亚麻ISSR的反应体系优化及引物筛选

对亚麻ISSR反应条件进行了优化。同时利用3个有代表性的亚麻样品从60条ISSR引物中筛选出13条多态性好、重复性高的引物,为亚麻的分子鉴定奠定了基础。     

利用ISSR分子标记检测老化扁蓿豆种质遗传完整性的变化

采用ISSR分子标记,对经老化处理的不同发芽率水平扁蓿豆种质进行遗传完整性分析。结果表明:用10个有效引物对4个不同发芽率居群进行PCR扩增,共检测到48个等位基因,每个位点的等位基因数为3～8个,平均为4.8个;老化处理群体的多态性位点百分率(PPB)、等位基因数(Na)、有效等位基因数(Ne)、基因多样性指数(H)、Shannon指数(I)等遗传参数,与对照群体的遗传参数相比都有所下降,表明老化处理后群体内的遗传多样性低于对照群体的遗传多样性;与对照群体相比,发芽率为20%时,各遗传参数呈极显著性差异,说明扁蓿豆种质更新发芽率临界值在20%～40%;ISSR分子标记可以作为老化扁蓿豆种子遗传完整性变化的检测方法,同时认为对于异质种质资源材料,低发芽率更新标准不利于种质资源遗传完整性的保持。     

新疆主栽苹果品种ISSR分析及富士芽变优系分子鉴定

以新疆主栽苹果品种及富士芽变优系为材料,进行了ISSR(Inter-simple sequence repeat)分析,构建了17个苹果品种的指纹图谱,并聚类分析确定不同苹果品种的亲缘关系。从100个ISSR引物中,共筛选出3个多态性好、稳定且重复性好的引物(UBC846、UBC881和UBC895)。17个苹果品种共在41个位点上扩增出条带,多态性条带百分率为77.2%。UBC895引物能够把富士芽变优系和长富2号品种清楚地区分开,构建的数字化指纹也可以清楚地鉴定其他苹果品种。聚类结果与传统的苹果谱系相一致,表现出ISSR分子标记在品种鉴定和亲缘关系分析中的稳定性和可行性。     

应用ISSR技术分析苦瓜种质资源的多态性

运用ISSR分子标记,对48个苦瓜样品的遗传多样性进行研究。从96条随机引物中筛选出14条ISSR引物,共扩增出181条带,其中多态性谱带为113条,多态性比率为60.32%。聚类分析结果表明:48个苦瓜品种之间的遗传相似系数在0.685～0.865。     

景宁木兰与木兰属其它植物之间亲缘关系与遗传基础研究

采用ISSR(Inter simple sequence repeat)分子标记研究景宁木兰(Magnolia sinostellata)与17个木兰科木兰属其它植物之间的亲缘关系,揭示景宁木兰的遗传基础。在本试验条件下,从120个ISSR引物中筛选出20个具有多态性的引物,共检测到178条清晰的条带,其中163条具有多态性,占91.60%。通过POPGENE 32软件分析,除了木兰亚属的山玉兰独自成一类外,符合赫钦生的木兰属分类系统:厚朴、广玉兰及其变种为木兰亚属一类,其余包括景宁木兰、天目木兰、武当木兰等为玉兰亚属一类;玉兰与辛夷的杂交种二乔木兰与辛夷的遗传距离最近,为0.231 4,与玉兰的遗传距离也只有0.316 7;木兰属内遗传距离最近的为天目木兰与日本辛夷,为2.232 1,遗传距离最远的为山玉兰与日本辛夷,为0.736 9;景宁木兰与本属其它植物间的遗传距离在0.278 2～0.610 9,其中与天目木兰的遗传距离最近,为0.278 2,与山玉兰的遗传距离最远,为0.610 9。这表明景宁木兰具有其特有的遗传基础,且与天目木兰亲缘关系最近。     

刚竹属14个品种遗传多样性的ISSR分析

利用ISSR分子标记手段,研究刚竹属部分竹子品种的遗传多样性。实验从50条ISSR引物中筛选出10条重复性好、条带清晰的引物进行PCR扩增。通过对75条谱带进行相似系数的聚类分析,结果同形态学分类结果基本吻合;运用特殊谱带,建立竹子品种的分子检索表,说明ISSR标记能够揭示材料间较高的遗传多样性。     

粤东地区橄榄种质资源遗传多样性的ISSR分析

该研究在于分析粤东地区橄榄种质资源的遗传多样性,从而为橄榄的保护和研究提供依据。利用ISSR分子标记对64份橄榄种质进行遗传多样性分析。共扩增得到128条谱带,其中多态性条带99条,多态性比率为77.34%。平均观察等位基因数为1.7734±0.4203、有效等位基因数1.4823±0.3888、Nei’s基因多样性0.2746±0.1975、Shannon’s信息指数0.4072±0.2721。64份种质的遗传相似性系数范围是0.5714~0.9379,平均遗传相似性系数为0.7793。聚类分析将64份种质分为5个类群,主坐标分析也将其分为5个类群。由结果可知,该地区橄榄种质总体遗传多样性水平较低,同时分子聚类分析结果与供试材料来源地无关。     

ISSR分子标记技术在桃品种鉴定中的应用

为了有效地鉴别桃苗木优良品种或类型,为桃种质资源研究和新品种保护提供理论基础,利用ISSR标记对28份桃种质进行了品种鉴定及亲缘关系检测。结果发现：23条ISSR引物共扩增出188条DNA片段,其中96条具有多态性,多态性频率为51.06%;材料间遗传相似系数GS变幅为0.84～0.97,平均为0.92。获得28份供试材料的指纹图谱,鉴定出9条特异出现DNA片段和11条特异缺失DNA片段。通过UPGMA聚类分析发现,供试材料中地理来源及部分农艺性状相近的品种或类型聚为小类,但大的聚类群没有明确的划分标识,可能与桃种质在地区间互换及所选择的ISSR标记有关。     

Somatic embryogenesis in Cymbopogon pendulus and evaluation of clonal fidelity of regenerants using ISSR marker

An efficient plant propagation system through somatic embryogenesis was established in Cymbopogon pendulus, an aromatic grass followed by analysis of genetic status of regenerants using ISSR markers. Optimum embryogenic callus induction was observed on MS basal medium supplemented with 13.57 μM 2,4-dicholorophenoxyacetic acid (2,4-D) with 8.88 μM N⁶-benzyladenine (BA). Subsequent culturing of embryogenic calli on MS medium containing 4.52 μM 2,4-D and 8.88–13.32 μM BA gave maximum number of somatic embryos. Addition of coconut water (CW) promoted induction, growth and differentiation of callus and somatic embryogenesis. Further development of embryos into plantlets was achieved on MS medium supplemented with lower concentration of biotin and calcium pantothenate (CaP) along with BA (4.44–13.32 μM) and kinetin (2.32–4.65 μM). The root meristems were established on half strength MS medium containing 2% sucrose and 2.46–9.84 μM Indole3-butyric acid (IBA) and successfully established in soil with 77.8% survival rate in field condition. Thirteen randomly selected regenerated clones were screened using six ISSR primers. Nine clones produced similar monomorphic amplification profiles while remaining clones showed minor variation with absence of certain parental bands and appearance of unique band. Majority of the regenerants maintained genetic fidelity with the generation of few variants as evidenced from similarity matrix estimates using Nei Li's coefficient of similarity data.     

Analysis of phenetic relationship between Citrus indica Tanaka and a few commercially important citrus species by ISSR markers

Analysis of polymorphism in ISSR amplicons revealed diverse genetic relationship between Citrus indica and five other citrus species. In a consensus UPGMA dendrogram, based on Nei and Li's distance matrices, the C. indica samples from three different sites of Meghalaya, India were clustered together with 99% bootstrap support. C. reticulata, C. sinensis, C. aurantifolia formed a cluster with 67% bootstrap separation. C. macroptera and C. maxima samples from two sites of Meghalaya formed separate clusters with respectively 100% and 98% bootstrap supports. Principal Components Analysis projected C. indica to be more closely related to C. aurantifolia than to the other citrus species of the study. PCA also exposed variations within the C. maxima and C. macroptera samples.