Analysis of genetic diversity for wild and captive green peafowl populations by random amplified polymorphic DNA technique

The genetic diversity of the populations for 14 wild green peafowls (Pavo muticus) and 18 captive green pea-fowls was investigated by using the technology of random amplified polymorphic DNA (RAPD). Totally 161 and 166 ampli-fied bands were obtained by using 23 arbitrary primers to amplify the genomic DNA of wild and captive green peafowls re-spectively. The results showed that the average relative genetic distance of the wild and captive green peafowls popula-tions was 0.0555 and 0.1355, respectively, and difference of the average relative genetic distances between the two popu-lations was 0.1635. The Shannon diversity index for the wild and captive green peafowl populations was 0.4348 and 1.0163, respectively, which means that there exists significant difference in genetic diversity between the two populations, and the genetic diversity of wild green peafowl was low. The two populations originated from two different families according to analysis by the UPGMA method. This research can provide the theoretical basis for supervising genealogies management of peafowl populations.

Use of random amplified polymorphic DNA （RAPD） analysis to detect jujube witches＇ broom phytoplasmas

Jujube witches＇ broom is a devastating disease of Ziziphusjujube that occurs in various jujube regions of China. Nucleic acid extracted from midribs of samples collected from three jujube varieties （＂Suanzao＂, ＂Lajiaozao＂ and ＂Langzao＂） from symptomatic and asymptomatic shoots were tested by random amplified polymorphic DNA analyses. Using 13 different 10 and 11-bp random primers the amplification of jujube DNA was achieved from all the samples; AMI4 primer provided amplification of specific DNA fragments of about 400 bp, only from samples collected from symptomatic plants. No genetic variations in these varieties were identified using the other 11 arbitrary primers; only with primer AL07 it was possible to differentiate ＂Langzao＂ from the other two varieties tested. All the experiments were repeated 2 times and the results were consistent. Compared with PCR analyses with phytoplasma-specific primers, RAPD techniques resulted to be an alternative rapid and sensitive method for detecting jujube phytoplasmas presence in different jujube varieties.

Authentication and Genetic Origin of Medicinal <i>Angelica acutiloba</i>Using Random Amplified Polymorphic DNA Analysis

Some Angelica species are used for medicinal purposes. In particular, the roots of Angelica acutiloba var. acutiloba and A. acutiloba var. sugiyamae, known as “Toki” and “Hokkai Toki”, respectively, are used as important medicinal materials in traditional Japanese medicine. However, since these varieties have recently outcrossed with each other, it is difficult to determine whether the Japanese Angelica Root material used as a crude drug is the “pure” variety. In this study, we developed an efficient method to authenticate A. acutiloba var. acutiloba and A. acutiloba var. sugiyamae from each other and from other Angelica species/varieties. The random amplified polymorphic DNA (RAPD) method efficiently discriminated each Angelica variety. A. acutiloba var. sugiyamae was identified via a characteristic fragment amplified by the decamer primer OPD-15. This fragment showed polymorphisms among Angelica species/varieties. The unique fragment derived from A. acutiloba var. sugiyamae was also found in one strain of A. acutiloba var. acutiloba, implying that this strain arose from outcrossing between A. acutiloba var. acutiloba and A. acutiloba var. sugiyamae. This RAPD marker technique will be useful for practical and accurate authentication among A. acutiloba var. acutiloba, A. acutiloba var. sugiyamae, and their adulterants.

Genetic Diversity Caused by Environmental Stress in Natural Populations of Niupidujuan as Revealed by RAPD Technique

Multiplex environmental factors are generally expected to have significant effects on genetic diversity of plant populations.In this study,randomly amplified polymorphic DNA（RAPD） technique was used to reveal the genetic diversity in the same species of four populations collected from Niupidujuan（Rhododendron chrysanthum） at different altitudes,an endangered species,endemic to Northeast China.Initially,twenty informative and reproducible primers were chosen for final RAPD analysis.A total of 152 clear bands were obtained,including 143 polymorphic ones.With the help of POPGENE software,the poly rate was calculated to be 94.07% and the evenness of amplified bands for every primer was 6.8.Additionally,the mean observed number of alleles was 1.7265 with an effective number of 1.3608.An examination of the gene indicated a diversity of 0.2162 with an information diversity index of 0.3313.For these data,the clustering blurred analysis was performed with the aid of NTSYS-pc software to define the Nei＇s gene diversity and the Shannon information diversity index of the four plant populations.The relationships between the genetic diversity indexes on the one hand and the geographic and climatic factors on the other hand were estimated by the Pearson correlation with SPSS 11.0 software.The results of the correlation analysis show that there were significant（P〈0.05） or highly significant（P〈0.01） correlations between each of the genetic diversity indexes and the different temperature which were mainly caused by the altitude different populations located.These data highlight the importance of native populations in shaping the spatial genetic structure in Niupidujuan.

RAPD Technique Used to Determine the Purity of Hybrid Hot Pepper Seeds

Two hybrid hot pepper varieties Xiangyan 5 and Xiangyan 10, and their parents were analyzed the polymerase chain reaction with MJ /PT 200 Peltrier Themal Cycler and DS 800 White-ultravilot Transilluminator to set up a RAPD system adaptable to the purity determination of the hy-brid seeds. Among the 39 random primers, 2 and 4 primers were found to be used effectively in Xiangyan 5 and Xiangyan 10 respectively.

三系杂交稻亲本随机扩增多态性DNA(RAPD)分析

选用９个随机引物对３１份杂交水稻亲本材料进行了ＲＡＰＤ分析，共检测到６０条多态性带。聚类分析结果表明，所有供试材料可以被明确地区分。在９个随机引物中，有８个具有较高的多态性检测能力。以这８个引物为基础，选用任两个引物即可在任一对材料中检测出多态性的频率在９６１３％以上，而选用任３个引物则该频率在９９２１％以上。这显示了运用ＲＡＰＤ鉴定稻种具有简便、灵敏、高效的优点，在鉴定杂交稻种的实践中有着良好的应用前景。

栽培稻和普通野生稻基因组的随机扩增多态性DNA（RAPD）初步分析

用３５个随机引物对５个籼稻品种、５个粳稻品种和２７份中国普通野生稻进行了ＲＡＰＤ分析。６０％以上的引物能在籼稻和粳稻基因组间显示差异；中国普通野生稻与栽培稻的差异主要表现在与籼稻的不同，绝大部分供试野生稻的ＲＡＰＤ带型与粳稻的相同。这说明多数中国普通野生稻偏粳，但也存在偏籼的普通野生稻。

RAPD分析氮离子注入甜菊种子后的幼苗基因组DNA变异

应用ＲＡＰＤ 技术检测经低能氮离子注入甜菊纯系种子引起的幼苗基因组ＤＮＡ 变异。筛选出ＯＰＪ系列中的１５ 种引物对实验及对照基因组ＤＮＡ 进行了ＰＣＲ 扩增，共获扩增片段１０３ 条，分子量在０．３ － ３ｋｂ 之间，其中５ 种引物ＯＰＪ－ １ ，７ ，９，１１ ，１２ 扩增出差异片段１２ 条。结果表明，低能氮离子注入甜菊种子可引起体内基因组ＤＮＡ 发生突变；ＲＡＰＤ 技术是检测基因组ＤＮＡ 发生诱变的一种简便、有效方法。本文同时探讨了离子强度和Ｔａｇ ＤＮＡ 聚合酶用量对甜菊ＲＡＰＤ 分析结果的影响，以及氮离子注入诱变效应的可能机制。

广东野百合DNA提取和RAPD条件的优化

以野百合(Lilium brownii)新鲜叶片、硅胶干燥叶片及鳞片为材料,研究了DNA的提取方法,并对影响随机扩增多态DNA(RAPD)反应的各因素进行了优化。建立了野百合RAPD的优化反应体系及程序,即在20μl反应体系中,含20 ng模板DNA,2.0 mmol/L Mg2+、0.2 mmol/L dNTPs、1.5 U Taq DNA聚合酶、0.3μmol/L随机引物S1519;扩增程序为:94℃预变性5 min,然后94℃30 s,38℃50 s,72℃1 min,35个循环,最后72℃延伸10 min,4℃保存。

西伯利亚蝗基因组DNA提取及RAPD分析条件的优化

以西伯利亚蝗Gomphocerus sibiricus(L.)为研究材料,利用改良的SDS法提取高质量的DNA,分别测试了dNTP浓度、镁离子浓度、TaqDNA聚合酶用量、模板DNA的量等因素对反应结果的影响。通过各因子的组合比较,建立了西伯利亚蝗RAPD优化体系:25μLPCR反应体系,10×buffer2·5μL;dNTP0·24mmol/L;MgCl22·0mmol/L;Taq DNA聚合酶1U;DNA模板45ng;引物30ng。扩增程序为:94℃预变性1min45s、94℃变性30s、35℃退火1min30s、72℃延伸2min,45个循环、72℃延伸10min。结果表明,利用优化的反应条件进行西伯利亚蝗基因组DNA分析,实验有着良好的重复性和稳定性。

辣椒细胞质雄性不育系与其保持系线粒体DNA的RAPD分析

以辣椒细胞质雄性不育系21A及其相应的保持系21B为试材，从苗龄10～15d的黄化苗中分离纯化线粒体并提取线粒体DNA。线粒体负染后电镜观察发现，分离纯化的线粒体形态完整，呈椭球形，杂质较少。电泳检测提取的线粒体DNA的琼脂糖，结果表明，线粒体DNA纯度较高，无降解现象，浓度为20～50ng／山，可用作RAPD分析。经初步筛选，100个RAPD随机引物中有8个引物表现出多态性，经再次重复筛选，获得了辣椒细胞质雄性不育系21A线粒体DNA与雄性不育性相关的RAPD标记CMSAG3430。

芸薹类蔬菜基因组DNA遗传多样性的RAPD分析

采用随机引物扩增多态性 DNA( RAPD)技术 ,对芸薹类 ( 2 n=2 0 )蔬菜作物的 3 3个品种进行了遗传多样性检测 .从 70个引物中筛选出 3 7个引物 ,共扩增出 3 53带 .其中 ,多态带有 2 60条 ,扩增片段长度大多数集中在 0 .9～ 1 .6kb之间 .不同引物的检测效率相差很大 .运用 5个引物扩增的 1 0条RAPD特征带 ,可以作为一组 DNA指纹 ,区分所有供试

丁不同群体间形态学差异与随机扩增多态DNA(RAPD)分析

采用聚类分析、主成分分析和判别分析3种数理统计方法,对我国新疆朱家湖丁群体7、3水库丁群体和从捷克引进我国的丁群体的可量性状和框架参数进行分析。聚类分析和主成分分析显示,朱家湖群体与73水库群体较为接近,而捷克群体与前两群体相距较远。判别分析亦可将捷克群体与前两群体分开,准确率达100%。从60个随机引物中筛选出的20个引物对丁朱家湖群体、73水库群体和捷克群体进行了随机扩增多态DNA(RAPD)分析。引物S440在捷克群体扩增出的450bp片段为该群体特有的标记片段。朱家湖群体、73水库群体、捷克群体多态位点比例分别为24%、22.67%和18.42%;群体内遗传相似度分别为0.8967、0.9035和0.9309;遗传多态度(π)分别为0.1539、0.1489和0.1142。表明朱家湖群体保持着较高的遗传变异。三群体间的遗传相似度为0.6868—0.9496,群体遗传分化系数(Fst)为0.048—0.238,分子方差分析发现群体内方差占总方差的83.96%,群体间的方差只占16.04%,由此推断三群体间遗传分化并不大。

克罗诺杆菌的随机多态性扩增DNA(RAPD)基因分型研究

研究了克罗诺杆菌标准菌株和样品中分离菌株的分子特征,为克罗诺杆菌的种间鉴别和分子溯源提供一种有效手段。采用随机引物扩增多态性DNA分析对38株菌株进行基因分型。筛选出1条随机引物,每一菌株可扩增出1～7条DNA片段,片段在400-3500bp,可将8株标准菌株的种间区分开来。以相似性50%为标准,38株克罗诺杆菌按照其遗传差异可以分为8个基因型类群。所建立的RAPD技术可用于克罗诺杆菌的种间鉴别和分子溯源研究。

用随机扩增多态DNA(RAPD)技术鉴别中药材天花粉及其类似品

目的：对来源于１３个种３个变种的天花粉及其类似品进行鉴别研究，并进行方法学的探讨。方法：应用随机扩增多态ＤＮＡ（ＲＡＰＤ）技术进行鉴别，采用聚类分析方法分析结果，并对药材贮存时间及产地对实验结果的影响进行探讨。结果：把天花粉正品与类似品有效地分成三大类。结论：认为在实验时采取设对照组，对结果采用聚类分析等方法，ＲＡＰＤ技术鉴别药材具有一定的可靠性和实用价值，这为解决粉末及破碎药材的鉴别提供了新的方法。

药用百合DNA提取和RAPD条件的优化

目的确定药用百合最佳随机扩增多态DNA(RAPD)分析方法。方法以药用百合新鲜鳞叶为材料,研究DNA的提取方法,并对影响RAPD反应的各因素进行优化。结果药用百合RAPD反应体系及程序为:在40μL反应体系中,含40 ng模板DNA,0.3μmol/L随机引物,0.2 mmol/L dNTPs,1.7 mmol/L Mg2+,3μL 10×PCRBuffer,1.5 U Taq酶;扩增程序为:第一步94℃预变性2 min,第二步94℃变性1 min,第三步36℃退火45 s,第四步72℃延伸90 s,返回至第二步重复40个循环,第六步72℃延伸10 min,最后4℃保存以备电泳。结论建立了药用百合RAPD的优化反应体系及程序。

用随机扩增多态性DNA(RAPD)技术区分不同地理株白纹伊蚊

目的 对来自北京、广州、成都、上海和日本的冲绳及长乐寺 6个不同地理株的 12只雄蚊进行随机扩增多态性DNA(RAPD)分析。方法 RAPD -PCR。结果 UPGMA法构建的系统聚类图表明 ,12只白纹伊蚊可归为明显不同的两个组 ,不同地理株之间存在着显著的遗传分化。结论 用RAPD法可以区分不同地理株的白纹伊蚊。

高粱DNA导入水稻的RAPD分子验证

对以密穗型高粱ＤＮＡ为供体、通过花粉管通道技术导入水稻所获得的水稻后代进行了ＲＡＰＤ分子验证．结果表明：从１２２个引物中找到９个引物检测出ＤＮＡ的多态性。

随机扩增多态性DNA(RAPD)技术用于青海田鼠鼠疫菌株亲缘性的研究

目的 对青海省称多县境内和四川省石渠县境内青海田鼠体内分离的 32株鼠疫耶尔森氏菌的基因进行分析 ,明确两地鼠疫耶尔森氏菌间的亲缘关系。方法 用随机扩增多态性DNA(RAPD)技术。结果 扩增产物在凝胶电泳上显示的条带 ,除 7株菌略有不同外 ,其余 2 5株均相同。TreeView[Win32 ]统计软件分析的结果也显示两地鼠疫耶尔森氏菌之间具有很近的亲缘关系。结论 青海省称多县和四川省石渠县青海田鼠体内分离到的鼠疫耶尔森氏菌在遗传学上属同一来源。

杂交粳稻亲本随机扩增多态性DNA(RAPD)分析(研究简报)

选用 6 0个随机引物对 15份杂交粳稻亲本材料进行了RAPD分析 ,其中 10种引物的扩增结果具有明显的多态性。在这 15份杂交粳稻亲本材料中共扩增得到 153条条带 ,其中的86条为多态性条带。聚类分析结果表明 ,所有供试材料可以被明确区分。在此基础上分析了这15个水稻品种的亲缘关系 ,对田间育种有一定的参考价值。