基于SSR分子标记的芋种遗传多样性分析

本研究利用SSR分子标记对来自于国家种质武汉水生蔬菜资源圃的110份芋种资源进行了遗传多样性分析.10对SSR引物在110份芋种资源中共扩增得到40条带,多态性百分率为100％,Shannon信息指数范围为0.390 5～1.426 8,反映了这110份芋种资源的遗传多样性程度较高.110份芋种资源遗传相似系数介于0.43～1,在遗传相似系数0.63处,聚类图将其分为6个类群.该研究结果为芋种资源的保护和利用奠定了基础.     

我国薯芋皂甙元的工业生产和资源的回顾与展望

本文对我国薯芋皂素(Diosgenin)的工业生产进行了概略回顾,对各种工艺流程进行了比较,并介绍了薯芋属植物的资源现状。     

基于ISSR标记的中国芋种质资源遗传多样性分析

本研究利用ISSR分子标记技术,对国家种质武汉水生蔬菜资源圃收集并保存的来自全国各地的72份芋种质资源进行遗传多样性检测。研究结果表明,13条ISSR引物在72份芋种质资源中共扩增出109条带,其中85条带具有多态性,平均多态性位点比率为78.51%;不同芋种质资源间遗传相似系数的变幅为0.56~1,说明ISSR标记能够揭示芋种质资源之间较高的遗传多样性;在遗传系数0.725处,72份芋种质资源被聚为3大类,为进一步研究芋种质资源分类、起源、保存和利用奠定了基础。     

206份芋种质资源品质性状分析

对206份芋种质资源品质性状进行了分析,结果表明:按球茎类型来分,干物质含量魁芋>多头芋>魁子兼用芋>多子芋。淀粉含量魁芋>魁子兼用芋>多头芋>多子芋,干物质和淀粉含量都以魁芋最高,分别为27.38%、20.21%,多子芋最低,分别为19.54%、13.35%。蛋白质含量多头芋>魁芋>多子芋>魁子兼用芋,以多头芋的含量最高为1.54%,其他类型相差不大。可溶性糖含量魁芋>魁子兼用芋>多子芋>多头芋,以魁芋的可溶性糖含量最高为1.21%,其他芋类型相差不大。按母芋芽色来分,多子芋中,干物质、淀粉和蛋白质含量红芽多子芋>白芽多子芋,可溶性糖含量白芽多子芋>红芽多子芋;多头芋中,干物质、淀粉、可溶性糖白芽多头芋>红芽多头芋,蛋白质含量红芽多头芋>白芽多头芋。在整个芋种质资源中,芋的干物质含量与淀粉含量(r=0.9583**)、干物质含量与蛋白质含量(r=0.5529**)、淀粉含量与蛋白质含量(r=0.5284**)呈极显著正相关,干物质含量与可溶性糖含量(r=-0.2934**)、淀粉含量与可溶性糖含量(r=-0.3391**)、蛋白质含量与可溶性糖含量(r=-0.4498**)均呈极显著负相关。此外本研究还按叶柄颜色类型对芋种质资源品质性状进行了分析。     

利用SRAP标记分析四川省芋种质资源遗传多样性

在分子水平研究四川省芋资源的遗传多样性和亲缘关系,为芋种质资源的分类、保护和有效利用遗传资源以及新品种选育提供依据。本研究利用SRAP分子标记技术,使用28对SRAP引物组合对65份四川省不同地区芋种质资源材料进行遗传多样性分析,采用NT-SYS 2.1统计软件对数据进行分析,建立树状聚类图。扩增出并检测到341条条带,平均每个引物组合扩增检测出12.18条带,多态性带251条,多态率73.6%。UPGMA树形图表明,所用的SRAP引物组合可以将65份材料分成5类,分别与这些材料在园艺分类学上按母芋和子芋的生长习性分类基本相符,与以芋叶心色斑颜色、叶柄中下部颜色、母芋芽色及母芋肉色4种形态性状组合描述具有相关性。研究表明,从四川省不同地区、不同生态环境下收集的不同类型芋种质资源间存在着较丰富的遗传多样性,SRAP分析聚类结果与主要形态学性状分类基本一致,可以解释芋栽培种的进化关系。     

芋(Colocasia esculenta)的民族植物学

应用民族植物学的基本原理和方法,选择中国云南和山东为试点,兼顾其他省区,开展芋[Colocasia esculenta(L.)Schott]的民族植物学研究.结果表明:在云南传统栽种芋的菜园和农地被高附加值的经济作物所代替,芋在不同民族家庭中的地位也从传统作为主食变成蔬菜或杂粮;在山东已形成芋的产业化、标准化生产的格局,芋在汉族农家经济中的地位得到提升.在云南分布有芋的野生近缘种、半栽培种、栽培品种,种质资源丰富;在山东未发现芋的野生类型,以旱芋类型的多子芋栽培品种为主.由于经济的发展和主流文化的影响,民间对芋的植物崇拜及植物崇拜文化丢失的速度大大加剧.在古朴的传统食芋文化中,蕴含着丰富的关于芋植物资源利用和保护的传统知识和朴素的科学内涵,需要进行深入的挖掘和探讨.     

芋螺毒素基因资源研究进展

芋螺毒素和微生物的次生代谢产物与植物的生物碱一样,具有生物多样性的特点。芋螺毒素特有的二硫键骨架和化学修饰后特异的空间结构,使其具有特异的稳定性和药理学活性。对芋螺毒素基因的分析和新型基因的克隆筛选,是深入研究各种受体、离子通道及其亚型,进而在克隆表达的靶受体上设计和筛选高效新药的前提。芋螺毒素基因资源的研究在芋螺毒素新基因及其编码产物毒素肽的发现与利用方面发挥了重要作用。现对该领域的新进展进行论述。     

RAPD在植物研究中的应用

RAPD作为现代生物科学研究的一项技术得到了广泛应用,在此介绍了RAPD的原理和主要技术步骤及其存在的优缺点,并介绍了RAPD在植物的系统分类、种质资源研究、性状育种、抗性育种、观赏植物育种五个方面的应用;同时,对RAPD在植物研究中的前景作了探讨。     

芋种质资源开花结实状况调查

对保存在国家种质武汉水生蔬菜资源圃中的芋属（Colocasia）种质资源的开花结实状况调查表明：（1）在自然条件下,所有滇南芋[Colocasia antiquorum (L.) Schott]（2n=2x）中的野芋[即原C. esculenta (L.) var. antiquorum]和花用芋（即原C. tonoimo Nakai）资源全部开花;芋[Colocasia esculenta (L.) Schott]开花状况表现为：魁芋（2n=2x）、多头芋（2n=3x）、魁子兼用芋（2n=3x）都不开花,国内多子芋中,紫柄多子芋（2n=3x）都不开花,乌绿柄多子芋（2n=3x）和绿柄多子芋（2n=3x）有部分品种开花,但开花的品种具有不确定性;来源于东南亚国家的多子芋中,绿柄多子芋（2n=3x）开花率高,花期长,另有1份紫柄多子芋（2n=3x）开花,与国内多子芋（2n=3x）差异较大。开花率表现为东南亚多子芋（2n=3x）等于或高于国内芋;国内芋中,二倍体大于三倍体。花期最长的是东南亚的绿柄多子芋（2n=3x）,国内芋中,二倍体的开花率大于三倍体,二倍体的花期长于三倍体。（2）乌绿柄多子芋和花用芋的芽色都为红色,属于短附属器,其余类型的芽色为白色,属于长附属器。（3）在自然条件下,仅有2份紫柄野芋种质结实,其它种质未见结实。（4）本文还对不同类型芋的始花期、末花期、花期、单株花序丛数、单个叶轴花序数、单株总花序数、佛焰苞管部长度、佛焰苞檐部长度、附属器长、雄花序长、中性花序长、雌花序长等进行了比较。     

芋ISSR-PCR扩增反应体系的建立

目的:获得清晰可靠、重复性较好的ISSR-PCR扩增反应体系,应用于芋种质资源进行遗传多样性的研究中.方法:以芋的幼叶提取基因组DNA为材料,采用正交试验设计L16(45),从模板DNA浓度、引物浓度、Mg2+浓度、dNTPs浓度及TaqDNA聚合酶的用量5因素4水平出发,构建芋最佳反应体系.结果:芋ISSR-PCR的最佳反应体系为:在25μl的反应体系中,40ng DNA模板、0.4μmol/L引物浓度、2.5 mmol/L Mg2+、0.2 mmol/L dNTPs、1 U Taq DNA聚合酶.结论:利用芋种质资源对最佳反应体系的验证,结果显示该反应体系具有扩增稳定性.     

RAPD分子标记在食用菌研究中的应用

RAPD分子标记以其简单、快速、经济等优点正在被广泛应用于食用菌的研究中。RAPD分子标记克服了传统的标记手段和形态分类学的缺点,在食用菌种质的鉴定和遗传多样性研究,在亲本和杂交种的鉴别以及在基因克隆与分离和遗传图谱的建立等研究中都起着重要的作用。RAPD分子标记为食用菌的深入研究提供了强有力的工具和手段,其应用和发展前景广阔。     

Studying genetic diversity in inbred sunflower lines by RAPD and isoenzyme analyses

Genetic diversity of 30 inbred sunflower lines was examined by RAPD and isozyme analyses. The inbred lines were shown to be highly polymorphic by RAPD markers. The line distribution on genetic similarity dendrograms based on the RAPD and isozyme data was analyzed. High effectiveness of RAPD analysis for differentiating genotypes of inbred sunflower lines was demonstrated.     

Studying Genetic Diversity in Inbred Sunflower Lines by RAPD and Isozyme Analyses

Genetic diversity of 30 inbred sunflower lines was examined by RAPD and isozyme analyses. The inbred lines were shown to be highly polymorphic by RAPD markers. The line distribution on genetic similarity dendrograms based on the RAPD and isozyme data was analyzed. High effectiveness of RAPD analysis for differentiating genotypes of inbred sunflower lines was demonstrated.     

RAPD Analysis of Pathogenic Variability in Ascochyta rabiei

Thirty Italian isolates of the phytopathogenic fungus Ascochyta rabiei (Pass.) Labr., the causal organism of Ascochyta blight on chickpea (Cicer arietinum L.), were analysed by a random oligonucleotide primer dependent polymerase chain, reaction (PCR) technique called random amplified polymorphic DNA analysis (RAPD) using three decamer primers. In previous investigations these isolates had been differentiated in six pathogenic groups. RAPD results were summarized in an analysis using the program PAUP. With each of the primers several amplification products were observed which were common to all isolates. The results of the RAPD analyses also showed that all isolates could be identified by a unique RAPD pattern. No correlation between RAPD patterns and the division of the isolates in pathogenic groups could be established. The application of the RAPD technique for cataloguing isolates and to obtain specific genetic markers for all isolates of the species Ascochyta rabiei is discussed.     

RAPD analysis of seed purity in a commercial hybrid cabbage (Brassica oleracea var. capitata) cultivar.

The use of random amplified polymorphic DNA (RAPD) markers for evaluating seed purity in a commercial F1-hybrid cabbage (Brassica oleracea var. capitata) cultivar is demonstrated. Genomic DNA isolated from single ungerminated seed was found to be suitable for RAPD analysis. DNA from F1-hybrid and its parental lines was subjected to RAPD screening with 36 random decamer arbitrary primers. A total of 241 scorable products were observed with 54 (22%) being polymorphic. The RAPD data showed that the parental lines of this commercial cabbage cultivar were not very closely related. Two primers were chosen for purity testing of the F1-hybrid seeds. The sib (inbred seed; seed from self-pollination of parental lines) contamination results obtained by RAPD analysis were comparable to the commonly used grow-out trial and isozyme analysis, hence showing that RAPD analysis can be used for seed purity testing of commercial hybrid cabbage seeds.     

DNA fragments of organellar origin in random amplified polymorphic DNA (RAPD) patterns of sugar beet (Beta vulgaris L.)

The technique of random amplified polymorphic DNA (RAPD) offers a broad range of applications in the investigation of plant genomes. A promising prospect is the use of RAPD products as genetic markers. We have investigated a possible organellar source of fragments in RAPD patterns of total DNA. Two nearly-isogenic lines of cytoplasmic male-sterile and male-fertile sugar beet (Beta vulgaris L.) were subjected to RAPD analysis with six different primers. Total, nuclear, mitochondrial (mt), and chloroplast (cp), DNA from each line were investigated. Reproducible DNA fingerprints could be obtained from both organellar DNAs. Differences in band patterns of mtDNA between cytoplasmic male-sterile and -fertile lines were observed with five out of six primers, whereas different cpDNA patterns were generated by one of the primers. Consequently, the RAPD technique can be used to discriminate between different cytoplasms. Clear evidence is provided for the organellar origin of fragments in genomic (total DNA) RAPD patterns. The consequences of these results for the interpretation of RAPD analyses are discussed.     

Qualitative and quantitative characterization of RAPD variation among snap bean (Phaseolus vulgaris) genotypes

Ten snap bean (Phaseolus vulgaris) genotypes were screened for polymorphism with 400 RAPD (random amplified polymorphic DNA) primers. Polymorphic RAPDs were scored and classified into three categories based on ethidium bromide staining intensity. An average of 5.19 RAPD bands were scored per primer for the 364 primers that gave scorable amplification products. An average of 2.15 polymorphic RAPDs were detected per primer. The results show that primer screening may reduce the number of RAPD reactions required for the analysis of genetic relationships among snap-bean genotypes by over 60%. Based on the analysis of the distribution of RAPD amplification, the same number of polymorphic RAPDs were amplified from different genotypes for all RAPD band intensity levels. A comparison of RAPD band amplification frequency among genotypes for the three categories of bands classified by amplification strength revealed a measurable difference in the frequencies of RAPDs classified as faint (weakly amplifying) compared to RAPD bands classified as bold (strongly amplifying) indicating a possible scoring error due to the underscoring of faint bands. Correlation analysis showed that RAPD bands amplified by the same primer are not more closely correlated then RAPD bands amplified by different primers but are more highly correlated then expected by chance. Pairwise comparisons of RAPD bands indicate that the distribution of RAPD amplification among genotypes will be a useful criterion for establishing RAPD band identity. For the average pairwise comparison of genotypes, 50% of primers tested and 15.8% of all scored RAPDs detected polymorphism. Based on RAPD data Nei's average gene diversity at a locus was 0.158 based on all scorable RAPD bands and 0.388 if only polymorphic RAPD loci were considered. RAPD-derived 1 relationships among genotypes are reported for the ten genotypes included in this study. The data presented here demonstrate that many informative, polymorphic RAPDs can be found among snap bean cultivars. These RAPDs may be useful for the unique identification of bean varieties, the organization of bean germplasm, and applications of molecular markers to bean breeding.     

Population Structure of Puccinia recondita in Western Europe During 1995, as Assessed by Variability in Pathogenicity and Molecular Markers

The population structure of Puccinia recondita f. sp. tritici (Prt) in western Europe was examined by assessing variability in pathogenicity and in randomly amplified polymorphic DNA (RAPD) among 61 single uredinial isolates. The isolates were chosen to represent pathotypes detected in a previous survey of pathogenic variability in the fungus in western Europe in 1995. Thirty‐five pathotypes were identified by assessing infection types produced by the 61 isolates on 24 differential lines, each with a single gene for resistance to Prt. In contrast, only 18 RAPD phenotypes were identified by scoring 19 polymorphic RAPD bands generated with eight RAPD primers. When analysed by cluster and bootstrap analyses, the pathogenicity and RAPD results revealed little evidence for robust distinct clusters among the isolates. Multiple isolates of several pathotypes collected from widely separated locations such as Belgium, Germany, France, Italy and Switzerland had the same RAPD phenotype, providing evidence of clonal migration over considerable distances in western Europe. Some variability (one or two band differences) was observed in RAPD phenotype within several pathotypes, indicating the possible occurrence of genetic changes independent of pathogenicity, and/or the independent development of pathotypes with different genetic backgrounds. Two groups of isolates identified in the 1995 survey, differentiated by pathogenicity for genes Lr3a, Lr3bg, Lr3ka and Lr30, were not distinguished by RAPD phenotype, indicating that the groups probably do not constitute separate lineages within the pathogen population. Little correlation was apparent between the polymorphisms observed in pathogenicity and RAPD phenotypes. The similarity in the genetic backgrounds of the isolates, as assessed by RAPD markers, suggest that the observed differences in pathogenicity may have arisen by selection for specific virulences corresponding to genes for resistance in wheat cultivars grown in the region. Three isolates of pathotype 3, restricted in its distribution to southern France during 1995, were distinct from all other isolates in RAPD phenotype. Circumstantial evidence suggests that this pathotype originated from northern Africa, and that it belongs to a group of leaf rust pathogens specialized to durum wheats.     

Utility of RAPD markers in identifying genetic linkages to genes of economic interest in peach

The identification of molecular markers linked to economically important traits for use in crop improvement is very important in long-lived perennial species. Three-hundred-and-sixty RAPD primers were used with bulked segregant analysis to identify markers linked to loci of specific interest in peach [(Prunus persica) L. Batch] and peach x almond [(Prunus dulcis) Batch] crosses. The traits analyzed included flesh color, adhesion, and texture; pollen fertility; plant stature; and three isozyme loci. The Mendelian behavior of the RAPD loci was established, and RAPD markers were mapped relative to the loci controlling flesh color, adhesion, and texture, and the isozyme loci Mdh-1, 6Pgd-2 and Aat-1, as well as the existing RFLP genetic linkage map constructed previously using a peach x almond F₂ population. This technique has facilitated rapid identification of RAPD and RFLP markers that are linked to the traits under study. Loci controlling these traits mapped predominantly to linkage groups 2 and 3 of the peach genetic linkage map. Linkages to genes with both dominant and co-dominant alleles were identified, but linkages to dominant genes were more difficult to find. In several crosses, RAPD marker bands proved to be allelic. One co-dominant RAPD formed a heteroduplex band in heterozygous individuals and in mixtures of alternate homozygotes. The Mendelian behavior of the RAPD loci studied was established and the results suggest that RAPD markers will be useful for plant improvement in peach.     

A Comparison of AFLP and RAPD Markers for Genetic Diversity and Cultivar Identification in Cotton

AFLP and RAPDmarkers were employed in sixteen diploid cotton (Gossypium sp) cultivars for genetic diversity estimation and cultivar identification. Polymorphism information content (PIC) and percent polymorphism were found to be more for AFLP markers as compared to RAPD markers. Average Jaccard’s genetic similarity index was found to be almost similar using either AFLP or RAPD markers. All the cultivars could be distinguished from one another using AFLP markers and also by the combined RAPD profiles. Cultivar identification indicators like resolving power, marker index and probability of chance identity of two cultivars suggested the usefulness of AFLP markers over the RAPD markers. AFLP and RAPD analyses revealed limited genetic diversity in the studied cultivars. Cluster analysis of both RAPD and AFLP data produced two clusters, one containing cultivars of G. herbaceum and another containing cultivars of G. arboreum species. Highly positive correlation between cophenetic matrices using RAPD and AFLP markers was observed. AFLP markers were found to be more efficient for genetic diversity estimation, polymorphism detection and cultivar identification.