多花黑麦草品种间杂交F2代分子标记遗传分析

运用序列相关扩增多态性(SRAP)和微卫星标记(SSR)技术对6个多花黑麦草亲本品种及其9个杂交F₂代组合80份材料进行遗传分析。筛选出具有多态性的15对SRAP引物、16对SSR引物,SRAP标记平均每对引物扩增出16.7条带,多态性位点百分率为68.88％。SSR标记平均每对引物扩增出14.75条带,多态性位点百分率为58.05％。通过聚类树状图分析得出结论,杂交F₂代内同一组合基本聚为一类,长江2号×剑宝和长江2号×阿伯德组合除外;父本和母本与其杂交F₂代遗传距离与聚类结果基本一致。同时还发现SRAP标记对杂交F₂代及亲代聚类分析可靠性高于SSR标记。     

An improved and efficient micropropagation of Eclipta alba through transverse thin cell layer culture and assessment of clonal fidelity using RAPD analysis

An improved and efficient in vitro regeneration system has been developed for Eclipta alba, a medicinally important plant, through transverse thin cell layer culture (tTCL). The transverse section of the nodal segment of field grown plants was used as tTCL explants for plant regeneration. Shoot multiplication from tTCL nodal explants was influenced by BAP and their interaction with Kin or NAA. MS medium containing 13.2 μM BAP and 4.6 μM Kin was most effective for shoot multiplication from tTCL nodal explants. Upon this medium, percent response for shoot proliferation was 100% with an average of 32.6 shoot buds per tTCL nodal explant. Regenerated shoots from tTCL nodal explants were rooted on the growth regulator free MS medium. The rooted plantlets were successfully acclimatized and established in soil with a survival frequency of 90-100%. Random amplified polymorphic DNA (RAPD) markers were used to evaluate the genetic fidelity of the micropropagated plants. RAPD profile analysis indicated that micropropagated plants were genetically similar to mother plant.     

First approach to studying the genetics of the meagre (Argyrosomus regius; Asso, 1801) using three multigene families

The meagre (Argyrosomus regius) is a commercially important species for fisheries, and aquaculture production has been increasing in recent years. However, this growing importance has not been matched by increased genetic knowledge of the species. For this reason, an initial approach has been made to understanding the genetics of the meagre, using three particular multigene families (45S and 5S ribosomal DNA and U2 snRNA gene) as both molecular and cytogenetic markers. Only one type of 5S rDNA and only one ITS‐1 (from 45S rDNA) were obtained, and these therefore could provide the basis for the development of genetic markers for this species. However, the U2 snRNA gene produced a multi‐band electrophoretic pattern, and some of these bands were demonstrated to be linked with the U1 snRNA gene. This linkage could also provide a good species‐specific marker which could be useful for tracing the evolutionary history of the Sciaenidae family. In this study, we have described for the first time the karyotype of the meagre: this is composed of 48 acrocentric chromosomes. Each of the three gene families were localized on different chromosome pairs, although the U2 snRNA gene probe was also located scattered throughout the genome of the meagre, as expected by the amplification pattern.     

大麦遗传多样性及连锁不平衡分析

为了合理评价引进种质资源,为大麦基因发掘及育种组合配置提供依据,选用分布于全基因组的64个SSR标记,对221份大麦材料进行了基因型分析。共检测到192个等位变异,变幅为2~7个;基因频率变异范围为0.0090~0.9729,平均0.3333;全部位点的基因多样性变化范围在0.0528~0.7807,平均0.4813;多态性信息含量(PIC)变异范围在0.0514~0.7464,平均0.4113。供试材料间遗传相似系数变幅为0.4844~0.9792,平均0.7023。221份材料被划分成两大群7个亚群,国内地方品种与1份北京品种为一大群,国内育种品种与所有国外引进品种为另一群。遗传结构分析与聚类结果基本一致,两大类群间的遗传距离为0.3358,且第二大群多样性比第一大群丰富。2016个SSR位点成对组合中,不论共线性组合还是非共线性组合,都存在一定程度的连锁不平衡(LD)。D′统计概率(P<0.01)支持的LD成对位点830个,占全部位点组合的41.2%,D′平均值为0.4,整体LD水平较高。栽培品种的LD水平高于地方品种,且现代遗传改良的目标性状集中于2H、4H、6H和7H染色体。     

苦荞SSR分子遗传图谱的构建及分析

构建苦荞遗传连锁图谱,为今后有关苦荞基因组结构、重要农艺性状QTL定位、分子标记辅助育种和基因克隆等研究工作奠定基础。以栽培苦荞‘滇宁一号’和苦荞野生近缘种杂交产生的119份F4代分离材料为作图群体,利用SSR分子标记来构建苦荞的分子遗传连锁图谱。本研究构建的连锁图谱包含15个连锁群,由89个标记组成,其中偏分离的标记有22个,占24.7%,每条连锁群上的标记在2~16之间。连锁群长度在6.9~165.8 cM的范围,覆盖基因组860.2 cM,总平均长度9.7 cM。本研究构建了首张苦荞SSR遗传连锁图谱,为苦荞QTL定位、基因克隆、遗传选育等研究奠定了基础。     

濒危树种金钱松RAPD体系的建立和遗传多样性分析

通过优化随机扩增多态性DNA（RAPD） 反应体系,对浙江省不同来源的金钱松Pseudolarix amabilis进行遗传多样性分析。结果表明：18条随机引物在62个样品中可检测到172个可重复位点,其中多态性位点有170个,多态性比率为99.2%。聚类分析的结果表明：同一来源金钱松材料聚为一类,说明金钱松天然群体的RAPD多态性分类和其地理分布有一定的关系,但也有特殊个体不一致,表明金钱松天然种群体存在较高的遗传多样性,并建议对金钱松进行就地保存和迁地保存。图2表3参31     

水稻正反交杂合体在不同海拔产生的F2群体的遗传分化研究

利用云南海拔2650 m高寒稻区种植的地方粳稻老品种小麻谷与具有籼稻细胞质背景的改良品系南34为亲本,基于SSR标记评价了在400、1860和2200 m 3个不同海拔下产生的正反交F2群体的遗传多样性与遗传分化。结果表明,F2(小麻谷×南34)群体的遗传多样性高于在同一海拔下产生的反交F2群体。2个组合在海拔400 m的海拔产生的群体中遗传多样性均最小,并随海拔的升高而增加,在海拔2200 m的海拔产生的群体中遗传多样性最丰富。正反交组合F2群体间的遗传分化高于同一组合在不同海拔下产生的F2群体之间的遗传分化。同一组合在不同海拔产生的F2群体间的遗传分化随产生群体的海拔差异增加而扩大。本研究在DNA水平上证实了细胞质和海拔明显影响水稻杂种后代群体的遗传多样性和遗传分化。     

玉米种子纯度鉴定中DNA提取方法的比较研究

分别以玉米干种子胚、幼芽和幼苗为材料提取DNA,并对所得DNA的纯度和质量进行检测,结果显示,利用玉米干种子胚提取的DNA质量及分子量大小与幼苗法相当,完全可以满足SSR分子标记对玉米杂交种纯度的鉴定。     

DNA分子标记在果树种质资源遗传多样性研究中的应用

简要介绍了DNA分子标记的主要类型、原理及特点,重点综述了RFLP、RAPD、AFLP、SSR、EST-SSR和SNP标记技术在果树种质资源遗传多样性研究中的应用现状。     

仁用杏微卫星非变性聚丙烯酰胺凝胶电泳体系的优化

[目的]优化仁用杏微卫星非变性聚丙烯酰胺凝胶电泳体系。[方法]以25份仁用杏和3份食用杏为材料,利用微卫星标记结合非变性聚丙烯酰胺凝胶电泳技术筛选出多态性高、可重复的SSR位点。并比较不同因子对电泳体系的影响,探索仁用杏微卫星非变性聚丙烯酰胺凝胶电泳的优化体系。[结果]仁用杏微卫星非变性聚丙烯酰胺凝胶电泳的优化体系为：聚丙烯酰胺凝胶的浓度为6%,丙烯酰胺与甲叉双丙烯酰胺的比例为29∶1,1000V下电泳2～3h,0.1%AgNO3染色15min后显影。[结论]该优化体系为仁用杏资源微卫星标记遗传分析奠定了技术基础。