Medicago Sativa Complex Research Articles

Karyotypic evolution of the Medicago complex: sativa-caerulea-falcata inferred from comparative cytogenetic analysis

BackgroundPolyploidy plays an important role in the adaptation and speciation of plants. The alteration of karyotype is a significant event during polyploidy formation. The Medicago sativa complex includes both diploid (2n = 2× = 16) and tetraploid (2n = 2× = 32) subspecies. The tetraploid M. ssp. sativa was regarded as having a simple autopolyploid origin from diploid ssp. caerulea, whereas the autopolyploid origin of tetraploid ssp. falcata from diploid form ssp. falcata is still in doubt. In this study, detailed comparative cytogenetic analysis between diploid to tetraploid species, as well as genomic affinity across different species in the M. sativa complex, were conducted based on comparative mapping of 11 repeated DNA sequences and two rDNA sequences by a fluorescence in situ hybridization (FISH) technique.ResultsFISH patterns of the repeats in diploid subspecies caerulea were highly similar to those in tetraploid subspecies sativa. Distinctly different FISH patterns were first observed in diploid ssp. falcata, with only centromeric hybridizations using centromeric and multiple region repeats and a few subtelomeric hybridizations using subtelomeric repeats. Tetraploid subspecies falcata was unexpectedly found to possess a highly variable karyotype, which agreed with neither diploid ssp. falcata nor ssp. sativa. Reconstruction of chromosome-doubling process of diploid ssp. caerulea showed that chromosome changes have occurred during polyploidization process.ConclusionsThe comparative cytogenetic results provide reliable evidence that diploid subspecies caerulea is the direct progenitor of tetraploid subspecies sativa. And autotetraploid ssp. sativa has been suggested to undergo a partial diploidization by the progressive accumulation of chromosome structural rearrangements during evolution. However, the tetraploid subspecies falcata is far from a simple autopolyploid from diploid subspecies falcata although no obvious morphological change was observed between these two subspecies.

Variation in Crown and Root Organic Reserves Among Lucerne Genotypes of Different Morphology and Flower Colour

AbstractPrevious evidence indicates that differences in the concentration of underground organic reserves can drive the survival and growing ability of lucerne under cold and defoliation stresses. In order to provide the selection process with further information on compounds that may influence plant performance under grazing, we assessed variations in cold‐season concentrations of nitrogen and carbon reserves on genotypes that had been identified for morphological features that possibly enhance grazing tolerance. The selected genotypes encompassed distinct morphological patterns (defined as ‘models’) and different taxa within the Medicago sativa complex, as shown by different flower colours. Crown concentrations of reserves were determined on 90 genotypes, whereas root concentrations were measured on a subsample of 15 genotypes. Wide intergenotypic variation was observed for all reserve substances. Comparisons among models and among flower colour classes highlighted the high concentrations of crown carbohydrates and root and crown‐soluble proteins of the model coded as ‘D1’, characterized by prostrate, rhizomatous habit and long dormancy, which largely corresponded to plants with yellow or variegated flowers, typical of ssp. falcata and × varia, respectively. There was a strong ‘flower colour × storage organ’ interaction for sugar concentration, and the results suggested a preferential compartment of sugars in the roots of purple‐flowered genotypes that belonged to the ssp. sativa. A rank correlation analysis indicated a positive relationship between persistence after two years under grazing of half‐sib progenies deriving from 19 genotypes out of the 90 and crown concentrations of carbohydrates of the 19 mother plants.

Volatiles from Medicago sativa complex flowers

Volatiles from flowers of several lucerne ( Medicago sativa complex) genotypes, belonging to different taxonomic groups and characterised by various flower colours, were isolated by Tenax-trapping and analysed by GC and GC-mass spectrometry. Thirty-three compounds were identified, including aldehydes, alcohols, esters, ketones, terpenes and furanoids. Trans-2-hexenal was the most abundant volatile in all genotypes. Ranking of individual volatile compounds was similar in all genotypes but absolute values (μg g −1 for each compound varied widely from genotype to genotype, being largely associated with flower colour intensity. In terms of total volatile quantity, the following ranking of flower colours was observed: dark green & violet > bright or light yellow. © 1997 Elsevier Science Ltd. All rights reserved

Cytological studies of the nucleolus organizing regions in the Medicago complex: sativa–coerulea–falcata

A cytological examination of the nucleolus organizing regions (NORs) of three species from the Medicago sativa complex was conducted to evaluate the structural and functional evolution of the ribosomal RNA (rRNA) loci that encode the 18S, 5.8S, and 26S rRNAs. Mitotic chromosomes in root-tip preparations from tetraploid M. sativa and diploids Medicago coerulea and Medicago falcata were visualized by four methods that provide new data. Fluorescent in situ hybridization using the M. sativa 18S gene as probe localized the structural rDNA to the constricted regions of the satellited chromosomes only. Chromomycin A3 (CMA3) staining and 4′,6-diamidino-2-phenylindole (DAPI) staining identified these chromosomal segments as the most GC-rich regions in the alfalfa karyotype. Medicago falcata exhibited fewer DAPI bands and chromocenters than did M. sativa and M. coerulea. Positive silver nitrate staining showed that all four rDNA regions in M. sativa (located in two chromosome pairs) and both rDNA sites in both diploid species remain transcriptionally active. Counts of nucleoli confirmed that all rDNA regions are independently capable of nucleolus organization. Thus, the number of active NORs in M. sativa is double the number found in M. coerulea or M. falcata. Consequently, if M. sativa originated from sexual hybridization of 2n gametes involving one or both diploid species, no major reorganization or loss of structural or functional rDNA loci has occurred. Key words : alfalfa evolution, CMA3 banding, DAPI banding, fluorescent in situ hybridization, silver nitrate staining.

Evaluation of a lucerne germplasm collection in relation to traits conferring grazing tolerance

A collection of 90 accessions of different germplasm groups from the Medicago sativa complex was evaluated at Lodi, Italy, for dry-matter yield, time to anthesis, height of regrowth, plant diameter, stem density, vegetative recovery after winter, and presence of deep-set crown and ability to spread with creeping roots or rhizomes, these latter characters possibly contributing to grazing tolerance. The germplasm groups proved rather distinct for these traits, but all included types potentially suitable for grazing. Interesting types combining deep-set crown and high yield occurred mainly in subsp. x varia and materials originated from artificial crosses involving at various extent the subspecies falcata, sativa and x varia. Though belonging to subsp. sativa, “mielga” wild populations from Spain had aerial traits similar to those of the long-dormant, low-yielding subsp. falcata. Deep-crowned plants had greater plant diameter than shallow-crowned plants, and could be categorised into four distinct top-growth models according to their habit and vigour. One of these models combined spreading ability with high yield. The about 150 plants selected for their combination of deep crown, wide diameter, and good yield were characterised by a branch-rooted system. There was general correspondence between top-growth and root morphology of each model. Of the four deep-crowned models, three had rhizomes, with wide range of underground development and morphology, while the fourth had creeping roots. The latter model was the most rare among the examined accessions, never appearing in either cultivated or wild subsp. sativa. The implications of the results on the development of grazing-tolerant lucerne varieties are discussed.

Genetic variation for disease and nematode resistances and forage quality in perennial diploid and tetraploid lucerne populations (Medicago sativa L.)

Twenty-five lucerne populations of the Medicago sativa complex, which were either diploid or tetraploid and wild or cultivated, were analysed for their resistance to four different fungal diseases and to stem nematode. Forage quality, including stem digestibility and saponin content, was also tested. Populations varied in susceptibility to the diseases caused by Colletotrichum trifolii, Verticillium albo-atrum, Sclerotinia trifoliorum and Pseudopezizza medicaginis, and to the nematode Ditylenchus dipsaci. Except for Sclerotinia rot, sativa and falcata subspecies differed in susceptibility, but this grouping of populations did not account for the full range of variation among them. However, the resistance to P. medicaginis was much lower in the sativa than in the falcata populations. Populations also varied significantly in stem fiber content and digestibility. Stem digestibility was negatively correlated to forage yield. Wild sativa and falcata populations had lower fiber content and higher digestibility than cultivated sativa populations. The medicagenic acid was the sapogenin responsible for the anti-nutritional effect of the lucerne measured by the yellow mealworm larvae Tenebrio molitor. The medicagenic acid content was lowest for the pure sativa populations, highest for the pure falcata populations, and intermediate for the French sativa varieties that have some traits originating from falcata germplasm. Some populations could be used in breeding programs to improve disease and nematode resistance, and forage quality.

Characterization of Meiotic Mutants Producing 4N Pollen and 2N Eggs in the Medicago Sativa Complex

Characterization of Meiotic Mutants Producing 4N Pollen and 2N Eggs in the Medicago Sativa Complex

Chromosome numbers of the Medicago sativa complex in Turkey

Turkish representatives of the Medicago sativa complex comprise three basic morphological classes distinguishable by the following: (1) purple flowers, coiled pods; (2) yellow flowers, uncoiled pods; (3) intermediates and recombinants between 1 and 2. Chromosome determinations were made for 329 plants representing 35 cultivated (alfalfa) and 87 wild populations. All cultivars collected fell into class 1 and were tetraploid (2n = 4x = 32). Wild plants representative of all three morphological classes were found at both the diploid (2n = 2x = 16) and tetraploid (2n = 4x = 32) level, and examples of hybrid zones between classes 1 and 2 were common at both levels. Most populations appeared to contain plants at the same ploidy level, but a few contained a mixture of both diploids and tetraploids, and at several locations diploid and tetraploid populations grew adjacent to each other. Tetraploids were widespread across Turkey whereas diploids were confined to the northeast region.