徐州、平山两地中华稻蝗和日本稻蝗的群体遗传关系分析

 应用RAPD技术分析了江苏徐州和河北平山两地混生的中华稻蝗（Oxya chinensis）和日本稻蝗(O.japonica) 的群体遗传关系。用10条随机引物从43头个体中产生125条清晰、稳定的谱带， 其多态性条带占99%。 Shannon信息指数表明，中华稻蝗遗传多样性高于日本稻蝗，分别为0.3432和0.2781。Nei's遗传距离显示：种群间遗传距离明显小于种间遗传距离；同一采集地点混居的中华稻蝗与日本稻蝗之间的遗传距离小于地理上存在较大跨度的两物种间的遗传距离，如：江苏徐州中华稻蝗和日本稻蝗之间的遗传距离为0.2411，河北平山中华稻蝗和日本稻蝗的遗传距离为0.2578，而江苏徐州中华稻蝗和河北平山日本稻蝗之间的遗传距离为0.2749，江苏徐州日本稻蝗和河北平山中华稻蝗之间的遗传距离为0.2982。基于Nei's遗传距离，分别用UPGMA和NJ法构建分子系统树，结果供试的43个个体分为两大聚类簇，江苏徐州和河北平山两地的中华稻蝗聚为一支，日本稻蝗聚为另一支。上述结果表明，RAPD分析方法可显示个体间的多态性，反映个体或物种间的细微差异，是区分近缘物种的有效分子标记。     

镉胁迫对青菜幼苗某些生理特性以及基因组多态性的影响

以青菜品种"苏州青"为试材,采用生理生化方法和随机扩增多态性DNA(RAPD)技术,研究了镉(Cd)胁迫对青菜幼苗相关生理特性以及基因组多态性的影响。结果表明:不同浓度(1、5、10、20、40 mg·L-1)的Cd处理20 d后,青菜幼苗的鲜重、根伸长受到显著抑制,叶片的叶绿素以及蛋白质含量下降,丙二醛含量和细胞膜透性增加。选用8条寡核苷酸引物(10 bp)对青菜幼苗基因组DNA进行RAPD扩增,对照组的RAPD图谱中可分辨出55条谱带,Cd胁迫后的RAPD图谱发生改变,呈现谱带的增加、缺失和荧光强度的改变,并且与Cd浓度剂量-效应相关。随着Cd浓度的增加,幼苗叶片基因组的模板稳定性逐步下降,并且RAPD多态性与青菜幼苗的生理指标具有显著的相关性。以上结果表明,利用RAPD技术获得的DNA多态性变化谱带可作为检测Cd对青菜毒性效应的生物标记物。     

中国西北内陆干旱区大叶白麻野生居群遗传多样性研究(英文)

[Objective] Study on the genetic diversity in wild populations of Poacynum hendersonii.[Method] Random amplified polymorphic DNA(RAPD)technique was employed to analyze the genetic diversity in five wild populations of P.hendersonii sampled from Xinjiang,Gansu and Qinghai provinces.[Result] Totally 165 clear and repeatable bands were generated in RAPD reaction by using 20 primers screened from 80 primers,of which 110 were polymorphic,accounting for 66.67%.At species level,Nei’s gene diversity index(H),Shannon’s information index(I)and genetic differentiation coefficient(Gst)were 0.220 5,0.304 7 and 0.908 2,respectively.P.hendersonii germplasm resources share a high level of genetic diversity,and genetic differentiation mainly exists among the populations.Results from genetic distances and cluster analysis showed that relationships among P.hendersonii populations were to some extent related with their geographical and climatic characters.[Conclusion] This study suggests that the conservation of P.hendersonii should focus on the protection of many populations,particularly the Qinghai population.     

中国西北内陆干旱区大叶白麻野生居群遗传多样性研究(英文)

[Objective] Study on the genetic diversity in wild populations of Poacynum hendersonii.[Method] Random amplified polymorphic DNA(RAPD)technique was employed to analyze the genetic diversity in five wild populations of P.hendersonii sampled from Xinjiang,Gansu and Qinghai provinces.[Result] Totally 165 clear and repeatable bands were generated in RAPD reaction by using 20 primers screened from 80 primers,of which 110 were polymorphic,accounting for 66.67%.At species level,Nei's gene diversity index(H),Shannon's information index(I)and genetic differentiation coefficient(Gst)were 0.220 5,0.304 7 and 0.908 2,respectively.P.hendersonii germplasm resources share a high level of genetic diversity,and genetic differentiation mainly exists among the populations.Results from genetic distances and cluster analysis showed that relationships among P.hendersonii populations were to some extent related with their geographical and climatic characters.[Conclusion] This study suggests that the conservation of P.hendersonii should focus on the protection of many populations,particularly the Qinghai population.     

Genetic Diversity in Wild Populations of Poacynum hendersonii from Inland Arid Regions of Northwest China

[Objective] Study on the genetic diversity in wild populations of Poacynum hendersonii.[Method] Random amplified polymorphic DNA(RAPD)technique was employed to analyze the genetic diversity in five wild populations of P.hendersonii sampled from Xinjiang,Gansu and Qinghai provinces.[Result] Totally 165 clear and repeatable bands were generated in RAPD reaction by using 20 primers screened from 80 primers,of which 110 were polymorphic,accounting for 66.67%.At species level,Nei's gene diversity index(H),Shannon's information index(I)and genetic differentiation coefficient(Gst)were 0.220 5,0.304 7 and 0.908 2,respectively.P.hendersonii germplasm resources share a high level of genetic diversity,and genetic differentiation mainly exists among the populations.Results from genetic distances and cluster analysis showed that relationships among P.hendersonii populations were to some extent related with their geographical and climatic characters.[Conclusion] This study suggests that the conservation of P.hendersonii should focus on the protection of many populations,particularly the Qinghai population.     

不同地理种源雷公藤的RAPD分析

应用RAPD技术,对25个不同地理种源的雷公藤进行了多态性分析。采用POP-GENE32软件对25个种源进行聚类分析,DPS(9.5)软件对所有个体进行聚类分析,并用UPGMA法对扩增DNA片段数据构建系统发育树。结果表明:多态性条带百分率为91.14%,Shannon's信息指数I=0.4472,Nei's基因多样性指数H=0.2973,观测等位基因数Na=1.911 4,有效等位基因数Ne=1.5120。这表明25个种源具有较高的遗传多样性水平,因此收集不同种源以保存尽可能多的遗传多样对于雷公藤的良种选育是非常有必要的。     

Study on Insecticidal Activities and Effect on Three Kinds of Enzymes by 5-Aminolevulinic Acid on Oxya chinensis

Insecticidal activities and effects on three enzymic activities caused by 5-aminolevulinic acid （ALA） on Oxya chinensis were studied. Fourth-instar nymphs of O. chinensis were treated with different doses ofALA （A1,250 mM; A2, 450 mM; A3,750 mM; A4, 1 000 mM）. Mortality and the activities of acetylcholinesterase （ACHE）, glutathione S-transferase （GSTs）, and glutathione peroxidase （GPx） were determinated. The mortality of O. chinensis rose with an increasing dose of ALA. The mortality of high-dose treatments A3 and A4 reached 66.19 and 80.21%, respectively. The value of LD50 was 3.61 （3.29-3.93） mg·g^-1 body weight （95% confidence interval）. Biochemical studies showed that the activities of AChE and GPx in the A4 treatment declined by 51.53 and 42.82% in the female, and 42.65 and 43.85% in the male compared to the control, respectively, and the degree of decline reached a significant level at P 〈 0.05. Meanwhile, the GSTs activities of O. chinensis enhanced with increasing dose of ALA. The GSTs activities of female and male O. chinensis in the A4 treatment remarkably increased by 171.05 and 97.42% compared to the control （P〈 0.05）. ALA had an obviously toxic effect on O. chinensis. Moreover, ALA caused the photoinactivation of AChE and GPx, which induced nerve transmission blocking and the capability to defend oxidation damage declining. Meanwhile, a high dose of ALA could activate GSTs, which caused a feedback inhibition of the insect to the phototoxic substance.     

Analysis of the Germplasm Resources and Genetic Relationships Among Hybrid Cymbidium Cultivars and Native Species with RAPD Markers

The random amplified polymorphic DNA （RAPD） marker was assessed to detect the genetic relationships among 48 hybrid Cymbidium cultivars from Japan, Korea, China, and USA, and 2 species of native Cymbidium. Twenty primers were screened from 100 random decamer primers, and a total of 258 DNA bands were amplified, 253 of which （98.1%） were polymorphic. The average number of polymorphic DNA bands amplified by each primer was 12.6. All cultivars were distinguishable when a number of primers were considered. Genetic similarities among the cultivars and species were estimated based on the amount of band sharing ranging from 0.364-0.817 with an average of 0.581. According to the data, a dendrogram of genetic relationship, which was constructed using the UPGMA method, showed that all the tested cultivars and native species were classified into five cluster groups with the similarity coefficient of 0.592. It revealed that the genetic relationships among tested accessions were to some extent related with their origin, flower colour, branch type, and genealogy. It further indicated that the RAPD technique is a useful tool for studying the genetic relationships among hybrid Cymbidium cultivars.     

三种重金属对中华稻蝗金属硫蛋白基因表达的影响

以栖息于稻田且主要取食水稻茎叶的中华稻蝗(Oxya chinensis)成虫为供试材料,经氯化镉、氯化铜和硫酸锌(Cd Cl2、Cu Cl2和Zn SO4)3种重金属急性染毒,采用RT-q PCR技术检测中华稻蝗2个金属硫蛋白基因(Oxya chinensis metallothionein,Oc MT1,Oc MT2)在精巢、卵巢和肌肉中的表达变化。结果表明,3种重金属均可诱导中华稻蝗MT基因不同程度的上调表达。经Cd急性处理后,3个器官组织中Oc MT1和Oc MT2表达水平在一定浓度范围内随着Cd浓度的增高而诱导表达上调;经Cu急性处理后,精巢和卵巢中Oc MT1在最低浓度时的诱导表达量最高,Oc MT2表达水平随着Cu浓度的增大而升高,肌肉中2个MT基因随着Cu浓度的增大而升高;经Zn处理后,Oc MT1和Oc MT2表达水平随着Zn浓度的升高而升高。由实验结果可以看出,3种重金属对中华稻蝗MT在不同器官组织均有诱导表达作用,但各浓度处理后其诱导表达水平不同,Oc MT对不同重金属的敏感性具有显著差异。     

五龙鹅豁眼性状遗传变异的RAPD分析

利用RAPD分子标记技术对五龙鹅的豁眼组和无豁眼组群体进行了种质鉴定和遗传分析,从130个10碱基随机引物中筛选出19个多态性高的引物,分别扩增出192条和168条DNA带,多态位点比率达84.90%和67.26%,平均每个引物扩增多态性带10.1条和8.8条,Shannon多样性指数为0.4610和0.3641,Nei's指数为0.3103和0.2462,豁眼组和无豁眼组群体间的平均遗传距离为0.1979和0.1603,群体内个体间的平均遗传相似度分别为0.8021和0.8397,选择引物扩增的特异性标记,可用来区分五龙鹅的豁眼组和无豁眼组群体.用SPSS软件对群体内个体间遗传相似度的方差分析结果表明,在无豁眼组群体内存在显著差异(P<0.05),而在豁眼组群体内则差异不显著,表明豁眼组的种质较纯.两组五龙鹅之间相似度为0.7901,遗传距离为0.2099,这与形态学分析的结果相一致.     

运用TRAP标记分析5个鲤群体的遗传多样性

运用TRAP标记技术,选取10个多态性较好的引物组合对荷包红鲤、黄河鲤、建鲤、兴国红鲤和黑龙江野鲤等5个鲤群体进行遗传多样性分析,其中固定引物是根据目标候选基因-GHR基因的序列设计。结果表明,共扩增出168个位点,其中多态性位点134个,平均多态位点比例为80.41%,平均多态性信息含量为0.29。分子变异方差分析(AMOVA)结果显示群体内的方差贡献率达96.97%,表明各个鲤群体内存在较大的遗传变异。种群再分效应固定指数(FST=0.030 26,P＜0.05)表明不同鲤群体间有显著的遗传分化。基于目标候选基因的TRAP标记对5个鲤群体进行聚类分析,结果表明建鲤和兴国红鲤首先聚成一类,然后黄河鲤和黑龙江野鲤聚为一类,再与荷包红鲤聚为一类。     

非洲桃花心木不同地理群体遗传多样性分析

应用SRAP 分子标记技术,对来自非洲的5 个不同地理群体非洲桃花心木的遗传多样性和遗传分化进行了分 析。结果表明:筛选出28 对SRAP 引物组合用于群体遗传分析,共扩增出条带77 条,其中多态性条带59 条,多态 性水平为76.62%;5 个群体遗传距离在0.036 1 ~ 0.131 7 之间,总的Nei爷s 基因多样性指数为0.209 3,总的 Shannon爷s 信息指数分别为0.330 0,遗传变异主要来自群体内个体间(70.04%),群体基因流(Nm)为1.185 8。      

安徽省辣椒疫霉菌株遗传多样性的RAPD分析

通过利用RAPD(Random Amplified Polymorphic DNAs)随机引物中筛选到的可扩增出清晰条带、主带明显、稳定的10条引物,对来自安徽合肥、淮南、和县、潜山、岳西、江苏南京和四川邛崃等县市的发病辣椒上分离鉴定获得23个辣椒疫霉菌(Phytophthora capsici)进行全基因组DNA RAPD标记遗传多样性分析。选用引物共标出DNA指纹图带113条,其中多态性条带96条,多态性检测率为84.96%,表明辣椒疫霉菌具有较丰富的遗传多样性。根据引物扩增的DNA指纹图谱,运用UPGMA法分析,以遗传相似系数0.7为阈值,供试菌株被划分为3个遗传聚类组,除部分相同地理来源的菌株被划分为同一组外,其他不同菌株间的遗传相似性与地理来源无直接相关性。     

新疆梨种质资源亲缘关系的ISSR和RAPD分析

[目的]探讨新疆梨种质资源亲缘关系以及遗传多样性,旨在为供试梨资源的分类提供科学依据.[方法]采用ISSR和RAPD分子标记对48份梨种质资源进行聚类分析和遗传关系研究.[结果]14条ISSR引物共扩增出113条清晰的谱带,其中101条显示多态性,平均每个引物扩增出7.2条多态性谱带.19条RAPD引物共扩增出163条清晰的谱带,其中多态性谱带138条,平均每个引物扩增出7.2条多态性谱带.基于ISSR和RAPD两种标记,利用UPGMA分别构建了48份梨资源的聚类树状图.ISSR和RAPD分别聚类以及两种标记混合聚类均将48份梨种质分为3类:第Ⅰ类群中包括1份种质;第Ⅱ类群中包括23份种质;第Ⅲ类群中包括24份种质.[结论]两种标记适合于梨种质资源亲缘关系和遗传多样性分析,可为梨资源的开发利用及新品种的选育提供科学依据.     

盾叶薯蓣居群亲缘关系的RAPD分析

用随机扩增多态DNA标记(RAPD)技术对华中农业大学薯蓣资源圃中收集的11个盾叶薯蓣居群及4个薯蓣属植物居群的混合样品进行了遗传多样性的检测。对于11个盾叶薯蓣居群,19个RAPD引物扩增出153条带,其中113条为多态带,占73.86%。UPGMA聚类分析结果揭示了各居群间的亲缘关系。     

神农架4个珙桐居群遗传多样性的RAPD分析

用随机扩增多态DNA（RAPD）技术对神农架地区4个居群28个珙桐（Davidia involucrate Baill.）个体进行了遗传多样性分析.16个10 bp寡核苷酸引物共检测到87个位点,多态位点49个,多态百分率56.3％,而居群平均多态位点百分率为30.2％.用POPGENE对数据进行了分析,结果显示:居群的平均Nei’s基因多样性为0.111 8,Shannon’s遗传多样性信息指数为0.165 1,总的居群基因多样性为0.169 2,基因分化系数为0.339 2.这表明神农架地区珙桐的遗传多样性较低,其遗传变异以居群内遗传变异为主.      

Research of Genetic Diversity in Seven Kobresia by AFLP in Tibetan Plateau

This work analyzed the genetic diversity of Kobresia accessions at the molecular level, and further obtained the necessary information for breeding and germplasm evaluation. Genomic DNA of Kobresia was amplified with four E＋3 and M＋3 primer combinations with AFLP （amplified fragment length polymorphism）. AFLP analysis produced 164 scorable bands, of which 154 （93.96%） were polymorphic. The mean Nei＇s gene diversity index （H） was 0.2430, and the Shannon＇s information index （I） was 0.4012, indicating the abundant genetic diversity of Kobresia. The 11 Kobresia accessions from Tibetan Plateau, China, can be classified into five groups after cluster analysis based on the UPGMA （unweigbted pair group method arithmetic average） method. In general, there was abundant genetic diversity among Kobresia accessions resources, and the genetic coefficient was unrelated to their geographic latitude. Natural habitats influenced genetic differentiation of Kobresia.     

我国近海中国对虾种群遗传差异的RAPD分析

应用随机扩增多态DNA（RAPD）技术对取自我国近海烟台、长岛、青岛、宁波等4个地方的20只中国对虾进行了种群内和种群间遗传差异分析。在使用的50个随机引物中，有9引物扩增出条带清晰的多态性片段。烟台、长岛、青岛、宁波等4个地理种群内的遗传距离分别为0．0154、0．0370、0．0462、0．0550，种群之间的遗传距离分别为0．0793、0．1139、0．0988、0．1485、0．0880、0．1260。UPGMA和NJ法构建的种群间分子系统树基本一致，结果表明不同地理种群这间存在一定程度的遗传差异。     

花椒种质资源的RAPD分析

利用30条RAPD引物对21份花椒种质资源的DNA进行了扩增,计算了样品间的遗传距离并进行了聚类分析。共扩增出257条谱带,其中多态性谱带186条(占72.37%),平均每条引物扩增多态位点6.2个。21份种质材料间均存在遗传差异,其中韩城红葡萄椒和韩城大红袍遗传距离最近,平均遗传距离为0.290 4;凤县野生构椒和韩城白葡萄椒的遗传距离最远,平均遗传距离为0.833 6;同一种质资源内个体间存在不同程度的差异,其中山东大红袍个体间差异最大,个体间平均遗传距离为0.087 2,但临夏一号和凤县大红袍2份种质内个体间均未检测到差异。当遗传距离取0.529 7~0.576 3时,21份花椒种质资源可以聚类为3组,当遗传距离取0.496 3~0.503 7时,可聚类为7组。RAPD多态性分析从分子水平上反映出了花椒种质资源复杂的遗传背景;为花椒品种的鉴定与分类研究提供了依据。     

应用RAPD标记技术研究大花蕙兰种质资源亲缘关系

利用RAPD标记技术对来自2个企业的18个大花蕙兰品种进行了检测。结果表明：从50个10碱基随机引物中筛选出15个多态性高、重复性好的引物,共扩增出92条DNA条带,其中90条为多态性带,占97.8%,平均每个引物扩增多态性带6条。18个种质之间的相似系数变化范围为0.1975～0.6704,平均相似系数为0.4761;遗传距离变化范围为0.3296～0.8025,平均遗传距离0.5239。基于RAPD的扩增结果建立了UPGMA亲缘关系聚类图,18份种质在遗传距离0.5239处被划分为4个类群。供试的18份种质间的遗传亲缘关系与其花色和花枝类型存在一定的相关性。