Evolutionary history of an Irano-Turanian cushion-forming legume (Onobrychis cornuta).

The Irano-Turanian region is one of the largest floristic regions in the world and harbors a high percentage of endemics, including cushion-like and dwarf-shrubby taxa. Onobrychis cornuta is an important cushion-forming element of the subalpine/alpine flora of the Irano-Turanian floristic region. To specify the genetic diversity among the populations of this species (including individuals of O. elymaitica), we employed nrDNA ITS and two noncoding regions of plastid DNA (rpl32-trnL(UAG) and trnT(UGU)-trnL(UAA)). The most striking feature of O. cornuta assemblages was the unexpectedly high nucleotide diversity in both the nDNA and cpDNA dataset. In the analyses of nuclear and plastid regions, 25 ribotypes and 42 haplotypes were found among 77 and 59 accessions, respectively, from Iran, Turkey, and Afghanistan. Network analysis of the datasets demonstrated geographic differentiation within the species. Phylogenetic analyses of all dataset retrieved O. cornuta as a non-monophyletic species due to the inclusion of O. elymaitica, comprising four distinct lineages. In addition, our analyses showed cytonuclear discordance between both nuclear and plastid topologies regarding the position of some O. cornuta individuals. The underlying causes of this inconsistency remain unclear. However, we speculate that chloroplast capture, incomplete lineage sorting, and introgression were the main reasons for this event. Furthermore, molecular dating analysis indicated that O. cornuta originated in the early Pliocene (around 4.8 Mya) and started to diversify throughout the Pliocene and in particular the Pleistocene. Moreover, O. elymaitica was reduced to a subspecific rank within the species.

In vitro Evaluation of Skin Related Enzyme Inhibitory Effect and Emulgel Formulation Development Studies of Onobrychis Argyrea subsp. Argyrea with Phytochemical Analysis.

Species of Onobrychis have been used to treat skin disorders such as wounds and cuts in folk medicine and Onobrychis argyrea subsp. argyrea (OA) commonly known as 'silvery sainfoin', is a member of this genus. In this study, it was aimed to investigate the skin-related biological activities and phytochemical characterization of OA. Moreover, an emulgel formulation was developed from the main methanolic extract of the plant (OAM). Initially, to identifiy of the active fractions, aerial parts of the plant material was extracted with methanol and fractionated by n-hexane, chloroform, ethyl acetate and n-butanol, respectively. Antioxidant activity was determined by CUPRAC, TOAC, FRAP and DPPH assays. Thereafter, the inhibition potential of OAM, novel formulation and all fractions was measured against elastase, collagenase, tyrosinase and hyaluronidase enzymes. OAM was analyzed and characterized by LC/MS-MS. The major bioactive flavonoids which are rutin and isoquercetin were measured and compared as qualitative and quantitative via high performance thin layer chromatography (HPTLC) analysis in OAM and fractions. The results showed that extracts of OA can be a potential cosmeceutical agent for skin related problems.

Inclusion of Sainfoin in the Concentrate of Finishing Lambs: Fatty Acid Profiles of Rumen, Plasma, and Muscle.

The effects of sainfoin inclusion (Onobrychis viciifolia) in the finishing concentrate for light lambs on the fatty acid (FA) composition of the ruminal digesta, plasma, and meat were evaluated. Twenty-six weaned male lambs were divided into three groups and fed individually ad libitum for 40 days with one of three concentrates differing in the level of sainfoin inclusion: 0% (0SF), 20% (20SF), and 40% (40SF). The rumen digesta showed an increase in C18:3 n-3 concentration and a decrease in C18:1 t10 concentration when sainfoin was included in the concentrate regardless of the level of inclusion. However, the highest C18:1 t11 and the lowest C18:2 n-6 proportions were obtained only in the 40SF rumen, showing a stronger t11 biohydrogenation pathway. In plasma, most effects were associated with changes in the levels of polyunsaturated FA (PUFA) n-3. The meat FA profile of 40SF lambs presented higher percentages of PUFA n-3 and CLA c9,t11 and a lower PUFA n-6/PUFA n-3 ratio compared with those from 0SF and 20SF diets because of the potentiation of the ruminal t11 pathway. Inclusions of 20 and 40% sainfoin both showed beneficial effects on meat quality; furthermore, these effects were most marked in the 40% sainfoin diet.

Chemical Composition and In Vitro Evaluation of Antioxidant and Antiproliferative Effects of Volatile Oils Hydrodistilled from Onobrychis carduchorum C.C. Towns., a Kurdish Traditional Plant.

The volatile oils hydrodistilled from the aerial parts and roots of O. carduchorum C.C Towns. (Fabaceae) have been chemically characterized for the first time. A total of 43 constituents with an abundance >0.03% were identified and quantified in the two oils by GC/MS and GC/FID analyses. They comprise 38 components (98.58%) of the oil isolated from the aerial parts (OCA) and 34 components (93.33%) of the oil from the roots (OCR). Six constituents, α-pinene (23.11 ± 0.1%), ß-elemene (17.33 ± 0.1%), 1,8-cineole (12.15 ± 0.2%), furfural (7.91 ± 0.1%), terpineol-4-ol (6.32 ± 0.2%), and limonene (4.13 ± 0.1%), accounted for about 75% of the total OCA oil. On the other hand, 1,8-cineole (15.79 ± 0.1%), furfural (10.44 ± 0.1%), ß-elemene (10.14 ± 0.2%), α-terpineol (7.74 ± 0.1%), linalool (7.45 ± 0.1%), and α-pinene (4.76 ± 0.1%) made up about 60% of the OCR oil. The IC50 values of the scavenging activities of the OCA and OCR oils towards the DPPH radical and H2O2 were 79.8 ± 0.5 and 153.3 ± 0.6 µg/mL and 394.09 ± 0.2 and 311.67 ± 0.4 µg/mL, respectively. In addition, in the MTS assay, the OCA and OCR oils showed significant antiproliferative effects against T47D, MDA-MB-453, BG-1, and A549 human cancer cells that were more powerful than those against two normal human cell lines, HEK-293 and HFF-1. The abundant presence of ß-elemene as an antiproliferative component of the two oils suggested the existence of a new chemotype of O. carduchorum.

Effect of sainfoin proanthocyanidins on milk fatty acids from ewes rearing suckling lambs.

Proanthocyanidins (PAC) can modulate the fatty acid (FA) profile of animal products and make them healthier for human consumption, but their effects are highly variable depending on several factors such as PAC chemical structure or dose. The present experiment aimed to evaluate the effect of PAC on the milk FA profile of Rasa Aragonesa ewes fed fresh sainfoin (PAC-containing forage legume) during the rearing period of suckling lambs (4 weeks postlambing). Twenty lactating ewes rearing a single male lamb were fed fresh sainfoin ad libitum plus 200 g/d of barley. Half the ewes were orally dosed with 100 mL of water (Sainfoin Group; n = 10) and the other half with polyethylene glycol (50 g PEG4000/100 mL water, Sainfoin + PEG Group; n = 10) to block PAC effects. Sainfoin and milk samples were collected weekly to determine their FA profile by gas chromatography. Fresh sainfoin contents of C18:3n-3 decreased from week 1 to week 2, C16:0 and C18:0 increased from week 1 to week 3, and C18:2n-6 did not change. Regarding milk FA, there were minor effects of PAC on milk-saturated FA. During the whole study, the presence of PAC increased C18:0, C18:2n-6, C18:3n-3 and total polyunsaturated FA (PUFA) n-6 and n-3, and decreased C18:1 t11, branched- and odd-chain FA. However, the decrease of milk concentrations of trans-monounsaturated FA, C18:1 t10, and total conjugated linoleic acid (CLA) and the increase of total PUFA due to the presence of PAC occurred only in week 1, while CLA c9,t11 was lower during weeks 1 and 2. The canonical analyses confirmed the differences between treatments in the FA profile of milk. Overall, the use of fresh sainfoin in the diet of lactating ewes resulted in a beneficial modification of the concentration of several milk FAs, suggesting some changes in ruminal biohydrogenation.

Sainfoin (Onobrychis spp.) crop ontology: supporting germplasm characterization and international research collaborations.

Sainfoin (Onobrychis spp.) is a perennial forage legume that is also attracting attention as a perennial pulse with potential for human consumption. The dual use of sainfoin underpins diverse research and breeding programs focused on improving sainfoin lines for forage and pulses, which is driving the generation of complex datasets describing high dimensional phenotypes in the post-omics era. To ensure that multiple user groups, for example, breeders selecting for forage and those selecting for edible seed, can utilize these rich datasets, it is necessary to develop common ontologies and accessible ontology platforms. One such platform, Crop Ontology, was created in 2008 by the Consortium of International Agricultural Research Centers (CGIAR) to host crop-specific trait ontologies that support standardized plant breeding databases. In the present study, we describe the sainfoin crop ontology (CO). An in-depth literature review was performed to develop a comprehensive list of traits measured and reported in sainfoin. Because the same traits can be measured in different ways, ultimately, a set of 98 variables (variable = plant trait + method of measurement + scale of measurement) used to describe variation in sainfoin were identified. Variables were formatted and standardized based on guidelines provided here for inclusion in the sainfoin CO. The 98 variables contained a total of 82 traits from four trait classes of which 24 were agronomic, 31 were morphological, 19 were seed and forage quality related, and 8 were phenological. In addition to the developed variables, we have provided a roadmap for developing and submission of new traits to the sainfoin CO.

Amino acid and fatty acid profiles of perennial Baki™ bean.

To realize the potential of sainfoins to contribute to sustainable agriculture and expand on demonstrated uses and benefits, de novo domestication is occurring to develop perennial Baki™ bean, the trade name used by The Land Institute for pulses (i.e., grain legumes) derived from sainfoins. The objective of this study was to characterize amino acid and fatty acid profiles of depodded seeds from commercial sainfoin (Onobrychis viciifolia) seed lots, and compare these results with data published in the Global Food Composition Database for Pulses. The fatty acid profile consisted primarily of polyunsaturated fatty acids (56.8%), compared to monounsaturated (29.0%) and saturated fatty acids (14.2%), and n-3 fatty acids (39.5%), compared to n-9 (28.4%) and n-6 (17.6%) fatty acids. The essential fatty acid linolenic acid (18,3 n-3) was the most abundant fatty acid (39.2%), followed by oleic acid (18,1 cis-9) (27.8%), and the essential fatty acid linoleic acid (18,2 n-6) (17.3%). The amino acid profile consisted primarily of the nonessential amino acids glutamic acid (18.3%), arginine (11.6%), and aspartic acid (10.8%), followed by the essential amino acids leucine (6.8%), and lysine (5.8%). Essential amino acid content met adult daily requirements for each amino acid. This indicates that sainfoin seeds may be a complete plant protein source. However, further research is necessary to better understand protein quality, defined by protein digestibility in addition to the amino acid profile. By demonstrating favorable fatty acid and amino acid profiles to human health, these results contribute to a growing body of evidence supporting the potential benefits of perennial Baki™ bean, a novel, perennial pulse derived from sainfoins.

The Chromosome Number and rDNA Loci Evolution in Onobrychis (Fabaceae).

The evolution of chromosome number and ribosomal DNA (rDNA) loci number and localisation were studied in Onobrychis Mill. Diploid and tetraploid species, as well as two basic chromosome numbers, x = 7 and x = 8, were observed among analysed taxa. The chromosomal distribution of rDNA loci was presented here for the first time using fluorescence in situ hybridisation (FISH) with 5S and 35S rDNA probes. Onobrychis species showed a high polymorphism in the number and localisation of rDNA loci among diploids, whereas the rDNA loci pattern was very similar in polyploids. Phylogenetic relationships among the species, inferred from nrITS sequences, were used as a framework to reconstruct the patterns of basic chromosome number and rDNA loci evolution. Analysis of the evolution of the basic chromosome numbers allowed the inference of x = 8 as the ancestral number and the descending dysploidy and polyploidisation as the major mechanisms of the chromosome number evolution. Analyses of chromosomal patterns of rRNA gene loci in a phylogenetic context resulted in the reconstruction of one locus of 5S rDNA and one locus of 35S rDNA in the interstitial chromosomal position as the ancestral state in this genus.

Analyses on Flavonoids and Transcriptome Reveals Key MYB Gene for Proanthocyanidins Regulation in Onobrychis Viciifolia.

Onobrychis viciifolia (sainfoin) is one of the most high-quality legume forages, which is rich in proanthocyanidins that is beneficial for the health and production of animals. In this study, proanthocyanidins and total flavonoids in leaves of 46 different sainfoin germplasm resources were evaluated, and it showed that soluble proanthocyanidin contents varied greatly in these sainfoin germplasm resources, but total flavonoids did not show significant difference. Transcriptome sequencing with high and low proanthocyanidins sainfoin resulted in the identification of totally 52,926 unigenes in sainfoin, and they were classed into different GOC categories. Among them, 1,608 unigenes were differentially expressed in high and low proanthocyanidins sainfoin samples, including 1,160 genes that were upregulated and 448 genes that were downregulated. Analysis on gene enrichment via KEGG annotation revealed that the differentially expressed genes were mainly enriched in the phenylpropanoid biosynthetic pathway and the secondary metabolism pathway. We also analyzed the expression levels of structural genes of the proanthocyanidin/flavonoid pathway in roots, stems, and leaves in the high proanthocyanidin sainfoin via RT-qPCR and found that these genes were differentially expressed in these tissues. Among them, the expression levels of F3'5'H and ANR were higher in leaves than in roots or stems, which is consistent with proanthocyanidins content in these tissues. Among MYB genes that were differentially expressed, the expression of OvMYBPA2 was relatively high in high proanthocyanidin sainfoin. Over-expression level of OvMYBPA2 in alfalfa hairy roots resulted in decreased anthocyanin content but increased proanthocyanidin content. Our study provided transcriptome information for further functional characterization of proanthocyanidin biosynthesis-related genes in sainfoin and candidate key MYB genes for bioengineering of proanthocyanidins in plants.

Comparative Metabolomic Analysis of Four Fabaceae and Relationship to In Vitro Nematicidal Activity against Xiphinema index.

The grapevine fanleaf virus (GFLV), responsible for fanleaf degeneration, is spread in vineyards by the soil nematode Xiphinema index. Nematicide molecules were used to limit the spread of the disease until they were banned due to negative environmental impacts. Therefore, there is a growing interest in alternative methods, including plant-derived products with antagonistic effects to X. index. In this work, we evaluated the nematicidal potential of the aerial parts and roots of four Fabaceae: sainfoin (Onobrychis viciifolia), birdsfoot trefoil (Lotus corniculatus), sweet clover (Melilotus albus), and red clover (Trifolium pratense), as well as that of sainfoin-based commercial pellets. For all tested plants, either aerial or root parts, or both of them, exhibited a nematicidal effect on X. index in vitro, pellets being as effective as freshly harvested plants. Comparative metabolomic analyses did not reveal molecules or molecule families specifically associated with antagonistic properties toward X. index, suggesting that the nematicidal effect is the result of a combination of different molecules rather than associated with a single compound. Finally, scanning electron microscope observations did not reveal the visible impact of O. viciifolia extract on X. index cuticle, suggesting that alteration of the cuticle may not be the primary cause of their nematicidal effect.

Analysis of genetic diversity among Onobrychis accessions with high agronomic performance by simple sequence repeat (SSR) markers.

BACKGROUND: Onobrychis viciifolia Scop. is a short-lived perennial cool-season legume used for forage production. It is a common native species in Asia Minor, especially in Turkey, the districts of the Caucasus, and the Caspian fringes. It can grow well in a broad range of climatic and soil types found in Asia, Europe, and North America. It is a non-bloating crop, making it suitable for use in both hay and pasture. METHODS AND RESULTS: The aim was to assess the diversity of the 83 sainfoin genotypes selected based on their high agronomic performance from a germplasm collection evaluated in the experimental field of Tekirdag Namik Kemal University, Turkey. Ten nuclear simple sequence repeat (nSSR) primers (OVK036, OVK046, OVK094, OVK101, OVK125, OVK161, OVK174, OVM033, OVM061, and OVM125) were used in the study. All nSSR loci were found to be polymorphic and totally 92 alleles were detected. The mean observed number of alleles per locus was calculated as 9.2. Among the genetic diversity parameters, Shannon Index (I = 0.375), unbiased genetic diversity value (uh = 0.243), and mean polymorphic information content (PIC = 0.240) were calculated. The genetic distance value varied between 0.43 and 0.95. Based on the dendrogram built by the UPGMA clustering method using genetic distance values, it was observed that the studied sainfoin genotypes were divided into two main clusters, whereas the STRUCTURE analysis results had high support for three clusters. CONCLUSIONS: The results obtained from this study provide important information on the genetic structures of the studied sainfoin genotypes and their genetic relationship. Therefore acquired genetic data will be useful in designing more efficient polycross nurseries, allowing open pollination of best performing and genetically diverse genotypes in the isolated conditions, which will increase genetic gain in sainfoin breeding programs.

Sainfoin in the Dams' Diet as a Source of Proanthocyanidins: Effect on the Growth, Carcass and Meat Quality of Their Suckling Lambs.

Sainfoin (Onobrychis viciifolia) is a forage legume with a medium content of proanthocyanidins (PAC), which may affect animal performance and product quality. The objective of the present study was to assess the effect of PAC from sainfoin fed to dams, using polyethylene glycol (PEG) as a blocking agent, on the performance and carcass and meat quality of their suckling male lambs. After lambing, twenty lactating dams were fed fresh sainfoin ad libitum plus 200 g per day of barley; ten were orally dosed with water (Sainfoin), and ten were dosed orally with a water dilution of 100 g PEG (Sainfoin + PEG). Their lambs (4.1 ± 0.64 kg at birth) suckled ad libitum until they reached the target slaughter weight of 10-12 kg. The presence of PAC in the dams' diet did not affect the growth, blood metabolites and carcass weight and fatness of the suckling lambs but decreased the lightness of caudal fat (p < 0.05) and increased the weight of the digestive compartments (p < 0.05). Regarding the meat characteristics, PAC only decreased polyphenols content (p < 0.05). In conclusion, the presence of PAC in the dams' diet had not significant effects on the performance and product quality of their suckling lambs.

The complete chloroplast genome of Onobrychis gaubae (Fabaceae-Papilionoideae): comparative analysis with related IR-lacking clade species.

BACKGROUND: Plastome (Plastid genome) sequences provide valuable markers for surveying evolutionary relationships and population genetics of plant species. Papilionoideae (papilionoids) has different nucleotide and structural variations in plastomes, which makes it an ideal model for genome evolution studies. Therefore, by sequencing the complete chloroplast genome of Onobrychis gaubae in this study, the characteristics and evolutionary patterns of plastome variations in IR-loss clade were compared. RESULTS: In the present study, the complete plastid genome of O. gaubae, endemic to Iran, was sequenced using Illumina paired-end sequencing and was compared with previously known genomes of the IRLC species of legumes. The O. gaubae plastid genome was 122,688 bp in length and included a large single-copy (LSC) region of 81,486 bp, a small single-copy (SSC) region of 13,805 bp and one copy of the inverted repeat (IRb) of 29,100 bp. The genome encoded 110 genes, including 76 protein-coding genes, 30 transfer RNA (tRNA) genes and four ribosome RNA (rRNA) genes and possessed 83 simple sequence repeats (SSRs) and 50 repeated structures with the highest proportion in the LSC. Comparative analysis of the chloroplast genomes across IRLC revealed three hotspot genes (ycf1, ycf2, clpP) which could be used as DNA barcode regions. Moreover, seven hypervariable regions [trnL(UAA)-trnT(UGU), trnT(GGU)-trnE(UUC), ycf1, ycf2, ycf4, accD and clpP] were identified within Onobrychis, which could be used to distinguish the Onobrychis species. Phylogenetic analyses revealed that O. gaubae is closely related to Hedysarum. The complete O. gaubae genome is a valuable resource for investigating evolution of Onobrychis species and can be used to identify related species. CONCLUSIONS: Our results reveal that the plastomes of the IRLC are dynamic molecules and show multiple gene losses and inversions. The identified hypervariable regions could be used as molecular markers for resolving phylogenetic relationships and species identification and also provide new insights into plastome evolution across IRLC.

Effects of feeding sainfoin proanthocyanidins to lactating ewes on intake, milk production and plasma metabolites.

There is increasing interest in using sainfoin (Onobrychis viciifolia) to feed sheep, but it contains proanthocyanidins (PACs), and the associated effects of PAC on sheep production are not well-known. The aim of the study was to assess the effect of the presence of PAC from sainfoin, through the inclusion of polyethylene glycol (PEG), on the intake and productive parameters of local ewes bearing one male lamb. For the experiment, 20 ewes and their newborn male lambs were placed in individual indoor cages. All ewes were fed ad libitum fresh sainfoin plus 200 g/d barley. Twice daily, half of the ewes were orally dosed with only water (Sainfoin Group; n = 10), and the other half were orally dosed with 100 g/d PEG 4000 per ewe (Sainfoin + PEG Group; n = 10). Sucking lambs were permanently housed with their dams until they reached 10-12 kg BW. The intake of sainfoin was recorded daily, and its chemical composition was analysed. Weekly, the BW, body condition score (BCS), milk yields and individual milk and blood samples were recorded. At the beginning and end of the experiment, faecal samples were collected from ewes and analysed for the anthelmintic role of PAC. The chemical composition, polyphenol content and antioxidant capacity of the diet and milk were analysed. The presence of PAC did not affect the intake, BW, BCS or milk yield of the dams (P > 0.05); however, all parameters were affected by the week of lactation (P < 0.05). Milk components were affected by the week of lactation (P < 0.001), but only the polyphenol and urea contents were reduced in the presence of PAC (P < 0.01). Similarly, the presence of PAC decreased the plasma urea concentration (P < 0.01) without effect on the rest of metabolites, polyphenols and antioxidant activity (P > 0.05). The presence of PAC had no effect on parasitism (P > 0.05). In conclusion, the presence of PAC had no relevant effects on milk production, although it affected protein metabolism, as indicated by the urea contents in milk and plasma.

Cattle manure compost and biochar supplementation improve growth of Onobrychis viciifolia in coal-mined spoils under water stress conditions.

Surface mining is a critical anthropogenic activity that significantly alters the ecosystem, while the use of appropriate revegetation techniques can be considered an important and feasible strategy in the way to improve the ecosystem services of degraded land. In the present study, we carried out a pot experiment to investigate the effects of three different variables on morpho-physiological and biochemical parameters of Onobrychis viciifolia to assess the capability of this species to be used for restoration purposes. Specifically, the variables studied were: (a) water (W) regime, working at five values as regards field capacity (FC) (i.e., 80% FC = highest, 72% FC = high, 60% FC = moderate, 48% FC = low, and 40% FC = very-low dose); and (b) rates of cattle manure compost (CMC) and wood biochar (BC) (weight/weight ratio), working at five rates (i.e., 4.0% = highest, 3.2% = high, 2.0% = moderate, 0.8% = low, and 0% = either no-CMC or no-BC dose). In addition, soil physical-chemical properties and enzyme activities were also investigated at the end of the experimental period. It was found that morphological growth attributes such as plant height, maximum root length, and dry biomass significantly increased with W, CMC and BC applications. Compared to control, moderate-to-high W, CMC and BC doses (W80CMC2BC2) increased net photosynthesis rate (by 42%), stomatal conductance (by 50%), transpiration rate (by 29%), water use efficiency (by 10%), chlorophyll contents (by 73%), carotenoid content (by 81%), leaf relative water content (by 33%) and leaf membrane stability index (by 30%). Under low-W content, the application of CMC and BC enhanced osmotic adjustments by increasing the content of soluble sugar and the activities of superoxide dismutase, catalase, peroxidase and ascorbate peroxidase, decreasing the oxidative stress, as verified by low levels of hydrogen peroxide, superoxide anion, malondialdehyde and proline contents in leaf tissues. Moreover, application of W, CMC and BC significantly improved soil water holding capacity, available nitrogen, phosphorus and potassium, urease and catalase activities, which facilitate plant growth. These results would aid in designing an appropriate strategy for achieving a successful revegetation of O. viciifolia, providing optimum doses of W (64% field capacity), CMC (2.4%) and BC (1.7%), with the final aim of reaching ecological restoration in arid degraded lands.

A Pilot Approach Investigating the Potential of Crop Rotation With Sainfoin to Reduce Meloidogyne enterolobii Infection of Maize Under Greenhouse Conditions.

Root-knot nematodes (RKNs) are one of the most important plant-parasitic nematodes of cereal crops in sub-Saharan Africa. This study was designed to evaluate the rotation effects of different cultivars of sainfoin (Esparsette, Perly, Taja and Visnovsky), soybean (DM-5953-RSF) and alfalfa (BAR 7) with maize (P-2432-R), on a Meloidogyne enterolobii population, compared to monoculture maize. The results showed that sainfoin (Perly and Esparsette) and alfalfa had significantly (P ≤ 0.05) lower numbers of M. enterolobii eggs and second stage juveniles (J2) compared to the monoculture maize in the first experiment. However, in the repeat experiment all treatments had significantly (P ≤ 0.05) lower numbers of eggs and J2 compared to monoculture maize. Rotation of sainfoin Esparsette/maize resulted in the lowest numbers of eggs and J2 (91 and 202, respectively) in the first and repeat experiments. Rotation of sainfoin Esparsette/maize reduced M. enterolobii population density by 81 and 60% in the first and repeat experiments, respectively, followed by alfalfa (54 and 43%, respectively). Ultimately, substantial variation was evident in terms of the efficacy of different sainfoin cultivars with regards to their effect on nematode reduction when used in rotation with maize.

The complete chloroplast genome of leguminous forage Onobrychis viciifolia.

Onobrychis viciifolia is mainly distributed in Europe and has been widely cultivated in North and Northwest of China. The complete chloroplast genome was sequenced using the Illumina Hiseq X-Ten platform. The genome lacks an inverted repeat (IR) region, containing 76 protein-coding genes, 30 tRNAs genes and 4 rRNAs. The overall GC content is 34.6%. A phylogenetic tree based on the whole chloroplast genomes of 14 species indicated that Onobrychis viciifolia belonged to the tribe Hydysareae in IRLC group of the subfamily Papilionoideae (Leguminosae), and it was sister to the genus Hedysarum.

The complete chloroplast genome sequence of Sainfoin (Onobrychis viciifolia).

Sainfoin (Onobrychis viciifolia) is one of the dominant legume forages distributed in Northern China. In our study, we assembled and annotated the structure of the complete chloroplast genome of sainfoin. The length of the circular genome is 122,102 bp. It contains 115 genes, including 79 protein-coding genes (68.7%), 31 tRNA genes (26.96%) and 5 rRNA genes (4.35%). The GC content of the total chloroplast genome of sainfoin is 34.58%. We construct the phylogenetic relationships between the chloroplast genome of sainfoin and the other 16 species by the Maximum likelihood (ML), and found sainfoin is most closely related to Hedysarum petrovii and Hedysarum taipeicum.

Effects of intrinsic environmental predictability on intra-individual and intra-population variability of plant reproductive traits and eco-evolutionary consequences.

BACKGROUND AND AIMS: It is widely accepted that changes in the environment affect mean trait expression, but little is known about how the environment shapes intra-individual and intra-population variance. Theory suggests that intra-individual variance might be plastic and under natural selection, rather than reflecting developmental noise, but evidence for this hypothesis is scarce. Here, we experimentally tested whether differences in intrinsic environmental predictability affect intra-individual and intra-population variability of different reproductive traits, and whether intra-individual variability is under selection. METHODS: Under field conditions, we subjected Onobrychis viciifolia to more and less predictable precipitation over 4 generations and 4 years. We analysed effects on the coefficient of intra-individual variation (CVi-i) and the coefficient of intra-population variation (CVi-p), assessed whether the coefficients of intra-individual variation (CsVi-i) are under natural selection and tested for transgenerational responses (ancestor environmental effects on offspring). KEY RESULTS: Less predictable precipitation led to higher CsVi-i and CsVi-p, consistent with plastic responses. The CsVi-i of all studied traits were under consistent stabilizing selection, and precipitation predictability affected the strength of selection and the location of the optimal CVi-i of a single trait. All CsVi-i differed from the optimal CVi-i and the maternal and offspring CsVi-i were positively correlated, showing that there was scope for change. Nevertheless, no consistent transgenerational effects were found in any of the three descendant generations, which contrasts with recent studies that detected rapid transgenerational responses in the trait means of different plant species. This suggests that changes in intra-individual variability take longer to evolve than changes in trait means, which may explain why high intra-individual variability is maintained, despite the stabilizing selection. CONCLUSIONS: The results indicate that plastic changes of intra-individual variability are an important determinant of whether plants will be able to cope with changes in environmental predictability induced by the currently observed climatic change.

High allelic diversity of the centromere-specific histone H3 (CENH3) in the legume sainfoin (Onobrychis viciifolia).

The centromere is a structurally and functionally specialized region on each eukaryotic chromosome and is essential for accurate and complete segregation of chromosomes during cell division. Centromeric nucleosomes differ from canonical nucleosomes by replacement of the histone H3 with its centromere-specific variant CENH3. CENH3 is essential for active centromeres in most eukaryotes. Homologs of CENH3 are identified in many organisms. Sainfoin (Onobrychis viciifolia) is an agriculturally important perennial forage and is a legume of the Fabaceae family. There is very limited information on the structure of the sainfoin genome and no data are available on its centromere structure. Here, we aim to characterize the sainfoin CENH3 homolog (OvCENH3). Using a sequence homology-based strategy with gene-specific primers, we were able to clone transcripts from sainfoin total RNA. The amplified clones were sequenced and compared by bioinformatics tools. Four distinct alleles of OvCENH3 were detected. Our study provides the first structural features on sainfoin centromeres with a possible allotetraploid origin for sainfoin. We discuss and compare our findings with that for other important legume species.