Improved drought tolerance of EMS mutagenized Alfalfa (Medicago sativa L.) mutants by in vitro screening at germination stage.
Ontology highlight
ABSTRACT:
SUBMITTER: Tiryaki I
PROVIDER: S-EPMC9325702 | biostudies-literature |
REPOSITORIES: biostudies-literature
Similar Datasets
Project description:Alfalfa (Medicago sativa L.) is an important legume forage crop with great economic value. However, as the growth of alfalfa is seriously affected by an inadequate supply of water, drought is probably the major abiotic environmental factor that most severely affects alfalfa production worldwide. In an effort to enhance alfalfa drought tolerance, we transformed the Arabidopsis Enhanced Drought Tolerance 1 (AtEDT1) gene into alfalfa via Agrobacterium-mediated transformation. Compared with wild type plants, drought stress treatment resulted in higher survival rates and biomass, but reduced water loss rates in the transgenic plants. Furthermore, transgenic alfalfa plants had increased stomatal size, but reduced stomatal density, and these stomatal changes contributed greatly to reduced water loss from leaves. Importantly, transgenic alfalfa plants exhibited larger root systems with larger root lengths, root weight, and root diameters than wild type plants. The transgenic alfalfa plants had reduced membrane permeability and malondialdehyde content, but higher soluble sugar and proline content, higher superoxide dismutase activity, higher chlorophyll content, enhanced expression of drought-responsive genes, as compared with wild type plants. Notably, transgenic alfalfa plants grew better in a 2-year field trial and showed enhanced growth performance with increased biomass yield. All of our morphological, physiological, and molecular analyses demonstrated that the ectopic expression of AtEDT1 improved growth and enhanced drought tolerance in alfalfa. Our study provides alfalfa germplasm for use in forage improvement programs, and may help to increase alfalfa production in arid lands.
| S-EPMC5733502 | biostudies-literature
Project description:Salinity tolerance-associated phenotypes of 35 EMS mutagenized wheat lines originating from BARI Gom-25 were compared. Vegetative growth was measured using non-destructive image-based phenotyping. Five different NaCl concentrations (0 to 160 mM) were applied to plants 19 days after planting (DAP 19), and plants were imaged daily until DAP 38. Plant growth, water use, leaf Na+, K+ and Cl- content, and thousand kernel weight (TKW) were measured, and six lines were selected for further analysis. In saline conditions, leaf Na+, K+, and Cl- content variation on a dry weight basis within these six lines were ~9.3, 1.4, and 2.4-fold, respectively. Relative to BARI Gom-25, two (OA6, OA62) lines had greater K+ accumulation, three (OA6, OA10, OA62) had 50-75% lower Na+:K+ ratios, and OA62 had ~30% greater water-use index (WUI). OA23 had ~2.2-fold greater leaf Na+ and maintained TKW relative to BARI Gom-25. Two lines (OA25, OA52) had greater TKW than BARI Gom-25 when grown in 120 mM NaCl but similar Na+:K+, WUI, and biomass accumulation. OA6 had relatively high TKW, high leaf K+, and WUI, and low leaf Na+ and Cl-. Phenotypic variation revealed differing associations between the parameters measured in the lines. Future identification of the genetic basis of these differences, and crossing of lines with phenotypes of interest, is expected to enable the assessment of which combinations of parameters deliver the greatest improvement in salinity tolerance.
| S-EPMC9570438 | biostudies-literature
Project description:In recent years, drought stress caused by global warming has become a major constraint on agriculture. The thiamine thiazole synthase (THI1) is responsible for controlling thiamine production in plants displaying a response to various abiotic stresses. Nonetheless, most of the THI1 activities in plants remain largely unknown. In this study, we extracted MsTHI1 from alfalfa and demonstrated its beneficial impact on improving the resistance of plants to stress conditions. The highest levels of MsTHI1 expression were identified in alfalfa leaves, triggered by exposure to cold, drought, salt, or alkaline conditions. The upregulation of MsTHI1 in drought-stressed transgenic plants resulted in enhanced accumulation of vitamin B1 (VB1), chlorophyll a (Chl a), chlorophyll b (Chl b), soluble protein, higher soil and plant analyzer development (SPAD) value, and the activity of peroxidase (POD), maintained Fv/Fm, and decreased lipid peroxidation. Moreover, overexpression of MsTHI1 upregulated the transcription of THI4, TPK1, RbcX2, Cu/Zn-SOD, CPK13, and CPK32 and downregulated the transcription of TH1 and CPK17 in transgenic alfalfa under drought stress. These results suggested that MsTHI1 enhances drought tolerance by strengthening photosynthesis, regulating the antioxidant defense system, maintaining osmotic homeostasis, and mediating plant signal transduction.
| S-EPMC9495609 | biostudies-literature
Project description:Melatonin (MT), polyamines (PAs), and ethylene have been suggested to play key roles in plant growth and development in response to environmental abiotic stresses. However, the effect of melatonin on polyamine and ethylene metabolism under waterlogging stress has rarely been elucidated. The main purpose of this study was to investigate the effect of melatonin pretreatment on waterlogging stress in alfalfa. The experiment was arranged into four treatment groups control with water pretreatment (CK-MT), control with melatonin pretreatment (CK+MT), waterlogging pretreated with water (WL-MT) and waterlogging pretreated with melatonin (WL+MT), with three replications. Six-week-old alfalfa seedlings were pretreated with 100 μM melatonin and exposed to waterlogging stress for 10 days. Plant growth rate, different physiological characteristics, and gene expression level were measured. Results showed that waterlogging induced melatonin accumulation, and melatonin pretreatment increased endogenous MT levels for the control and water-logged plants. Waterlogging stress caused a significant reduction in plant growth, chlorophyll content, photochemical efficiency (Fv/Fm) and net photosynthetic rate (Pn), while also causing increased leaf electrolyte leakage (EL) and malondialdehyde (MDA) content. Pretreatment with melatonin alleviated the waterlogging-induced damage and reduction in plant growth, chlorophyll content, Fv/Fm and Pn. Waterlogging stress significantly increased leaf polyamines (Put, Spd, Spm) and ethylene levels, and the increased PAs and ethylene levels are coupled with higher metabolic enzymes and gene expressions. While pretreatment with melatonin further increased Put, Spd and Spm levels, it also decreased ethylene levels under waterlogging, and those increased PAs levels or decreased ethylene levels are regulated by the metabolic enzymes and gene expressions. The results in this study provide more comprehensive insight into the physiological and molecular mechanisms of melatonin-improved waterlogging tolerance in alfalfa. Furthermore, they suggested that melatonin improved waterlogging tolerance in alfalfa at least partially by reprogramming ethylene and PA biosynthesis, attributable to the increased PAs and decreased ethylene levels, which leads to more enhanced membrane stability and photosynthesis as well as less leaf senescence caused by ethylene.
| S-EPMC6367245 | biostudies-literature
Project description:Salinity is one of major abiotic stresses limiting alfalfa (Medicago sativa L.) production in the arid and semi-arid regions in US and other counties. In this study, we used a diverse panel of alfalfa accessions previously described by Zhang et al. (2015) to identify molecular markers associated with salt tolerance during germination using genome-wide association study (GWAS) and genotyping-by-sequencing (GBS). Phenotyping was done by germinating alfalfa seeds under different levels of salt stress. Phenotypic data of adjusted germination rates and SNP markers generated by GBS were used for marker-trait association. Thirty six markers were significantly associated with salt tolerance in at least one level of salt treatments. Alignment of sequence tags to the Medicago truncatula genome revealed genetic locations of the markers on all chromosomes except chromosome 3. Most significant markers were found on chromosomes 1, 2, and 4. BLAST search using the flanking sequences of significant markers identified 14 putative candidate genes linked to 23 significant markers. Most of them were repeatedly identified in two or three salt treatments. Several loci identified in the present study had similar genetic locations to the reported QTL associated with salt tolerance in M. truncatula. A locus identified on chromosome 6 by this study overlapped with that by drought in our previous study. To our knowledge, this is the first report on mapping loci associated with salt tolerance during germination in autotetraploid alfalfa. Further investigation on these loci and their linked genes would provide insight into understanding molecular mechanisms by which salt and drought stresses affect alfalfa growth. Functional markers closely linked to the resistance loci would be useful for MAS to improve alfalfa cultivars with enhanced resistance to drought and salt stresses.
| S-EPMC4923157 | biostudies-literature
Project description:Reducing lignin concentration in lignocellulosic biomass can increase forage digestibility for ruminant livestock and saccharification yields of biomass for bioenergy. In sorghum (Sorghum bicolor (L.) Moench) and several other C4 grasses, brown midrib (bmr) mutants have been shown to reduce lignin concentration. Putative bmr mutants isolated from an EMS-mutagenized population were characterized and classified based on their leaf midrib phenotype and allelism tests with the previously described sorghum bmr mutants bmr2, bmr6, and bmr12. These tests resulted in the identification of additional alleles of bmr2, bmr6, and bmr12, and, in addition, six bmr mutants were identified that were not allelic to these previously described loci. Further allelism testing among these six bmr mutants showed that they represented four novel bmr loci. Based on this study, the number of bmr loci uncovered in sorghum has doubled. The impact of these lines on agronomic traits and lignocellulosic composition was assessed in a 2-yr field study. Overall, most of the identified bmr lines showed reduced lignin concentration of their biomass relative to wild-type (WT). Effects of the six new bmr mutants on enzymatic saccharification of lignocellulosic materials were determined, but the amount of glucose released from the stover was similar to WT in all cases. Like bmr2, bmr6, and bmr12, these mutants may affect monolignol biosynthesis and may be useful for bioenergy and forage improvement when stacked together or in combination with the three previously described bmr alleles.
| S-EPMC4232537 | biostudies-literature
Project description:The toxicity of aluminum (Al) in acidic soils poses a significant limitation to crop productivity. In this study, we found a notable increase in DnaJ (HSP40) expression in the roots of Al-tolerant alfalfa (WL-525HQ), which we named MsDjB4. Transient conversion assays of tobacco leaf epidermal cells showed that MsDjB4 was targeted to the membrane system including Endoplasmic Reticulum (ER), Golgi, and plasma membrane. We overexpressed (MsDjB4-OE) and suppressed (MsDjB4-RNAi) MsDjB4 in alfalfa hairy roots and found that MsDjB4-OE lines exhibited significantly better tolerance to Al stress compared to wild-type and RNAi hairy roots. Specifically, MsDjB4-OE lines had longer root length, more lateral roots, and lower Al content compared to wild-type and RNAi lines. Furthermore, MsDjB4-OE lines showed lower levels of lipid peroxidation and ROS, as well as higher activity of antioxidant enzymes SOD, CAT, and POD compared to wild-type and RNAi lines under Al stress. Moreover, MsDjB4-OE lines had higher soluble protein content compared to wild-type and RNAi lines after Al treatment. These findings provide evidence that MsDjB4 contributes to the improved tolerance of alfalfa to Al stress by facilitating protein synthesis and enhancing antioxidant capacity.
| S-EPMC10421020 | biostudies-literature
Project description:An adenylate cyclase activity in Medicago sativa L. (alfalfa) roots was partially characterized. The enzyme activity remains in the supernatant fluid after centrifugation at 105,000 g and shows in crude extracts an apparent Mr of about 84,000. The enzyme is active with Mg2+ and Ca2+ as bivalent cations, and is inhibited by EGTA and by chlorpromazine. Calmodulin from bovine brain or spinach leaves activates this adenylate cyclase.
| S-EPMC1148778 | biostudies-other
Project description:BackgroundSoil salinity is an important factor affecting growth, development, and productivity of almost all land plants, including the forage crop alfalfa (Medicago sativa). However, little is known about how alfalfa responds and adapts to salt stress, particularly among different salt-tolerant cultivars.ResultsAmong seven alfalfa cultivars, we found that Zhongmu-1 (ZM) is relatively salt-tolerant and Xingjiang Daye (XJ) is salt-sensitive. Compared to XJ, ZM showed slower growth under low-salt conditions, but exhibited stronger tolerance to salt stress. RNA-seq analysis revealed 2237 and 1125 differentially expressed genes (DEGs) between ZM and XJ in the presence and absence of salt stress, among which many genes are involved in stress-related pathways. After salt treatment, compared with the controls, the number of DEGs in XJ (19373) was about four times of that in ZM (4833). We also detected specific differential gene expression patterns: In response to salt stress, compared with XJ, ZM maintained relatively more stable expression levels of genes related to the ROS and Ca2+ pathways, phytohormone biosynthesis, and Na+/K+ transport. Notably, several salt resistance-associated genes always showed greater levels of expression in ZM than in XJ, including a transcription factor. Consistent with the suppression of plant growth resulting from salt stress, the expression of numerous photosynthesis- and growth hormone-related genes decreased more dramatically in XJ than in ZM. By contrast, the expression levels of photosynthetic genes were lower in ZM under low-salt conditions.ConclusionsCompared with XJ, ZM is a salt-tolerant alfalfa cultivar possessing specific regulatory mechanisms conferring exceptional salt tolerance, likely by maintaining high transcript levels of abiotic and biotic stress resistance-related genes. Our results suggest that maintaining this specific physiological status and/or plant adaptation to salt stress most likely arises by inhibition of plant growth in ZM through plant hormone interactions. This study identifies new candidate genes that may regulate alfalfa tolerance to salt stress and increases the understanding of the genetic basis for salt tolerance.
| S-EPMC5815232 | biostudies-literature
Project description:BackgroundSoil pollution by heavy metals increases the bioavailability of metals like hexavalent chromium (Cr (VI)), subsequently limiting plant growth and reducing the efficiency of phytoremediation. Plant growth-promoting rhizobacteria (PGPR) have substantial potential to enhance plant growth as well as plant tolerance to metal stress. The aim of this research was to investigate Cr (VI) phytoremediation enhancement by PGPR.ResultsThe results showed that the 27 rhizobacterial isolates studied were confirmed as Cr (VI)-resistant PGPR, by using classical biochemical tests (phosphate solubilization, nitrogen fixation, indole acetic acid, exopolysaccharides, hydrogen cyanide, siderophores, ammonia, cellulase, pectinase, and chitinase production) and showed variable levels of Cr (VI) resistance (300-600 mg/L). The best four selected Cr (VI)-resistant PGPR (NT15, NT19, NT20, and NT27) retained most of the PGP traits in the presence of 100-200 mg/L concentrations of Cr (VI). The inoculation of Medicago sativa with any of these four isolates improved the shoot and root dry weight. The NT27 isolate identified using 16S rDNA gene sequence analyses as a strain of Pseudomonas sp. was most effective in terms of plant growth promotion and stress level decrease. It increased shoot and root dry weights of M. sativa by 97.6 and 95.4%, respectively, in the presence of Cr (VI) when compared to non-inoculated control plants. It also greatly increased chlorophyll content and decreased the levels of stress markers, malondialdehyde, hydrogen peroxide, and proline. The results of the effect of Pseudomonas sp. on Cr content and bioaccumulation factor (BAF) of the shoots and roots of M. sativa plants showed the increase of plant biomass concomitantly with the increase of Cr root concentration in inoculated plants. This would lead to a higher potential of Cr (VI) phytostabilization.ConclusionsThis study demonstrates that the association M. sativa-Pseudomonas sp. may be an efficient biological system for the bioremediation of Cr (VI)-contaminated soils.
| S-EPMC8494867 | biostudies-literature