Background: Sainfoin (Onobrychis viciifolia Scop) is not only a high-quality legume forage, but also a nectar-producing plant. Therefore, the flower color of sainfoin is an important agronomic trait, but the factors affecting its flower phenotype are still unclear. To gain insights into the regulatory networks associated with metabolic pathways of coloration compounds (flavonoids or anthocyanins) and identify the key genes, we conducted a comprehensive analysis of the phenotype, metabolome and transcriptome of WF and AF of sainfoin. Results: Delphinidin, petunidin and malvidin derivatives were the main anthocyanin compounds in the AF of sainfoin. These substances were not detected in the WF of sainfoin. The transcriptomes of WF and AF in sainfoin at the S1 and S3 stages were obtained using the Illumina HiSeq4000 platform. Overall, 10,166 (4273 upregulated and 5893 downregulated) and 15,334 (8174 upregulated and 7160 downregulated) DEGs were identified in flowers at S1 and S3 stages, respectively (WF-VS-AF). KEGG pathway annotations showed that 6396 unigenes were annotated to 120 pathways and contained 866 DEGs at S1 stages, and 6396 unigenes were annotated to 131 pathways and included 1546 DEGs at the S3 stage. Nine DEGs belonging to the “flavonoid biosynthesis”and “phenylpropanoid biosynthesis” pathways involved in flower color formation were identified and verified by RT-qPCR analyses. Among these DEGs, 4CL3, FLS, ANS, CHS, DFR and CHI2 exhibited downregulated expression, and F3H exhibited upregulated expression in the WF compared to the AF, resulting in a decrease in anthocyanin synthesis and the formation of WF in sainfoin. Conclusions: This study is the first to use transcriptome technology to study the mechanism of white flower formation in sainfoin. Our transcriptome data will be a great enrichment of the genetic information for sainfoin. In addition, the data presented herein will provide valuable molecular information for genetic breeding and provide insight into the future study of flower color polymorphisms in sainfoin.

Phosphite (Phi), the reduced form of phosphate (Pi), is characterized by its stability, high solubility, efficient transport, resistance to fixation in soil, and widespread occurrence in natural environments. Although Phi exhibits greater suitability than Pi as a soil fertilizer, it cannot be metabolized by plants. In agricultural applications, Phi serves as a bio-stimulant, fungicide, herbicide, and has other purposes. As a bio-stimulant, Phi has been shown to promote plant growth, enhance stress resistance, and improve fruit quality. Additionally, when used as a fungicide or pesticide, it effectively inhibits the growth of phytopathogens in various crop species. The discovery of the phosphite dehydrogenase (ptxD) gene in microorganisms has significantly expanded the potential applications of Phi, including its use as a herbicide, phosphatic fertilizer, and a selectable chemical for generating marker-free transgenic plants. Therefore, the dual fertilization and weed control system of ptxD/Phi facilitates the utilization of Phi as the sole phosphorus source while concurrently suppressing the evolution of herbicide-resistant weeds in the future. Notably, ptxD also acts as an ideal selectable marker because its resistant is specific to Phi, thereby eliminating the risk of false positive clones. The application of Phi provides a promising strategy for addressing phosphorus resource shortages and improving the efficiency of phosphatic fertilizers in agriculture. Furthermore, Phi is considered an environmentally friendly fertilizer, as it contributes to the mitigation of eutrophication. In prospect, Phi is anticipated to play a significant role as a chemical fertilizer that promotes the sustainable development of agriculture. In this review, we provide a comprehensive analysis of the functional mechanisms of Phi and its current applications in agriculture, with the aim of offering deeper insights into its potential benefits and practical utility.

Alfalfa (Medicago sativa L.) is one of the most widely planted forages due to its useful characteristics. Although alfalfa spatial distribution is an important source of basic data, manual surveys incur high survey costs, require large workloads and confront difficulties in collecting data over large areas; remote sensing compensates for these shortcomings. In this study, the time-series variation characteristics of different vegetation types were analyzed, and the extraction method of alfalfa mapping was established according to different spatial- and temporal-resolution remote sensing data. The results provided the following conclusions: (1) when using the wave peak and valley number of normalized difference vegetation index (NDVI) curves, in the study area, the number of wave peak needed to be greater than 2 and the number of wave valley needed to be greater than 1; (2) 91.6% of alfalfa sampling points were extracted by moderate resolution imaging spectroradiometer (MODIS) data using the wave peak and valley method, and 5.0% of oats sampling points were extracted as alfalfa, while no other vegetation types met these conditions; (3) 85.3% of alfalfa sampling points were identified from Sentinel-2 multispectral instrument (MSI) data using the wave peak and valley method; 6.0% of grassland vegetation and 8.7% of oats satisfied the conditions, while other vegetation types did not satisfy this rule; and (4) the temporal phase selection was very important for alfalfa extraction using single-time phase remote sensing images; alfalfa was easily separated from other vegetation at the pre−wintering stage and was more difficult to separate at the spring regreening stage due to the variability in the alfalfa overwintering rate; the overall classification accuracy was 92.9% with the supervised classification method using support vector machine (SVM) at the pre-wintering stage. These findings provide a promising approach to alfalfa mapping using different remote sensing data.

Psathyrostachys juncea is a long-lived perennial Gramineae grass with dense basal tillers and soft leaves. It is used widely in cold and dry areas of Eurasia and North America to establish grazing pasture and is even used as an ideal plant for revegetation and ecological restoration. Plant architecture, especially tillering traits, is critical for bunch grasses in breeding programs, and these traits in plants are mostly quantitative traits. In this study, the genetic diversity, population structure, and linkage disequilibrium of 480 individual lines were analyzed using 127 pairs of the EST-SSR marker, and a significant association between ten plant-architecture-related traits of P. juncea and molecular markers was found. The results of the genetic diversity analysis showed that the number of observed alleles was 1.957, the number of effective alleles was 1.682, Shannon’s information index was 0.554, observed heterozygosity was 0.353, expected heterozygosity was 0.379, and the polymorphism information content was 0.300. A total of 480 individual lines were clustered into five groups based on population genetic structure, principal coordinate analysis, and unweighted pair group method with arithmetic mean analysis (UPGMA). The linkage disequilibrium coefficient (r²) was between 0.00 and 0.68, with an average of 0.04, which indicated a relatively low level of linkage disequilibrium among loci. The results of the association analysis revealed 55 significant marker–trait associations (MTA). Moreover, nine SSR markers were associated with multiple traits. This study provides tools with promising applications in the molecular selection and breeding of P. juncea germplasm.

Stems are more important to forage quality than leaves in alfalfa. To understand lignin formation at different stages in alfalfa, lignin distribution, anatomical characteristics and transcriptome profile were employed using two alfalfa cultivars. The results showed that the in vitro true digestibility (IVTD) of stems in WL168 was significantly higher than that of Zhungeer, along with the significantly lower neutral detergent fiber (NDF), acid detergent fiber (ADF) and lignin contents. In addition, Zhungeer exhibited increased staining of the xylem areas in the stems of different developmental stages compared to WL168. Interestingly, the stems of WL168 appeared intracellular space from the stage 3, while Zhungeer did not. The comparative transcriptome analysis showed that a total of 1993 genes were differentially expressed in the stem between the cultivars, with a higher number of expressed genes in the stage 4. Of the differentially expressed genes, starch and sucrose metabolism as well as phenylpropanoid biosynthesis pathways were the most significantly enriched pathways. Furthermore, expression of genes involved in lignin biosynthesis such as PAL, 4CL, HCT, CAD, COMT and POD coincides with the anatomic characteristics and lignin accumulation. These results may help elucidate the regulatory mechanisms of lignin biosynthesis and improve forage quality in alfalfa.