Molecular genetics and phenotypic assessment of foxtail millet (Setaria italica (L.) P. Beauv.) landraces revealed remarkable variability of morpho-physiological, yield, and yield‐related traits

Foxtail millet (Setaria italica (L.) P. Beauv.) is highly valued for nutritional traits, stress tolerance and sustainability in resource-poor dryland agriculture. However, the low productivity of this crop in semi-arid regions of Southern India, is further threatened by climate stress. Landraces are valuable genetic resources, regionally adapted in form of novel alleles that are responsible for cope up the adverse conditions used by local farmers. In recent years, there is an erosion of genetic diversity. We have hypothesized that plant genetic resources collected from the semi-arid climatic zone would serve as a source of novel alleles for the development of climate resilience foxtail millet lines with enhanced yield. Keeping in view, there is an urgent need for conservation of genetic resources. To explore the genetic diversity, to identify superior genotypes and novel alleles, we collected a heterogeneous mixture of foxtail millet landraces from farmer fields. In an extensive multi-year study, we developed twenty genetically fixed foxtail millet landraces by single seed descent method. These landraces characterized along with four released cultivars with agro-morphological, physiological, yield and yield-related traits assessed genetic diversity and population structure. The landraces showed significant diversity in all the studied traits. We identified landraces S3G5, Red, Black and S1C1 that showed outstanding grain yield with earlier flowering, and maturity as compared to released cultivars. Diversity analysis using 67 simple sequence repeat microsatellite and other markers detected 127 alleles including 11 rare alleles, averaging 1.89 alleles per locus, expected heterozygosity of 0.26 and an average polymorphism information content of 0.23, collectively indicating a moderate genetic diversity in the landrace populations. Euclidean Ward’s clustering, based on the molecular markers, principal coordinate analysis and structure analysis concordantly distinguished the genotypes into two to three sub-populations. A significant phenotypic and genotypic diversity observed in the landraces indicates a diverse gene pool that can be utilized for sustainable foxtail millet crop improvement

Antarctic Moss Multiprotein Bridging Factor 1c Overexpression in Arabidopsis Resulted in Enhanced Tolerance to Salt Stress

Polytrichastrum alpinum is one of the moss species that survives extreme conditions in the Antarctic. In order to explore the functional benefits of moss genetic resources, P. alpinum multiprotein-bridging factor 1c gene (PaMBF1c) was isolated and characterized. The deduced amino acid sequence of PaMBF1c comprises of a multiprotein-bridging factor (MBF1) domain and a helix-turn-helix (HTH) domain. PaMBF1c expression was induced by different abiotic stresses in P. alpinum, implying its roles in stress responses. We overexpressed PaMBF1c in Arabidopsis and analyzed the resulting phenotypes in comparison with wild type and/or Arabidopsis MBF1c (AtMBF1c) overexpressors. Overexpression of PaMBF1c in Arabidopsis resulted in enhanced tolerance to salt and osmotic stress, as well as to cold and heat stress. More specifically, enhanced salt tolerance was observed in PaMBF1c overexpressors in comparison to wild type but not clearly observable in AtMBF1c overexpressing lines. Thus, these results implicate the evolution of PaMBF1c under salt-enriched Antarctic soil. RNA-Seq profiling of NaCl-treated plants revealed that 10 salt-stress inducible genes were already up-regulated in PaMBF1c overexpressing plants even before NaCl treatment. Gene ontology enrichment analysis with salt up-regulated genes in each line uncovered that the terms lipid metabolic process, ion transport, and cellular amino acid biosynthetic process were significantly enriched in PaMBF1c overexpressors. Additionally, gene enrichment analysis with salt down-regulated genes in each line revealed that the enriched categories in wild type were not significantly overrepresented in PaMBF1c overexpressing lines. The up-regulation of several genes only in PaMBF1c overexpressing lines suggest that enhanced salt tolerance in PaMBF1c-OE might involve reactive oxygen species detoxification, maintenance of ATP homeostasis, and facilitation of Ca2+ signaling. Interestingly, many salt down-regulated ribosome- and translation-related genes were not down-regulated in PaMBF1c overexpressing lines under salt stress. These differentially regulated genes by PaMBF1c overexpression could contribute to the enhanced tolerance in PaMBF1c overexpressing lines under salt stress

Salt and drought stress responses in cultivated beets (Beta vulgaris L.) and wild beet (Beta maritima L.)

Cultivated beets, including leaf beets, garden beets, fodder beets, and sugar beets, which belong to the species Beta vulgaris L., are economically important edible crops that have been originated from a halophytic wild ancestor, Beta maritima L. (sea beet or wild beet). Salt and drought are major abiotic stresses, which limit crop growth and production and have been most studied in beets compared to other environmental stresses. Characteristically, beets are salt- and drought-tolerant crops; however, prolonged and persistent exposure to salt and drought stress results in a significant drop in beet productivity and yield. Hence, to harness the best benefits of beet cultivation, knowledge of stress-coping strategies, and stress-tolerant beet varieties, are prerequisites. In the current review, we have summarized morpho-physiological, biochemical, and molecular responses of sugar beet, fodder beet, red beet, chard (B. vulgaris L.), and their ancestor, wild beet (B. maritima L.) under salt and drought stresses. We have also described the beet genes and noncoding RNAs previously reported for their roles in salt and drought response/tolerance. The plant biologists and breeders can potentiate the utilization of these resources as prospective targets for developing crops with abiotic stress tolerance

Salt and drought stress-mitigating approaches in sugar beet (Beta vulgaris L.) to improve its performance and yield

Main conclusion: Although sugar beet is a salt- and drought-tolerant crop, high salinity, and water deprivation significantly reduce its yield and growth. Several reports have demonstrated stress tolerance enhancement through stress-mitigating strategies including the exogenous application of osmolytes or metabolites, nanoparticles, seed treatments, breeding salt/drought-tolerant varieties. These approaches would assist in achieving sustainable yields despite global climatic changes. Abstract: Sugar beet (Beta vulgaris L.) is an economically vital crop for ~ 30% of world sugar production. They also provide essential raw materials for bioethanol, animal fodder, pulp, pectin, and functional food-related industries. Due to fewer irrigation water requirements and shorter regeneration time than sugarcane, beet cultivation is spreading to subtropical climates from temperate climates. However, beet varieties from different geographical locations display different stress tolerance levels. Although sugar beet can endure moderate exposure to various abiotic stresses, including high salinity and drought, prolonged exposure to salt and drought stress causes a significant decrease in crop yield and production. Hence, plant biologists and agronomists have devised several strategies to mitigate the stress-induced damage to sugar beet cultivation. Recently, several studies substantiated that the exogenous application of osmolytes or metabolite substances can help plants overcome injuries induced by salt or drought stress. Furthermore, these compounds likely elicit different physio-biochemical impacts, including improving nutrient/ionic homeostasis, photosynthetic efficiency, strengthening defense response, and water status improvement under various abiotic stress conditions. In the current review, we compiled different stress-mitigating agricultural strategies, prospects, and future experiments that can secure sustainable yields for sugar beets despite high saline or drought conditions

Development of SNP Markers for White Immature Fruit Skin Color in Cucumber (Cucumis sativus L.) Using QTL-seq and Marker Analyses

Despite various efforts in identifying the genes governing the white immature fruit skin color in cucumber, the genetic basis of the white immature fruit skin color is not well known. In the present study, genetic analysis showed that a recessive gene confers the white immature fruit skin-color phenotype over the light-green color of a Korean slicer cucumber. High-throughput QTL-seq combined with bulked segregation analysis of two pools with the extreme phenotypes (white and light-green fruit skin color) in an F2 population identified two significant genomic regions harboring QTLs for white fruit skin color within the genomic region between 34.1 and 41.67 Mb on chromosome 3, and the genomic region between 12.2 and 12.7 Mb on chromosome 5. Further, nonsynonymous SNPs were identified with a significance of p < 0.05 within the QTL regions, resulting in eight homozygous variants within the QTL region on chromosome 3. SNP marker analysis uncovered the novel missense mutations in Chr3CG52930 and Chr3CG53640 genes and showed consistent results with the phenotype of light-green and white fruit skin-colored F2 plants. These two genes were located 0.5 Mb apart on chromosome 3, which are considered strong candidate genes. Altogether, this study laid a solid foundation for understanding the genetic basis and marker-assisted breeding of immature fruit skin color in cucumber

Development of SNP Markers for White Immature Fruit Skin Color in Cucumber (<i>Cucumis sativus</i> L.) Using QTL-seq and Marker Analyses

Despite various efforts in identifying the genes governing the white immature fruit skin color in cucumber, the genetic basis of the white immature fruit skin color is not well known. In the present study, genetic analysis showed that a recessive gene confers the white immature fruit skin-color phenotype over the light-green color of a Korean slicer cucumber. High-throughput QTL-seq combined with bulked segregation analysis of two pools with the extreme phenotypes (white and light-green fruit skin color) in an F2 population identified two significant genomic regions harboring QTLs for white fruit skin color within the genomic region between 34.1 and 41.67 Mb on chromosome 3, and the genomic region between 12.2 and 12.7 Mb on chromosome 5. Further, nonsynonymous SNPs were identified with a significance of p Chr3CG52930 and Chr3CG53640 genes and showed consistent results with the phenotype of light-green and white fruit skin-colored F2 plants. These two genes were located 0.5 Mb apart on chromosome 3, which are considered strong candidate genes. Altogether, this study laid a solid foundation for understanding the genetic basis and marker-assisted breeding of immature fruit skin color in cucumber

An insight into the abiotic stress responses of cultivated beets (Beta vulgaris L.)

Cultivated beets (sugar beets, fodder beets, leaf beets, and garden beets) belonging to the species Beta vulgaris L. are important sources for many products such as sugar, bioethanol, animal feed, human nutrition, pulp residue, pectin extract, and molasses. Beta maritima L. (sea beet or wild beet) is a halophytic wild ancestor of all cultivated beets. With a requirement of less water and having shorter growth period than sugarcane, cultivated beets are preferentially spreading from temperate regions to subtropical countries. The beet cultivars display tolerance to several abiotic stresses such as salt, drought, cold, heat, and heavy metals. However, many environmental factors adversely influence growth, yield, and quality of beets. Hence, selection of stress-tolerant beet varieties and knowledge on the response mechanisms of beet cultivars to different abiotic stress factors are most required. The present review discusses morpho-physiological, biochemical, and molecular responses of cultivated beets (B. vulgaris L.) to different abiotic stresses including alkaline, cold, heat, heavy metals, and UV radiation. Additionally, we describe the beet genes reported for their involvement in response to these stress conditions

Cloning and Functional Characterization of a Vacuolar Na⁺/H⁺ Antiporter Gene from Mungbean (<i>VrNHX1</i>) and Its Ectopic Expression Enhanced Salt Tolerance in <i>Arabidopsis thaliana</i>

<div><p>Plant vacuolar NHX exchangers play a significant role in adaption to salt stress by compartmentalizing excess cytosolic Na⁺ into vacuoles and maintaining cellular homeostasis and ionic equilibrium. We cloned an orthologue of the vacuolar Na⁺/H⁺ antiporter gene, <i>VrNHX1</i> from mungbean (<i>Vigna radiata</i>), an important Asiatic grain legume. The <i>VrNHX1</i> (Genbank Accession number JN656211.1) contains 2095 nucleotides with an open reading frame of 1629 nucleotides encoding a predicted protein of 542 amino acids with a deduced molecular mass of 59.6 kDa. The consensus amiloride binding motif (⁸⁴LFFIYLLPPI⁹³) was observed in the third putative transmembrane domain of <i>VrNHX1</i>. Bioinformatic and phylogenetic analysis clearly suggested that <i>VrNHX1</i> had high similarity to those of orthologs belonging to Class-I clade of plant NHX exchangers in leguminous crops. <i>VrNHX1</i> could be strongly induced by salt stress in mungbean as the expression in roots significantly increased in presence of 200 mM NaCl with concomitant accumulation of total [Na⁺]. Induction of <i>VrNHX1</i> was also observed under cold and dehydration stress, indicating a possible cross talk between various abiotic stresses. Heterologous expression in salt sensitive yeast mutant AXT3 complemented for the loss of yeast vacuolar <i>NHX1</i> under NaCl, KCl and LiCl stress indicating that <i>VrNHX1</i> was the orthologue of <i>ScNHX1</i>. Further, AXT3 cells expressing <i>VrNHX1</i> survived under low pH environment and displayed vacuolar alkalinization analyzed using pH sensitive fluorescent dye BCECF-AM. The constitutive and stress inducible expression of <i>VrNHX1</i> resulted in enhanced salt tolerance in transgenic <i>Arabidopsis thaliana</i> lines. Our work suggested that <i>VrNHX1</i> was a salt tolerance determinant in mungbean.</p></div

Total intracellular ion estimation in yeast strains W303-1B, AXTYES2.0 and AXTVrNHX1.

<p>Yeast cells were grown in APG medium (pH 4.0) with 1 mM KCl supplemented in presence (stressed) or absence of 75 mM NaCl (unstressed) and harvested at a cell density of 0.3. Total intracellular, vacuolar and cytoplasmic Na⁺ and K⁺ content was determined as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0106678#s2" target="_blank">materials and methods</a> section. Data are means of 3 independent events (n = 3) and standard errors are plotted in the graph. Statistically significant values at P≤0.05 are indicated as “*”, using Bonferroni analysis.</p

The phylogenetic tree for plant Na⁺/H⁺ antiporters was generated using MEGA4: Tree Explorer software.

<p>The evolutionary history was inferred using the neighbor-joining method and analyzed using bootstrap analysis with 500 replicates. Branches corresponding to partitions reproduced in less than 50% bootstrap replicates are collapsed. The tree is drawn to scale, with branch lengths in the same units as those of the evolutionary distances used to infer the phylogenetic tree. The evolutionary distances were computed using the Poisson correction method and are in the units of the number of amino acid substitutions per site. The GenBank Accession numbers for NHX proteins used are: VrNHX1 (AEO50758.1), VuNHX1 (AEO72079.2), GmNHX1 (AAY430061.1), CkNHX1 (ABG89337.1), MsNHX1 (AAS84487.1), CaNHX1 (ADL28385.1), TrNHX1 (ABV00895.1), LtNHX1 (ACE78322.1), AhNHX1 (ADK74832.1), AtNHX1 (NM_122597.2).</p