海南岛红树植物海桑遗传多样性的ISSR分析

海桑 (Sonneratia caseolaris)是海桑科红树植物 ,在我国仅天然分布于海南万宁、琼海、文昌等地。采用 ISSR分子标记技术对所有天然种群和海南东寨港红树林自然保护区引种的人工种群共 4个种群 86个个体进行了遗传变异分析。 11个引物共扩增出 2 39条带 ,其中 194条具多态性 ,多态位点百分率为 81.17%。在种群水平上多态位点百分率 4 0 .5 9%～ 5 0 .2 1% ,平均值为4 5 .71%。 Nei的基因多样性、Shannon信息指数在物种水平上分别为 0 .2 10 0和 0 .32 5 6 ,在种群水平上平均值分别为 0 .14 6 8和0 .2 2 10。 Nei的遗传分化系数 Gst和 AMOVA分析表明种群间已发生了较高的遗传分化。种群间的遗传一致度为 0 .90 11。估测的种群间的基因流为 0 .5 787。依据 Nei的遗传距离对不同种群进行 U PGMA聚类 ,聚类结果为横山种群 (HS)和东寨港种群(DZG)聚在一起 ,万宁种群 (WN)和琼海种群 (QH)聚为一类。Mantel检验表明遗传距离与地理距离之间有一定的正相关 ,但不显著。种群遗传多样性与环境因子间的相关性分析表明 :海桑种群遗传多样性水平与各环境因子间相关性均不显著。因东寨港引种种群的遗传多样性明显低于天然种群 ,为保护遗传多样性 ,应加强对琼海、万宁种群的就地保护和迁地保护工作。

19份苜蓿种质遗传多样性的ISSR分析

采用ISSR分子标记技术对19份国内外苜蓿种质的遗传多样性进行检测分析。12对ISSR引物在供试苜蓿材料中共获得有效扩增位点71个,其中多态性位点69个,多态性位点百分率为96.25%。引物的有效等位基因数、Nei′s基因多样性指数、Shannon′s信息指数和多态性信息含量平均分别为1.50、0.30、0.46和0.33。供试苜蓿材料的遗传相似系数介于0.296～0.887,平均为0.617,表明被研究苜蓿材料遗传异质性较好。聚类分析结果显示,供试材料在遗传相似系数0.522处可被划分为两大类,第1类群包括的苜蓿种质数达到17个,占到所有供试苜蓿种质总数的89.47%,第2类群包含的苜蓿种质数只有2个。主成分分析获得了与聚类分析基本一致的结论。

鳡遗传多样性的ISSR分析

利用ISSR分子标记技术对长江下游苏州段野生和野生F1代人工养殖的2个鳡群体遗传多样性进行分析研究。从77个ISSR引物中筛选出4个引物对鳡2个群体48个样品进行扩增,得到41个清晰的扩增位点。鳡野生和野生F1代人工养殖2个群体的多态位点比率和群体内遗传多样性指数分别为21.95%、17.07%,0.0724、0.0426;前者遗传多样性较后者略高。基因分化系数Gst和群体内遗传多样性指数估算分析均显示2个鳡群体之间出现一定遗传分化。鳡UPGMA系统树有较明显的歧化,表现出一定的遗传趋异。结果分析表明,鳡群体的遗传多样性相对贫乏;野生F1代人工养殖群体尚未形成自己独立的遗传结构,但2个群体间已经产生了一定的遗传分化,经过较多世代的人工繁育有可能形成自己独立而稳定的遗传结构。

雅鲁藏布江河谷丝须蒟蒻薯遗传多样性的初步研究

分布于雅鲁藏布江河谷的丝须蒟蒻薯(Taccaintegrifolia)与其在东南亚的主要分布区具有明显的间断分布格局。为了探讨地理隔离对其居群遗传结构和遗传多样性的影响,我们应用ISSR分子标记方法对采自西藏墨脱的3个丝须蒟蒻薯居群共65个个体进行了遗传多样性和居群遗传结构分析,并与马来西亚Seremban的1个居群(19个个体)进行了比较。19个ISSR引物共扩增到165个位点,其中111个为多态位点,占67.68%。丝须蒟蒻薯在物种水平上的遗传多样性虽然不低(PPB=67.68%,HT=0.185,Hsp=0.292),但在居群内的遗传多样性却非常低(PPB=12.81%,HE=0.065,Hpop=0.044)。与马来西亚居群(Ma)(PPB=31.71%)相比,墨脱的3个居群遗传多样性极低(PPB分别为3.66%,8.54%,7.32%)。Ma居群与墨脱居群相隔2000km以上,两个地区间的遗传分化程度很大(GST=0.777,FST=0.9206),而墨脱的3个居群间(0.28%)及居群内(7.94%)的遗传分化却非常低(P<0.001)。居群间极其有限的基因流(Nm=0.1435)可能是由于该物种是以自交为主的种类、种子散布很困难、居群间的隔离、生境的破碎化等原因所致。总之,雅鲁藏布江河谷地区特殊的地形和地貌以及与其他热带地区的地理隔离是造成丝须蒟蒻薯在这一地区遗传多样性极低的可能原因。

ISSR Analysis on Genetic Diversity of New Oil Palm Germplasm

[Objective]The aim of this study was to analyze the genetic diversity of new oil palm germplasm from DNA molecular level and provides a theoretical basis for the development and utilization of new oil palm germplasm. [Method]The genetic diversity of 22 new oil palm germplasm from Hainan and Yunnan were analyzed by ISSR technique. [Result]A total number of 201 bands were amplified with 23 ISSR primers,and polymorphic bands accounted for 41.8%. 22 oil palm varieties were divided into two groups by cluster analysis：Group I included new varieties of R1-R12 from Hainan and V20 from Yunnan,while group II included new varieties of V13-V22 from Yunnan,except V20. [Conclusion]New breeding oil palm germplasm from Hainan and Yunnan may come from different countries or regions.

ISSR Analysis of Genetic Relationship of Germplasm Resource in Prunus mume Sibe. et Zucc

Using ISSR technique to analyze the diversity and genetic relationship of germplasm resources in 39 Prunus mume Sibe. et Zucc., the result showed that 10 primers were screened with high resolution from 51 primers, 120 fragments were amplified, of which 98 were polymorphic loci, accounting for 81.67% of total. Tested materials were divided into 3 classes, as was fundamentally accorded with the traditional classification base on horticulture. There was no obvious difference in geographic relationship among the clustering results.

Genetic Variability of Astragalus sinicus L. Based on ISSR Markers

Genetic relationships among 22 accessions of Astragalus sinicus L. col ect-ed from different provinces of China were analyzed by inter-simple sequence repeat （ISSR） markers. The results showed that 10 highly reproducible ISSR fragments a-mong 40 primers were screened. Using these primers, a total of 684 ISSR frag-ments from 500 to 3 000 bp were amplified, and 59.2% of them showed polymor-phic by unweighted pair-group method with arithmetic （UPGMA） analysis. It revealed that the 22 accessions had a similarity range from 0.63 to 0.95, and existed biolog-ical diversities. Based on cluster and principal coordinate analyses, al accessions could be divided into four distinct groups.

The loss of genetic diversity during captive breeding of the endangered sculpin, Trachidermus fasciatus, based on ISSR markers: implications for its conservation

Abstract Inter-simple sequence repeat （ISSR） markers were used to determine the genetic variation and genetic differentiation of cultured and wild populations of Trachidermus fasciatus, an endangered catadromous fish species in China. Six selected primers were used to amplify DNA samples from 85 individuals, and 353 loci were detected. Relatively low genetic diversity was detected in the cultured population （the percentage of polymorphic loci PPL=73.80%, Nei＇s gene diversity h--0.178 2, Shannon information index I=0.276 9）. However, the genetic diversity at the species level was relatively high （PPL-91.78%; h = 0.258 3, I= 0.398 6）. The UPGMA tree grouped together the genotypes almost according to their cultured and wild origin, showing distinct differences in genetic structure between wild and cultured populations. The pairwise F^t values confirmed significant genetic differentiation between wild and cultured samples. The cultivated population seems to be low in genetic diversity as a result of detrimental genetic effects in the captive population. The results suggest that ISSR markers are effective for rapid assessment of the degree of diversity of a population, thus giving important topical information relevant to preserving endangered species.

Analysis of genetic relationship in 12 species of Section Strobus with ISSR markers

Genetic relationship of 12 species of Section Strobus was analyzed with ISSR markers. 117 loci were detected with 12 ISSR primers. Percentage of polymorphic bands （PPB） varied from 5.93% to 19.92%. P. pumila had the highest levels of genetic differentiation and P. flexilis had lowest. Total genetic diversity （Hr） of 12 species in Section Strobus was 26.21%, of which intraspecific genetic diversity （Hs） was 7.66%, and interspecific genetic diversity （DST） was 18.55%, and the genetic variation in interspecies accounted for 70.78% of the total genetic diversity. According to the cluster results of genetic distance, the 12 species were classified into two groups. The first group included P. griffithii, P.armandi, P. fenzeliana, P. kwangtungensis, P. strobus, P. monticola and P. wangii. The second group included P. albicaulis, P pumila, P. flexilis, P. sibirica and P koraiensis.

ISSR Analysis on Genetic Diversity of Local Varieties of Chinese Hu mulberry(Morus L.)

[Objective] The aim was to study on the genetic diversity of local varieties of Chinese Hu mulberry (Morus L.). [Method] The genetic diversity of 141 copies of Hu mulberry varieties was analyzed by ISSR molecular markers. [Result] 12 ISSR primers had amplified a total of 90 amplified,of which 57 bands were polymorphic,and the polymorphic rate was 63.33%. The genetic similarity coefficients of 141 Hu mulberry germplasm resources varied from 0.633 3 to 1.000 0 with the average of 0.483 35,indicating that there was difference on genetic diversity among different varieties of Hu mulberries. A dendrogram of all 141 Hu mulberry varieties based on the genetic similarity coefficients using ISSR molecular markers was generated by UPGMA cluster method. Clustering of the 141 Hu mulberry varieties did not correspond with the conventional classification involving differences in style,leaf,branch,fruit and other morphological or agronomical characters. [Conclusion] Four subgroups clearly represented the genetic relationships in the 141 accessions which were benefit for the variety improvement and germplasm resource conservation.

Genetic diversity analysis with ISSR PCR on green algae Chlorella vulgaris and Chlorella pyrenoidosa

In the present study, genetic polymorphism and diversity in unicellular clones of Chlorella vulgaris Beijerinck and Chlorella pyrenoidosa Chick were studied with Inter Simple Sequence Repeats PCR （ISSR PCR）. Samples including four clones of C. vulgaris and three clones of C. pyrenoidosa were purified by single-clone-choice method. For four C. vulgaris unicellular clones, the total number of the bands scored for 18 primers was 298; and the number of the polymorphic bands was 118, of which 39.6% were polymorphic. The size of PCR products ranged from 200 to 2 500 bp. The total number of bands scored for 18 primers, the number of polymorphic bands and the percentage of three C. pyrenoidosa unicellular clones was 194.83 and 30.8%, respectively. POPGENE analysis show that the average Nei genetic diversity （h^＊） and Shannon index of diversity （I^＊） in the four C. vulgaris unicellular clones was 0.2181 and 0.3208, respectively, which is slightly higher than those of the three C. pyrenoidosa unicellular clones （0.190 3 and 0.274 8）, which agreed with the percentage of polymorphic bands in the mixed samples of the two species. The results suggest that ISSR is a useful method to Chlorella for intra-species genetic analysis.

Genetic Diversity Analysis of Balsam Pear(Momordica charantia L.) by ISSR Marker

[Objective] The aim was to explore the relationship between the classification and regional distribution of balsam pear. [Method] In the research, 30 varieties of balsam pear were analyzed by ISSR marker. [Result] The research showed that172 bands were amplified by 18 primers, in which, 132 bands were polymorphism and the polymorphic proportion was 76.7%. All of the samples can be divided into four categories by UPGMA analysis. [Conclusion] The results indicated that the classification of balsam pear is similar as varieties' regional distribution, which will be useful for genetic relationships analysis and parents selection in hybridization breeding.

Assessment of Genetic Diversity of Argania spinosa L. Growing in Arid and Semi-arid Areas of Morocco as Revealed by Inter-Simple Sequence Repeats

Argania spinosa L. （Sapotaceae family）, endemic to Morocco, is a multipurpose tree with an important ecological and socio-economical interest. Nevertheless, the sustainability of the agro-forestry system is now threatened by overgrazing and over-exploitation leading to an alarming decline in the number of trees. In the present study, inter-simple sequence repeats （ISSR） markers were used to evaluate the genetic variation and to assess the genetic diversity distribution within and among 17 argan populations growing naturally under semi-arid and arid climate. Thus, a total of seven primers generated 260 well-defined bands, with an average of 37.14 bands per primer. Studied populations showed a relatively high level of genetic diversity at species level and low level of genetic diversity at population level. The percentage of polymorphic bands, Nei＇s gene diversity and Shannon＇s information index at population and species level were 30.15%, 0.164, 0.217 and 98.8%, 0.172, 0.293, respectively. A relative low level of genetic differentiation （Gst = 0.39） was in agreement with the results obtained from the analysis of molecular variance （AMOVA）. Estimated gene flow was Nm = 0.781 among populations. Mantel test revealed a non-significant correlation between genetic and geographic distance （r = 0.065, P 〈 0.05）, suggesting that the geographic distribution is not the major factor that shaped the present population genetic structure. The results can help preserve A. spinosa L. tree as an endangered species, however, refining these finding using co-dominant markers is recommended in order to establish conservation strategies.

Genetic diversity of oil-tea camellia germplasms revealed by ISSR analysis

Genetic diversity of 51 oil-tea camellia germplasms was analyzed using the optimized inter-simple sequence repeat （ISSR）-PCR reaction system with 22 primers screened from a set of 100 ISSR primers. The results showed that 493 discernible loci with distinct elec- trophoretic bands were obtained, of which, 478 loci （96.78%） were polymorphic. This indicated that oil-tea germplasms possess abundant genetic diversities. By clustering analysis performed using softwares of NTSYS 2.10 and Winboot, 51 oil-tea germplasms were divided into two groups： Group I had 48 lines of Camellia oleifera Abel, while Group II had three C. oleifera Abel related species and their similarity coefficient was 0.62. Group I was further divided into Group I-1 and Group I-2, and their similarity coefficient （Gs） was 0.634. All members of Group I-1 originated from Hunan Province, while Group I-2 included the rest of Hunan lines and those originated from other regions of China. Analyzed by software POPGENE 1.32, the Shannon＇s information index （I＊） of genetic polymorphism was 0.3852, the genetic diversity among different region popula- tions （Ht） was 0.2537, the genetic diversity within populations （Hs） was 0.15545, the dif- ferentiation coefficient of genetic diversity among populations （Gst） was 0.3967, and the gene flow among populations （Nm＊） was 0.8262. The Nei＇s genetic distances between the Hunan population and the populations originated from other regions of China implied that geographic isolation strongly influenced genetic differentiation among populations. Meanwhile, seedling rootstock grafting and high grafting for tree crown produced genetic variations among clonal offsprings.