2014年-2016年广西地区猪繁殖与呼吸综合征病毒Nsp2和ORF5基因的遗传多样性分析

为了解广西地区猪繁殖与呼吸综合征病毒(PRRSV)流行毒株的遗传进化情况,对2014年-2016年来自广西各地的部分PRRSV阳性病料进行Nsp2和ORF5基因的扩增和测序分析.结果获得34个Nsp2基因序列和45个ORF5基因序列,均属于美洲型毒株.Nsp2基因间核苷酸序列的同源性为91.8％~100％,与PRRSV美洲型毒株VR-2332、CH-1a、JXA1及NADC30株核苷酸序列的同源性分别为81.3％~84.3％、88.9％~92.1％、94.3％~99.3％和73.5％~75.1％,而与PRRSV欧洲型毒株LV株核苷酸序列的同源性为51.5％~53.2％.ORF5基因间核苷酸序列的同源性为82.8％~100％,与PRRSV美洲型毒株VR-2332、CH-1a、JXA1及NADC30株核苷酸序列的同源性分别为83.7％~99.5％、85％~95％、83.8％~99.7％和83.2％~86.4％,而与PRRSV欧洲型毒株LV株核苷酸序列的同源性为62.4％~64.5％.基于Nsp2和ORF5基因推导的氨基酸序列绘制的遗传进化树中,广西地区的毒株主要分布在以JXA1为代表的Ⅳ亚群.表明当前广西PRRSV流行毒株以JXA1株为代表的高致病性美洲型毒株为主,各毒株Nsp2和ORF5基因序列存在一定的差异,尚未发现欧洲型毒株和美洲型NADC30类毒株.     

苎麻茎秆机械分离过程力学建模与试验分析

现有苎麻茎秆机械分离技术主要核心工作部件是剥麻辊结构,试验以该核心工作部件为基础,构建了新型茎秆分离台架。苎麻茎秆分离过程是由折断和麻皮麻骨分离瞬时完成的,以此推导了辊齿对麻秆作用的折断能和分离能的关系表达式,分析了结构参数对茎秆分离的影响;在此基础上,对主要部件剥麻辊以辊转速、辊半径、辊齿数、啮合深度为试验因素,设置了L9(34)正交试验。结果表明,各因素对剥净率影响的高低排序为剥麻辊转速、啮合深度、剥麻辊齿数、剥麻辊半径,较优组合为剥麻辊转速900 r/min、啮合深度5 mm、剥麻辊齿数20个、剥麻辊半径120 mm,此时剥净率为98.42%。试验揭示了苎麻茎秆机械分离的机理,优化了剥麻技术的工艺参数,为剥麻技术的开发提供了技术支撑。     

基于GBS技术的中华绒螯蟹的遗传特征分析

[目的]研究中华绒螯蟹经过多年的人工养殖、跨流域引种和增殖放流等活动可能会对其遗传特性造成的影响。[方法]基于下一代GBS测序技术,利用SNP标记对辽河家系、辽河野生、长江群体、海南群体、黄河群体进行遗传特征分析。[结果]经过条件为测序深度4×、Miss0.5、次要等位基因频率(MAF)0.05的过滤后,共获得652 746个高质量的SNP位点用于群体的遗传分析,各群体平均观测杂合度(Ho)为0.084 4~0.124 9,平均期望杂合度(He)为0.097 4~0.200 3,群体平均π值(Pi)为0.158 3~0.254 4,群体Fi的平均测量值(Fis)为0.054 2~0.238 43。两两群体间遗传分化指数Fst值结合系统发生树,可以看出,长江群体与海南群体的遗传距离较近,分化较小,表示海南的中华绒螯蟹苗种可能来自长江流域。[结论]该研究揭示了不同水系中华绒螯蟹的遗传差异,探讨了其遗传多样性水平,为中华绒鳌蟹资源的合理开发利用与保护提供理论依据。     

Genetic Variation Analysis of Mitochondrial cox1 and nad4 Genes of Ascaridia galli

To study the genetic variations of Ascaridia galli (A. galli) and the phylogentic relationships with other Ascaridata, fragment of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene and complete sequence of NADH dehydrogenase subunit 4 (nad4) gene of 15 adult A. galli isolated from Xichang city in Sichuan province were amplified by PCR, then analyzed the sequences. NJ trees and Bayes trees based on pcox1 and pnad4 genes were reconstructed. The partial sequences of cox1 gene and complete sequence of nad4 gene were 1 152 and 1 236 bp, respectively. There were 33 and 40 variable sites in the pcox1 and nad4 gene sequences, the intraspecific sequence variations within A. galli were 0 to 2.1% for pcox1 gene, 0 to 2.3% for nad4 gene. 8 and 11 haplotypes were detected in pcox1 and nad4 genes, the global haplotype diversity were 0.733±0.124 and 0.933±0.054, the nucleotide diversities were 0.00433±0.00153 and 0.00541±0.00157, and the average genetic distances were 0.004 and 0.005, respectively. Phylogenetic trees based on pcox1 and pnad4 genes with Neighbour-Joining and Bayesian analysis method revealed that all A. galli were clustered in one clade, and they were more closely related to A. columbae than any other members of Ascaridata. These findings indicated that 15 isolates of A. galli from Xichang city kept low genetic variation, nad4 gene was more suitable and effective than cox1 gene as molecular marker for detecting genetic variation of A. galli.     

Evaluation of Associative Effects of Alfalfa Meal and Ammoniated Corn Straw by Gas Production Technique in vitro

This experiment was conducted to determine the best roughage combination of alfalfa meal (AM) and ammoniated corn straw (ACS),so as to improve the efficiency of roughage utilization and reduce the cost of feeding. The AM was mixed with ACS in the proportion of 100:0,80:20,60:40,50:50,40:60,20:80 and 0:100 with 3 replicates per group. The accumulated gas production (GP) at fermentation for 3,6,12,24,48 h,and the changes of pH,dry matter disappearance rate (DMD),ammonia nitrogen (NH₃-N) concentration, microbial protein (MCP) and volatile fat acids (VFA) concentration,and their single factor associative effects index (SFAEI) and multiply factors associative effects index (MFAEI) were determined by gas production technique in vitro after fermentation for 48 h. The results showed as follows:The accumulated gas production of AM20:ACS80 group was higher than that of the other groups,and the DMD of AM0:ACS100 group was significantly lower than other groups (P<0.05).The NH₃-N concentration of AM20:ACS80 and AM0:ACS100 groups was significantly higher than the other groups (P<0.05).The MCP concentration was highest in AM20:ACS80 group,and the concentration of acetic acid and TVFA in AM20:ACS80 group was the highest (P<0.05),and the acetic acid/propionic acid ratio in all groups was more than 3,which indicating that it belonged to the acetic acid fermentation type.The pH during the fermentation ranged from 6.69 to 6.85.The results of MFAEI and SFAEI of each combination indicated that only the AM20:ACS80 group showed the positive associative effects. In conclusion,the 20:80 combination ratio for AM and ACS showed the best associative effects.     

Isolation,Identification and Genetic Evolution Analysis of Mink Serratia marcescens

In order to determine drug resistance, pathogenicity and genetic of the pathogenic bacteria in mink of Heilongjiang province, three strains were isolated from dead minks and identified. Based on colony morphology, microscopic characteristics, biochemical test and 16S rRNA sequencing. The antibiotic susceptibility to 24 antibiotics were detected by using standard K-B paper method, and virulence assay, phylogenetic analysis were carried out at the same time. The result showed that the isolates were highly tolerance to antibacterials, could cause 100% death of mice in experimental group and were closely related to strains from activated sludge and volcanic deposits. It showed that Serratia marcescens infection of mink existed in Heilongjiang province. Strains resistance phenomenon was serious and had a strong pathogenicity, and might be originated from the environment. It was the first time that the pathogenic Serratia marcescens isolated from the mink was reported. The results provided information for the detection, identification, diagnosis, treatment and prevention of related diseases.     

Isolation and Identification of Variant HeNZK-2014 of Pseudorabies Virus and Sequence Analysis of gE Gene

In order to learn the situation of pig pseudorabies virus (PRV) variant in this study, tissue samples such as lymph nodes which were collected from clinical pigs with suspected PRV infection were identified by PCR. PRV positive sample were inoculated on PK-15 cells after grinding and degerming, with further experiment including virus isolation, plague purification, PCR and IFA identification, TCID₅₀ confirmed by Reed-Muench method, inoculation test and observation of clinical symptoms in mice. The gE gene of purified PRV and brain tissue samples of dead mice were identified by sequencing analysis. The results showed that the virus grown on PK-15 cells could produce typical cytopathic effect (CPE) after 24 h; After three rounds of plaque purification,the isolate was PRV positive identified by PCR and IFA, and nominated as HeNZK-2014; The TCID₅₀ of the isolate was 10^-9.77/0.1 mL; The virus in 1×10⁸ TCID₅₀ inoculation was able to cause itching, tearing, death in infected mice, and PRV could be detected in tissues of dead mice; The molecular genetic variation analysis of gE gene by PCR amplification and clone sequencing indicated that the gE gene from brain tissue of infected mice shared 100.0% homology with HeNZK-2014, and located in a relatively independent branch with newly pandemic isolates in recent years after 2011, but was far from the classical strains before 2011, and both had two insertion of aspartic acid (D) at sites of 48 and 496 amino acids, which were considered to be the typical characteristics of PRV variants. This study successfully isolated a PRV variant, which laid a foundation for further research on vaccine development, prevention and control against PRV variants.     

Etiological Investigation of Diarrhea in Newborn Piglets and Molecular Characterization and Genetic Variation Analysis of ORF3 Gene of PEDV in North Guangdong

In order to understand the main pathogen of newborn piglets diarrhea in North Guangdong region, 31 diarrhea samples were collected from six pig farms in North Guangdong, the pathogen of porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV) and porcine rotavirus (PoRV) were detected by Real-time RT-PCR, meanwhile, ORF3 gene of PEDV amplified from positive samples were sequenced and analyzed. Pathogen detection results showed that 83.87%(26/31) samples were positive for PEDV, all herds and samples were negative for TGEV and PoRV. The sequence analysis revealed that the ORF3 gene of 5 epidemic strains of PEDV were all 675 bp, homologies of nucleotides were 98.7% to 100.0%, and homologies with reference sequences of nucleotides were 94.5% to 100.0%, and some gene mutation in common nucleotides site. The results of gene phylogenetic trees showed that PEDV could be divided into two groups, PEDV genetic relationship between field strains in North Guangdong and some regions in China, Southeast Asia, North America, Europe from 2013 to 2015 was closer, classed as a gene subgroup, from 2011 to 2012 main epidemic strains in our country and vaccine strains classed as other two gene subgroups. These results indicated that PEDV infection was the main pathogen of newborn piglets diarrhea in North Guangdong region, as time passed, the PEDV epidemic strains gene presented a tendency of evolution and variation.     

植物多倍体的形成及其二倍化机制

大约70%的开花植物在进化史上都经历过多倍化的过程,多倍体植物通常具有较强的可塑性,容易形成新物种。而新形成的多倍体往往不能稳定存在,其基因组需要再经历一次二倍化的过程,从而在基因水平上和细胞学水平上更接近二倍体。对多倍体二倍化的机制和相关研究的未来发展情况进行了探讨。     

山西花生地方品种芽期耐寒性鉴定及SSR遗传多样性

低温寒害是引起花生产量和品质下降的主要因素之一, 培育和种植高产稳产耐寒性强的品种是降低低温寒害的理想途径。然而高耐寒性种质的缺乏和耐寒性鉴定的困难, 是限制耐寒性育种取得突破的主要原因。本研究对72份山西花生地方品种进行芽期耐寒性鉴定, 以其相对发芽率和相对发芽指数作为耐寒性评价指标, 初步将其分为高耐寒、耐寒、中感、敏感、高感5级。为了解山西花生地方品种耐寒性遗传多样性, 合理、高效利用耐寒型花生资源, 用多态性好的90对SSR引物评价了不同耐寒性花生品种, 结果显示, 参试品种遗传多样性存在较大差异, 在遗传距离为0.4时, 被聚类为三大类群。3份高耐寒品种和7份耐寒品种分别聚类到不同的3个类群中, 说明耐寒性花生遗传多样性丰富。