45 份木豆种质资源物候期及形态多样性分析

对收集的45 份木豆(Cajanus cajan)种质资源进行物候期和形态性状指标的观测, 并以此为基础进行各指标的变异系数、相关性分析和聚类分析, 分析木豆种质资源的遗传多样性, 筛选优异种质资源。结果表明: 木豆种质资源形态性状多样性丰富, 45 份种质资源的8 个数量遗传性状变异系数为3.73%-17.84%, 花轴长度与中央小叶宽、中央小叶叶长、旗瓣大小相关性显著(P<0.05)。基于8 个形态指标的聚类分析将45 份种质划分为三大类群, 各类群间形态性状差异明显, 群体表现跟地域来源间没有直接关系, 第一、第二类群可以进一步考察作为育种材料。     

木豆不同品种和叶龄对叶片氨基酸形成的影响和聚类分析

为了解木豆(Cajanus cajan)叶片的氨基酸含量的变化,对19份种质资源和1～3叶龄的木豆叶片氨基酸含量进行研究。结果表明,不同种质资源和叶龄的木豆叶片中均含有17种氨基酸,必需氨基酸以苏氨酸、异亮氨酸和赖氨酸的含量最高,分别为0.67%、0.70%和0.48%,非必需氨基酸以丝氨酸、酪氨酸和组氨酸的含量最高,分别为0.69%、0.66%和0.44%。除赖氨酸和丙氨酸外,其余15种氨基酸含量在木豆叶片中总体上随着叶龄的增加呈现上升趋势。不同种质资源的木豆叶片中总氨基酸含量差异明显。聚类分析可将19个木豆种质资源划分为低氨基酸型(I)、中氨基酸型(II)、中高氨基酸型(III)、高氨基酸型(IV)等4大类型。这为高品质木豆叶茶的加工与高氨基酸木豆品种选育提供了理论依据。     

木豆种质资源形态与农艺性状的多样性分析

为挖掘优异木豆(Cajanus cajan)种质资源,对10份木豆种质资源的10个质量性状和18个数量性状的遗传多样性进行了研究,并对其农艺性状进行了聚类分析。结果表明,质量性状的遗传多样性指数均较大,以鲜荚色(1.9219)的最高,其次是旗瓣点缀色、鲜籽粒颜色和干籽粒底色,多样性指数均为1.4855;再次为干籽粒色斑、干籽粒脐环色和有无种阜,均为0.8813;最小的是小叶叶形、旗瓣底色和株型,均为0.7219。聚类分析可将10份木豆种质资源划分为中茎稀疏型、中茎密生型和粗茎密生型3大类型。这为木豆品种选育提供了科学依据。     

木豆组织培养与植株再生

植物名称:木豆(Cajanus Cajan)。材料类别:上胚轴、子叶、幼叶。培养条件:诱导愈伤组织的培养基用B_5基本培     

木豆皮、茎、叶成分分析

在雨水充足的高地上,木豆(Cajanus cajan L.)生长9个月即成熟。木豆与黄麻有许多相似之处,它的皮比黄麻薄些,但用水浸泡后,也可制取纤维或用于造纸工业。木豆种子的含水乙醇提取物具有胞毒活性和低血糖活性。木豆叶的水提物可用来治疗黄疸、协助引产以及阻止子宫出血。目前,科研人员已经从木豆的叶、皮和根中提取分离出一些化合物单     

云南栽培种及野生种芋头的AFLP指纹分析

目的:分析云南芋头种质资源遗传多样性。方法:应用扩增片段长度多态性(AFLP)指纹技术,用3对AFLP引物对采集自云南省的9份芋头栽培品种及1份野生品种进行研究,分离AFLP多态性条带。结果:共分离到60个AFLP多态性条带,AFLP多态位点百分率为96.77%,云南芋头种质资源在DNA分子水平上表现出丰富的遗传多样性。聚类分析将10份芋头品种分为2组,遗传距离为0.101～0.908。结论:AFLP指纹技术是筛选品种间差异基因的有效方法,研究结果为云南省芋头品种鉴定、遗传相关性分析、特殊功能基因的分离等工作提供了一定的理论基础。     

树豆叶二苯乙烯类成分的甲醚化和体外抗肿瘤活性

为获得具有抗肿瘤活性的甲醚化先导化合物,对树豆(Cajanus cajan)叶中的二苯乙烯类成分进行甲醚化,并测定其对人肿瘤细胞的细胞毒性。将木豆素C、树豆酮酸A、Cajanotone和木豆素与碘甲烷-碳酸钾反应制备O-甲基产物,通过波谱数据分析产物结构分别鉴定为:2-异戊烯基-3,5-二甲氧基二苯乙烯(1)、树豆酮酸A甲醚(2)、5-O-methylcajanotone(3)和3-O-甲基木豆素(4),其中化合物3是新化合物。肿瘤细胞增殖抑制实验(CCK-8法)结果表明,木豆素C对乳腺癌MDA-MB-231、宫颈癌HeLa、肝癌Hep G2、结肠癌SW480及3种非小细胞肺癌细胞(A549, NCI-H460和NCI-H1299)的半数抑制浓度IC50分别为14.4、16.1、19.6、17.4和25.7～29.6μmol L~(–1);木豆素对宫颈癌和结肠癌之外的5种肿瘤细胞有弱抑制作用,IC50为44.9～78.3μmol L~(–1);而对照的3,4′,5-三甲氧基二苯乙烯(三-O-甲基白藜芦醇)对乳腺癌MDA-MB-231、宫颈癌HeLa、结肠癌SW480和肝癌Hep G2细胞有较强抑制作用(IC50为3.0～14.5μmol L~(–1))。树豆叶二苯乙烯甲醚化衍生物1～4对7种肿瘤细胞系无明显细胞毒活性。     

贵州三叶木通遗传多样性及亲缘关系分析

为科学保存和利用贵州三叶木通资源,采用AFLP和ISSR分子标记技术对贵州省部分县(市)的31份三叶木通资源进行遗传多样性及亲缘关系分析。结果显示:(1)两种标记方法分别通过8对引物扩增出9 252条、11 643条DNA谱带,多态性条带分别为8 818、11 643,扩增多态率分别为95.31%和100%,说明供试种质资源遗传多样性较丰富。(2)AFLP和ISSR方法所得遗传相似系数区间分别为0.762 3~0.893 5和0.828 9~0.917 1,说明不同种质间存在一定的遗传差异。(3)聚类分析表明,在遗传相似系数0.85处,31份三叶木通种质资源分别被聚为5组(AFLP)和3组(ISSR),其中26号(7-4-中1)、27号(8-1-1)、28号(8-3-中)和30号(9-3-中1)4个资源及31号(9-4-1)和29号(9-2-S1)2个资源在两组分析方法中分别处于同一亚组内,说明其为亲缘关系较近的样本,且样本4号(2-2-N1)、8号(3-1-S1)和样本26号(7-4-中1)、27号(8-1-1)两两相似系数最大,可能是同一来源的种质。     

花生种质资源的根系性状研究及AFLP分析

对22份花生种质资源进行根系性状和AFLP分析研究.结果表明,珍珠豆型花生种质资源具有根干重小、根基粗小和主侧根干重比小等特点;龙生型花生种质资源具有根干重大、根基粗大和主侧根干重比大等特点.用根系性状和AFLP标记都能将22份花生品种分为两大组,但是AFLP标记聚类结果比根系性状的聚类结果更能反映花生品种亲源关系和地域生态上的差异.     

111份大薯种质资源遗传多样性的AFLP分析

采用AFLP分子标记方法对收集于6省不同地区的111份大薯种质资源进行遗传多样性分析。筛选到的8个AFLP引物组合扩增到1291个位点,其中1286个是多态性位点。利用多态性信息含量(PIC)、标记指数(MI)和解析强度(RP)分析不同引物组合的标记效率,获得引物的PIC平均值为0.22,MI平均值为35.67,RP平均值为50.50,表明引物扩增位点的高多态性和对大薯种质资源具有强辨别能力,其中引物E-AAC/CAG-M(PIC 0.24、MI 38.56、RP 56.35)具有较高的标记效率。111份大薯种质的遗传相似系数(GS)在0.30~0.82之间,平均为0.58,表明大薯种质资源的遗传相似性较低。采用UPGMA对大薯种质进行聚类分析,遗传相似系数在0.54时,111份材料被划分为4个类群和3个单独的分支,不同地区来源的大薯材料在聚类图中没有明显界限。     

Characterization of Kenyan Isolates of Fusarium udum from Pigeonpea [Cajanus cajan (L.) Millsp.] by Cultural Characteristics, Aggressiveness and AFLP Analysis

Isolates of Fusarium udum from pigeonpea (Cajanus cajan) plants with wilt symptoms were collected from various districts in Kenya and were characterized using cultural characteristics, aggressiveness and amplified fragment length polymorphism (AFLP). The 56 isolates of F. udum showed a high level of variability in aerial mycelia growth, pigmentation and radial mycelia growth (colony diameter) on potato dextrose agar. The aggressiveness of 17 isolates of F. udum on seven pigeonpea varieties varied and five aggressive groups were observed in the present study. There were no relationships among cultural characteristics and aggressiveness. AFLP analysis of the 56 isolates was tested for genetic variability using seven primer combinations. A total of 326 fragments was generated of which 121 were polymorphic. Ten AFLP groups were identified among the Kenyan isolates and, although they were not genetically distinct, six AFLP subgroups were genetically distinct. AFLP had no relationship with cultural characteristics, aggressiveness and geographical origin of the isolates. This is the first report on the study of genetic variability of F. udum using DNA analysis.     

Genetic patterns of domestication in pigeonpea (Cajanus cajan (L.) Millsp.) and wild Cajanus relatives

Pigeonpea (Cajanus cajan) is an annual or short-lived perennial food legume of acute regional importance, providing significant protein to the human diet in less developed regions of Asia and Africa. Due to its narrow genetic base, pigeonpea improvement is increasingly reliant on introgression of valuable traits from wild forms, a practice that would benefit from knowledge of its domestication history and relationships to wild species. Here we use 752 single nucleotide polymorphisms (SNPs) derived from 670 low copy orthologous genes to clarify the evolutionary history of pigeonpea (79 accessions) and its wild relatives (31 accessions). We identified three well-supported lineages that are geographically clustered and congruent with previous nuclear and plastid sequence-based phylogenies. Among all species analyzed Cajanus cajanifolius is the most probable progenitor of cultivated pigeonpea. Multiple lines of evidence suggest recent gene flow between cultivated and non-cultivated forms, as well as historical gene flow between diverged but sympatric species. Evidence supports that primary domestication occurred in India, with a second and more recent nested population bottleneck focused in tropical regions that is the likely consequence of pigeonpea breeding. We find abundant allelic variation and genetic diversity among the wild relatives, with the exception of wild species from Australia for which we report a third bottleneck unrelated to domestication within India. Domesticated C. cajan possess 75% less allelic diversity than the progenitor clade of wild Indian species, indicating a severe "domestication bottleneck" during pigeonpea domestication.     

Differential inhibition of Helicoverpa armigera gut proteinases by proteinase inhibitors of pigeonpea (Cajanus cajan) and its wild relatives

The seeds of 36 pigeonpea [Cajanus cajan (L) Millsp.] cultivars, resistant and susceptible to pests and pathogens and 17 of its wild relatives were analysed for inhibitors of trypsin, chymotrypsin, and insect gut proteinases to identify potential inhibitors of insect (Helicoverpa armigera) gut enzymes. Proteinase inhibitors (PIs) of pigeonpea cultivars showed total inhibition of trypsin and chymotrypsin, and moderate inhibition potential towards H. armigera proteinases (HGP). PIs of wild relatives exhibited stronger inhibition of HGP, which was up to 87% by Rhynchosia PIs. Electrophoretic detection of HGPI proteins and inhibition of HGP isoforms by few pigeonpea wild relative PIs supported our enzyme inhibitor assay results. Present results indicate that PIs exhibit wide range of genetic diversity in the wild relatives of pigeonpea whereas pigeonpea cultivars (resistant as well as susceptible to pests and pathogens) are homogeneous. The potent HGPIs identified in this study need further exploration for their use in strengthening pigeonpea defence against H. armigera.     

Development of simple sequence repeat (SSR) markers for the assessment of gene flow and genetic diversity in pigeonpea (Cajanus cajan)

Pigeonpea (Cajanus cajan) is an important subsistence crop in India where traditional landraces and improved hybrids are grown alongside each other. Gene flow may result in genetic erosion of these landraces and their wild relatives, whilst transgene escape from future genetically engineered varieties is another potential hazard. To assess the impact of these factors gene flow needs to be measured. A set of 10 simple sequence repeat markers have been developed, which exhibit polymorphism across a range of pigeonpea varieties. Use of these markers also offers an efficient system for the assessment of genetic diversity within populations of pigeonpea.     

Genetic fingerprinting of pigeonpea [Cajanus cajan (L.) Millsp.] and its wild relatives using RAPD markers

Randomly amplified polymorphic DNA (RAPD) markers were used for the identification of pigeonpea [Cajanus cajan (L.) Millsp.] cultivars and their related wild species. The use of single primers of arbitrary nucleotide sequence resulted in the selective amplification of DNA fragments that were unique to individual accessions. The level of polymorphism among the wild species was extremely high, while little polymorphism was detected within Cajanus cajan accessions. All of the cultivars and wild species under study could be easily distinguished with the help of different primers, thereby indicating the immense potential of RAPD in the genetic fingerprinting of pigeonpea. On the basis of our data the genetic relationship between pigeonpea cultivars and its wild species could be established.NCL Communication No. 6062     

The first draft of the pigeonpea genome sequence

Pigeonpea (Cajanus cajan) is an important grain legume of the Indian subcontinent, South-East Asia and East Africa. More than eighty five percent of the world pigeonpea is produced and consumed in India where it is a key crop for food and nutritional security of the people. Here we present the first draft of the genome sequence of a popular pigeonpea variety ??Asha??. The genome was assembled using long sequence reads of 454 GS-FLX sequencing chemistry with mean read lengths of >550?bp and >10-fold genome coverage, resulting in 510,809,477?bp of high quality sequence. Total 47,004 protein coding genes and 12,511 transposable elements related genes were predicted. We identified 1,213 disease resistance/defense response genes and 152 abiotic stress tolerance genes in the pigeonpea genome that make it a hardy crop. In comparison to soybean, pigeonpea has relatively fewer number of genes for lipid biosynthesis and larger number of genes for cellulose synthesis. The sequence contigs were arranged in to 59,681 scaffolds, which were anchored to eleven chromosomes of pigeonpea with 347 genic-SNP markers of an intra-species reference genetic map. Eleven pigeonpea chromosomes showed low but significant synteny with the twenty chromosomes of soybean. The genome sequence was used to identify large number of hypervariable ??Arhar?? simple sequence repeat (HASSR) markers, 437 of which were experimentally validated for PCR amplification and high rate of polymorphism among pigeonpea varieties. These markers will be useful for fingerprinting and diversity analysis of pigeonpea germplasm and molecular breeding applications. This is the first plant genome sequence completed entirely through a network of Indian institutions led by the Indian Council of Agricultural Research and provides a valuable resource for the pigeonpea variety improvement.     

Low level of genetic diversity in cultivated Pigeonpea compared to its wild relatives is revealed by diversity arrays technology

Understanding the distribution of genetic diversity among individuals, populations and gene pools is crucial for the efficient management of germplasm collections and breeding programs. Diversity analysis is routinely carried out using sequencing of selected gene(s) or molecular marker technologies. Here we report on the development of Diversity Arrays Technology (DArT) for pigeonpea (Cajanus cajan) and its wild relatives. DArT tests thousands of genomic loci for polymorphism and provides the binary scores for hundreds of markers in a single hybridization-based assay. We tested eight complexity reduction methods using various combinations of restriction enzymes and selected PstI/HaeIII genomic representation with the largest frequency of polymorphic clones (19.8%) to produce genotyping arrays. The performance of the PstI/HaeIII array was evaluated by typing 96 accessions representing nearly 20 species of Cajanus. A total of nearly 700 markers were identified with the average call rate of 96.0% and the scoring reproducibility of 99.7%. DArT markers revealed genetic relationships among the accessions consistent with the available information and systematic classification. Most of the diversity was among the wild relatives of pigeonpea or between the wild species and the cultivated C. cajan. Only 64 markers were polymorphic among the cultivated accessions. Such narrow genetic base is likely to represent a serious impediment to breeding progress in pigeonpea. Our study shows that DArT can be effectively applied in molecular systematics and biodiversity studies.Electronic Supplementary Material Supplementary material is available to authorised users in the online version of this article at .     

An intra-specific consensus genetic map of pigeonpea [Cajanus cajan (L.) Millspaugh] derived from six mapping populations

Pigeonpea (Cajanus cajan L.) is an important food legume crop of rainfed agriculture. Owing to exposure of the crop to a number of biotic and abiotic stresses, the crop productivity has remained stagnant for almost last five decades at ca. 750?kg/ha. The availability of a cytoplasmic male sterility (CMS) system has facilitated the development and release of hybrids which are expected to enhance the productivity of pigeonpea. Recent advances in genomics and molecular breeding such as marker-assisted selection (MAS) offer the possibility to accelerate hybrid breeding. Molecular markers and genetic maps are pre-requisites for deploying MAS in breeding. However, in the case of pigeonpea, only one inter- and two intra-specific genetic maps are available so far. Here, four new intra-specific genetic maps comprising 59-140 simple sequence repeat (SSR) loci with map lengths ranging from 586.9 to 881.6?cM have been constructed. Using these four genetic maps together with two recently published intra-specific genetic maps, a consensus map was constructed, comprising of 339 SSR loci spanning a distance of 1,059?cM. Furthermore, quantitative trait loci (QTL) analysis for fertility restoration (Rf) conducted in three mapping populations identified four major QTLs explaining phenotypic variances up to 24?%. To the best of our knowledge, this is the first report on construction of a consensus genetic map in pigeonpea and on the identification of QTLs for fertility restoration. The developed consensus genetic map should serve as a reference for developing new genetic maps as well as correlating with the physical map in pigeonpea to be developed in near future. The availability of more informative markers in the bins harbouring QTLs for sterility mosaic disease (SMD) and Rf will facilitate the selection of the most suitable markers for genetic analysis and molecular breeding applications in pigeonpea.     

Genetics of a new male-sterility locus in pigeonpea (Cajanus cajan [L.] Millsp.).

A natural male-sterile mutant was found in the population of a short-duration pigeonpea (Cajanus cajan[L.] Millsp.) cultivar ICPL 85010. This mutant is characterized by light yellow anthers of reduced size that are devoid of pollen grains. This mutant was crossed with two pigeonpea cultivars to study its inheritance. The F1, F2, and test cross data of the two crosses suggested that this male sterility trait is genetic in origin and is controlled by a single recessivegene. The F1 (mutant x ICPL 85010) plants were crossed with translucent (ms1) and arrowhead type (ms2) genetic male steriles reported earlier to study their allelic relationships. Segregation in the three-way cross F1 and F2 populations revealed that the mutant male-sterile gene was nonallelic to ms1 and ms2 loci and it is designated ms3. The new male sterility sources in pigeonpea will help in producing high-yielding hybrids and populations in diverse phenological groups.