波尔山羊杂交后代及其亲本随机扩增多态DNA研究

利用随机扩增多态DNA技术研究了波尔山羊、唐山奶山羊、青龙本地山羊以及波尔山羊与这两个山羊群体杂交后代共计128个山羊个体的随机扩增多态DNA。结果表明：（1）总群体平均遗传多样性指数（Hsp）为0.6974,群体遗传分化指数为0.9706,山羊群体间平均遗传距离指数（0.1314～0.2052）明显大于群体内的相应值（0.0582～0.1440）,上述结果说明,所研究山羊群体不仅具有较为丰富的遗传多样性,而且其核基因组遗传变异主要存在于群体间。（2）山羊群体间的分子聚类关系与各群体间的亲缘关系基本一致。 Abstract:The random amplified polymorphic DNA（RAPD）of 128 individuals was studied,which were from Boer goat, Tangshan dairy goat,Qinglong native goat and their hybrids crossbred with Boer goat.The average index of genetic polymorphism for whole population（Hsp）and the index of genetic differentiation were 0.6974 and 0.9706,respectively.The average index of genetic distance between populations（0.1314～0.2052）was significantly higher than that within populations（0.0582～0.1440）.All of these indicated that the genetic polymorphism was not only abundant,but also the genetic variation was mainly existed between goat populations.The molecular dendrogram among goat populations was in accord with their genetic relationship.     

乌羊和小香羊的RAPD分析

用40条多态引物对乌羊、小香羊、南江黄羊、黑山羊、川东白山羊、波尔山羊和马头山羊7个品种(或群体)进行RAPD分析,其中28条引物扩增出多态性谱带,并用于进一步对12只乌羊个体和12只小香羊个体基因组进行扩增.扩增产物以1.5%琼脂糖凝胶(含0.5μg/ml溴化乙锭)电泳分离.Nei氏公式计算品种间的遗传距离指数和品种内的遗传相似指数,NJ法构建系统聚类图.结果表明:乌羊和川东白山羊间的遗传距离最小,亲缘关系较近,而小香羊与各品种间的遗传距离都较大,亲缘关系较远.乌羊群体及小香羊品种都具有一定的遗传稳定性。 Abstract:A total of 40 primers generated polymorphic loci were used in random amplified polymorphism DNA(RAPD)analysis in seven goat breeds(or populations)including Wu goat,Small-Xiang goat,Nanjiang Brown goat,Black goat,Chuandong white goat,Boer goat,Matou goat.28 of them were selected for further amplification in individuals of 12 Wu goats and 12 Small-Xiang goats because of their different amplified patterns among seven goat breeds.Amplification products were separated by 1.5% agarose gel(contain 0.5μg/ml ethidium bromide)electrophoresis.Genetic distance indexes among breeds and genetic similarity indexes within breed were calculated by Nei,and the phylogenetic tree was constructed by NJ method.The results indicate that the genetic distance index between Wu goat population and Chuandong white goat breed is smallest,therefore Wu goat population is closely related to Chuandong white goat breed.While the genetic distance indexes between Small-Xiang goat breed and any other goat breeds are all big,so Small-Xiang goat breed is far related to any other goat breeds.The results also indicate that the genetic stability either among the Wu goat population or the Small-Xiang goat breed reaches some extent.     

Classification of ecological types of the Chinese yak

To investigate the genetic resources and to clarify the classification status of the Chinese yak, the Maiwa yak, Jiulong yak, Datong yak, and Tianzhu White yak were intensively studied using microsatellite DNA, RAPD (random amplified polymorphic DNA), and AFLP (Amplified Fragment Length Polymorphism) DNA markers on the basis of the previous study work of the authors' on chromosomes and blood protein polymorphisms of the Chinese yak. The overall results showed that: (1) the genetic distance based on allele frequencies at the microsatellite loci between the Maiwa yak and Jiulong yak was the largest (1.506), and as expected, the two populations of the Maiwa yak had the smallest genetic distance (1.062). The four yak breeds were clustered into two groups: the first group comprised the Jiulong yak, and the second group comprised the remaining three breeds; (2) the genetic distance between the Datong yak and Tianzhu White yak was the smallest (0.0336), whereas the genetic distance between the Jiulong yak and Maiwa yak was the largest (0.0414) based on the data from the RAPD and RFLP analyses. The four yak breeds were again clustered into two groups with the Datong yak and Tianzhu White yak grouped together first, followed by the Maiwa yak; (3) analyses of the chromosomes and blood protein polymorphisms showed a similar pattern of relationship among the breeds. Therefore, it was concluded that the Chinese yak could be considered to be of two types with the Jiulong yak and Maiwa yak as their representatives, respectively. This conclusion was in accordance with the classification proposed later by professor Cai L and his colleagues, according to which the Chinese yak was classified into two types, the ‘Qinghai-Tibetan Plateau yak’ and the ‘Hengduan Mountain Alpine yak’, although this conclusion differed from the other classifications. Considering the geographic distribution, ecological factors, history and development of the Chinese yak, it was believed that this classification is reasonable.     

用4个微卫星标记分析7个绵羊群体之间的遗传关系

分析了4个微卫星基因座BM143、OarHH35、OarAE101、BMS2508在7个绵羊群体（小尾寒羊、湖羊、乌珠穆沁羊、萨福克羊、多赛特羊、夏洛来羊、多赛特公羊×小尾寒羊母羊F1代杂种羊）286只绵羊中的遗传多态性。结果表明,这4个微卫星标记在7个绵羊群体中的等位基因数分别为9、11、14和9,其多态信息含量/有效等位基因数/杂合度分别为0.7073/3.7231/0.7314、0.8267/6.4399/0.8447、0.5743/2.5178/0.6028、0.6172/3.0712/0.6744,其中OarHH35的遗传变异最大,OarAE101最小。7个绵羊群体中小尾寒羊的遗传变异最大,湖羊的最小。基于Nei氏DA距离和DS标准遗传距离,采用UPGMA方法构建了系统发生树。该发生树将中国地方品种（小尾寒羊、乌珠穆沁羊、湖羊）和法国的夏洛来羊归为一类,将F1杂种羊、英国品种（萨福克羊和多赛特羊）归为另一类。绵羊微卫星基因分型技术为检查品种（群体）之间的遗传关系提供了一个有用的工具。 Abstract：The genetic polymorphisms of four microsatellite loci BM143,OarHH35,OarAE101,and BMS2508 were analyzed in 286 sheep of seven sheep populations (Small Tail Han sheep, Hu sheep, Ujumqin sheep, Suffolk sheep, Dorset sheep, Charolais sheep, F1 of Dorset♂ × Small Tail Han sheep♀). The numbers of alleles for BM143,OarHH35,OarAE101,and BMS2508 are 9, 11, 14 and 9 in seven sheep populations, respectively. The polymorphism information content/number of effective alleles/ heterozygosity of BM143,OarHH35,OarAE101 and BMS2508 were 0.7073/3.7231/0.7314, 0.8267/6.4399/0.8447,0.5743/2.5178/0.6028,0.6172/3.0712/0.6744 in 286 sheep, respectively. The results revealed the greatest genetic variation at OarHH35 locus and the lowest at OarAE101, the greatest genetic variation in Small Tail Han sheep and the lowest in Hu sheep among seven sheep populations. In the unweighted pair group method with arithmetic mean (UPGMA) dendrograms based on Nei's DA distance and Nei's DS standard genetic distance, the Chinese native breeds (Small Tail Han sheep, Ujumqin sheep, Hu sheep) were grouped together, then with Charolais sheep. The F1 crossbred sheep, and the two British native sheep (Suffolk sheep, Dorset sheep) also clustered together. Microsatellite genotyping in sheep provided a useful tool for examining the genetic relationships among breeds(populations).     

三品系小型猪35个微卫星基因座的遗传学研究

利用35个微卫星基因座对中国三个品系的小型猪（贵州小型香猪、广西巴马小型猪、版纳小耳猪近交系）进行了遗传检测。计算出三个小型猪品系个体样本在35个微卫星基因座的纯合率,并对其进行t检验。计算出各品系的平均杂合度,多态信息含量（PIC）及品系间的遗传距离,并进行了系统聚类。结果表明三个品系的小型猪其基因纯合率均较高,其中版纳小耳猪近交系的基因纯合率最高;PIC和平均杂合度均较低;贵州小型香猪和广西巴马小型猪亲缘关系较近,并均与版纳小耳猪近交系的亲缘关系略远。 Abstract：The polymorphism of 35 microsatellites in the three miniature pig breeds in China(Guizhou miniature pig, Guangxi Bama miniature pig, Banna miniature pig inbred) was analysed. Rates of homozygote for 35 microsatellite loci in three miniature pig breeds were calculated,and t-test for them were performed. Mean heterozygosity and polymorphism information content(PIC) were calculated for all breeds, and genetic distances between these breeds were estimated. The dendrograms were obtained based on genetic distances. The results suggest that rates of homozygote in the three breeds are all high, and that is the highest in Banna miniature pig inbred. The results also suggest that polymorphism information content and mean heterozygosity in all the three breeds are low, and the genetic relationship between Guizhou miniature pig and Guangxi Bama miniature pig is closer than their relationship with Banna miniature pig inbred.     

利用RAPD标记分析鸵鸟种群的遗传变异

为探讨分子遗传标记在鸵鸟(Struthiocamelus)育种中的应用,利用RAPD技术分析了我国养殖的三个主要鸵鸟品种内和品种间的遗传变异.蓝颈鸵鸟、澳洲灰和非洲黑三个品种在品种内和品种间显示出低水平的变异.品种间的遗传距离较好地说明了三个品种的育种改良过程,同时预示出用澳洲灰与其他品种鸵鸟进行杂交育种的可行性.选择的20条在鸡中能稳定扩增的RAPD引物其75%能在鸵鸟基因组中扩增出产物,显示出两种禽类DNA具有同源性,为今后利用鸡的微卫星引物对鸵鸟进行基因组扫描(genomescan)分析奠定基础。 Abstract:To evaluate the molecular genetic markers on ostrich (Struthio camelus)breeding,we have analyzed the genetic variability of three main ostrich breeds by using RAPD.The studies show that the Blue,Australia Grey and African Black have low genetic variability.The genetic distance between breeds clearly illustrates the breeding history,at the same time,it also showes the possibility that cross breeding can be applied to Australia Grey and the breeds.75 percent selected RAPD primers working well in chicken could stably amplify products in ostriches,which implied DNA identity between chicken and ostrich.The work provided a foundation for genomic scanning.     

Analysis of genetic diversity and phylogenetic relationships among red jungle fowls and Chinese domestic fowls

Genetic diversity and phylogenetic relationships among 568 individuals of two red jungle fowl subspe- cies (Gallus gallus spadiceus in China and Gallus gallus gallus in Thailand) and 14 Chinese domestic chicken breeds were evaluated with 29 microstaellite loci, the genetic variability within population and genetic differentiation among population were estimated, and then genetic diversity and phylogenetic relationships were analyzed among red jungle fowls and Chinese domestic fowls. A total of 286 alleles were detected in 16 population with 29 microsatellite markers and the average number of the alleles observed in 29 microsatellite loci was 9.86±6.36. The overall expected heterozygosity of all population was 0.6708±0.0251, and the number of population deviated from Hardy-Weinberg equilibrium per locus ranged from 0 to 7. In the whole population, the average of genetic differentiation among population, measured as FST value, was 16.7% (P<0.001), and all loci contributed significantly (P<0.001) to this differentiation. It can also be seen that the deficit of heterozygotes was very high (0.015) (P<0.01). Reynolds' distance values varied between 0.036 (Xiaoshan chicken-Luyuan chicken pair) and 0.330 (G. gallus gallus-Gushi chicken pair). The Nm value ranged from 0.533 (between G. gallus gallus and Gushi chicken) to 5.833 (between Xiaoshan chicken and Luyuan chicken). An unrooted consensus tree was constructed using the neighbour-joining method and the Reynolds' genetic distance. The heavy-body sized chicken breeds, Luyuan chicken, Xiaoshan chicken, Beijing Fatty chicken, Henan Game chicken, Huainan Partridge and Langshan chicken formed one branch, and it had a close genetic relationship between Xiaoshan chicken-Luyuan chicken pair and Chahua chicken-Tibetan chicken pair. Chahua chicken and Tibetan chicken had closer genetic relationship with these two subspecies of red jungle fowl than other domestic chicken breeds. G. gallus spadiceus showed closer phylogenetic relationship with Chinese domestic chicken breeds than G. gallus gallus. All 29 microstaellite loci in this study showed high levels of polymorphism and significant genetic differentiation was observed among two subspecies of red jungle fowl and 14 Chinese domestic chicken breeds. The evolutional dendrogram is as follows: evolutional breeds→primitive breeds (Chahua chicken and Tibetan)→red jungle fowl in China (G. gallus spadiceus)→red jungle fowl in Thailand (G. gallus gallus). The results supported the theory that the domestic fowls might originate from different subspecies of red jungle fowl and Chinese domestic fowls had independent origin.     

RAPD分析在绢丝昆虫亲缘关系研究中的应用Ⅱ.柞蚕品种间的遗传差异

采用RAPD技术,对5个柞蚕品种的遗传差异进行比较研究.结果表明,所采用的40个随机引物中,有27个引物扩增谱带清晰且重复性较好,扩增总片段数253条,单个引物的扩增片段数在4～16之间,片段大小在0.33～3.0kb之间.不同柞蚕品种间的遗传差异较小,遗传距离(D)在0.066～0.1659之间,根据D值,由UPGMA聚类分析软件绘制了它们的分子进化树。 Abstract:Random amplified Polymorphic DNA (RAPD) was used to analyze the genetic diversity among Antheraea pernyi.The genetic variance of five Antheraea pernyi was studied.The result showed that:27 of 40 arbitrary primers could amplify clear and repeating bands.A total of 262 fragments were obtained.Each primer gave 4~16 bands and the average was 9.7.The length of the band was 0.33~3.0kb.The D value between different breeds of Antheraea pernyi was 0.066~0.1659.The D value was used to construct a dendrogram by UPGMA.     

赣中南花猪随机扩增多态DNA与群体遗传关系的研究

用RAPD技术检测了泰和冠朝猪、永丰藤田花猪、瑞金三花猪、兴国茶园猪、上犹花猪、万安花猪、乐安花猪等7个赣中南花猪地方类群基因组混合DNA的多态性,经80个随机引物扩增筛选,16个引物产生了共44个多态标记。遗传距离指数计算结果显示：泰和冠朝猪和瑞金三花猪亲缘关系最近,而兴国茶园猪与乐安花猪遗传距离最远。各类群的UPMGA和NJ聚类分析结果与采用生化、免疫和细胞遗传标记分析的结论不尽相同,在此基础上,结合现行分类方法,认为各受试猪群可暂时归并为赣中南花猪类型,但对泰和冠朝猪和万安花猪的品种归属性应作进一步分析。 Abstract: Random amplified polymorphic DNAs (RAPDs) were used to investigate bulked genomic DNA polymorphism of seven pig populations including Taihe Guanchao,Yongfeng Tengtian Spotted,Ruijin Spotted,Xingguo Chayuan,Shangyou Spotted,Wanan Spotted,and Lean Spotted pig.Eighty random primers were screened,of them,16 primers generated polymorphic markers with total number of 44.The genetic distance index matrix indicated that the genetic relationship between Taihe Gaunchao and Ruijin Spotted pig population was the closest,while that between Xingguo Chayuan and Laan Spotted pig population was the farthest.Our molecular phylogenetic trees constructed by UPMGA and NJ methods were in general consistent with that based on biochemical,immunogenetic and cellular genetic markers except for differences in some populations.It was inferred from this study that all pig populations examined could be temporary classified into one breed,however,the population genetic character of Taihe Guanchao and Wanan Spotted pig populations should be further evaluated.     

3个山羊群体中4个微卫星DNA多态性及其与杂种优势的关系

利用4个微卫星标记（OarFCB11,OarAE101,McM218,McM38）对波尔山羊、太行山羊和河北奶山羊的等位基因频率、群体多态信息含量、有效等位基因数、杂合度和遗传距离进行了遗传检测,并测定了波尔山羊与河北奶山羊及太行山羊的杂交效果。结果表明：4个微卫星标记在波尔山羊、太行山羊和河北奶山羊3个品种中存在多态性,可以用于山羊遗传多样性的评估;从不同品种来看,太行山羊的遗传变异程度最大,而波尔山羊的遗传变异程度相对较小;波尔山羊与河北奶山羊的遗传距离大于与太行山羊,波尔山羊与河北奶山羊的杂种优势高于与太行山羊,与实际杂种优势测定结果相符。 Abstract: Gene frequency, polymorphism information contents, number of effective alleles, heterozygosity and genetic distances were studied in Boer goat, Taihang goat and Hebei dairy goat using four microsatellite markers（OarFCB11,OarAE101,McM218,McM38）. The crossing effects on Hebei dairy goat and Taihang goat with Boer goat were tested. The results indicated that there are genetic polymorphisms at four microsatellite markers in three goat breeds. Four microsatellite markers can be used for genetic diversity evaluation in goat breeds. The genetic variability of Taihang goat is the highest, and Boer goat is the lowest in three goat breeds. Genetic distances between Boer goat and Hebei dairy goat is bigger than that between Boer goat and Taihang goat. The heterosis between Boer goat and Hebei dairy goat is higher than that between Boer goat and Taihang goat. It accords with testing results on actual heterosis.     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.