SRAP标记在植物遗传多样性中的应用进展

该文综述了SRAP标记在植物种类遗传多样性国内近几年的应用进展,以便为今后的研究提供相应的理论依据.同时,并讨论了一些SRAP标记在植物种类遗传多样性应用中尚存在的问题.     

SRAP分子标记分析西瓜遗传多态性

目的:探讨西瓜遗传多样性和遗传基础。方法:采用SRAP分子标记对西瓜品种D1、D2、D3、H1、H2、H3、M1、M2、M3、m1、m2、m3的多态性进行了分析。结果:每对引物组合产生13~25对比较清晰的扩增带.8对引物组合共产生131条扩增带。平均每对引物组合产生16.375条。8对引物组合共产生多态性带37条,每对引物组合产生3~7条,平均4.625条。每对引物组合产生的多态性带的比例为16.666%~38.464%,平均为28.675%。另外,对银染过程进行了优化。结论:SRAP标记多态性还是较高的,可以适于分析西瓜等遗传差异小的作物。     

棉花遗传多样性SCoT和SRAP标记的研究及比较分析

利用SCoT和SRAP两种分子标记技术对30份彩色棉与白色棉种质资源,进行遗传多样性研究。用29对SRAP引物组合和26个SCoT引物分别对供试棉花的基因组DNA进行扩增。SCoT引物共扩增出163条带,多态性比率为61.96%,遗传相似系数GS值变化范围为0.5405～0.9972。SRAP引物组合共扩增条带1067条,多态性比率仅为14.1%,遗传相似系数GS值变化范围为0.5415～0.9109。两种标记系统得到了相似但并不完全相同的聚类图,2种标记方法间存在显著相关性(r=0.5518,P<0.05)。结果表明,SRAP与SCoT标记均适用于棉花种质的遗传多样性分析,且SCoT的标记指数MI高于SRAP标记,为SCoT这种新兴的标记技术在棉花育种中的应用提供了重要的依据。     

SSR标记在辣椒DUS测试中的应用研究北大核心CSCD

为了评价SSR分子标记在辣椒(Capsicum annuum L.)品种特异性、一致性、稳定性(DUS)测试中的应用潜力,采用聚丙烯酰胺凝胶电泳技术和毛细管电泳技术,对辣椒SSR引物进行筛选,采用入选的30对核心引物,构建130份不同来源品种的指纹图谱。30对引物共检测到等位基因163个,每对引物等位基因变幅为2～15,平均等位基因数为5.43,多态信息量(PIC)的变异范围为0.17～0.76,平均为0.43。聚类分析表明,该套引物可以将所有参试品种区分开来,遗传基础相近的品种优先聚在一起。130个品种的遗传距离为0.01～0.91。有20对品种遗传距离小于0.10,其中3对品种有1个位点的差异,采用差异位点数判断品种特异性更适合目前辣椒品种现状。采用10对引物对品种0313963-4进行了一致性检测,综合一致性比率R值为96.5%,低于DUS测试中对表型性状一致性的要求。因此SSR标记直接用于辣椒DUS判定还需进一步研究,但是可用于近似品种筛选。     

利用SRAP标记分析中国野生石蒜的遗传多样性

采用SRAP（Sequence-related amplified polymorphism,序列相关扩增多态性）标记对中国13个省24份野生石蒜（Lycoris radiata）资源94个样品进行了检测。10个引物组合共扩增出218条带,其中173条为多态性条带,多态性百分比达79.36%。石蒜的观测等位基因数（na）、有效等位基因数（ne）、基因多样性指数（h）、Shannon信息指数（I）分别为1.7936、1.4131、0.2415和0.3664。石蒜不同种源间的遗传分化系数（Gst）达0.9547、基因流（Nm）仅0.0136,表明种源间遗传分化显著,遗传变异主要存在于种源间。根据Nei′s遗传距离对24份种源进行UPGMA聚类,所有石蒜种源聚成两大类,第I大类由7份种源组成,除江苏连云港的石蒜（JS3）外,均来自我国西南或西北地区;其余的17份种源构成第II大类,它们遍及华南、华中和华东地区;各大类中的分支结果与野生石蒜外部形态及生长发育习性有一定联系。将石蒜种源的遗传多样性与其所处的经度、纬度、海拔、年均降雨量、年均温等进行相关性分析,结果显示它们之间的相关性均不显著,即石蒜对环境依赖性小,能分布在各种生境中。根据以上结果,我们认为野生石蒜具有较丰富的遗传多样性,而种源间遗传分化显著的原因主要是基因流的隔离。研究结果对我国的野生石蒜资源的开发利用与保护有重要意义。     

SRAP标记及其在植物遗传育种中的应用

SRAP标记是基于选择性扩增开放性阅读框的新型分子标记,具有简便、高效、重复性好、高共显性等优点,在植物育种中已经得到广泛应用.本文介绍了SRAP标记基本原理和特点,对SRAP标记在遗传多样性研究、遗传连锁图谱构建、比较基因组以及分子标记辅助选择育种等方面的应用进行了综述.     

利用SRAP标记分析水稻新品系遗传多样性

为了更加准确地了解水稻新品系的亲缘关系,本研究选用分辨力强、多态性较高的20对SRAP引物标记对42个水稻新品系进行了遗传多样性分析。结果表明,20对引物扩增出204个等位点,其中,有135个多态等位点,多态百分率为66.2%;每对引物分别检测出等位点变异2～9个,平均值为6.75。供试水稻新品系间的遗传差异系数为0.02～0.29。用欧式平方距离对42个水稻新品系的聚类分析表明,当遗传相似性为0.11时可将42个水稻新品系可聚为7大类群。研究结果在分子水平上进一步了解水稻新品系的遗传差异,为水稻育种工作中新品系的淘汰与推广提供了有效的补充依据。     

基于SRAP标记的山药种质资源遗传多样性分析

目的:研究我国山药种质资源遗传多样性,为合理利用资源和开展选育种工作提供理论依据。方法:以国内94份山药种质资源为材料,采用SRAP标记并通过NTSYS2.10软件进行SHAN聚类分析、PROJECTION主成分分析;利用POPGENE软件估算遗传多样性参数。结果:从49对SRAP引物中筛选出30对能产生稳定清晰可辨的扩增产物的引物,共扩增出754条DNA带,其中多态性条带616条,占总条带的81.7%。聚类结果表明:当遗传相似系数(GS)为0.822时,可将94份资源分为5类:第Ⅰ类20份、第Ⅱ类43份、第Ⅲ类7份、第Ⅳ类3份、第Ⅴ类21份。第Ⅰ、Ⅱ、Ⅲ、Ⅴ类分别为薯蓣、褐苞薯蓣、山薯和参薯。主成分分析结果显示:第一与第二主成分可解释88.34%(82.10%和6.24%)的遗传总变异。遗传多样性参数分析表明:比较5个遗传多样性参数值,5个群体的遗传多样性水平表现为Ⅰ>Ⅴ>Ⅲ>Ⅱ>Ⅳ,第Ⅰ类(薯蓣)遗传多样性水平高;山药遗传群体间遗传分化系数为51.88%,大部分差异存在于群体之间,群体间遗传分化高。结论:山药种质资源丰富且群体遗传分化高,有利于山药新品种的选育。SRAP标记可有效应用于山药种质资源的鉴别和遗传多样性分析。通过DNA指纹鉴定技术鉴别山药品种具有重要性与紧迫性。     

新型标记SRAP在棉花F2分离群体及遗传多样性评价中的适用性分析

将新型分子标记SRAP(Sequence-related Amplified Polymorphism)应用于棉花的遗传研究,并建立了完整的PCR反应体系,此体系稳定可靠、扩增效果好、可重复性强。采用30个SRAP引物组合对海岛棉品种“Pima90”和陆地棉品种“邯郸208”进行比较扩增,29个引物组合可以获得多态性扩增,显示了较高的多态性。对上述两个品种的F2群体进行检测,共产生149个多态性条带,平均每个组合产生5．14个,单引物组合最多可产生13个多态性条带。用SRAP标记对11份陆地棉材料进行遗传多样性检测,30个引物组合中15个组合有多态性,得到22个多态性条带,显示了较高的多态性比率。研究结果表明,SRAP标记可在棉花分子生物学领域中广泛应用。     

SRAP分子标记在园林植物遗传育种中的应用

相关序列扩增多态性(sequence-related amplified polymorphism,SRAP)分子标记作为一种新型的分子标记技术,以其多态性高、稳定性高、重复性好、操作简便、效率高及成本低等优点,已经在园林植物中广泛应用.简单介绍SRAP标记的原理和特点,综述其目前在遗传多样性评析、亲缘关系分析、遗传图谱构建、种质资源鉴定及基因克隆与基因定位等方面的研究进展,并对该标记在园林植物的遗传育种的应用前景进行了展望.     

Detection of genetic diversity using RAPD-PCR and sugar analysis in watermelon [Citrullus lanantus (Thunb.) Mansf.] germplasm

RAPD (random amplified polymorphic DNA) markers generated by 15 arbitrary decamers were used to determine the frequency of DNA polymorphism in 39 watermelon [Citrullus lanantus (Thunb.) Mansf.] germplasms. Of the 15 primers tested, all except 1 (primer 275) directed the amplification of polymorphic products. A total of 162 amplification products were generated across all 39 genotypes. Among the 162 fragments, 35 (21%) appeared to be reliable polymorphic markers. The mean value by marker difference in this comparison was 0.24, and the highest, 0.69. Eight RAPD markers could be utilized in the unique variety discrimination 8 watermelon genotypes. From the phenograms constructed by UPGMA based on the comparison of RAPD markers, four clusters were resolved. Each group was also characterized and identified with morphological and genetic characteristics for each genotype. The free sugars of the edible parts of watermelons were analyzed by HPLC (high-performance liquid chromatography). Results from the phylogenetic analysis of band sharing data were consistent with sweetness as measured by HPLC. In conclusion, RAPD assays can be used for providing alternative markers for identifying genotypes and quantitative characteristics in watermelon.     

基于SSR标记的荷花品种遗传多样性及群体结构分析

采用SSR标记技术对42个荷花品种( Nelumbo spp.)的基因组DNA进行扩增,在此基础上,对供试品种进行UPGMA聚类分析、群体结构分析和主坐标分析( PCoA)。结果表明：采用17对SSR引物从42个荷花品种的基因组DNA中扩增出77个位点,多态性位点百分率为88.31%;每对引物可扩增出1~9个多态性位点。根据Nei's遗传距离,供试的42个荷花品种可被分成Ⅰ和Ⅱ两组,分别包含3和39个品种;在Nei's遗传距离0.150处,Ⅱ组被进一步分成Ⅱa、Ⅱb和Ⅱc 3个亚组,分别包含3、16和20个品种。群体结构分析结果表明：组分概率高于等于0.80时,供试的42个荷花品种被分成Pop1、Pop2和混合群3个亚群,分别包含17、16和9个品种。 PCoA分析结果表明：在F1水平上,供试的42个荷花品种被分成2个部分;其中,Pop1亚群的品种均分布在第二和第三象限,而Pop2亚群的品种则分布在第一和第四象限。总体来看,聚类分析、群体结构分析和PCoA分析的结果基本一致。综合分析结果表明：玉组包含美洲黄莲( N. lutea Pers.)品种‘艾江南',且与传统中国莲( N. nucifera Gaertn.)品种的亲缘关系最远,故认为该组为美洲黄莲;Ⅱ组为中国莲,其中,Ⅱc亚组以传统中国莲品种为主,而Ⅱb亚组则偏重于美洲黄莲。总体上看,供试的42个荷花品种主要被分为中国莲和美洲黄莲两组,而中美杂交莲并没有独立成组,其成因有待进一步研究。     

Comparison of the genetic diversity of common wild rice (Oryza rufipogon Griff.) and cultivated rice (O. sativa L.) using RFLP markers

Forty fourth single-copy RFLP markers were used to evaluate the genetic diversity of 122 accessions of common wild rice (CWR, Oryza rufipogon Griff.) and 75 entries of cultivated rice (Oryza sativa L. ) from more than ten Asian countries. A comparison of the parameters showing genetic diversity, including the percentage of polymorphic loci (P), the average number of alleles per locus (A), the number of genotypes (Ng), the average heterozygosity (Ho) and the average genetic multiplicity (Hs) of CWR and indica and japonica subspecies of cultivated rice from different countries and regions, indicated that CWR from China possesses the highest genetic diversity, followed by CWR from South Asia and Southeast Asia. The genetic diversity of CWR from India is the second highest. Although the average gene diversity (Hs)of the South Asian CWR is higher than that of the Southeast Asian CWR, its percentage of polymorphic loci (P), number of alleles (Na) and number of genotypes (Ng) are all smaller. It was also found that the genetic diversity of cultivated rice is obviously lower than that of CWR. At the 44 loci investigated, the number of polymorphic loci of cultivated rice is only 3/4 that of CWR, while the number of alleles, 60%, and the number of genotypes is about 1/2 that of CWR. Of the two subspecies studied, the genetic diversity of indica is higher than that of japonica. The average heterozygosity of the Chinese CWR is the highest among all the entries studied. The average heterozygosity of CWR is about two-times that of cultivated rice. It is suggested that during the course of evolution from wild rice to cultivated rice, many alleles were lost through natural and human selection, leading to the lower heterozygosity and genetic diversity of the cultivated rice. Received: 19 May 1999 / Accepted: 26 April 2000     

Development and characterization of ten new microsatellite markers in a mangrove tree species Bruguiera gymnorrhiza (L.)

Bruguiera gymnorrhiza is an ecologically and somewhat economically important mangrove tree species. We isolated 10 polymorphic microsatellite loci from B. gymnorrhiza using a dual‐suppression polymerase chain reaction (PCR) technique. These loci provided microsatellite markers with polymorphism of two to five alleles per locus within 216 individuals from nine natural populations of B. gymnorrhiza on Iriomote Island, the Sakishima Islands, Japan. The expected and observed heterozygosities ranged from 0.220 to 0.720 and from 0.104 to 0.447, respectively.     

A validated HPLC method for the determination of dimethyl‐4,4′‐dimethoxy‐5,6,5′,6′‐dimethylene dioxybiphenyl‐2,2′‐dicarboxylate (DDB) with fluorescence detection in raw material and pill form: application to an in vitro dissolution test and a content uniformity test

A simple, sensitive and rapid HPLC method with fluorescence detection for the determination of dimethyl‐4,4′‐dimethoxy‐5,6,5′,6′‐dimethylene dioxybiphenyl‐2,2′‐dicarboxylate (DDB) in the raw material and pill form was developed. Liquid chromatography was performed on a C₁₈ column (250 × 4.6 mm i.d., 5 µm particle size), the mobile phase consisted of methanol and 0.05 M sodium dihydrogen phosphate buffer (80 : 20, v/v), and the apparent pH of the mobile phase was adjusted to 3. The fluorescence detector was operated at excitation/emission wavelengths of 275/400 nm. The proposed method allows the determination of DDB within concentration range 0.1–1.5 µg/mL with a limit of detection of 0.032 µg/mL, a limit of quantification of 0.097 µg/mL and a correlation coefficient of 0.9997. The proposed method has been successfully applied for the analysis of DDB in its pills with a percentage recovery of 98.45 ± 0.32. The method was fully validated according to ICH guidelines. Moreover, the high sensitivity of the method permits its use in an in vitro dissolution test for DDB under simulated intestinal conditions. In addition, the proposed method was extended to a content uniformity test according to USP guidelines. Copyright © 2014 John Wiley & Sons, Ltd.     

In vitro culture of lavenders (Lavandula spp.) and the production of secondary metabolites

Lavenders (Lavandula spp., Lamiaceae) are aromatic ornamental plants that are used widely in the food, perfume and pharmaceutical industries. The large-scale production of lavenders requires efficient in vitro propagation techniques to avoid the overexploitation of natural populations and to allow the application of biotechnology-based approaches for plant improvement and the production of valuable secondary metabolites. In this review we discuss micropropagation methods that have been developed in several lavender species, mainly based on meristem proliferation and organogenesis. Specific requirements during stages of micropropagation (establishment, shoot multiplication, root induction and acclimatization) and requisites for plant regeneration trough organogenesis, as an important step for the implementation of plant improvement programs, were revised. We also discuss different methods for the in vitro production of valuable secondary metabolites, focusing on the prospects for highly scalable cultures to meet the market demand for lavender-derived products.     

The physiological and morphological characteristics of Neoplectana carpocapsae (Nematoda,Steinernematidae) in two insect hosts

Morphometry, catalytic (redox potential) activity, and body surface potential of two infective-stage juvenile populations of Neoaplectana carpocapsae, obtained from Sitophilus granarius adults and Galleria mellonella larvae, were compared. The relationship among these three morphological and physiological parameters and the degree of pathogenicity (=infectivity) and incidence of the nematode infection (=extensity) in Achroia grisella caterpillars, Trogoderma granarium larvae, and Tribolium confusum adults was discussed.     

熏烤肉制品中苯并芘的危害及控制措施

本文从食品安全的角度出发,对熏烤肉制品中的有害物质苯并(a)芘的产生,对人体的危害作用,以及控制熏烤肉制品中苯并(a)芘残留的方法进行了初步的研究.这对提高熏烤肉制品品质和对苯并(a)芘进行深入的研究具有重要意义.