Alfalfa species Medicago sativa L. (MS) and Medicago falcata L. (MF), globally prominent perennial leguminous forages, hold substantial economic value. However, our comprehension of the molecular mechanisms governing their resistance to cold stress remains limited. To address this knowledge gap, we scrutinized and compared MS and MF cold-stress responses at the molecular level following 24 h and 120 h low-temperature exposure (4 °C). Our study revealed that MF had superior physiological resilience to cold stress compared with MS, and its morphology was healthier under cold stress, and its malondialdehyde content and superoxide dismutase activity increased, first, and then decreased, while the soluble sugar content continued to accumulate. Transcriptome analysis showed that after 120 h of exposure, there were different gene-expression patterns between MS and MF, including 1274 and 2983 genes that were continuously up-regulated, respectively, and a total of 923 genes were included, including star cold-resistant genes such as ICE1 and SIP1. Gene ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses revealed numerous inter-species differences in sustained cold-stress responses. Notably, MS-exclusive genes included a single transcription factor (TF) gene and several genes associated with a single DNA repair-related pathway, whereas MF-exclusive genes comprised nine TF genes and genes associated with 14 pathways. Both species exhibited high-level expression of genes encoding TFs belonging to AP2-EREBP, ARR-B, and bHLH TF families, indicating their potential roles in sustaining cold resistance in alfalfa-related species. These findings provide insights into the molecular mechanisms governing cold-stress responses in MS and MF, which could inform breeding programs aimed at enhancing cold-stress resistance in alfalfa cultivars.

Assessing the nutritional quality of silage maize (Zea mays L.) hinges largely on its oil content, a complex quantitative trait influenced by multiple genes. Mining candidate genes within oil content-related quantitative trait loci (QTLs) can provide genetic resources and a theoretical foundation for cultivating high-oil silage maize varieties. This study employed 274 doubled haploid (DH) lines derived from the parental lines BY4944 and DNF34-2 to perform main gene plus polygene mixed genetic analysis and complex interval mapping (CIM), with the goal of pinpointing oil content-related QTLs and genes distributed across the Z. mays L. genome. Leveraging 5400 single nucleotide polymorphism (SNPs), a high-resolution silage maize genetic linkage map covering 3864.51 cM was constructed with an average interval between markers of 0.74 cM. Analysis of the map revealed 13 oil content-related QTLs. The most significant large-effect QTL (qOIL-1-1), located on chromosome 1 within the region spanning 240.93 Mb to 256.57 Mb, exhibited a logarithm of odds (LOD) score of 3.34 and explained 5.06% of oil content-related phenotypic variation. Within these QTLs, 617 genes were annotated. Through transcriptome analysis combined with quantitative real-time polymerase chain reaction (RT-qPCR), five candidate genes potentially associated with oil content were predicted and subsequently validated within these genetic loci. This research underscores the potential of identifying candidate genes to enhance breeding efforts aimed at augmenting oil content, thereby advancing animal husbandry practices.

Alfalfa (Medicago sativa L.), a perennial leguminous herb, exhibits robust cold and saline-alkali tolerance. In this study, two alfalfa cultivars, LJ and 218TR, were treated with saline-alkali stress, cold stress, and combined saline-alkali-cold stress , and p henotype, physiology , key metaboli te and stress-responsive genes were analyzed . The results showed m alondialdehyde, soluble sugar, proline content, and the activities of phenylalanine ammonia-lyase (PAL), superoxide dismutase, catalase, and peroxidase initially increased under individual stresses but declined after combined stress . T he maximum photochemical efficiency of photosystem II and chlorophyll content declined under individual and combined stresses. The staining of leaves revealed that combined stress induced significantly higher cell mortality and accumulation of superoxide anio n compared to individual stresses. LJ exhibited superior resistant to saline-alkali, cold, and combined stress compared to 218TR . Metabolite analysis showed salicylic acid (SA) in two alfalfa wa s the most responsive metabolite to combined stress. The isochorismate synthase (ICS) and PAL as critical genes for SA biosynthesis were up-regulated in expression under single or combined stress, and promote d SA accumulation , thereby improving alfalfa resilience to combined saline-alkali-cold stress. This study elucidates the physiological and molecular mechanisms underlying alfalfa’s response to combined saline-alkali and cold stress, providing a theoretical basis for breeding stress-tolerant cultivars.

Heavy metal contamination is a serious environmental problem worldwide, with substantial negative ecological and economic effects. Serine hydroxymethyltransferase (SHMT) is a key metabolic and photorespiratory enzyme in plant cells, and it is also involved in stress responses. In this study, LcSHMT4 was isolated from sheepgrass (Leymus chinensis (Trin.) Tzvel) after transcriptome sequence analysis. The transcript levels of LcSHMT4 in sheepgrass seedlings increased under Cd and Mn stresses, and subcellular localization analysis in tobacco leaves revealed that its encoded protein localizes at the mitochondria. Transgenic yeast and rice lines overexpressing LcSHMT4 showed increased tolerance to Cd and Mn, compared with that of their controls. In addition, compared with the control, transgenic rice overexpressing LcSHMT4 accumulated more Cd and Mn in brown rice grains. The transcript levels of genes encoding Cd or Mn transporters were increased in the LcSHMT4-overexpressing transgenic rice lines. We speculate that LcSHMT4 may enhance Cd and Mn tolerance by increasing the activities of antioxidant enzymes and the glutathione content and increase heavy metal accumulation by inducing the expression of genes encoding transporters. These results highlight useful genetic resources and provide a theoretical basis for further research on heavy metal tolerance and the phytoremediation of heavy-metal-contaminated soil.