中国特有种爆杖花的微卫星分子标记开发与评价

爆杖花（Rhododendron spinuliferum）是中国西南地区特有的观赏和药用植物。为了研究爆杖花和碎米花之间的杂交物种形成过程,本研究利用FIASCO方法对爆杖花进行微卫星引物开发,从100对引物中筛选出28个微卫星标记,其中22个为多态。利用爆杖花两个居群共24个个体对22个多态性位点进行分析,结果显示：每个位点具有2~5个等位基因,平均为34个,其观测杂合度和期望杂合度分别为0083~0792和0153~0744。对筛出的28个微卫星标记在碎米花的两个自然居群中也做了检测,结果显示：有22个微卫星标记成功扩增,其中20个有多态性;每个多态位点有2~6个等位基因,平均为32个,其观测杂合度和期望杂合度分别为0000~0833和0117~0736。开发的微卫星标记可用于爆杖花及其近缘物种的居群遗传学分析和杂交物种形成研究。     

白条草蜥微卫星多态性的分离和鉴定

白条草蜥（Takydromus wolteri）是一种年产多窝卵的蜥蜴。为了对其婚配制度、同一雌性个体所产卵的窝内和窝间的父权状况、种群的遗传结构和物种的进化历史等研究内容进行探讨,本研究筛选出白条草蜥的9个具有高度多态性的微卫星位点。微卫星位点筛选自包含(AC)n 和(ATAG)n重复片度的微卫星富集文库。在白条草蜥安徽滁州种群的16~32个个体中对上述位点进行检测后发现,上述座位的等位基因数目范围为12~20个,期望杂合度范围为0.894~0.955,观测杂合度范围0.565~0.938,表明这些微卫星标记具有良好的遗传多样性,它们将在白条草蜥的种群遗传结构、基因流水平、种群分化和婚配制度的研究中发挥重要作用。     

七种菊科植物SSR标记在大黄橐吾和奇形橐吾杂交区类群中的通用性检测

牦牛山橐吾(Ligularia×maoniushanensis)是大黄橐吾(L.duciformis)和奇形橐吾(L.paradoxa)的一个自然杂交种。为了研究大黄橐吾和奇形橐吾杂交区类群的杂交关系和遗传结构,本研究利用两个杂交区杂交种及亲本的94个个体,从来自橐吾属、大吴风草属、千里光属和向日葵属7个种中已开发的145对核SSR引物进行筛选。结果表明在所研究的三个类群中至少能在其中一个类群中成功扩增的位点有54个,都能扩增的位点17个。在这17个位点中,牦牛山橐吾中11个位点具多态性,每个位点有2~14个等位基因,平均为4.8个,观测杂合度和期望杂合度分别为0.053~1.000和0.051~0.887;大黄橐吾中9个位点有多态性,每个位点有3~12个等位基因,平均为7.4个,观测杂合度和期望杂合度分别为0.200~0.971和0.283~0.844;奇形橐吾中10个位点有多态性,每个位点具4~15个等位基因,平均为7.6个,观测杂合度的值为0.025~0.974,期望杂合度的值为0.097~0.846。以上结果对大黄橐吾和奇形橐吾杂交区类群的杂交程度和遗传结构的进一步研究有重要作用。     

中国特有种爆杖花的微卫星分子标记开发与评价

爆杖花（Rhododendron spinuliferum）是中国西南地区特有的观赏和药用植物。为了研究爆杖花和碎米花之间的杂交物种形成过程,本研究利用FIASCO方法对爆杖花进行微卫星引物开发,从100对引物中筛选出28个微卫星标记,其中22个为多态。利用爆杖花两个居群共24个个体对22个多态性位点进行分析,结果显示：每个位点具有2～5个等位基因,平均为3．4个,其观测杂合度和期望杂合度分别为0．083～0．792和0．153～0．744。对筛出的28个微卫星标记在碎米花的两个自然居群中也做了检测,结果显示：有22个微卫星标记成功扩增,其中20个有多态性;每个多态位点有2。6个等位基因,平均为3．2个,其观测杂合度和期望杂合度分别为0．000～0．833和0．117～0．736。开发的微卫星标记可用于爆杖花及其近缘物种的居群遗传学分析和杂交物种形成研究。     

山羊群体遗传多样性的微卫星分析

为了保护和合理利用我国地方山羊品种遗传资源提供理论基础,本研究利用国际农粮组织和国际家畜研究所推荐的10对微卫星引物,结合荧光标记PCR,检测了中国9个地方山羊品种和1个引进山羊品种的遗传多样性。所研究的10个品种中7个呈现出高度多态,3个呈现出中度多态。并共检测到119个等位基因,有效等位基因数在1.4641~9.2911之间,座位平均杂合度在0.2618~0.7672之间,品种平均杂合度在0.5196~0.7024之间,其中SRCRSP23位点和河西绒山羊(HXR)平均杂合度最高。聚类关系(NJ和UPGMA)和主成分分析结果与其起源、育成历史及地理分布基本一致。     

梨小食心虫微卫星标记的扩增稳定性及多态性分析

【目的】筛选适合我国梨小食心虫Grapholita molesta种群遗传学研究的微卫星位点,并依据所筛选的微卫星位点进行梨小食心虫地理种群的遗传多样性分析。【方法】利用欧洲梨小食心虫和苹果蠹蛾Cydia pomonella种群中已报道的11个微卫星位点, 分析各位点在我国12个种群257头梨小食心虫样本中的扩增稳定性,再进行其多态性分析,筛选适合的位点,然后进行种群遗传多态性分析。【结果】在分析的11个微卫星位点中, 位点Gm01, Gm03, Gm04和Cyd15无法稳定扩增; 位点Gm05扩增成功率较低, 位点Gm07遗传多态性较低; 而位点Gm02, Gm06, Gm08, Gm09和Gm10等扩增效果稳定且遗传多态性丰富。这5个稳定扩增的微卫星位点平均等位基因数量（N_A）为7.417~12.500, 平均观察杂合度（H_o）为0.366~0.655, 平均期望杂合度（H_e）为0.642~0.846, 多态信息含量（PIC）为0.800~0.935。【结论】本研究成功筛选出位点Gm02, Gm06, Gm08, Gm09和Gm10等5个微卫星位点。基于这5个微卫星位点标记的结果显示, 山东和陕西不同梨小食心虫地理种群均具有丰富的遗传多样性。 这5个位点可以适用于我国梨小食心虫种群的进一步遗传分析研究。     

白条草蜥多态性微卫星位点的分离与鉴定

白条草蜥(Takydromus wolteri)是一种年产多窝卵的蜥蜴。为了对其婚配制度、同一雌性个体所产卵的窝内和窝间的父权状况、种群的遗传结构和物种的进化历史等研究内容进行探讨,本研究筛选出白条草蜥的9个具有高度多态性的微卫星位点。微卫星位点筛选自包含(AC)n和(ATAG)n重复片度的微卫星富集文库。在白条草蜥安徽滁州种群的16～32个个体中对上述位点进行检测后发现,上述座位的等位基因数目范围为12～20个,期望杂合度范围为0.894～0.955,观测杂合度范围0.565～0.938,表明这些微卫星标记具有良好的遗传多样性,它们将在白条草蜥的种群遗传结构、基因流水平、种群分化和婚配制度的研究中发挥重要作用。     

基于SLAF-seq技术的沙带鱼(Lepturacanthus savala)微卫星标记开发以及在近缘种中的通用性

沙带鱼是带鱼科中一种重要的海洋经济鱼类,广泛分布于印度-西太平洋海域。近年来,过度捕捞和渔业生态环境变化使沙带鱼的自然资源和种群结构受到了严重影响。因此,为了制定出科学合理的资源保护和管理措施,急需开发出大量多态的沙带鱼微卫星位点以促进其种群遗传学研究。本研究采用SLAF-seq技术,首次在沙带鱼基因组DNA中筛选出28个多态性微卫星位点。每个位点的等位基因数为2~18,平均值9.11;观测杂合度和期望杂合度分别为0.077~0.871(平均值0.634)和0.151~0.942(平均值0.711);各位点多态信息含量为0.137~0.919,平均值0.670。经过Bonferroni校正后,有3个位点显著偏离哈迪-温伯格平衡(HWE),并显示出较高的近交系数和无效等位基因频率,其余25个位点均符合HWE。这些多态性微卫星DNA标记可为沙带鱼的种群遗传资源研究提供有效的分子标记和技术支撑。此外,跨物种扩增结果表明,有19个微卫星标记在4种带鱼属鱼类中具有较好的通用性,这些标记可为阐明带鱼科属间或种间的系统进化关系提供新的研究手段和视角。     

跨种扩增筛选大鳄龟微卫星标记及遗传多样性分析

跨种扩增是一种能够快速、有效地获得物种微卫星标记的方法。本研究利用在近缘种中已发表的微卫星DNA引物,对大鳄龟（Macroclemys temminckii）进行跨种PCR扩增,在合成的69对引物中获得8对具有多态性的微卫星位点。对PCR扩增产物进行统计,得出观测杂合度（H_o）的范围是0.041 7~0.954 5,平均为0.384 8;期望杂合度（H_E）的范围为0.041 7~0.811 8,平均为0.479 1;多态信息含量范围为0.040 0~0.759 2,平均为0.423 2;经过卡方检验后,部分微卫星位点符合哈迪-温伯格平衡。总体来说,这些位点是研究大鳄龟遗传结构的良好分子标记。     

基于粪便DNA的岩羊微卫星引物的筛选及个体识别

贺兰山保护区岩羊Pseudois nayaur种群数量近年来增长很快,为了解贺兰山保护区的岩羊种群的遗传健康状况,从而更好地保护和管理岩羊种群,需要对岩羊种群的遗传多样性进行研究。在其近缘物种中选出36对微卫星引物对收集到的岩羊粪便DNA样品进行PCR扩增。结果发现,有9对引物在岩羊粪便DNA中扩增出了多态性位点。各位点基因杂合度介于0.26~0.95之间,平均杂合度为0.48,各位点多态信息含量介于0.23~0.68之间,平均多态性为0.48。     

Genetic Diversity Analysis of Nelumbo Accessions Based on EST SSR Markers

EST-SSR markers were applied to estimate the genetic diversity for 30 accessions of Nelumbo nucifera, 6 accessions of Nelumbo lutea and 14 hybrids between these two species. The 52 of 123 EST SSR markers (423%) were screened and then applied to amplify the 50 Nelumbo accessions. A total of 177 alleles were identified, and the number of alleles per locus and Polymorphic Information Content (PIC) value varied from 2 to 8 with an average of 34 alleles and from 063 (NNFB 1059) to 091 (NNFB 750) with an average value of 079, respectively. Jaccard similarity coefficients of the amplification results were analyzed by NTSYS pc2.11 software and the genetic similarity coefficient was from 024 to 086. The clustering dendrogram constructed by UPGMA method indicated that 50 accessions of Nelumbo could be divided into four major groups at the similarity coefficient of 037. Group I and group II included Nnucifera; group III included the majority of Asian American lotus hybrids; and group IV included Nlutea. In addition, the Asian American hybrids were closer to Nnucifera based on genetic relationship, which is consistent with the traditional classification result and the previous reports.     

ISSR and SRAP Markers Reveal Genetic Diversity and Population Structure of an Endangered Slipper Orchid,Paphiopedilum micranthum (Orchidaceae)

Paphiopedilum micranthum is an endangered pink slipper orchid mainly distributed in the limestone areas of southwestern China. Wild populations of this species have been seriously threatened by excessive collections, rampant smuggling for export, and habitat destruction. We used 15 ISSR markers and 11 SRAP markers to investigate the genetic diversity and structure of 15 natural populations. A high degree of diversity was observed at the species level (ISSR: PPB=9166%, He=03839; SRAP: PPB=9929%, He=02806). Certain degree of genetic differentiation among populations (ISSR: Gst=02577; SRAP: Gst=02383) was detected maybe caused by low gene flow (ISSR: Nm=07201; SRAP: Nm=07991). Consistent with the results of Principal Coordinate Analysis, the UPGMA dendrogram analysis divided the 15 populations into two main clades. In addition to geographic distance, the difference in elevation was another natural factor contributing to this differentiation. Knowledge about genetic diversity and structure gained from our study will be beneficial for the development of reasonable and efficient strategies to conserve this endangered species.     

The Transferability of Nuclear Microsatellite Markers in Four Castanopsis Species to Castanopsis tibetana (Fagaceae)

Castanopsis tibetana is a typical and dominant component in subtropical evergreen broad leaved forests of China. However, its genetic diversity and long term persistence have been jeopardized by habitat fragmentation. To examine its genetic diversity and genetic structure, cross species amplification of 51 microsatellite loci which are derived from four congeneric species, ie., Castanopsis chinensis, Csclerophylla, Ccuspidata and Csieboldii, were tested in Ctibetana.Thirty five loci were successfully amplified in Ctibetana, among them 11 were polymorphic across four wild populations of Ctibetana (24 individuals in total). Genetic diversity of four Ctibetana populations (Jiangxi) was relatively low. The number of alleles (NA) ranged from 2 to 7 (average 45) and the ranges of observed (HO) and expected (HE) heterozygosities were 0505 and 0471,respectively. The results will facilitate the study and conservation of genetic diversity of Ctibetana.     

Systematic Position of Allium macrostemon Based on nrDNA ITS and cpDNA trnL-F Sequence Data

Allium macrostemon is an important medicinal and edible plant. Its systematic position and taxonmical classification remain controversial to date. To explore this issue, we reconstructed phylogenetic relationships among Amacrostemon and other related taxa using nrDNA ITS and cpDNA trnL F markers. The phylogenetic trees derived from Bayesian inference and maximum parsimony analysis showed that Amacrostemon is monophyletic, and has a close relationship with some species of polyphyletic sections Caerulea and Pallasia instead of sections Codonoprasum and Allium. The including of Amacrostemon within section Allium was not supported by both molecular data and morphological characters of spathe, filaments and ovary. Allium macrostemon should be included in a new section, however further studies using additional samples (especially those from Central Asia) is necessary. In addition, we also provided a discussion on the phylogenetic relationships among four original plants (Amacrostemon, Achinense, Acaeruleum and Aneriniflorum) of Allii Macrostemonis Bulbus and systematic position of partial species of section Pallasia.     

Genetic Diversity of Global Aromatic Rice Varieties

Genetic diversity of 434 rice accessions collected from 16 countries, including 345 fragrance rice varieties and 89 non fragrance rice varieties, have been analyzed. A total of 573 alleles were detected by using 77 simple sequence repeats (SSR) primer pairs covering all rice 12 chromosomes. The value of allelic polymorphism information content (PIC) ranged from 0090 to 0845, with an average of 0516 per locus; Gene diversity (GD) varied from 0091 to 0859, with an average of 0573; The mean value of major allele frequencies (MAF) was 0540, covering from 0251 to 0953. In addition, 434 rice accessions are divided into three sub populations by cluster and population structure analysis, and FST between sub populations showed a mean of -0116, ranging from -0623 to 0494; The score of genetic distance calculated by Nei′s method appeared from 0207 to 0355. Major allele frequencies within each sub population distributed from 0408 to 0746, and gene diversity level from 0354 to 0699, while PIC from 0320 to 0658. Sequencing 6 mitochondrion genes in 18 rice varieties exhibited no different in 5 genes, whereas Mit4 contains a 3 SNPs in the gene body, which acts as an important marker to understanding the relationship between Indica/Japonica differentiation and the evolution of fragrant gene. Finally, genetic diversity and mitochondrion gene sequencing would help to know about the origin of fragrant resource and benefit rice breeding.     

利用微卫星标记研究鸭品种的遗传多样性

利用32个微卫星标记研究了5个福建省地方鸭品种（金定鸭、连城白鸭、莆田黑鸭、山麻鸭和番鸭）和2个对照品种（北京鸭和卡基康贝尔鸭）的遗传多样性和种质特性.结果表明：32个微卫星标记在7个鸭品种中检测到371个等位基因,平均每个基因座有11.719个等位基因,平均多态信息含量为0.522,有效等位基因数在5.141～6961,基因杂合度变化范围在0.512～0.700,杂合度较高;基于Nei氏标准遗传距离(D_S)和D_A遗传距离,采用UPGMA方法构建了系统发生树,将金定鸭、莆田黑鸭、连城白鸭、山麻鸭和卡基康贝尔鸭聚为一类,而北京鸭和番鸭各自聚为一类.所得聚类图与种群的真实关系较为符合.     

The Transferability of SSR Markers in Seven Asteraceae Species to Taxa in the Natural Hybrid Zone between Ligularia duciformis and Lparadoxa

Ligularia × maoniushanensis (Asteraceae) is a natural hybrid species of Lduciformis and Lparadoxa. In order to study hybridization and genetic structure of taxa in the hybrid zones between Lduciformis and Lparadoxa, cross species SSR primers were screened using 94 individuals of two hybrid zones’ hybrids and its parents from 145 nuclear SSR primers which had been developed from Ligularia, Farfugium, Senecio and Helianthus. The results indicated that 54 loci were successfully amplified at least one taxon of three taxa. A total of 17 loci were successfully amplified at three taxa, of which 11 were polymorphic in L. × maoniushanensis, and the number of alleles (NA) ranged from 2 to 14 (average 48) and the ranges of observed (HO) and expected (HE) heterozygosities were 0053~1000 and 0051~0887, respectively. Of which 9 were polymorphic in Lduciformis, and the number of alleles (NA) ranged from 3 to 12 (average 74) and the ranges of observed (HO) and expected (HE) heterozygosities were 0200~0971 and 0283~0844, respectively. Of which 10 were polymorphic in Lparadoxa, the number of alleles (NA) ranged from 4 to 15 (average 7.6) and the ranges of observed (HO) and expected (HE) heterozygosities were 0025~0974 and 0097~0846, respectively. The results will facilitate the further analysis of genetic structure and hybrid extent about taxa in hybrid zone between Lduciformis and Lparadoxa.     

虾池常见微藻种群温度、盐度和氮、磷含量生态位

就温度、盐度、氮磷含量设计试验,对4种虾池常见微藻种群新月菱形藻、啮蚀隐藻、微绿球藻、蛋白核小球藻的生态位进行研究.结果显示：啮蚀隐藻在温度和盐度资源上的生态位宽度值最大,分别达0.980和0.988,新月菱形藻在氮、磷含量资源上的生态位宽度值最大,达0.990,蛋白核小球藻在各资源的生态位宽度值均最小,平均只有0.926;啮蚀隐藻与蛋白核小球藻在温度和盐度资源上的生态位重叠值最小,分别为0.809和0.702,啮蚀隐藻与新月菱形藻在氮、磷含量资源上的生态位重叠值最小,只有0.829,蛋白核小球藻与微绿球藻在温度、盐度和氮、磷含量资源上的生态位重叠值均最大,分别为0.986、0.974和0.989.表明在养殖水环境调控过程中,啮蚀隐藻适宜与新月菱形藻、微绿球藻或蛋白核小球藻共同培育,而蛋白核小球藻和微绿球藻对资源的竞争性明显,不适合同时引入同一池塘进行培育.     

食物密度对镜湖萼花臂尾轮虫不同克隆生活史特征的影响

对镜湖萼花臂尾轮虫（Brachionus calyciflorus）夏季种群内4个生化遗传特征上互不相同的克隆（克隆A、B、C和D）在4个斜生栅藻（Scenedesmus obliquus）密度（1.0×10⁶、2.0×10⁶、4.0×10⁶和8.0×10⁶ cells·L^-1）下的生活史特征进行了研究.结果表明：食物密度对各克隆轮虫的存活率和繁殖率均有不同的影响.4克隆中克隆C的世代时间最短,克隆B的世代时间、生命期望和平均寿命最长,克隆A的后代混交雌体百分率最高;净生殖率和个体适合度在4克隆间无显著差异.镜湖萼花臂尾轮虫在2.0×10⁶ cells·L^-1的食物密度下净生殖率最低;在1.0×10⁶cells·L^-1的食物密度下平均寿命和生命期望最短,而后代混交雌体百分率却最高;在8.0×10⁶cells·L^-1的食物密度下种群内禀增长率最高,平均寿命和生命期望最长;在高食物密度（4.0、8.0×10⁶cells·L^-1）下个体适合度较大.克隆C的个体适合度在密度为3.9×10⁶cells·L^-1时最小,而克隆D的个体适合度在食物密度为6.34×10⁶cells·L^-1时最大.食物密度的变化可能是7月份之后镜湖萼花臂尾轮虫从水环境中消失的原因,而4克隆轮虫个体适合度的相似性则可能是镜湖轮虫共存于同一水体的原因之一.