Variable salinity responses of 12 alfalfa genotypes and comparative expression analyses of salt-response genes

Twelve alfalfa genotypes that were selected for biomass under salinity, differences in Na and Cl concentrations in shoots and K/Na ratio were evaluated in this long-term salinity experiment. The selected plants were cloned to reduce genetic variability within each genotype. Salt tolerance (ST) index of the genotypes ranged from 0.39 to 1. The most salt-tolerant genotypes SISA14-1 (G03) and AZ-90ST (G10), the top performers for biomass, exhibited the least effect on shoot number and height. SISA14-1 (G03) accumulated low Na and Cl under salinity. Most genotypes exhibited a net reduction in shoot Ca, Mg, P, Fe, and Cu, while Mn and Zn increased under salinity. Salinity reduced foliar area and stomatal conductance; while net photosynthetic rate and transpiration were not affected. Interestingly, salinity increased chlorophyll and antioxidant capacity in most genotypes; however neither parameter correlated well to ST index. Salt-tolerant genotypes showed upregulation of the SOS1, SOS2, SOS3, HKT1, AKT1, NHX1, P5CS1, HSP90.7, HSP81.2, HSP71.1, HSPC025, OTS1, SGF29 and SAL1 genes. Gene expression analyses allowed us to classify genotypes based on their ability to regulate different components of the salt tolerance mechanism. Pyramiding different components of the salt tolerance mechanism may lead to superior salt-tolerant alfalfa genotypes.EEA Santiago del EsteroFil: Sandhu, Devinder. USDA. Agricultural Research Service. US Salinity Lab; Estados UnidosFil: Cornacchione, Monica. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Santiago del Estero; ArgentinaFil: Ferreira, Jorge F.S. USDA. Agricultural Research Service. US Salinity Lab; Estados UnidosFil: Suarez, Donald L. USDA. Agricultural Research Service. US Salinity Lab; Estados Unido

Expression of the High-Affinity K+ Transporter 1 (PpHKT1) Gene From Almond Rootstock ‘Nemaguard’ Improved Salt Tolerance of Transgenic Arabidopsis

Soil salinity affects plant growth and development, which directly impact yield. Plants deploy many mechanisms to cope with, or mitigate, salt stress. One of such mechanism is to control movement of ions from root to shoot by regulating the loading of Na+ in the transpiration stream. The high-affinity K+ transporter 1 (HKT1) is known to play a role in the removal of Na+from the xylem and bring it back to the root. As almond is a salt-sensitive crop, the rootstock plays an important role in successful almond cultivation in salt-affected regions. We currently lack knowledge on the molecular mechanisms involved in salt tolerance of almond rootstocks. In this study, we complemented the Arabidopsis athkt1 knockout mutant with HKT1 ortholog (PpHKT1) from the almond rootstock ‘Nemaguard’. Arabidopsis transgenic lines that were generated in athkt1 background with the constitutive promoter (PpHKT1OE2.2) and the native promoter (PpHKT1NP6) were subjected to different salt treatments. Both transgenic lines survived salt concentrations up to 120 mM NaCl, however, the mutant athkt1 died after 18 days under 120 mM NaCl. At 90 mM NaCl, the dry weight of athkt1 decreased significantly compared to the transgenic lines. Both transgenic lines showed significantly longer lateral roots compared to the athkt1 mutant at 80 mM NaCl treatment. The transgenic lines, PpHKT1OE2.2 and PpHKTNP6 had lower electrolyte leakage and higher relative water content compared to athkt1, suggesting that transgenic plants coped well with increased salt concentration by maintaining the integrity of the membranes. The expression analyses showed that PpHKT1 was induced in PpHKT1OE2.2 and PpHKTNP6 lines under salt treatment, which confirmed that both over-expression and native expression of PpHKT1 in the Arabidopsis mutant can complement salt tolerance function

The Genetics of Fertility in Soybean

Male and female reproductive structures play an important role in seed development in plants. Abnormalities in male or female reproductive structures can lead to sterility. In soybean, Glycine max (L.) Merr., about 75 sterility mutants have been identified and most of them have been mapped to chromosomes. Mapping results have shown that some chromosomal regions are hotspots for fertility genes. Fine mapping of some of the male-sterile, female-fertile mutants and male-sterile, femalesterile mutants resulted in identification of candidate genes for fertility. Sequence comparisons further helped in locating a few putative candidates. A CACTA- like transposable element that is responsible for reversion from sterility-to-fertility has been identified, and complete association between the presence of a transposon and sterility also has been shown. Several studies are underway that are using transformation sequences to clone fertility genes. Cloning and characterization of genes involved in male sterility and female sterility will help us recognize molecular mechanisms controlling sterility and help us understand the reproductive biology of soybean. This advancement of knowledge will assist in the development of a stable sterility system in soybean that can be utilized for hybrid seed production

Systemic acquired resistance in soybean is regulated by two proteins, Orthologous to Arabidopsis NPR1

<p>Abstract</p> <p>Background</p> <p>Systemic acquired resistance (SAR) is induced in non-inoculated leaves following infection with certain pathogenic strains. SAR is effective against many pathogens. Salicylic acid (SA) is a signaling molecule of the SAR pathway. The development of SAR is associated with the induction of pathogenesis related (<it>PR</it>) genes. Arabidopsis <it>non-expressor </it>of <it>PR1 </it>(<it>NPR1</it>) is a regulatory gene of the SA signal pathway <abbrgrp><abbr bid="B1">1</abbr><abbr bid="B2">2</abbr><abbr bid="B3">3</abbr></abbrgrp>. SAR in soybean was first reported following infection with <it>Colletotrichum trancatum </it>that causes anthracnose disease. We investigated if SAR in soybean is regulated by a pathway, similar to the one characterized in Arabidopsis.</p> <p>Results</p> <p>Pathogenesis-related gene <it>GmPR1 </it>is induced following treatment of soybean plants with the SAR inducer, 2,6-dichloroisonicotinic acid (INA) or infection with the oomycete pathogen, <it>Phytophthora sojae</it>. In <it>P. sojae</it>-infected plants, SAR was induced against the bacterial pathogen, <it>Pseudomonas syringae </it>pv. glycinea. Soybean <it>GmNPR1-1 </it>and <it>GmNPR1-2 </it>genes showed high identities to Arabidopsis <it>NPR1</it>. They showed similar expression patterns among the organs, studied in this investigation. <it>GmNPR1-1 </it>and <it>GmNPR1-2 </it>are the only soybean homologues of <it>NPR1</it>and are located in homoeologous regions. In <it>GmNPR1-1 </it>and <it>GmNPR1-2 </it>transformed Arabidopsis <it>npr1-1 </it>mutant plants, SAR markers: (i) <it>PR-1 </it>was induced following INA treatment and (ii) <it>BGL2 </it>following infection with <it>Pseudomonas syringae </it>pv. tomato (<it>Pst</it>), and SAR was induced following <it>Pst </it>infection. Of the five cysteine residues, Cys⁸², Cys¹⁵⁰, Cys¹⁵⁵, Cys¹⁶⁰, and Cys²¹⁶involved in oligomer-monomer transition in NPR1, Cys²¹⁶ in GmNPR1-1 and GmNPR1-2 proteins was substituted to Ser and Leu, respectively.</p> <p>Conclusion</p> <p>Complementation analyses in Arabidopsis <it>npr1-1 </it>mutants revealed that homoeologous <it>GmNPR1-1 </it>and <it>GmNPR1-2 </it>genes are orthologous to Arabidopsis <it>NPR1</it>. Therefore, SAR pathway in soybean is most likely regulated by <it>GmNPR1 </it>genes. Substitution of Cys²¹⁶residue, essential for oligomer-monomer transition of Arabidopsis NPR1, with Ser and Leu residues in GmNPR1-1 and GmNPR1-2, respectively, suggested that there may be differences between the regulatory mechanisms of GmNPR1 and Arabidopsis NPR proteins.</p

Elemental Analysis of Nanomaterial Using Photon-Atom Interaction Based EDXRF Technique

Presence of trace amount of foreign impurities (both metallic and non-metallic) in standard salts used for sample preparation and during the synthesis process can alter the physical and chemical behavior of the pure and doped nano-materials. Therefore, it becomes important to determine concentration of various elements present in synthesized nano-material sample. In present work, the elemental and compositional analysis of nano-materials synthesized using various methods has been performed using photon-atom interaction based energy dispersive x-ray fluorescence (EDXRF) technique. This technique due to its multielement analytical capability, lower detection limit, capability to analyze metals and non-metals alike and almost no sample preparation requirements can be utilized for analysis of nano-materials. The EDXRF spectrometer involves a 2.4 kW Mo anode x-ray tube (Pananalytic, Netherland) equipped with selective absorbers as an excitation source and an LEGe detector (FWHM = 150 eV at 5.895 keV, Canberra, US) coupled with PC based multichannel analyzer used to collect the fluorescent x-ray spectra. The analytical results showed good agreements with the expected values calculated on the basis of the precursor used in preparation of nano-materials

Strategies for combating plant salinity stress: the potential of plant growth-promoting microorganisms.

Global climate change and the decreasing availability of high-quality water lead to an increase in the salinization of agricultural lands. This rising salinity represents a significant abiotic stressor that detrimentally influences plant physiology and gene expression. Consequently, critical processes such as seed germination, growth, development, and yield are adversely affected. Salinity severely impacts crop yields, given that many crop plants are sensitive to salt stress. Plant growth-promoting microorganisms (PGPMs) in the rhizosphere or the rhizoplane of plants are considered the second genome of plants as they contribute significantly to improving the plant growth and fitness of plants under normal conditions and when plants are under stress such as salinity. PGPMs are crucial in assisting plants to navigate the harsh conditions imposed by salt stress. By enhancing water and nutrient absorption, which is often hampered by high salinity, these microorganisms significantly improve plant resilience. They bolster the plants defenses by increasing the production of osmoprotectants and antioxidants, mitigating salt-induced damage. Furthermore, PGPMs supply growth-promoting hormones like auxins and gibberellins and reduce levels of the stress hormone ethylene, fostering healthier plant growth. Importantly, they activate genes responsible for maintaining ion balance, a vital aspect of plant survival in saline environments. This review underscores the multifaceted roles of PGPMs in supporting plant life under salt stress, highlighting their value for agriculture in salt-affected areas and their potential impact on global food security

Evaluation of spontaneous generation of allelic variation in soybean in response to sexual hybridization and stress

Intra-cultivar variation reported in pure lines of soybean has been hypothesized to result from genetic mechanisms contributing to de novo genetic variation. In this study we have detected allele switching by following segregation pattern of Aconitase-4 isozyme in sexual crosses and pure lines. In sexual crosses, one F2 plant showed switch at the Aconitase- 4 (Aco4) locus from the expected heterozygous genotype Aco4-ac to Aco4-ab. In the pure lines grown in a honeycomb planting design and treated with an accelerated aging test, multiple cases of allele switching were detected at the Aco4 locus. Both single and double switches were detected that were stable and heritable. These findings indicate that the generation of endogenous variation continues in pure lines as a result of intrinsic genetic mechanisms. With a long-term goal of understanding the genetic nature of the changes, we genetically mapped the Aco4 gene to a 3.3 cM region on Chromosome 11. The corresponding physical region is ~293 kb with 39 predicated genes. Of these, Glyma.11g080600 is of particular interest, as it shows 93% and 88% identity to Medicago truncatula and Arabidopsis aconitase genes, respectively. Further characterization of the soybean Aco4 gene may shed light on genetic mechanisms responsible for allele switching

Candidate Gene Identification for a Lethal Chlorophyll-Deficient Mutant in Soybean

Chlorophyll-deficient mutants have been studied persistently to understand genetic mechanisms controlling metabolic pathways. A spontaneous chlorophyll-deficient lethal mutant was observed in self-pollinated progeny of a soybean cultivar “BSR 101”. Observed segregation patterns indicated single-gene recessive inheritance for this lethal-yellow mutant. The objectives of this investigation were to develop a genetic linkage map of the region containing the lethal-yellow (YL_PR350) gene and identify putative candidate genes for this locus. The YL_PR350 gene was mapped to chromosome 15 and is flanked by BARCSOYSSR_15_1591 and BARCSOYSSR_15_1597. This region physically spans ~153 kb and there are 14 predicted genes that lie in this region. The predicted gene Glyma.15g275900 is an excellent candidate for the YL_PR350 gene as it is homologous to an Arabidopsis gene, At3g08010, which codes for a chloroplast-localized protein (ATAB2) involved in the biogenesis of Photosystem I and II. This thylakoid membrane protein is crucial for photosynthesis in Arabidopsis. Future characterization of the candidate gene may enhance our knowledge about photosynthesis, a complex metabolic process critical for sustainability of plants

The Male Sterility Locus ms3 Is Present in a Fertility Controlling Gene Cluster in Soybean

Soybean [Glycine max (L.) Merr.] is self-pollinated. To produce large quantities of hybrid seed, insect-mediated cross-pollination is necessary. An efficient nuclear male-sterile system for hybrid seed production would benefit from molecular and/or phenotypic markers linked to male fertility/sterility loci to facilitate early identification of phenotypes. Nuclear male-sterile, female-fertile ms3 mutant is a single recessive gene and displays high outcrossed seed set with pollinators. Our objective was to map the ms3 locus. A segregating population of 150 F2 plants from Minsoy (PI 27890) × T284H, Ms3ms3 (A00-68), was screened with 231 simple sequence repeat markers. The ms3 locus mapped to molecular linkage group (MLG) D1b (Gm02) and is flanked by markers Satt157 and Satt542, with a distance of 3.7 and 12.3 cM, respectively. Female-partial sterile-1 (Fsp1) and the Midwest Oilseed male-sterile (msMOS) mutants previously were located on MLG D1b. msMOS and Fsp1 are independent genes located very close to each other. All 3 genes are located in close proximity of Satt157. We believe that this is the first report of clustering of fertility-related genes in plants. Characterization of these closely linked genes may help in understanding the evolutionary relationship among them

Using Microsatellites to Understand the Physical Distribution of Recombination on Soybean Chromosomes

Soybean is a major crop that is an important source of oil and proteins. A number of genetic linkage maps have been developed in soybean. Specifically, hundreds of simple sequence repeat (SSR) markers have been developed and mapped. Recent sequencing of the soybean genome resulted in the generation of vast amounts of genetic information. The objectives of this investigation were to use SSR markers in developing a connection between genetic and physical maps and to determine the physical distribution of recombination on soybean chromosomes. A total of 2,188 SSRs were used for sequence-based physical localization on soybean chromosomes. Linkage information was used from different maps to create an integrated genetic map. Comparison of the integrated genetic linkage maps and sequence based physical maps revealed that the distal 25% of each chromosome was the most marker-dense, containing an average of 47.4% of the SSR markers and 50.2% of the genes. The proximal 25% of each chromosome contained only 7.4% of the markers and 6.7% of the genes. At the whole genome level, the marker density and gene density showed a high correlation (R2) of 0.64 and 0.83, respectively with the physical distance from the centromere. Recombination followed a similar pattern with comparisons indicating that recombination is high in telomeric regions, though the correlation between crossover frequency and distance from the centromeres is low (R2 = 0.21). Most of the centromeric regions were low in recombination. The crossover frequency for the entire soybean genome was 7.2%, with extremes much higher and lower than average. The number of recombination hotspots varied from 1 to 12 per chromosome. A high correlation of 0.83 between the distribution of SSR markers and genes suggested close association of SSRs with genes. The knowledge of distribution of recombination on chromosomes may be applied in characterizing and targeting genes