花生微卫星标记的研究进展

近年来花生微卫星标记的开发取得了一定的进展, 初步揭示了花生在DNA水平上的遗传多样性。花生微卫星标记的开发途径主要包括通过构建小片段基因组文库开发基因组SSR标记, 根据花生EST序列开发EST-SSR标记, 根据豆科植物序 列信息和SSR标记开发花生SSR标记, 将SSR标记与其它分子标记结合开发新的DNA标记, 以及基于SSR核心序列开发ISSR标记。花生微卫星标记主要应用于遗传多样性研究、遗传图谱与品种指纹图谱构建以及分子标记辅助育种等领域。本文综述了花生SSR标记开发研究的进展及应用。     

花生微卫星标记的研究进展

近年来花生微卫星标记的开发取得了一定的进展,初步揭示了花生在DNA水平上的遗传多样性。花生微卫星标记的开发途径主要包括通过构建小片段基因组文库开发基因组SSR标记,根据花生EST序列开发EST-SSR标记,根据豆科植物序列信息和SSR标记开发花生SSR标记,将SSR标记与其它分子标记结合开发新的DNA标记,以及基于SSR核心序列开发ISSR标记。花生微卫星标记主要应用于遗传多样性研究、遗传图谱与品种指纹图谱构建以及分子标记辅助育种等领域。本文综述了花生SSR标记开发研究的进展及应用。     

花生抗青枯病分子标记研究

利用抗、感青枯病的花生品种为素本配制杂交组合中花5号×远杂9102,构建重组近交系,以其F6为研究材料,分析青枯痛抗性遗传规律,结果表明,花生青枯病抗性是由两时主效基因控制的遗传,并且主效基因的遗传力较高,为84%;同时采用AFLP技术和BSA分析方法,获得两个与花生青枯病抗性连锁的分子标记,标记与抗性间的遗传距离分别为8.12cM和11.46cM.利用获得的分子标记对抗、感青枯病的花生种质进行了分子鉴定,证实了标记P3M59与膏枯病抗性的符合率为70%,标记PIM58的符合率为50%,从而为花生青枯病抗性辅助选择育种提供理论基础.     

基于简化基因组的花生InDel标记开发和功能解析

InDel在基因组中的分布密度仅次于SNP,可作为动植物群体遗传分析、分子辅助育种等研究领域的有效分子标记。花生是世界范围内重要的油料作物之一。目前,花生栽培种全基因组已经公布,为准确挖掘花生基因组信息提供了重要参考。本研究通过169份花生核心种质的GBS(Genotyping-by-sequencing)测序和比对,共获得大小分布在1～14 bp范围内的10401个InDels。染色体Arahy.16上分布的InDels最多,达741个;而在染色体Arahy.08上分布的最少,有263个。参考基因组注释信息,仅有1167个InDels分布在功能基因相关区域。经GO注释,InDel分子功能主要包括催化活性(catalytic activity)和结合(binding);生物过程主要涉及代谢过程(metabolic process)、单组织过程(single-organism process)和细胞过程(cellular process)。经KEGG通路分析发现,InDels所在的基因区域的功能主要与代谢相关。本研究开发出全基因组水平的InDel标记,并做了相应功能分类和注释,为进一步分子验证和利用提供丰富的基因资源。     

SSR分子标记检测出的花生类型内遗传变异

花生是我国重要的食用油和蛋白质来源作物,鉴定其DNA分子多态性对品种改良和资源评价具有重要的意义。从已公布的花生Genomic-SSR和EST-SSR引物中筛选出34对引物,用来分别鉴定花生4大类型各24份共96份品种资源的分子变异,其中龙生型资源全部来自广西,普通型资源中有11份从国外引进,有13份来自广西和国内其他省市,多粒型资源只有两份来自中国,其他22份分别来自印度、美国和非洲等地,珍珠豆型资源中有22份是来自中国各地的育成品种或农家品种,有2份来自国外。研究结果为:分别有10～16对SSR引物能在4大类型花生资源中扩增出多态性DNA片段;这些多态性SSR引物都具有多位点特性;首次为SSR分子标记设立了一个新的评价指标——区别指数,多态性SSR引物的区别指数最高达0.992;资源间的平均遗传距离,多粒型为0.59,普通型为0.48,珍珠豆型为0.38,龙生型为0.17。根据遗传距离采用最长距离法对4大类型花生资源分别进行了聚类分析,构建了资源间的遗传关系图,花生4大类型可进一步分成不同类群,资源间的亲缘关系与其来源相关。观察到PM15和PMc297的扩增产物具有类型特异性,PM15能在龙生型、普通型和多粒型花生资源中扩增出多态性条带,而在珍珠豆型花生中扩增条带完全相同,PMc297也有相似的扩增结果。由于在多粒型花生资源中检测出的遗传多样性最丰富,研究结果支持西班牙专家Krapovickas 1994年公布的花生栽培种分类系统。总之在花生4大类型内资源中能检测出丰富的SSR分子标记,开发出更多的SSR分子标记将能充分揭示花生分子水平的变异,从而使花生遗传图谱构建、分子标记辅助育种成为可能。     

花生黄曲霉侵染抗性的SCAR标记

利用与花生黄曲霉侵染抗性基因紧密连锁的AFLP标记 “E45/M53-440”, 经PAGE凝胶电泳后回收、克隆、测序, 并根据测序结果设计PCR特异引物, 通过对PCR条件的优化, 成功地将AFLP标记“E45/M53-440”转化为实验结果稳定, 操作更简单的SCAR标记“AFs-412”, 标记与花生黄曲霉侵染抗性间的遗传距离为6.5 cM。利用获得的SCAR标记对抗、感黄曲霉的花生种质资源进行了分子鉴定, 结果表明标记与抗性鉴定结果具有较高的一致性, 证实了该标记应用于研究群体之外的育种潜力。SCAR标记的建立为开展花生黄曲霉侵染抗性的标记辅助选择育种提供了简便实用的鉴定技术。     

花生EST数据库的分析、评价和应用前景

花生是我国重要的油料和经济作物。花生产业的发展对我国国民经济具有重要战略意义。随着分子生物学的发展,植物基因工程和功能基因组学的先进技术将有效推动花生种质创新和科技进步。分析了现有的花生EST数据,结合其他作物功能基因组学的最新研究进展,深入探讨了花生EST数据资源在基因克隆、分子标记开发及表达谱研究等方面的利用价值,并在分析花生EST数据库特点的基础上,展望了新一代测序技术在花生中的应用前景。为更好的利用花生EST数据库提供参考。     

向日葵分子生物学研究进展

向日葵是一种营养价值极高的资源植物,向日葵的研究和生产在当前我国中西部大开发的战略中具有重要的意义。本文对近年来国内外向日葵分子生物学研究的最新进展,在蛋白质、酶、基因及基因工程、分子标记等方面进行了综述。特别对生化标记、分子标记技术在向日葵研究上的应用及所取得的成果作了重点介绍,并对今后向日葵研究工作进行了展望。     

卡瓦胡椒RAPD反应体系的建立

卡瓦胡椒RAPD分子标记的研究,目前国内外尚未见有报道。该试验通过CTAB法提取卡瓦胡椒基因组DNA,通过对模板DNA用量、Mg^2＋浓度、退火温度、电泳上样量等几个单因子试验来建立RAPD稳定扩增体系和反应条件,RAPD扩增结果重复性好,稳定可靠,为卡瓦胡椒RAPD分子标记的研究打下基础。     

西瓜甜瓜质量性状的分子标记与定位研究进展

分子标记在作物育种、资源遗传多样性分析、遗传图谱构建等方面有着广泛的应用,常见的分子标记方法主要有RFLP、RAPD、SSR、AFLP、SCAR、CAPS、SNP等.迄今为止,国内外学者已经利用分子标记技术对西瓜和甜瓜的20多个质量性状进行了标记和定位.本文主要对西瓜和甜瓜质量性状的分子标记和定位研究进展进行了综述,发现RAPD标记使用最多,而更具优势的SSR标记使用较少;对抗病虫等育种相关的基因研究较多,且多数标记遗传距离偏大,达不到图位克隆的要求,而对西瓜和甜瓜遗传基础相关的基因研究较少.     

Taxonomic relationships among Arachis sect. Arachis species as revealed by AFLP markers.

Cultivated peanut, Arachis hypogaea L., is a tetraploid (2n = 4x = 40) species thought to be of allopolyploid origin. Its closest relatives are the diploid (2n = 2x = 20) annual and perennial species included with it in Arachis sect. Arachis. Species in section Arachis represent an important source of novel alleles for improvement of cultivated peanut. A better understanding of the level of speciation and taxonomic relationships between taxa within section Arachis is a prerequisite to the effective use of this secondary gene pool in peanut breeding programs. The AFLP technique was used to determine intra- and interspecific relationships among and within 108 accessions of 26 species of this section. A total of 1328 fragments were generated with 8 primer combinations. From those, 239 bands ranging in size from 65 to 760 bp were scored as binary data. Genetic distances among accessions ranged from 0 to 0.50. Average distances among diploid species (0.30) were much higher than that detected between tetraploid species (0.05). Cluster analysis using different methods and principal component analysis were performed. The resulting grouping of accessions and species supports previous taxonomic classifications and genome designations. Based on genetic distances and cluster analysis, A-genome accessions KG 30029 (Arachis helodes) and KSSc 36009 (Arachis simpsonii) and B-genome accession KGBSPSc 30076 (A. ipaensis) were the most closely related to both Arachis hypogaea and Arachis monticola. This finding suggests their involvement in the evolution of the tetraploid peanut species.     

Genetic diversity of cultivated and wild-type peanuts evaluated with M13-tailed SSR markers and sequencing

Thirty-one genomic SSR markers with a M13 tail attached were used to assess the genetic diversity of the peanut mini core collection. The M13-tailed method was effective in discriminating almost all the cultivated and wild accessions. A total of 477 alleles were detected with an average of 15.4 alleles per locus. The mean polymorphic information content (PIC) score was 0.687. The cultivated peanut (Arachis hypogaea L.) mini core produced a total of 312 alleles with an average of 10.1 alleles per locus. A neighbour-joining tree was constructed to determine the interspecific and intraspecific relationships in this data set. Almost all the peanut accessions in this data set classified into subspecies and botanical varieties such as subsp. hypogaea var. hypogaea, subsp. fastigiata var. fastigiata, and subsp. fastigiata var. vulgaris clustered with other accessions with the same classification, which lends further support to their current taxonomy. Alleles were sequenced from one of the SSR markers used in this study, which demonstrated that the repeat motif is conserved when transferring the marker across species borders. This study allowed the examination of the diversity and phylogenetic relationships in the peanut mini core which has not been previously reported.     

Development and use of single nucleotide polymorphism markers for candidate resistance genes in wild peanuts (Arachis spp)

The cultivated peanut (Arachis hypogaea L.) is an allotetraploid of recent origin, with an AABB genome and low genetic diversity. Perhaps because of its limited genetic diversity, this species lacks resistance to a number of important pests and diseases. In contrast, wild species of Arachis are genetically diverse and are rich sources of disease resistance genes. Consequently, a study of wild peanut relatives is attractive from two points of view: to help understand peanut genetics and to characterize wild alleles that could confer disease resistance. With this in mind, a diploid population from a cross between two wild peanut relatives was developed, in order to make a dense genetic map that could serve as a reference for peanut genetics and in order to characterize the regions of the Arachis genome that code for disease resistance. We tested two methods for developing and genotyping single nucleotide polymorphisms in candidate genes for disease resistance; one is based on single-base primer extension methods and the other is based on amplification refractory mutation system-polymerase chain reaction. We found single-base pair extension to be an efficient method, suitable for high-throughput, single-nucleotide polymorphism mapping; it allowed us to locate five candidate genes for resistance on our genetic map.     

The phylogenetic relationship of possible progenitors of the cultivated peanut

The cultivated peanut (Arachis hypogaea L.) is an allotetraploid composed of A and B genomes. The phylogenetic relationship among the cultivated peanut, wild diploid, and tetraploid species in the section Arachis was studied based on sequence comparison of stearoyl-ACP desaturase and oleoyl-PC desaturase. The topology of the trees for both fatty acid desaturases displayed two clusters; one cluster with A genome diploid species and the other with B genome diploid species. The two homeologous genes obtained for each of the two fatty acid desaturases from the tetraploid species A. hypogaea and A. monticola were separated into the A and B genome clusters, respectively. The gene phylogenetic trees showed that A. hypogaea is more closely related to the diploid species A. duranensis and A. ipaensis than to the wild tetraploid species A. monticola, suggesting that A. monticola is not a progenitor of the cultivated peanut. In addition, for the stearoyl-ACP desaturase, the A. duranensis sequence was identical with one of the sequences of A. hypogaea and the A. ipaensis sequence was identical with the other. These results support the hypothesis that A. duranensis and A. ipaensis are the most likely diploid progenitors of the cultivated tetraploid A. hypogaea.     

iTRAQ-Based Proteomic Reveals Cell Cycle and Translation Regulation Involving in Peanut Buds Cold Stress

Russian Journal of Plant Physiology - Cold stress is one major threats to field crops. The cold tolerant ability is a key limiting factor for the popularization of peanut (Arachis hypogaea L.) in...     

强光下高温与干旱胁迫对花生光系统的伤害机制

为探讨高温和干旱胁迫对花生光合系统的不同影响机制,以鲁花14为试材进行高温(42 ℃)强光(1200 μmol · m^-2 · s^-1)(HH)、干旱(PEG6000,30%)强光(1200 μmol · m^-2 · s^-1)(DH)和强光(1200 μmol · m^-2 · s^-1)胁迫(NH)处理,以未处理为对照(CK)的实验。与CK及NH处理相比,HH和DH的最大光化学效率(Fv/Fm)和820 nm光吸收大幅下降,叶绿素荧光动力学曲线上J点相对荧光(Vj)上升,单位面积内吸收的光量子(ABS/CSm)、单位面积内反应中心捕获的光量子(TRo/CSm)和单位面积内有活性的反应中心的数目(RC/CSm)均出现大幅下降,而PSⅡ的关闭程度(1-qP)明显升高,依赖于叶黄素循环的非辐射能量耗散(NPQ)升高,同时超氧化物歧化酶(SOD)活性出现下降,丙二醛(MDA)和膜透性增加,这些结果表明,HH和DH胁迫引起了花生叶片的严重光抑制,但快速叶绿素荧光诱导动力学曲线中均没有出现K点,表明花生叶片光合系统放氧复合体(OEC)对高温和干旱胁迫不敏感,光合系统Ⅱ(PSⅡ)反应中心的受体侧更容易受到高温和干旱的影响,而对花生光系统造成严重破坏的主要原因则是过剩光能的积累,一方面虽然叶黄素循环可以耗散部分能量,但不是全部;另一方面水-水循环受到高温和干旱的影响不能有效起到能量消耗的作用,造成活性氧的大量积累。HH和DH处理对花生光系统造成的伤害相似,但DH处理对花生光系统的伤害程度大一些,强光下,高温和干旱对花生叶片的伤害位点及破坏机制却较为相似。     

Heat-stable Proteins and Acquisition of Desiccation Tolerance in Peanut Seeds

The acquisition and induction of desiccation tolerance associated with the expression of heat-stable proteins in the developing peanut (Arachis hypogaea L. ) seeds were studied. Desiccation tolerance of peanut seeds was achieved during 45 to 65 DAP (days after pegging) embryogenesis, while a set of low molecular weight (9 to 15.5 kD) heat-stable polypeptides was preferentially expressed. Slow drying regime applied in vitro to 25 and 35 DAP peanut embryos induced desiccation tolerance and the expression of the same subset of polypeptides. Mature drying treatment enhanced the ability of 65 DAP peanut embryos to withstand fast drying, also increased the heat stability of arachins, the major peanut storage protein, which was heat labile during 45 to 65 DAP embryogenesis. It was concluded that the heat-stable proteins may contribute to desiccation tolerance of the peanut seeds, and the low molecular weight heat-stable polypeptides may confer nonspecifieally heat tolerance on peanut storage proteins which were normally heat labile.     

Phosphorus-fertilisation has differential effects on leaf growth and photosynthetic capacity of Arachis hypogaea L.

Plant and Soil - The objectives of this study were to assess how Arachis hypogaea L. (peanut or groundnut) responds to different P supplies in terms of growth and photosynthesis, and to determine...     

花生种子耐脱水力的获得与热稳定蛋白的关系

花生（ＡｒａｃｈｉｓｈｙｐｏｇａｅａＬ．）种子的耐脱水能力在果针入土后４５ｄ以后的胚胎发育期逐渐增加,与一组９～１５．５ｋＤ低分子量热稳定蛋白的丰富表达有关。缓慢干燥可以诱导不耐脱水的果针入土后２５ｄ及３５ｄ花生胚获得耐脱水能力并同时诱导胚轴表达这组热稳定蛋白。成熟脱水促进花生胚耐脱水能力的获得,并增加了花生球蛋白的热稳定性。