Genetic Diversity Associated with Eco-geographical Parameters, Morphological Characteristics and Soil Analyzes of Common Vetch (Vicia sativa L.) in Algeria

Background: In the present study, we have set a goal, to study the genetic variability and phylogenetic affinities of 26 spontaneous Vicia sativa L. accessions collected in Algeria, as part of the recovery and enhancement of the local genetic heritage and its integration into the breeding and varietal creation programs. Methods: Morphological traits, performance parameters and phenological stages of each accession were tracked during an experimental test in pots under greenhouse conditions from December 29, 2019 to June 1, 2020. An eco-geographical characterization of samples was also carried out through geographical coordinates. On the other side and for the first time in Algeria, soil analyses of each site were undertaken. Result: The phylogenetic study of accessions confirm the taxonomic complexity of the group V. sativa L. According to the principal component analyses and kruskal-Wallis test, the most segregating variables with the highest values and the largest K variance ratios are most of the attributes associated with the flower, pod and seed. The association of the two dissimilarity matrix carried out by a Mantel test between the morphological and eco-geographic parameters showed a significant effect with a p value = 0.004 at the threshold of alpha=0.05 and a correlation coefficient r (AB) = 0.156.

An Efficient Root Transformation System for Recalcitrant Vicia sativa

Common vetch (Vicia sativa) is a multi-purpose legume widely used in pasture and crop rotation systems. Vetch seeds have desirable nutritional characteristics and are often used to feed ruminant animals. Although transcriptomes are available for vetch, problems with genetic transformation and plant regeneration hinder functional gene studies in this legume species. Therefore, the aim of this study was to develop a simple, efficient and rapid hairy root transformation system for common vetch to facilitate functional gene analysis. At first, we infected the hypocotyls of 5-day-old in vitro or in vivo, soil-grown seedlings with Rhizobium rhizogenes K599 using a stabbing method and produced transgenic hairy roots after 24 days at 19 and 50% efficiency, respectively. We later improved the hairy root transformation in vitro by infecting different explants (seedling, hypocotyl-epicotyl, and shoot) with R. rhizogenes. We observed hairy root formation at the highest efficiency in shoot and hypocotyl-epicotyl explants with 100 and 93% efficiency, respectively. In both cases, an average of four hairy roots per explant were obtained, and about 73 and 91% of hairy roots from shoot and hypocotyl-epicotyl, respectively, showed stable expression of a co-transformed marker β-glucuronidase (GUS). In summary, we developed a rapid, highly efficient, hairy root transformation method by using R. rhizogenes on vetch explants, which could facilitate functional gene analysis in common vetch.

Determination of salt tolerance levels and genetic relationships of Vicia sativa cultivars using gene targeted functional markers

The objectives of the present study were to determine salt tolerance levels of 12 different common vetch (Vicia sativa L.) cultivars at germination stage in the presence of 250 mM NaCl and to reveal genetic relationships based on gene targeted functional markers (GTFMs) associated with salt tolerance. The results revealed the presence of a significant genetic variation among the cultivars although s alt stress significantly reduced all germination parameters tested. The cultivar Ozveren was the most salt tolerant with 20.1% reduction in final germination percentage compared to control seeds while cultivars Alınoglu, Ayaz and Bakir did not germinate. The maximum delays in germination rate (G50 = 3.78 days) and synchrony (G10-90 = 3.45 days) were obtained from the cultivars Urkmez and Ozveren, respectively. The GTFMs provided a total of 53.1% polymorphism. The primers of MtSOS2 gene gave the highest numbers of alleles per primer pair while the highest polymorphism rate (77.8%) was obtained from the MtP5CS gene. The first three components of principal component analysis explained 57.63% of total variation. This study concluded that the cultivars determined to be salt tolerant and sensitive at germination stage distributed into three main clades determined by UPGMA analysis while the GTFMs associated with salt tolerance successfully determined the genetic relationships of common vetch cultivars.

Potential of Sunflower-Legume Intercropping: A Way Forward in Sustainable Production of Sunflower in Temperate Climatic Conditions

The ongoing climate change, coupled with the transformations of cultivation practices and land use in sole crop production, may cause a significant decline in sunflower yield stability. Considering that the sunflower is the third oilseed in the world oilseed market, with 45 million tons of grain per year, and that it takes the fourth position in the vegetable oil market, the implementation of sustainable growing technology in sunflower production is a necessity. Considering the above, the goal of the research was to analyze and recommend sustainable sunflower production technology in intercropping systems. A four-year trial was conducted in Serbia’s agroecological conditions (45∘34′23.2′′ N 19∘86′18.9′′ E) using a split-plot design. Two oil-type hybrids and one confectionary-type hybrid were intercropped with common vetch, red clover, and alfalfa. Common vetch × sunflower intercropping resulted in the decrease in almost all sunflower traits. Moreover, sunflower × alfalfa intercropping proved to be the most appropriate. The yield of NS Gricko and Rimi PR were statistically on the same level with sole cropping, while alfalfa biomass had better results after alfalfa was intercropped with NS Gricko, as compared to sole cropping. In view of the prevailing belief that yields are more stable in intercropping than in sole cropping, further research is needed in this respect, in addition to further research of the time and method of sowing.

Genome survey sequencing of common vetch (Vicia sativa L.) and genetic diversity analysis of Chinese germplasm with genomic SSR markers

Background Common vetch (Vicia sativa L.) is an annual legume with excellent suitability in cold and dry regions. Despite its great applied potential, the genomic information regarding common vetch currently remains unavailable. Methods and results In the present study, the whole genome survey of common vetch was performed using the next-generation sequencing (NGS). A total of 79.84 Gbp high quality sequence data were obtained and assembled into 3,754,145 scaffolds with an N50 length of 3556 bp. According to the K-mer analyses, the genome size, heterozygosity rate and GC content of common vetch genome were estimated to be 1568 Mbp, 0.4345 and 35%, respectively. In addition, a total of 76,810 putative simple sequence repeats (SSRs) were identified. Among them, dinucleotide was the most abundant SSR type (44.94%), followed by Tri- (35.82%), Tetra- (13.22%), Penta- (4.47%) and Hexanucleotide (1.54%). Furthermore, a total of 58,175 SSR primer pairs were designed and ten of them were validated in Chinese common vetch. Further analysis showed that Chinese common vetch harbored high genetic diversity and could be clustered into two main subgroups. Conclusion This is the first report about the genome features of common vetch, and the information will help to design whole genome sequencing strategies. The newly identified SSRs in this study provide basic molecular markers for germplasm characterization, genetic diversity and QTL mapping studies for common vetch.

Chromosome-level assembly of the common vetch reference genome (Vicia sativa)

Background: Vicia sativa L. (Common Vetch, n = 6) is an annual, herbaceous, climbing legume that is distributed in tropical, sub-tropical and temperate climates. Originating in the Fertile Crescent of the Middle East, V. sativa is now widespread and grows in the Mediterranean basin, West, Central and Eastern Asia, North and South America. V. sativa is of economic importance as a forage legume in countries such as Australia, China, and the USA and contributes valuable nitrogen to agricultural rotation cropping systems. To accelerate precision genome breeding and genomics-based selection of this legume, we here present a chromosome-level reference genome sequence for V. sativa. Results: We applied a combination of long-read Oxford Nanopore sequencing, short-read Illumina sequencing, and high-throughput chromosome conformation data (CHiCAGO and Hi-C) analysis to construct a chromosome-level genome of V. sativa. The chromosome-level assembly of six pseudo-chromosomes has a total genome length of 1.9 gigabases (Gb) with a median contig length of 684 kb. Benchmarking Universal Single-Copy Orthologs (BUSCO) of the assembly demonstrated a very high completeness of 98 % of the dicotyledonous orthologs. RNA-seq analysis and gene modelling enabled the annotation of 58,415 protein-coding genes. Conclusions: The high-quality chromosome-level genome assembly of V. sativa will provide novel insights into vetch genome evolution and be a valuable resource for genomic breeding, genetic diversity and for understanding adaption to diverse arid environments.

Potential of Sunflower-Legume Intercropping: A Way Forward in Sustainable Production of Sunflower in Temperate Climatic Conditions

Changing climate conditions coupled with the transformations of cultivation practices and land use in sole crop-based sunflower production may significantly decline yield stability of this oilseed crop. Given that sunflower takes the third place in the world oilseed market, with 45 million tons per year, and in the fourth place in vegetable oil production, it is necessary to adapt production technologies toward sustainable agriculture. Considering that, the goal of the research was to analyze and beneficial sustainable production technology of sunflower in intercropping systems. A four-year trial was conducted in Serbia’s agroecological rain-fed conditions (45°34’23.2"N 19°86’18.9"E) using a split-plot design. Two oil types and one confectionary sunflower hybrid were intercropped with common vetch, red clover and alfalfa. Analyses showed that intercropping of sunflower with common vetch resulted in the decrease in almost all sunflower trait values. Also, sunflower × alfalfa intercropping provided to be the most appropriate. The yield of NS Gricko and Rimi PR were statistically on the same level with sole cropping, while alfalfa biomass had better results when intercropped with NS Gricko as compared to sole cropping. Concerning the general belief that yields are more stable in intercropping than in sole crop, further research in this respect is needed, in addition to the research of time and method of sowing.

Physiological Adaptation And Transcriptome Changes In The Plateau Plant Common Vetch (Vicia Sativa L.) In Response To The Plain Environment

Common vetch (Vicia sativa L.) is an annual herb with high nutritional value, strong adaptability and cold tolerance. It is one of the forage varieties widely planted in the construction of artificial grassland in Qinghai Tibet Plateau. In order to reveal the molecular regulation mechanism of common vetch introduced into plain, physiological and transcriptome analysis of common vetch seedlings in Plateau and plain environment were carried out. In the plain environment, the leaf structure and some physiological indexes of common vetch can adapt to the plain environment gradually and keep stable. However, the maximum photochemical quantum yield (fv/fm) and chlorophyll content (Chl) of PSII fluctuated and could not keep stable. Further transcriptome sequencing showed that there were many different genes involved in photosynthesis pathway, photosynthetic antenna protein pathway, carbon dioxide fixation pathway in photosynthetic organisms and porphyrin and chlorophyll metabolism pathway in plain environment. Similarly, TF analysis showed that MYB, NAC, AP2-EREBP and Orphans were the main transcription factors involved in the adaptation of common vetch to the changes of plain environment. These results may explain that the main reason why the common vetch is not suitable for the plain environment is the difference of the light intensity between the plain and the plateau. These findings provide a theoretical basis for scientific introduction and breeding of new varieties from plateau to plain.