11个猕猴桃品种间的遗传多样性分析

利用随机扩增多态性DNA标记(RAPD)技术对采自四川省猕猴桃科研基地的11个猕猴桃品种进行了遗传多样性分析.通过引物筛选,从15个RAPD引物中扩增出135条带,其中100条为多态带,占74.07%.UPGMA聚类分析结果揭示了各品种间的亲缘关系.RAPD标记说明猕猴桃品种间遗传距离与地理分布有一定关系,地理位置较近的材料能聚在一起.分子标记可用于猕猴桃种质资源的分类、鉴定及良种选育.图2表2参15     

与甘蔗抗黑穗病基因连锁的RAPD标记筛选

为了寻找与由黑粉菌(Ustilago sictaminea Syd．)引起的甘蔗黑穗病抗病基因连锁的分子标记，从而借助分子标记辅助选择(MAS)手段提高抗病育种效率，用甘蔗抗病亲本Co1001为母本，用感病亲本Ya71-374为父本，配置杂交组合创制抗病性分离群体为材料，采用人工接种新植蔗，田问种植方法鉴定分离个体新植和宿根的抗病性，借助RAPD和集群分离分析法(BSA)，筛选出一个与抗病基因连锁的多态性片段，并在抗、感池中的个体和池外部分个体上得到验证，该片段大小约520bp．对该片段的进一步克隆、测序和转换研究还在进行中，研究结果为抗病育种的MAS奠定了基础．图2表3参12     

利用RAPD技术分析兰属（Cymbidium）品种间的亲缘关系

本文利用RAPD技术来检测兰属13个品种间的亲缘关系。通过对55种10个碱基随机引物的筛选,其中4种引物能得到重复性、稳定性较高扩增产物,四种引物共扩增出93条多态性带,多态率为100％,根据RAPD标记进行邻体聚类分析,表明兰属各种间的亲缘关系与形态学分类结果不完全一致。实验结果认为RAPD技术可用于兰属植物的系统学研究。图2表3参13     

家蚕斑纹限性品种SC_1的RAPD差异DNA片段的克隆及其酶切图谱分析

利用RAPD技术对家蚕限性普斑品种SC1的雌雄体分别进行PCR扩增通过120种引物的筛选,得到1个雌体特异的RAPDDNA片段将该片段克隆到pUC19质粒,井进行了限制性内切酶图谱分析.     

不同DNA分子标记在云杉属植物遗传多样性研究中的应用

综述国内外近20a来在DNA分子水平上埘云杉属植物遗传多样性的研究进展，简述用RELP、RAPD、SCAR、AFLP、SSR、EST、STS以及SNP标记对于云杉属核DNA、线粒体DNA、叶绿体DNA和rDNA多态性进行的研究以及在云杉属品种鉴定、遗传作图、遗传多样性、进化及分子生态学研究中的应用，参85     

RAPD扩增中商品酶体系对扩增产物的影响

随机选用了10条RAPD引物，采用3种商品TapDNA聚合酶体系，以5个不同物种的动植物总DNA为模板，进行RAPD扩增，实验发现，在对同一模板、采用同一引物进行的RAPD扩增中，仅仅因为使用了不同的商品TaqDNA聚合酶，获得的扩增产物差别极大；不同商品来源的PCR反应缓冲体系，对扩增产物也有一定影响，但相对较小，这说明，不同的商品酶体系对RAPD扩增产物影响很大，因此，影响RAPD扩增产物稳定性的一个关键因素是商品TaqDNA聚合酶本身；研究中前后一致地使用同一商品TaqDNA聚合酶体系，是成功进行RAPD分析的基础。     

基于RAPD技术检测不同烃链长度咪唑基离子液体对日本三角涡虫的遗传毒性

RAPD标记通常用于遗传作图、分类和系统发育研究,该技术也可用于检测DNA损伤或突变。本研究利用RAPD技术检测了4种不同烃链长度的咪唑基离子液体对日本三角涡虫(Dugesia japonica)的遗传毒性。在20条随机引物中共筛选出11条用于RAPD扩增,其中对照组共扩增出65条带。与对照组相比,离子液体处理组的RAPD图谱变化表现在条带亮度增强或减弱以及新带的出现和正常带的消失。各处理组多态性带及变异带的总数随离子液体烃链碳原子数增加而增加,而基因组模板稳定性则随离子液体烃链长度增加而降低,这说明咪唑基离子液体对涡虫有遗传毒性,且随着离子液体烃链长度增加其对涡虫的遗传毒性增强。同时研究结果也证明RAPD分析是一种检测环境污染物对动物DNA损伤的灵敏方法。     

糯稻和非糯稻蜡质基因的新STS分子标记

通过对前人公布的糯性水稻蜡质基因全序列测定结果的分析,根据糯性水稻与非糯性水稻蜡质基因序列位点的差异,设计了两个基于PCR扩增反应的可特异识别糯性水稻的显性和共显性STS分子标记;同时在前人公布的非糯性水稻蜡质基因的不同类型(Wxa和Wxb)的全序列比对分析的基础上,选取了两种基因型特定的差异位点,设计了两个基于PCR扩增反应用于特异识别这两种Wx基因的显性STS分子标记,并用新建立的标记对相应的水稻基因型进行了检测.图2表2参10     

胡枝子叶片转录组特征分析及其EST-SSR标记开发（英文）

胡枝子(Lespedeza bicolor)系豆科胡枝子属植物,性耐旱,是防风固沙及水土保持的优良植物.报道胡枝子的转录组序列,对其进行注释,并开发一系列EST-SSR标记用于进化研究.主要研究结果如下:(1)采用二代测序技术对胡枝子叶片进行转录组测序,得到120 913 unigenes,其平均长度为608 bp,N50的长度是978 bp.(2)分别在KEGG和KOG等数据库中对unigenes进行注释,总共注释72 613(60.05%)unigenes的功能.(3)筛选识别13 551个潜在的EST-SSR标记;根据重复单位的核苷酸数目以及重复次数的多少,从中选择173个EST-SSR标记进行引物设计.(4)对胡枝子和其他9种近缘种植物进行PCR扩增实验,最后得到56对引物可全部成功扩增出条带并表现出一定的多态性.本研究获得了较高质量的胡枝子转录组数据库,可进一步用于比较和功能基因组研究以及胡枝子及其近缘类群的基因表达研究;开发的EST-SSR标记将提供一个强大的工具,可用于研究遗传多样性和种群结构、构建DNA指纹图谱数据库、生成遗传图谱、预测分子标记辅助育种和保存遗传信息.     

水葫芦内生细菌脂肪酸生物标记特性研究（Characteristics of PLFA Biomarkers for the Endophyte Bacteria inside Eichhornia crassipe（Mart.）Solms）

应用基于微生物细胞脂肪酸成分鉴定的全自动微生物鉴定系统,鉴定水葫芦内生细菌25株,分属15个属.共检测到27个脂肪酸生物标记(PLFAs),这些生物标记分为4种类型,即(1)高频次分布:在25株细菌中出现13～22次,属于细菌总体类群(general)的生物标记.(2)中频次分布:在25株细菌中出现5～7次,可以用于代表细菌属类群(genus)识别生物标记.(3)低频次分布:在细菌中的分布概率较小,可以用于指示特定细菌种间差异的生物标记.(4)微频次分布:仅在一种细菌种类出现,是细菌种(species)特征生物标记.利用脂肪酸生物标记分析同属细菌不同种的差异,可将微杆菌属分为2类,第1类包括菌株9Microbacterium barkeri和11Microbacterium imperiale,这两个菌株17:0ISO和14:0ISO的脂肪酸生物标记含量均为0.第2类包括10Microbacterium esteraromaticum、12Microbacterium lacticum和13Microbacterium liquefaciens,这3个菌株均含有17:0ISO脂肪酸生物标记而区别于第一类菌株.利用脂肪酸生物标记的差异对25株内生细菌进行聚类分析,可将水葫芦内生细菌类群分为4类.不同植物内生菌群落中存在着特异的生物标记,利用特征脂肪酸生物标记的分析方法,分析水葫芦内生细菌的群体特性,对于植物内生菌微生物群落的研究具有重要意义.     

铜绿假单胞菌增强型绿色荧光蛋白标记的研究

根据增强型绿色荧光蛋白(EGFP)基因设计了上下游引物P1和P2,通过对铜绿假单胞菌菌株X3绿色荧光蛋白标记研究,构建了铜绿假单胞菌标记系统,获得带有EGFP标记的基因工程菌X9,为进行相关的跟踪研究创造了条件.该菌株绿色荧光标记性能稳定.通过转化子基因组DNAPCR和斑点杂交检测,外源EGFP基因存在于转化子染色体上. 图 6参 14     

RAPD markers suggest genotypic effects on forager specialization in a eusocial wasp

Genetic variability within insect societies may provide a mechanism for increasing behavioral diversity among workers, thereby augmenting colony efficiency or flexibility. In order to assess the possibility that division of labor has a genetic component in the eusocial wasp Polybia aequatorialis, I asked whether the genotypes of workers within colonies correlated with behavioral specialization. Workers specialized by foraging for one of the four materials (wood pulp, insect prey, nectar, or water) gathered by their colonies. I collected foragers on 2 days from each of three colonies and identified the material the foragers were carrying when collected. I produced random amplified polymorphic DNA (RAPD) markers from the genomic DNA of these foragers and estimated genotypic similarity of foragers based on sharing of variable RAPD marker bands. Contingency tests on 20 variable loci per colony showed statistically significant (P <0.05) biases in RAPD marker frequencies among forager types in the three colonies. Patterns of association of RAPD marker bands with specializations were constant in two colonies, but changed between collection days in one colony. RAPD marker biases suggest that division of labor among workers includes a genetic component in P. aequatorialis. Colony-level selection on variation in division of labor is a possible factor favoring the evolutionary maintenance of high genotypic variability (low relatedness) in epiponine wasp colonies and in other eusocial insects. Received: 18 July 1995/Accepted after revision: 1 October 1995     

Application of random amplified polymorphic DNA (RAPD) markers to evaluate intraspecific genetic variation in red mullet (Mullus barbatus)

The random amplified polymorphic DNA (RAPD) technique was used to evaluate genetic affinities among eight red mullet (Mullus barbatus L., 1758) samples from the Mediterranean Sea. Twenty-nine random primers were used. Despite the variability which was found within samples, no specific RAPD marker for the discrimination of the populations was detected. The data analysis revealed that the genetic diversity among populations is positively related to their geographic distances. The results of this study indicate that the estimated intraspecific variation may be more␣pronounced with RAPD markers than with allozymes when the two approaches are applied on the same populations. Received: 7 November 1997 / Accepted: 30 May 1998     

Acclimation of the European sea bass to freshwater: monitoring genetic changes by RAPD polymerase chain reaction to detect DNA polymorphisms

We investigated the use of the polymerase chain reaction (PCR) and the associated random amplification of polymorphic DNA (RAPD) technique to study variation in samples of the sea bass, Dicentrarchus labrax, before and after acclimation to freshwater. Acclimation trials were repeated twice, in 1989 and 1990, for two samples originating from the same broodstock (individuals from different localities along the Italian coasts), with overall mortality rates averaging 94 and 75% in the 2 yr, respectively. Analyses are based on 126 polymorphic RAPD markers, scored in at least 39 individuals for each of the starting and acclimated samples in both years. Analysis of RAPD patterns revealed high levels of DNA polymorphism in both 1989 and 1990 starting samples. The acclimated samples maintained similar polymorphism levels. Shifts in marker frequencies between starting and acclimated samples occurred in both years. A correspondence analysis carried out on multimarker individual profiles suggests that the two starting samples resulted from uneven sampling of the same heterogenous broodstock. This analysis clearly separates RAPD phenotypes from starting and acclimated samples in both years and identifies the RAPD markers responsible for such displacement. Patterns of RAPD variation are compared with previous allozymic studies carried out on the same samples. A major difference between the two studies was the number of markers available. Fewer allozyme loci were studied than were RAPD markers. The cause of repeated shifts for allozyme alleles in replicate experiments were almost certainly due to selection, while statistical chance could explain the repeated shift of only one out of more than 100 RAPD markers. We have shown that RAPD analysis, if carried out carefully, is quite reproducible and sensitive enough to reveal high levels of variation among individuals from the same broodstock. A major drawback of this approach is the still unclear inheritance patterns of RAPD polymorphisms. The use of multivariate analyses is suggested as a possible alternative to traditional population genetics techniques to analyze patterns of variation in the absence of a precise genetic interpretation.     

朱■的随机扩增多态DNA分析与种内亲缘关系研究

利用随机扩增多态ＤＮＡ技术对朱（Ｎｉｐｐｏｎｉａｎｉｐｐｏｎ）８个个体进行了随机扩增多态ＤＮＡ分析．用２０个１０ｂｐ的随机引物对每只朱的基因组ＤＮＡ进行扩增,共得到１６８个扩增片段,其中共有片段为１０２个．根据聚类分析所得到的树状图确定了８只朱的亲缘关系,这为进一步构建全部朱个体的谱系关系图打下了基础,有利于制定更有效的朱保护计划     

朱鹮的随机扩增多态DNA分析与种内亲缘关系研究

利用随机扩增多态DNA技术对朱鹮 ( Nipponia nippon ) 8个个体进行了随机扩增多态DNA分析. 用20个10bp的随机引物对每只朱鹮的基因组DNA进行扩增,共得到168个扩增片段,其中共有片段为102个. 根据聚类分析所得到的树状图确定了8只朱鹮的亲缘关系,这为进一步构建全部朱鹮个体的谱系关系图打下了基础,有利于制定更有效的朱鹮保护计划.     

玉米种子的DNA指纹计算机化鉴定

对１２个优良玉米自效系的ＤＮＡ进行了ＲＡＰＤ分析,共筛选了２５０个Ｏｐｅｒｏｎ引物,其中１２个引物共扩增出５９个多态性产物,根据这５９个多态性ＲＡＰＤ产物,对１２个自交系进行了聚类分析,把它们分为３个群,结果与根据系谱法的聚类结果一致。经过优选,用ＯＰＮ－１１扩增出的１１种ＤＮＡ带,建立了这１２个自交系的ＲＡＰＤ－ＤＮＡ指纹图谱。在该图谱呼弱个自交系都具有特异的ＤＮＡ指纹,可以与其它自效纱相区别。     

Population genetic structures of the fissiparous seastar Coscinasterias acutispina in the Sea of Japan

The morphological characteristics and the population genetic structures of the fissiparous seastar Coscinasterias acutispina were investigated for eight sites in the Sea of Japan in order to clarify the presence of sexual and asexual reproduction. Morphological observation based on arm length showed that fission was common at all eight sites examined, indicating the likely production of clonal individuals. A random amplified polymorphic DNA (RAPD) marker was used to detect clones arising by fission and to assess gene flow among sites. A simulation approach using RAPD data revealed the presence of clonal individuals at almost all sites, suggesting the existence of asexual reproduction. The result of phylogenetic analysis according to RAPD genotype showed no relationship between genetic and geographic distances. Considering the limited movement ability of seastar species during the adult phase, these observations suggest the existence of marked gene flow among sites, due to dispersal of planktonic larvae produced by sexual reproduction. These observations suggest that multi-locus genotypic compositions depend on the relative amounts of recruitment from sexual and asexual reproduction in each population.