虹鳟、山女鳟及其杂交子代（虹鳟♀×山女鳟♂）的微卫星分析

摘 要：利用虹鳟（♀）和山女鳟（♂）进行种间杂交,获得了90.00%的受精率,80.52%的发眼率,90.68%的孵化率和30.68%的鱼苗成活率。运用13个微卫星分子标记对杂交亲本与杂交子代进行了分子遗传机制的研究,结果表明：(1) 在13个微卫星位点中,3个位点只在虹鳟中得到扩增产物,6个位点扩增出虹鳟和山女鳟清晰的差异条带,另外4个位点在双亲中没有扩增出显著差异条带;(2) 双亲遗传分化显著,虹鳟和山女鳟存在杂交现象,虹鳟和山女鳟杂交子代的遗传符合孟德尔遗传规律,属两性融合生殖,是真正意义上的杂交种。 (3) 杂交后代与虹鳟和山女鳟的遗传相似性系数分别为0.4617和0.5965,遗传距离分别为0.7729和0.5168, 表明杂交F1与两亲本的遗传差异不是对等的, 而是偏向父本一方,UPGMA系统树也同样证明了这一点。本研究结果将为虹鳟、山女鳟两种鱼的杂交育种工作的进一步开展提供依据。     

人工控制自然交尾条件下中国对虾父本的微卫星识别

人工控制自然交尾条件下,在保持中国对虾雌、雄5∶1的交配比率下,获得了两尾交尾并产生子代的雌虾,同时有4个疑似父本需要识别.利用3个微卫星引物及其组成的三重PCR技术对4个疑似父本进行了鉴别,准确的找到了相应的与雌虾交尾的雄虾.这为建立半同胞家系提供了一种新的技术和思路.     

家畜基因组遗传多态标记—微卫星标记研究进展(下)

３微卫星标记的应用自微卫星被发现以来就受到了家畜遗传育种家们的高度重视,并且充分认识到了把它作为畜种的基因组变异研究所具有的巨大的潜在价值。现在它已被十分成功地作为富含信息、又容易获得的基因标记,广泛用于构建物种的遗传连锁图,基因鉴定（基因诊断）、物...     

Genetic characterisation of oak seedlings, epicormic, crown and micropropagated shoots from mature trees by RAPD and microsatellite PCR

DNA was isolated from seedlings of Quercus robur, collected from a single provenance, and from epicormic, crown shoots and in vitro shoots from a single tree of Q. petraea using a CTAB method of extraction. DNA was obtained in sufficient quantity and purity, from 13 out of 30 seedlings, and from all isolations from epicormic and in vitro shoots (2.5–10.0 μg/g fresh/ weight). Smearing was minimised at a primer concentration of 0.12 μM with Taq polymerase at 0.5 unit/reaction. Nine primers produced 142 bands, 28 of which were polymorphic. A similarity index showed that 11 seedlings were closely related with high coefficients (0.85–0.90), but each could be identified from another using only 9 primers (OPA-02 and -05, OPG-04 and -05, OPE-01, -02, -03, -08, -09). DNA was isolated from crown, epicormic and in vitro leaves originating from a single 150-yr old tree of Q. petraea and analysed by randomly amplified polymorphic DNA (RAPD) and microsatellites. With each primer, a characteristic RAPD pattern was obtained, and it was common to all six epicormic shoots derived from different parts of a single branch of this tree; also to the shoots from the crown of the same tree with OPE1 OPA-05, OPA-08, OPA-01, OPA-02, OPA-04, OPA-05, OPG-02, OPG-10, OPE-12. Similarly, the RAPD pattern obtained from shoot cultures in vitro, derived from individual nodes of epicormic shoots produced by six different branch segments, were uniform for each of 15 primers. This work was repeated using microsatellite PCR. Three microsatellite loci AG16, AG 1/2 and AG 1/5 were amplified by PCR. It showed a uniformity of these microsatellite loci in shoots from the crown of the tree, and from epicormic shoots cultures derived from six different sections of branch.     

Molecular identification of Pm12-carrying introgression lines in wheat using genomic and EST-SSR markers

The traditional process of obtaining maize hybrids involves the generation of inbred lines through successive generations of selfing and subsequent testcrosses in order to identify the best combining ability by allelic complementation. A fast alternative to obtain inbred lines is to induce the formation of haploids followed by chromosome doubling. However, even with the aid of haploid-inducing genetic sources, this strategy has not been widely used in maize breeding programs, partly due to difficulties inherent to haploid generation and identification. In order to evaluate the possibility of using dihaploids to generate homozygous maize tropical lines, we used the androgenetic haploid inducer line W23 as a female parent in crosses with the tropical single-cross hybrid BRS1010. Within the progeny of these crosses, 462 seeds were phenotypically selected as putative haploids by the purple-colored endosperm and colorless embryo conditioned by the R1-nj gene. Among these, only four individuals were confirmed as being haploids using SSR markers, chromosome counting and flow cytometry, showing that the phenotypic marker was not efficient in detecting haploids in the tropical maize genotype used. All four haploids as well as some diploid plants presented reduced size, corroborating the difficulties for haploid identification by phenotypic evaluation. Genetic diversity analysis revealed by SSR markers divided the haploids in two groups represented by flint and dent maize inbred lines, which could be helpful in identifying complementary dihaploid lines. The present article demonstrates that a combination of haploid production and SSR fingerprinting is a feasible strategy for maize hybrid development in tropical germplasm.     

棉花微卫星DNA扩增产物检测方法的优化研究

以棉花两个多标记基因系F582和F586及其后代为材料,对微卫星DNA的PCR扩增产物检测方法进行了优化研究。结果表明：检测微卫星DNA,聚丙烯酰胺银染灵敏度高于琼脂糖EB染色;聚丙烯酰胺凝胶的浓度需随着待测SSR序列的片段大小而作适当调整,一般情况下聚丙烯酰胺凝胶的浓度以6％为宜,但当待检测的DNA片段小到100bp～250bp的区域范围时,凝胶的浓度需提高到8％。胶板样品上样量以3μl为宜。在显色液预冷(约10℃)的前提下,显色的时间应控制在4min之内,以便得到的胶板DNA条带强度适中、对比度好。     

我国苏南地区美洲鲥7个养殖群体的遗传多样性分析

为探讨苏南地区美洲鲥（Alosa sapidissima）养殖群体的种质资源现状及遗传多样性水平,本研究采用15个微卫星标记和线粒体D-loop序列,对7个不同群体：镇江丹徒（Dtq）、镇江扬中（Yzq）、苏州张家港（Zjg）、苏州相城（Xcq）、南通中洋（Zyq）、常州滆湖（Ghq）和常州武进（Czq）共计210尾个体,进行了群体多样性分析。结果显示,15个SSR位点中除Asa-12外,其余位点均表现为高度多态性（PIC > 0.5）。其中,7个群体期望杂合度He为0.615～0.758,多态信息含量PIC为0.568～0.723,两者均以Zjg群体最高。D-loop序列共检测到32个变异位点,定义了20个单倍型,其中Zjg群体单倍型最多（11个）。7个群体的单倍型多样性、核苷酸多样性指数分别为0.618～0.945、0.003～0.008。基于SSR和D-loop序列的遗传距离分析,发现Ghq群体和Zyq群体的Nei’s遗传距离（0.058）和K2P遗传距离（0.003）最近,低于其他群体间的遗传距离（分别为0.073～0.397和0.003～0.006）。综合分析,我们发现7个美洲鲥群体的遗传多样性较为丰富,而群体间遗传变异程度不高,基因流Nm > 1和镶嵌式排列的个体进化树也证实7个群体的亲缘关系较近。基于本研究,可初步了解苏南地区鲥的种质现状,为后续进一步开展育种工作奠定理论基础。     

Evaluation of genetic architecture and mutation drift equilibrium of Marathwada buffalo population in Central India

In the present study, microsatellite data on 24 loci were generated and utilized to evaluate the genetic architecture and mutation drift equilibrium of Marathwada buffaloes, a Central Indian population maintained under low input system. Sufficient allelic diversity was observed with a total of 109 alleles across different loci. The genetic diversity analysis of Marathwada buffaloes displayed moderate level of within breed variability in terms of mean number of alleles per locus (4.48) and heterozygosity values (Ho = 0.532, He = 0.624). The studied Indian buffalo population showed considerable heterozygote deficiency (F_IS = 0.138) and deviation from HWE at many investigated loci. Three quantitative tests viz. sign test, standardized difference test and Wilcoxon sign rank test and a qualitative test for mode shift distortion of allelic frequencies were employed to evaluate mutation drift equilibrium under three different models of microsatellite evolution. The population was found to deviate significantly under IAM and TPM, while it was reverse under SMM. The qualitative test for mode shift supported the results under SMM indicating the absence of genetic bottleneck in the recent past in Marathwada buffaloes.     

微卫星DNA标记在番茄亲子鉴定中的应用

采用242对微卫星DNA标记引物分析了29份番茄自交系的遗传多态性,从中筛选出扩增带型清晰、稳定的34对引物,34对引物的平均等位基因数为4.53,平均多态信息量(PIC)为0.514 1,累积鉴别力(CDP)达0.999 999.利用其中的11对SSR引物对杂交种‘浙杂210’、母本‘01-199-1-1-0’及7个...