INTRODUCTION:Increased consumption of a high-calorie diet results in the development of metabolic disorders and associated neurological impairments, ultimately leading to cognitive decline. Metabolic disturbances such as hyperglycaemia, systemic insulin resistance, and hyperhomocysteinemia (HHcy) are linked to neurodegenerative diseases. These conditions share a common pathogenic feature of increased susceptibility to protein misfolding and aggregation, which triggers endoplasmic reticulum (ER) stress. Vitamin B12 is vital for brain health, and its deficiency is associated with HHcy and ER stress. This study investigated the effect of vitamin B12 supplementation on diet-induced metabolic disorder-associated ER stress and neurobehavioral outcomes in mice. METHODS:Two-month-old C57BL/6J male mice were randomly assigned to three groups and fed with respective diets for 8 months: Control (C) group (B12: 25 μg/kg diet), HFHS (high-fat, high-sucrose diet; B12: 25 μg/kg), and HFHS+B12 (B12: 50 μg/kg). RESULTS:Fasting blood glucose, glucose tolerance, triglycerides, and total cholesterol were similar across all groups. The HFHS diet led to body weight gain, higher insulin levels, elevated homocysteine (Hcy), and HOMA-IR compared to C group. However, B12 supplementation to HFHS mice significantly reduced Hcy levels compared to HFHS group. Additionally, the HFHS diet resulted in ER stress, neuronal apoptosis and astrogliosis in the cerebral cortex. However, B12 supplementation to HFHS group mitigated ER stress, protected against neuronal cell death, and reduced astrogliosis. Moreover, B12 supplementation enhanced neurotrophic support and reduced anxiety-like behaviour in HFHS+B12 group. CONCLUSIONS:Overall, these findings suggest that B12 supplementation confers protection against Hcy-induced ER stress and apoptosis, highlighting its potential as a neuroprotective agent.
Purpose:Diabetic retinopathy (DR), a severe microvascular complication of both type 1 and type 2 diabetes, is one of the leading causes of blindness. Prolonged hyperglycemia leads to vascular endothelial changes, inflammation, neovascularization, and apoptosis through multiple mechanisms, including increased aldose reductase (AR) activity and formation of advanced glycation end products, which contribute to the development of DR. Based on our previous studies with various functional foods that showed AR inhibition and prevented the formation of advanced glycation end products, in this study, a functional food (FF) mix was formulated and investigated its efficacy against DR progression in a rat model. Methods:An FF mix was prepared with powders of amla pericarp, turmeric rhizome, ginger rhizome, cinnamon bark, and black pepper seeds in a specific proportion. Two-month-old Sprague-Dawley rats were grouped into control (C), streptozotocin-induced diabetes (D), and diabetes fed with FF at two levels (FF1, 0.85 g/100 g diet; FF2, 4.25 g/100 g diet) for 6 months from the induction of diabetes. At the end of the experiment, electroretinography was performed, and the eyes were dissected after the animals were sacrificed. A set of eyes was formalin-fixed for histology and immunohistochemistry examination, and the retina from the remaining eyes was used for immunoblotting analysis. Results:Supplementation of FF mix in the diet to diabetic rats has improved retinal function (electroretinography), as well as prevented histomorphological changes and loss of photoreceptor cells (rhodopsin), compared to untreated diabetic rats. Further, FF mix ameliorated hyperglycemia-induced angiogenesis (vascular endothelial growth factor, hypoxia-inducible factor 1α) and gliosis (glial fibrillary acidic protein) in the diabetic rats, accompanied by decreased inflammation (phosphorylated nuclear factor κB, tumor necrosis factor α, monocyte chemoattractant protein 1) and apoptosis (Bax, Bcl2, caspase3, and caspase12). Conclusions:This study illustrates the potential of an FF mix, attributed to the synergistic effects of its components, alleviating the progression of diabetic retinopathy by reducing diabetes-induced hypoxia, gliosis, and inflammation, while also inhibiting apoptosis in retinal cells.
Diabetic nephropathy (DN) is a chronic microvascular complication of diabetes mellitus, characterized by glomerulomegaly, podocytopathy, and proteinuria. Among the many molecular mechanisms, accumulation of advanced glycation end-products (AGEs) due to non-enzymatic glycation and sorbitol accumulation due to increased aldose reductase (AR) activity are implicated in DN. We previously identified some functional foods and their bioactive molecules for inhibitory potential against AGE formation and, AR activity. Based on those studies, in this study, we formulated a functional food (FF) mix- composed of amla, turmeric, cinnamon, ginger, and black pepper in a specific proportion and tested its efficacy against DN in a rat model. Two-month-old Sprague Dawley rats were grouped into control (C), streptozotocin-induced diabetes (D), and diabetes treated with FF at two doses (FF1-0.85 g and FF2-4.25 g/ 100 g diet). The animals were maintained for 20 weeks on respective diets after the induction of diabetes. Elevated serum albumin, creatinine, and urea were observed in the untreated diabetic group compared to the control. These changes were significantly ameliorated by FF supplementation. FF2 showed better efficacy than FF1 in preventing proteinuria, as reflected in the albumin and creatinine ratio. Further, FF prevented diabetes induced AGE accumulation, inflammation, and activation of the polyol pathway in the kidney. The FF decreased the expression of TGF-beta in the diabetic kidney and prevented fibrotic changes. Most importantly, FF prevented the depletion of podocyte slit diaphragm proteins and histological changes. These results provide a mechanistic basis of FF and its potential against progression of DN in a rat model.
The current study was designed to test a functional food (FF) mixture containing aldose reductase inhibitors and antiglycation bioactive compounds for suppressing the onset and progression of cataracts in a diabetic rat model. Two -month -old Sprague Dawley rats were grouped as control (C), diabetes untreated (D), and diabetic rats treated with FF at two doses (FF1 = 1.35 g and FF2 = 6.25 g/100g of diet). Diabetes was induced by a single injection of streptozotocin. The FF is a mixture of amla, turmeric, black pepper, cinnamon, ginger, and fenugreek added to the rodent diet. The status of cataracts was monitored weekly by a slit lamp examination for 20 weeks, after which animals were sacrificed to collect eye lenses. Feeding FF1 and FF2 to diabetic rats yielded a significant anti -hyperglycaemic effect and marginally prevented body weight loss. FF delayed cataract progression, and FF2 showed better efficacy than FF1. FF prevented the loss of lens crystallins and their insolubilization in diabetic rats. The antioxidant potential of FF was evident with the lowered protein carbonyls, lipid peroxidation, and prevention of altered antioxidant enzyme activities induced by diabetes. These studies demonstrate the efficacy of plant -derived dietary supplements against the onset and progression of cataracts in a well -established rat model of diabetic eye disease.
Aging-related muscle atrophy/sarcopenia is the most common type of muscle impairment that affects the quality of life. In the current study, we examined the effect of a functional food mixture of amla, turmeric, black pepper, cinnamon, and ginger on D-galactose-induced muscle alterations in rats. Wistar rats were randomly divided into three groups: Control (C), D-galactose (G), and D-galactose + functional food mixture intervention (G + I). Rats in group-G and -G + I were injected with D-galactose (300 mg/kg/day) for 90 days. After 3 months of the experimental period, the rats were sacrificed to collect gastrocnemius muscle. Group-G rats showed elevated levels of inflammatory cytokines (TNF alpha and NF-kB), atrogenes (atrogin-1 and MuRF1), decreased insulin/IGF1 signaling (decreased AKT phosphorylation), altered mitochondrial dynamics (increased fission and decreased fusion proteins), increased apoptotic mediators (Bax/Bcl-2, and caspase-3), and decreased muscle cell cross-sectional area when compared with group-C (p < 0.05). Interestingly, supplementation with the functional food mixture prevented galactose-induced alterations in the muscle. The observed anti-inflammatory, insulin-sensitizing, mitochondria-protective, and antiapoptotic effects of the functional food could be the underlying mechanisms in displaying positive effects against galactose-induced muscle atrophy and, hence, may be useful for the prevention of age-related muscle disorders.
Background: Ageing entails a gradual decline of organ structure and function, in-cluding the kidneys. Chronic kidney disease is associated with cardiovascular complications, hyperlipidemia, and metabolic bone disease. Functional foods with antioxidant, anti-inflammatory, and anti-apoptotic properties protect kidney functions by influencing metabo-lism and immunity. Objective: We studied the prophylactic effect of a functional food mix [amla, cinnamon, ginger, turmeric, and black pepper] on D-galactose-induced renal ageing. Methods: Six-month-old female Wistar rats were divided into Control, D-galactose, and D-galactose + functional food groups and maintained for 90 days. The plasma lipid profile and renal function tests were assayed using spectrophotometry. The protein markers of the endo-plasmic reticulum [ER] stress, inflammation, and apoptosis were analysed by immunoblot-ting. Results: The results illustrate that the functional food prevented D-galactose-induced histo-logical alterations, dyslipidemia, podocin loss, renal dysfunction, ER stress [GRP78, pIRE1, and CHOP], inflammation [NF-κB, and TNFα], and apoptosis [Bax, and caspase-3] in the kidney. Conclusion: The antiglycation, antioxidant, and anti-inflammatory roles of functional food are the basis for preventing D-galactose-induced renal injury. Hence, it could be a prophy-lactic measure in individuals with chronic kidney disease.
Pearl millet [Pennisetum glaucum (L) R. Br.] is an important cereal crop of the semiarid tropics, which can withstand prolonged drought and heat stress. Considering an active involvement of the aquaporin (AQP) genes in water transport and desiccation tolerance besides several basic functions, their potential role in abiotic stress tolerance was systematically characterized and functionally validated. A total of 34 AQP genes from P. glaucum were identified and categorized into four subfamilies, viz., plasma membrane intrinsic proteins (PIPs), tonoplast intrinsic proteins (TIPs), nodulin-26-like intrinsic proteins (NIPs), and small basic intrinsic proteins (SIPs). Sequence analysis revealed that PgAQPs have conserved characters of AQP genes with a closer relationship to sorghum. The PgAQPs were expressed differentially under high vapor pressure deficit (VPD) and progressive drought stresses where the PgPIP2;6 gene showed significant expression under high VPD and drought stress. Transgenic tobacco plants were developed by heterologous expression of the PgPIP2;6 gene and functionally characterized under different abiotic stresses to further unravel their role. Transgenic tobacco plants in the T2 generations displayed restricted transpiration and low root exudation rates in low- and high-VPD conditions. Under progressive drought stress, wild-type (WT) plants showed a quick or faster decline of soil moisture than transgenics. While under heat stress, PgPIP2;6 transgenics showed better adaptation to heat (40°C) with high canopy temperature depression (CTD) and low transpiration; under low-temperature stress, they displayed lower transpiration than their non-transgenic counterparts. Cumulatively, lower transpiration rate (Tr), low root exudation rate, declined transpiration, elevated CTD, and lower transpiration indicate that PgPIP2;6 plays a role under abiotic stress tolerance. Since the PgPIP2;6 transgenic plants exhibited better adaptation against major abiotic stresses such as drought, high VPD, heat, and cold stresses by virtue of enhanced transpiration efficiency, it has the potential to engineer abiotic stress tolerance for sustained growth and productivity of crops.
Potassium (K + ) is the most abundant cation that plays a crucial role in various cellular processes in plants. Plants have developed an efficient mechanism for the acquisition of K + when grown in K + deficient or saline soils. A total of 47 K + transport gene homologs (27 HAKs, 4 HKTs, 2 KEAs, 9 AKTs, 2 KATs, 2 TPCs, and 1 VDPC) have been identified in Sorghum bicolor . Of 47 homologs, 33 were identified as K + transporters and the remaining 14 as K + channels. Chromosome 2 has been found as the hotspot of K + transporters with 9 genes. Phylogenetic analysis revealed the conservation of sorghum K + transport genes akin to Oryza sativa . Analysis of regulatory elements indicates the key roles that K + transport genes play under different biotic and abiotic stress conditions. Digital expression data of different developmental stages disclosed that expressions were higher in milk, flowering, and tillering stages. Expression levels of the genes SbHAK27 and SbKEA2 were higher during milk, SbHAK17 , SbHAK11 , SbHAK18 , and SbHAK7 during flowering, SbHAK18 , SbHAK10 , and 23 other gene expressions were elevated during tillering inferring the important role that K + transport genes play during plant growth and development. Differential transcript expression was observed in different tissues like root, stem, and leaf under abiotic stresses such as salt, drought, heat, and cold stresses. Collectively, the in-depth genome-wide analysis and differential transcript profiling of K + transport genes elucidate their role in ion homeostasis and stress tolerance mechanisms.
Acute lung injury (ALI), is a severe inflammatory lung disease. We tested the prophylactic effect of a functional food mix comprising three anti-inflammatory plant products: turmeric, amla, and black pepper (TAB) against lipopolysaccharide (LPS)-induced ALI in rats. Two-month-old male Wistar rats were randomly divided into three groups: control (C), LPS (5 mg/kg), and LPS with TAB (TAB). After 6 h of LPS injection, the rats were sacrificed by cervical decapitation to collect the lung tissue. Results showed that TAB partially ameliorated LPS-induced increase in circulating inflammatory cytokines (TNFα and IL6) and significantly prevented lung histopathological changes. TAB also suppressed LPS-activated ER stress markers (GRP78, pIRE1, and CHOP) and apoptotic markers (caspase-3 and − 12) in the lung. The anti-inflammatory effects of the TAB support its potential use as an adjuvant to mitigate ALI. Importantly, TAB’s ingredients have been used for centuries as part of the diet with limited or no toxic effects.
Systematic genome-wide analysis of Sorghum bicolor revealed the identification of a total of 48 homologous genes comprising 21 proline-rich proteins (PRPs) and 27 hybrid proline-rich proteins (HyPRPs). Comprehensive scrutiny of these gene homologs was conducted for gene structure, phylogenetic investigations, chromosome mapping, and subcellular localization of proteins. Promoter analysis uncovered the regions rich with phosphorous- (BIHD), ammonium-, sulfur-responsive (SURE), and iron starvation-responsive (IRO2) along with biotic, abiotic, and development-specific cis-elements. Further, PRPs exhibit more methylation and acetylation sites in comparison with HyPRPs. miRNAs have been predicted which might play a role in cleavage and translation inhibition. Several of the SbPRP genes were stimulated in a tissue-specific manner under drought, salt, heat, and cold stresses. Additionally, exposure of plants to abscisic acid (ABA) and zinc (Zn) also triggered PRP genes in a tissue-dependent way. Among them, SbPRP17 has been found upregulated markedly in all tissues irrespective of the stress imposed. The expressions of SbHyPRPs, especially SbHyPRP2, SbHyPRP6, and SbHyPRP17 were activated under all stresses in all three tissues. On the other hand, SbHyPRP8 (root only) and SbHyPRP12 (all three tissues) were highly responsive to cold stress and ABA while SbHyPRP26 was induced by drought and Zn in the stem. Taken together, this study indicates the critical roles that SbPRPs and SbHyPRPs play during diverse abiotic stress conditions and notably the plausible roles that these genes play upon exposure to zinc, the crucial micronutrient in plants.
Abstract Acute lung injury (ALI), is a severe inflammatory lung disease. We tested the prophylactic effect of a functional food comprising three different anti-inflammatory plant products: turmeric, amla, and black pepper (TAB) against lipopolysaccharide (LPS)-induced ALI in rats. Two-month-old male Wistar rats were randomly divided into three groups: control (C), LPS (5mg/kg), and LPS with TAB (TAB). After 6 hours of LPS injection, the rats were sacrificed by cervical decapitation to collect the lung tissue. Results showed that TAB partially ameliorated LPS-induced circulating inflammatory cytokines (TNFα and IL6) and significantly prevented lung histopathological changes. TAB also suppressed LPS-activated ER stress markers (GRP78, pIRE1, and CHOP) and apoptotic proteins (caspase-3 and -12) in the lung. The anti-inflammatory effects of the TAB support its potential use at least as an adjuvant to mitigate ALI. Importantly, the ingredients of TAB are in use for centuries as part of their diet with limited or no toxic effects identified.
Genes encoding bacterial cold shock proteins A (CspA, 213 bp) and B (CspB, 216 bp) were isolated from Escherichia coli strain K12, which showed 100% homology with gene sequences isolated from other bacterial species. In silico domain, analysis showed eukaryotic conserved cold shock domain (CSD) and ribonuclease-binding domain (RBD) indicating that they bind to RNA and are involved in temperature stress tolerance. Overexpression of these two genes in E. coli resulted in higher growth in presence of 200 mM NaCl and 300 mM mannitol. Western blot confirmed the translational products of the two genes. Seedlings of indica rice were transformed with Agrobacterium tumefaciens containing pCAMBIA1301 CspA and CspB genes. Transgene integration was confirmed by beta-glucuronidase (GUS) histochemical assay, polymerase chain reaction (PCR) amplification, and gene copy number by Southern blotting. Chlorophyll, proline, Na+, and K+ contents were higher in transgenics exposed to 150 mM NaCl and drought (imposed by withholding water) stresses during floral initiation stage. Catalase (CAT), superoxide dismutase (SOD), and guaiacol peroxidase (GPX) activities increased, while malondialdehyde (MDA) content was low in transgenics. Transgenics displayed increased root, shoot, and panicle lengths, root dry mass, and a distinct stay-green (SGR) phenotype. Higher transcript levels of CspA, CspB, SGR, chlorophyllase, isopentenyl adenine transferase 1 (IPT1), 9-cis-epoxycarotenoid dioxygenase (NCED), SOD, and sirtuin 1 (SIRT1) genes were observed in transgenics compared to wild type plants (WT) under multiple stresses. Present work indicates that bacterial chaperone proteins are capable of imparting SGR phenotype, salt and drought stress tolerance alongside grain improvement.
It is important to analyse the degree of genetic variation existing within the genome to extend the genetic base of linseed/flaxseed accessions in order to preserve, evaluate and use genetic resources accurately and successfully. The main aim of the current investigation was to evaluate the scope and spread of genomic variation across different linseed accessions by employing molecular markers (RAPD). The genomic DNA of 12 linseed accessions was amplified with 16 decamer RAPD primers that generated 81 total bands, among which 75 bands were polymorphic and 6 bands were monomorphic. Polymorphic band numbers varied from least 2 (OPS-11) to highest 10 (OPS-07). The magnitude of polymorphism ranged from 75% to 100% among all accessions with a mean of 93.15 % across all the accessions. The value of Polymorphic Information Content (PIC) varied from 0.133% to 0.708% with a mean of 0.45% for each primer. The maximum PIC value (0.708) was found with the primer OPS-07 and (0.702) with OPM-13. The primer OPS-03 showed the minimum PIC value (0.133). Two main different clusters -I and -II were seen in the cluster analysis depending on RAPD data. Cluster-II comprises one accession (IC 564585) that was the highly varied accession, whereas Cluster-I comprises of some sub clusters with all the remaining accessions. The Jaccard's similarity coefficient varied from 8.2 to 96.3%. The accessions BHU-A and BHU-B had the highest genetic similarity (96.3%), followed by BHU-B and IC 564605 (96.2%). More divergent accessions were discovered to be IC 564585, IC 564616, IC 564631, IC 564622, and IC 564630. The current investigation provides innovative knowledge to breeders on the germplasm of linseed that would be employed in subsequent research to improve linseed genotypes.
BACKGROUND:Sorghum, the C4 dry-land cereal, important for food, fodder, feed and fuel, is a model crop for abiotic stress tolerance with smaller genome size, genetic diversity, and bio-energy traits. The heat shock proteins/chaperonin 60s (HSP60/Cpn60s) assist the plastid proteins, and participate in the folding and aggregation of proteins. However, the functions of HSP60s in abiotic stress tolerance in Sorghum remain unclear. METHODS:Genome-wide screening and in silico characterization of SbHSP60s were carried out along with tissue and stress-specific expression analysis. RESULTS:A total of 36 HSP60 genes were identified in Sorghum bicolor. They were subdivided into 2 groups, the HSP60 and HSP10 co-chaperonins encoded by 30 and 6 genes, respectively. The genes are distributed on all the chromosomes, chromosome 1 being the hot spot with 9 genes. All the HSP60s were found hydrophilic and highly unstable. The HSP60 genes showed a large number of introns, the majority of them with more than 10. Among the 12 paralogs, only 1 was tandem and the remaining 11 segmental, indicating their role in the expansion of SbHSP60s. Majority of the SbHSP60 genes expressed uniformly in leaf while a moderate expression was observed in the root tissues, with the highest expression displayed by SbHSP60-1. From expression analysis, SbHSP60-3 for drought, SbHSP60-9 for salt, SbHSP60-9 and 24 for heat and SbHSP60-3, 9 and SbHSP10-2 have been found implicated for cold stress tolerance and appeared as the key regulatory genes. CONCLUSION:This work paves the way for the utilization of chaperonin family genes for achieving abiotic stress tolerance in plants.
Phosphorus is considered to be one of the least plant macronutrients available in the soil. This mineral element plays an important role in plant growth and development and also represents essential constitutes of nucleic acids, phosphoproteins, phospholipids, and ATP. Various environmental factors in the form of abiotic stresses adversely affect crop production in a significant manner. It is well established that phosphate stress tolerance is regulated using transcription factors (TFs) and microRNAs that enable plants to cope with unfavorable conditions. This chapter gives an overview of the role of important genes-encoding TFs belonging to four multigene families (WRKY, MYB, ZFP, and ARF). Since, these four specific families of TFs and their regulatory partners, i.e., noncoding, RNAs have gained widespread attention on account of their critical role in Pi-stress tolerance in plants; there is a growing interest in the use of genomic candidates for developing Pi-stress-tolerant plants.
Lysine (Lys) is indispensable nutritionally, and its levels in plants are modulated by both transcriptional and post-transcriptional control during plant ontogeny. Animal glutamate receptor homologs have been detected in plants, which may participate in several plant processes through the Lys catabolic products. Interestingly, a connection between Lys and serotonin metabolism has been established recently in rice. 2-Aminoadipate, a catabolic product of Lys appears to play a critical role between serotonin accumulation and the color of rice endosperm/grain. It has also been shown that expression of some lysine-methylated proteins and genes encoding lysine-methyltransferases (KMTs) are regulated by cadmium even as it is known that Lys biosynthesis and its degradation are modulated by novel mechanisms. Three complex pathways co-exist in plants for serine (Ser) biosynthesis, and the relative preponderance of each pathway in relation to plant development or abiotic stress tolerance are being unfolded slowly. But the phosphorylated pathway of L-Ser biosynthesis (PPSB) appears to play critical roles and is essential in plant metabolism and development. Ser, which participates indirectly in purine and pyrimidine biosynthesis and plays a pivotal role in plant metabolism and signaling. Also, L-Ser has been implicated in plant responses to both biotic and abiotic stresses. A large body of information implicates Lys-rich and serine/arginine-rich (SR) proteins in a very wide array of abiotic stresses. Interestingly, a link exists between Lys-rich K-segment and stress tolerance levels. It is of interest to note that abiotic stresses largely influence the expression patterns of SR proteins and also the alternative splicing (AS) patterns. We have checked if any lncRNAs form a cohort of differentially expressed genes from the publicly available PPSB, sequence read archives of NCBI GenBank. Finally, we discuss the link between Lys and Ser synthesis, catabolism, Lys-proteins, and SR proteins during plant development and their myriad roles in response to abiotic stresses.
Late embryogenesis abundant (LEA) proteins, the space fillers or molecular shields, are the hydrophilic protective proteins which play an important role during plant development and abiotic stress. The systematic survey and characterization revealed a total of 68 LEA genes, belonging to 8 families in Sorghum bicolor. The LEA-2, a typical hydrophobic family is the most abundant family. All of them are evenly distributed on all 10 chromosomes and chromosomes 1, 2, and 3 appear to be the hot spots. Majority of the S. bicolor LEA (SbLEA) genes are intron less or have fewer introns. A total of 22 paralogous events were observed and majority of them appear to be segmental duplications. Segmental duplication played an important role in SbLEA-2 family expansion. A total of 12 orthologs were observed with Arabidopsis and 13 with Oryza sativa. Majority of them are basic in nature, and targeted by chloroplast subcellular localization. Fifteen miRNAs targeted to 25 SbLEAs appear to participate in development, as well as in abiotic stress tolerance. Promoter analysis revealed the presence of abiotic stress-responsive DRE, MYB, MYC, and GT1, biotic stress-responsive W-Box, hormone-responsive ABA, ERE, and TGA, and development-responsive SKn cis-elements. This reveals that LEA proteins play a vital role during stress tolerance and developmental processes. Using microarray data, 65 SbLEA genes were analyzed in different tissues (roots, pith, rind, internode, shoot, and leaf) which show clear tissue specific expression. qRT-PCR analysis of 23 SbLEA genes revealed their abundant expression in various tissues like roots, stems and leaves. Higher expression was noticed in stems compared to roots and leaves. Majority of the SbLEA family members were up-regulated at least in one tissue under different stress conditions. The SbLEA3-2 is the regulator, which showed abundant expression under diverse stress conditions. Present study provides new insights into the formation of LEAs in S. bicolor and to understand their role in developmental processes under stress conditions, which may be a valuable source for future research.
Members of the plant Heme Activator Protein (HAP) or NUCLEAR FACTOR Y (NF-Y) are trimeric transcription factor complexes composed of the NF-YA, NF-YB and NF-YC subfamilies. They bind to the CCAAT box in the promoter regions of the target genes and regulate gene expressions. Plant NF-Ys were reported to be involved in adaptation to several abiotic stresses as well as in development. In silico analysis of Sorghum bicolor genome resulted in the identification of a total of 42 NF-Y genes, among which 8 code for the SbNF-YA, 19 for SbNF-YB and 15 for the SbNF-YC subunits. Analysis was also performed to characterize gene structures, chromosomal distribution, duplication status, protein subcellular localizations, conserved motifs, ancestral protein sequences, miRNAs and phylogenetic tree construction. Phylogenetic relationships and ortholog predictions displayed that sorghum has additional NF-YB genes with unknown functions in comparison with Arabidopsis. Analysis of promoters revealed that they harbour many stress-related cis-elements like ABRE and HSE, but surprisingly, DRE and MYB elements were not detected in any of the subfamilies. SbNF-YA1, 2, and 6 were found upregulated under 200 mM salt and 200 mM mannitol stresses. While NF-YA7 appeared associated with high temperature (40°C) stress, NF-YA8 was triggered by both cold (4°C) and high temperature stresses. Among NF-YB genes, 7, 12, 15, and 16 were induced under multiple stress conditions such as salt, mannitol, ABA, cold and high temperatures. Likewise, NF-YC 6, 11, 12, 14, and 15 were enhanced significantly in a tissue specific manner under multiple abiotic stress conditions. Majority of the mannitol (drought)-inducible genes were also induced by salt, high temperature stresses and ABA. Few of the high temperature stress-induced genes are also induced by cold stress (NF-YA2, 4, 6, 8, NF-YB2, 7, 10, 11, 12, 14, 16, 17, NF-YC4, 6, 12, and 13) thus suggesting a cross talk among them. This work paves the way for investigating the roles of diverse sorghum NF-Y proteins during abiotic stress responses and provides an insight into the evolution of diverse NF-Y members.
The SPX gene family, ubiquitous in all vascular plants, plays a critical role in plant development and growth as well as in response to phosphorus stress. Based on genomic census, 46 TaSPX genes were identified in the wheat genome. All of them are evenly distributed on 13 of the 21 wheat chromosomes and chromosome 7A contains the largest members. As many as 57 gene specific SSRs were discovered among genomic sequences of identified TaSPXs. MicroRNA target analysis revealed that TaSPX genes were targeted by 9 different miRNAs including tae-miR1120a, tae-miR1120b-3p, tae-miR1120c-5p, tae-miR1122b-3p, tae-miR1122c-3p, tae-miR1130a, tae-miR1130b-3p, tae-miR1137a, and tae-miR1137b-5p. Expression profiles derived from transcriptome data and real-time quantitative PCR revealed that TaSPX genes were significantly induced by Pi starvation. The modeled 3D structure of wheat SPX proteins shared high level of homology with template structures, providing information to understand their functions at proteomic level. We have also refined the modeled 3D structures on 10 ns using molecular dynamics simulations for conformational stability. The discovered members of SPX gene family and their targeting miRNAs may provide resource for genetic improvement and promote P use efficiency in cereals.