别对黄地老虎转录组的SSR和 SNP位点特征信息进行分析。【结果】黄地老虎转录组数据库包含66 469条Unigene序列。利用MISA对Unigene序 列进行搜索，得到SSR位点4 438个，分布于4 048条Unigene序列上，占总序列的6.09%。其中，以单碱基和三碱 基重复为主，分别占总重复类型的54.73%和29.29%。A/T基元是黄地老虎SSR的优势基元。SSR基元包含4~24次 重复，以5次重复（21.67%）为主。SSR位点序列长度为12~127 bp，包含3 493个中等多态性位点、820个高度多 态性位点。利用GATK对Unigene序列SNP位点进行搜索，得到转换类型（237 619个）和颠换类型（133 529个） 共371 148个。C/T占SNP位点总数的18.61%，比率最高；G/A位居其次（18.28%），均属于转换类型。转换类型 （64.02%）显著高于颠换类型（35.98%）。【结论】黄地老虎转录组数据库中SSR位点分布频率较高、种类较 多且多态性丰富，转换类型是主要的SNP变异类型，可为今后开展黄地老虎种群遗传结构与分化、遗传进化关 系、迁飞规律及该害虫综合防治等研究提供重要科学依据。     

基于12S rRNA和16S rRNA序列的龟鳖类系统进化特征研究

【目的】研究龟鳖类的系统进化关系,为龟鳖类资源的保护和合理开发利用提供理论依据。【方法】采用PCR扩增和测序的方法,获得小鳄龟（Chelydra serpentina）的12S rRNA和16S rRNA序列,分别结合NCBI中其他龟鳖的同源性序列进行比对分析;基于Kimura双参数模型计算龟鳖类种间、属间、科间的遗传距离;采用邻接（NJ）法、最大简约（MP）法和最大似然（ML）法构建分子系统进化树。【结果】经比对后得到394 bp的12S rRNA一致序列和544 bp的16S rRNA一致序列,二者合并得到938 bp的联合序列。其中,可变位点327个,序列总变异率为34.9%,简约信息位点222个,单变异多态位点105个。T、C、A、G的平均含量分别为23.6%、24.1%、33.5%和18.7%,A+T含量为57.1%,G+C含量为42.8%。在327个可变位点中,转换数为61,颠换数为24,转换/颠换比率（R）为2.54。拟水龟属间的遗传距离为0.021~0.060,平均为0.467;淡水龟科8属之间的遗传距离为0.022~0.110,平均为0.0724;曲颈龟亚目7科（除平胸龟属外）间的遗传距离为0.071~0.123,平均为0.105。分子系统进化树显示,木纹龟属首先和陆龟科聚在一起,然后再与淡水龟科汇聚;中华花龟、大头乌龟与拟水龟属的成员相互镶嵌,聚成一支;鳄龟科、海龟科、棱皮龟科聚为一支;动胸龟科独成一支。【结论】应将龟亚科、淡水龟亚科提升为龟科、淡水龟科,并将大头乌龟、中华花龟并入拟水龟属,而平胸龟应从鳄龟科中分离出来,独立自成平胸龟科。     

我国四大海域三疣梭子蟹线粒体控制区基因片段序列比较分析

通过对我国四大海域野生三疣梭子蟹(Portunus trituberculatus)9个群体线粒体DNA控制区基因片段进行扩增和测序,得到长度为530 bp的片段.分析结果表明:83条序列T、C、A、G4种核苷酸的平均含量分别为36.6%、16.1%、36.6%、10.7%.共检测到91个变异位点,其中缺失/插入位点2个,转换位点76个,颠换位点3个,转换、颠换同时存在位点10个.83个个体具有66种单倍型(haplotype),单倍型比率为79.5%,对9个群体用MEGA3.1构建NJ分子树,聚类结果显示黄海、东海、渤海6个群体之间的相对遗传距离比较近,聚在了一起,南海3个群体相对遗传距离比较近,单独聚在一起.我国四大海域的野生三疣梭子蟹不同群体间有一定遗传分化.     

我国四大海域三疣梭子蟹线粒体控制区基因片段序列比较分析

通过对我国四大海域野生三疣梭子蟹(Portunus trituberculatus)9个群体线粒体DNA控制区基因片段进行扩增和测序,得到长度为530 bp的片段.分析结果表明:83条序列T、C、A、G4种核苷酸的平均含量分别为36.6%、16.1%、36.6%、10.7%.共检测到91个变异位点,其中缺失/插入位点2个,转换位点76个,颠换位点3个,转换、颠换同时存在位点10个.83个个体具有66种单倍型(haplotype),单倍型比率为79.5%,对9个群体用MEGA3.1构建NJ分子树,聚类结果显示黄海、东海、渤海6个群体之间的相对遗传距离比较近,聚在了一起,南海3个群体相对遗传距离比较近,单独聚在一起.我国四大海域的野生三疣梭子蟹不同群体间有一定遗传分化.     

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。