The genus Medicago, with about 87 species, includes the model legume species M. truncatula, and a number of important forage species such as M. sativa (alfalfa), M. scutellata (snail medic), and M. lupulina (black medic). Relationships within the genus are not yet sufficiently resolved, contributing to difficulty in understanding the evolution of a number of distinguishing characteristics such as aneuploidy and polyploidy, life history, structure of cotyledons, and number of seeds per fruit. •Phylogenetic relationships of 70-73 species of Medicago and its sister genus Trigonella (including Melilotus) were reconstructed from nucleotide sequences of the plastid trnK/matK region and the nuclear-encoded GA3ox1 gene (gibberellin 3-β-hydroxylase) using maximum parsimony and Bayesian inference methods. •Our results support certain currently recognized taxonomic groups, e.g., sect. Medicago (with M. sativa) and sect. Buceras. However, other strongly supported clades-the "reduced subsection Leptospireae clade" that includes M. lupulina, the "polymorpha clade" that includes M. murex and M. polymorpha and the "subsection Pachyspireae clade" that includes M. truncatula-each of which includes species presently in different subsections of sect. Spirocarpos, contradict the current classification. •These results support the hypothesis that some characters considered important in existing taxonomies, for example, single-seeded fruits that have arisen more than once in both Medicago and Trigonella, are indeed homoplastic. Others, such as the 2n = 14 chromosome number, have also arisen independently within the genus. In addition, we demonstrate support for the utility of GA3ox1 sequences for phylogenetic analysis among and within closely related genera of legumes.

为了了解不同耐盐紫花苜蓿（Medicago sativa L.）的遗传多样性以及对耐盐紫花苜蓿品种改良，本试验筛选出9对简单重复序列（Simple Sequence Repeats，SSR）引物对10份耐盐、敏盐材料进行SSR多态性分析。并利用MEGA6.0软件对其进行了聚类分析。结果表明：10份紫花苜蓿品种中平均多态位点百分率为47.1%~71.8%；平均有效等位基因观察数与等位基因观察数的比为0.680~0.890；平均杂合度为0.759~0.828；平均香农指数为1.753~1.976；平均多态信息含量为0.735~0.811；遗传分化系数为0.100。利用IBM SPSS Statistics 19软件进行综合评价，根据综合评价值对10个紫花苜蓿品种内的遗传变异程度大小排序，排列顺序为： "巨能" > "骑士T" > "巨能7号" > "Asi" > "兴平" > "阿迪娜" > "秘鲁" > "抗旱15" > "草原3号" > "国产苜蓿"；利用MEGA6.0软件对Nei遗传距离进行UPGMA聚类分析可知：5份敏盐材料始终聚为一类；"巨能"与"巨能7号"聚为一类；"阿迪娜"和"抗旱15"又聚为一类，说明他们之间有较近的亲缘关系。

分别以早熟低产和晚熟高产苜蓿单株为父母本,通过人工杂交构建了四倍体紫花苜蓿(Medicago Sativa)F1遗传作图群体,采用单因子变量分析法,以降落式PCR和常规PCR结合的反应程序,建立了适宜于紫花苜蓿的分子标记扩增体系;应用130对SSR引物进行筛选,获得60对引物在父母本间存在多态性而被用于绘制遗传连锁图。采用PAGE电泳分析,对作图群体进行基因型分析。通过TetraploidMap软件对60个SSR标记进行连锁作图分析,有44个标记可以定位在8个连锁群上,占总标记数的33.8%,覆盖遗传距离979 cM,两标记间平均图距为22.25 cM,初步构建了四倍体紫花苜蓿遗传图谱的框架图,还需要进一步添加标记数量增大其饱和度,为重要性状的QTL定位奠定基础。

Expressed sequence tags (ESTs) are important resources for gene discovery and molecular marker development. From over 147,000 ESTs of Medicago truncatula, we have identified 4,384 ESTs containing perfect simple sequence repeats (EST-SSR) of di-, tri-, tetra- or pentanucleotides. Six hundred sixteen primer pairs (PPs) were designed and screened over a panel of eight genotypes representing six Medicago spp. and subspecies. Nearly, 74% (455) of the PPs produced characteristic SSR bands of expected size length in at least one Medicago species. Four hundred six (89%) of these 455 PPs produced SSR bands in all eight genotypes tested. Only 17 PPs were M. truncatula -specific. High levels of polymorphism (>70%) were detected for these markers in alfalfa, M. truncatula, and other annual medics. About 48% of the reported markers are part of gene transcripts linked to putative functions. Our results indicate that the SSR markers developed from M. truncatula ESTs are valuable genetic markers for the Medicago genus. These markers will be useful in establishing the genomic relationships of M. truncatula to important forage legume crops such as alfalfa and other annual medics.

Medicago species are important genetic sources for forage crops and nitrogen sources for various ecosystems. The ongoing genome sequencing of the model legume, Medicago truncatula, provides a wealth of genetic markers potentially useful for characterizing the population genetic structure and evolutionary history, and the potential of the wild Medicago species. Here we tested the PCR amplification of 92 microsatellites developed from M. truncatula in six other Medicago species, and found that the cross-species transferability, ranging from 53.26% to 61.96%, is comparable with those reported in other angiosperm genera. This article thus reports a number of microsatellites that are potentially useful for large-scale ecological and evolutionary genetic studies of wild Medicago species.© 2009 Blackwell Publishing Ltd.

Advances in deciphering the functional architecture of eukaryotic genomes have been facilitated by recent breakthroughs in sequencing technologies, enabling a more comprehensive representation of genes and repeat elements in genome sequence assemblies, as well as more sensitive and tissue-specific analyses of gene expression. Here we show that PacBio sequencing has led to a substantially improved genome assembly of Medicago truncatula A17, a legume model species notable for endosymbiosis studies, and has enabled the identification of genome rearrangements between genotypes at a near-base-pair resolution. Annotation of the new M. truncatula genome sequence has allowed for a thorough analysis of transposable elements and their dynamics, as well as the identification of new players involved in symbiotic nodule development, in particular 1,037 upregulated long non-coding RNAs (lncRNAs). We have also discovered that a substantial proportion (~35% and 38%, respectively) of the genes upregulated in nodules or expressed in the nodule differentiation zone colocalize in genomic clusters (270 and 211, respectively), here termed symbiotic islands. These islands contain numerous expressed lncRNA genes and display differentially both DNA methylation and histone marks. Epigenetic regulations and lncRNAs are therefore attractive candidate elements for the orchestration of symbiotic gene expression in the M. truncatula genome.

Alfalfa (Medicago sativa) is one of the most important forage crops in the world; however, its molecular genetics and breeding research are hindered due to the lack of a high-quality reference genome. Here, we report a de novo assembled 816-Mb high-quality, chromosome-level haploid genome sequence for 'Zhongmu No.1' alfalfa, a heterozygous autotetraploid. The contig N50 is 3.92 Mb, and 49 165 genes are annotated in the genome. The alfalfa genome is estimated to have diverged from M. truncatula approximately 8 million years ago. Genomic population analysis of 162 alfalfa accessions revealed high genetic diversity, weak population structure, and extensive gene flow from wild to cultivated alfalfa. Genome-wide association studies identified many candidate genes associated with important agronomic traits. Furthermore, we showed that MsFTa2, a Flowering Locus T homolog, whose expression is upregulated in salt-resistant germplasms, may be associated with fall dormancy and salt resistance. Taken together, these genomic resources will facilitate alfalfa genetic research and agronomic improvement.Copyright © 2020 The Author. Published by Elsevier Inc. All rights reserved.

Microsatellites are a widely-used marker system in plant genetics and forensics. The development of reliable microsatellite markers from resequencing data is challenging.We extended MISA, a computational tool assisting the development of microsatellite markers, and reimplemented it as a web-based application. We improved compound microsatellite detection and added the possibility to display and export MISA results in GFF3 format for downstream analysis.MISA-web can be accessed under http://misaweb.ipk-gatersleben.de/. The website provides tutorials, usage note as well as download links to the source code.scholz@ipk-gatersleben.de.© The Author(s) 2017. Published by Oxford University Press.

Expressed sequence tag (EST) projects have generated a vast amount of publicly available sequence data from plant species; these data can be mined for simple sequence repeats (SSRs). These SSRs are useful as molecular markers because their development is inexpensive, they represent transcribed genes and a putative function can often be deduced by a homology search. Because they are derived from transcripts, they are useful for assaying the functional diversity in natural populations or germplasm collections. These markers are valuable because of their higher level of transferability to related species, and they can often be used as anchor markers for comparative mapping and evolutionary studies. They have been developed and mapped in several crop species and could prove useful for marker-assisted selection, especially when the markers reside in the genes responsible for a phenotypic trait. Applications and potential uses of EST-SSRs in plant genetics and breeding are discussed.

Repetitive sequences have been proposed to be recombinogenic elements in eukaryotic chromosomes. We tested whether dinucleotide repeats sequences are preferential sites for recombination because of their high affinity for recombination enzymes. We compared the kinetics of the binding of the scRad51, hsRad51 and ecRecA proteins to oligonucleotides with repeats of dinucleotides GT, CA, CT, GA, GC or AT. Since secondary structures in single-stranded DNA (ssDNA) act as a barrier to complete binding we measured whether these oligonucleotides are able to form stable secondary structures. We show that the preferential binding of recombination proteins is conserved among the three proteins and is influenced mainly by secondary structures in ssDNA.