pGH和IGF-Ⅰ双基因共表达载体的构建及转双基因猪的获得与检测

旨在构建可高效表达pGH基因和IGF-Ⅰ基因的双基因共表达载体,制备转双基因(pGH+IGF-Ⅰ)猪,以期探索pGH基因和IGF-Ⅰ基因对猪生长发育的影响,为节粮型高瘦肉率新品种猪的培育奠定理论基础。从长白猪耳样中提取总RNA,经反转录RT-PCR获得pGH基因不含终止密码子的编码序列和IGF-Ⅰ基因完整的编码序列,经酶切连接克隆至pc DNA3.1(+)真核表达载体上,构建pc DNA3.1(+)-pGH-IGF-Ⅰ双基因共表达载体。将其转染PK15细胞,Q-PCR检测2个目的基因在PK15细胞中的表达情况。将构建的双基因共表达载体用纳米材料包裹后转染长白猪精子,采用精子载体法制备转双基因猪。PCR及测序鉴定转双基因阳性个体,Q-PCR检测2个目的基因在转双基因猪体内的表达情况。PCR及测序鉴定追踪检测转双基因猪体内pGH基因和IGF-Ⅰ基因的稳定情况。RT-PCR及测序结果表明,成功克隆了长白猪的pGH基因和IGF-Ⅰ基因的编码序列。酶切和测序分析表明成功构建了双基因真核共表达载体,转染PK15细胞后,Q-PCR检测表明,pGH基因和IGF-Ⅰ基因均在mRNA水平成功表达。母猪妊娠获得13头仔猪,经PCR及测序检测,其中4头仔猪为转双基因阳性,转双基因阳性率为30.76%。Q-PCR检测外源pGH基因与IGF-Ⅰ基因在转双基因猪体内成功表达。1~7月龄均可检测到外源pGH基因与IGF-Ⅰ基因,证明2个外源基因在转双基因猪体内稳定存在,并未随着生长而丢失。在转双基因公猪的精液中均能检测到2个外源基因,证明外源基因存在稳定传代的可能。     

长白和大白猪主要生长性状的遗传参数估计

旨在分析母猪的出生年份、出生季节、初生重、开测日龄等固定效应对长白、大白猪主要生长性状的影响,并对目标生长性状进行遗传参数估计（遗传力、遗传方差、表型相关和遗传相关）,为猪的遗传改良提供基本依据。本试验利用GLM模型分析试验猪群（398头长白猪和1 176头大白猪）的固定效应对猪生长性状的影响,并采用多性状动物模型对目标性状进行遗传参数估计。目标生长性状包括达100 kg体重日龄（age to 100 kg,AGE）、达100 kg背膘厚（backfat to 100 kg,BF）、100 kg平均日增重（average daily gain to 100 kg,ADG）。研究表明,在大白和长白猪中,猪的出生年、出生季、初生重以及开测日龄对生长性状均具有极显著的影响（P<0.001）;长白猪的AGE、ADG和BF的遗传力分别为0.321、0.327和0.324,大白猪对应性状的遗传力分别为0.454、0.469和0.408;长白猪的ADG和AGE之间的遗传相关、表型相关分别为-0.990、-0.995,大白猪的ADG和AGE之间的遗传相关、表型相关分别为-0.993、-0.998,均呈现较强的负相关。长白、大白猪的生长性状（AGE、ADG、BF）均属于中等遗传力性状,其出生年份、出生季节、初生重和开测日龄对猪的生长性状影响较大。在遗传参数估计分析时,提高样本数量并提升表型数据质量,可以增加遗传参数估计的可靠性。本研究中的生长性状遗传参数估计结果较为可靠,可为后续的遗传改良提供参考。     

Genetic analysis for sow stayability at different parities in purebred Landrace and Large White pigs

Genetic parameters for sow stayability were estimated from farrowing records of 10,295 Landrace sows and 8192 Large White sows. The record for sow stayability from parity k to parity k + 1 (k = 1, …, 6) was 0 when a sow had a farrowing record at parity k but not at parity k + 1, and 1 when a sow had both records. Heritability was estimated by using single-trait linear and threshold animal models. Genetic correlations among parities were estimated by using two-trait linear–linear and single-trait random regression linear animal models. Genetic correlations with litter traits at birth were estimated by using a two-trait linear–linear animal model. Heritability estimates by linear model analysis were low (0.065–0.119 in Landrace & 0.061–0.157 in Large White); those by threshold model analysis were higher (0.136–0.200 & 0.110–0.283). Genetic correlations among parities differed between breeds and models. Genetic correlation between sow stayability and number born alive was positive in many cases, implying that selection for number born alive does not reduce sow stayability. The results seem to be affected by decisions on culling made by farmers.     

基于SNP标记的爆裂玉米农家品种遗传多样性

为了评价广西爆裂玉米农家品种的遗传多样性,利用56K SNP芯片技术对中国广西45个爆裂玉米农家品种、中国2个爆裂玉米杂交种和6个南美爆裂玉米种质进行全基因组扫描,获得多态性标记14 338个,基因分型为6 种类型。其中A/G类型最多,为5 805个;A/T类型最少,为149个。1号染色体的多态性SNP位点最多,为2 302个;10号染色体最少,为848个。45个农家品种间平均遗传相似系数为0.62,变幅为0.41~0.99,总体遗传相似度高。聚类分析将参试品种划分为三大类群,相同来源的农家品种大多聚在一起;主成分分析显示大部分农家品种聚在一起,与杂交品种和南美品种之间没有交集,表明农家品种遗传相似性较高,与杂交品种和南美品种遗传差异较大。     

利用MFLP方法分析我国小麦地方品种及黄淮麦区主栽品种的遗传多样性

为明确小麦地方品种和生产主栽品种间的亲缘关系以及MFLP分子标记技术在小麦品种遗传多样性研究中的有效性,利用MFLP标记技术对24个地方品种和12个来自河北、山东和河南的生产主栽品种基因组DNA进行遗传多样性分析,并利用NTSYS pc 2.10软件对试验数据进行非加权组法（UPGMA）聚类研究。结果表明,5对MFLP引物共扩增出279 条具有特异性的多态性谱带。主栽品种间遗传相似系数为0.6989～0.8746,其遗传距离比较近;而地方品种间及地方品种与生产品种间遗传多样性差异较大。利用34个MFLP指纹图谱标记位点编制了分子检索表,能成功区分36个小麦品种。研究结果还表明,MFLP分子标记技术可有效地应用于小麦品种间的亲缘关系和遗传多样性研究中。     

Maintaining Genetic Resources of Peach Palm (<Emphasis Type="Italic">Bactris gasipaes</Emphasis> Kunth): The Role of Seed Migration and Swidden-fallow Management in Northeastern Peru

Knowledge of the effects of farmer practices on population genetic parameters of peach palm (Bactris gasipaes Kunth) is relevant to the improvement and conservation of the palm’s genetic resources. Microsatellite markers were used to assess genetic diversity and population structure of peach palm in swidden-fallow agroforestry systems in northeastern Peru. The study covered eight communities, comprising two study areas 160 km apart – one occupied by indigenous Amerindians and the other by mixed race campesinos. Simultaneous analysis of an ex situ peach palm germplasm collection provided a means to compare population genetic parameters. Farmers who were surveyed on seed selection practices for peach palm reported that an average of only four palms (4.3 for campesino and 1.5 for indigenous populations) were used to provide seed for the establishment of the forest gardens sampled. As expected, inbreeding coefficients observed within communities were relatively high (f = 0.105 − 0.210), however, observed heterozygosities within communities were also high (0.625–0.741). A metapopulation approach was used to describe migration within and among regions, implying a hierarchical structure of gene flow which maintains relatively high levels of genetic diversity. Seed migration was found to occur over longer distances (≤600 km) and at a higher frequency (46% of palms sampled) in the indigenous study area, and a proportionally greater number of alleles was found (49 vs. 43 over three loci) with twice as many private alleles occurring only in the indigenous populations. The farmers’ practice of preserving remnant palms through successive swidden generations may have contributed to the maintenance of alleles by reducing the severity of founder effects. Although the campesino study area exhibited a significant (20% of the variation; p < 0.01) isolation-by-distance relationship across 35 km distance, in general, both study populations had relatively limited genetic structure (θ = 0.012–0.03), which is believed to have resulted from the exchange of seeds over long distances and periods of time.     

Establishment of Chinese soybean Glycine max core collections with agronomic traits and SSR markers

It is very important to efficiently study and use genetic diversity resources in crop breeding and sustainable agriculture. In this study, different sampling methods and sample sizes were compared in order to optimize the strategies for building a rationally sized core collection of Chinese soybean (Glycine max). The diversity in the core collection captured more than 70% of that in the pre-core collection, no matter what sampling methods were used, at a sampling proportion of 1%. Core collections established with both simple sequence repeat (SSR) marker data and agronomic traits were more representative than those chosen on an independent basis. An optimal sampling method for a soybean core collection was determined, in which strategy ‘S’ (allocating accessions to clusters according to the proportion of square root of the original sample size within each ecotype) was used based on SSR and agronomic data. Curve estimation was used to estimate the allelic richness of the entire Chinese soybean germplasm and a minimum sample size for a core collection, on which a sampling proportion of about 2% was determined to be optimal for a core collection. Further analysis on the core collection with fourteen agronomic traits and allelic constitution at 60 SSR loci suggested that it highly represented the entire collections both on genetic structure and diversity distribution. This core collection would provide an effective platform in proper exploitation of soybean germplasm resources for the study of complex traits and discovering important novel traits for crop genetic development.     

长白猪主要选育性状间的典型相关分析

对长白猪的3组14个性状进行了典型相关分析。其中生长速度包含4个性状,体尺包含6个性状,膘厚和外貌评分等包含4个性状。结果表明,生长速度与体尺性状的第一和第二典型相关系数达极显著水平,分别为0.647 8和0.353 1,占总相关的96.52%。生长速度与膘厚和外貌评分等性状的第一和第二典型相关同样达极显著水平,分别为0.925和0.241 7,占总相关的99.9%。体尺与膘厚和外貌评分等性状只有第一个典型相关系数达极显著水平,其典型相关系数为0.328 7,占总相关的75.59%。分析还表明在这些性状中起主要作用的性状有X1（二月龄体重）、X3（测定结束体重）、X7（胸围）、X10（腿臀围）、X11（背膘厚）、X12（校正膘厚）。     

长白猪肝脏、肾脏和淋巴结的显微形态及糖原PAS反应研究

[目的]对商品长白猪的组织形态学进行研究,了解商品长白猪宰前的生理状态。[方法]采集长白猪的肝脏、肾脏和淋巴结3种组织器官并制成石蜡切片,采用H.E染色方法对其组织结构进行研究。[结果]肾脏和肝脏的组织结构形态正常,肾脏和肝脏的细胞核染色明显,淋巴结的皮质和髓质分界不清晰,周围组织没有明显的淋巴索,淋巴窦较小。糖原PAS染色结果表明肝脏中的糖原颗粒较少,分布不均匀,而肾脏中糖原或糖蛋白多集中于血管球和肾小囊壁上。[结论]该研究可为今后进一步的商品长白猪理论研究奠定基础。     

民猪与长白猪主要组织器官生长规律研究

选择民猪和长白猪为研究对象,利用异速生长式分析两个品种1～8月龄组织器官的早熟顺序。结果表明,民猪脂肪沉积较早,在2.5～6月龄间沉积大量的脂肪,生长发育全期腹内脂肪的沉积能力突出,而长白猪在6月龄后沉积脂肪速度加快;两个品种各脂肪组织占胴体比率在6～8月龄维持不变。民猪肉、脂、骨、皮早熟性顺序为:骨骼>肌肉>皮肤>脂肪,而长白猪为:骨骼>皮肤>肌肉>脂肪;两品种猪脂肪组织早熟性顺序均为:肠系膜脂肪>皮下脂肪>肾周脂肪>大网膜脂肪。民猪内脏器官早熟性顺序为:小肠>肾>胰>肝>脾>肺>心>胃>大肠;长白猪内脏器官早熟性顺序为:小肠>肾>肝>胰>心>脾>胃>肺>大肠。