Identification and characterization of differentially expressed genes in the rice root following exogenous application of spermidine during salt stress.

Salinity is a major limiting factor in crop production. Exogenous spermidine (spd) effectively ameliorates salt injury, though the underlying molecular mechanism is poorly understood. We have used a suppression subtractive hybridization method to construct a cDNA library that has identified up-regulated genes from rice root under the treatment of spd and salt. Total 175 high-quality ESTs of about 100-500 bp in length with an average size of 200 bp are isolated, clustered and assembled into a collection of 62 unigenes. Gene ontology analysis using the KEGG pathway annotation database has classified the unigenes into 5 main functional categories and 13 subcategories. The transcripts abundance has been validated using Real-Time PCR. We have observed seven different types of post-translational modifications in the DEPs. 44 transmembrane helixes are predicted in 6 DEPs. This above information can be used as first-hand data for dissecting the administrative role of spd during salinity.

Nanoparticles regulate redox metabolism in plants during abiotic stress within hormetic boundaries.

Abiotic stress management remains under scrutiny because of the unpredictable nature of climate, which undergoes abrupt alterations. Population pressure, loss of cultivable lands, environmental pollution and other anthropogenic disturbances add to the problem and grossly hinder ongoing management strategies. This has driven increasing effort to find better performing, eco-friendly and reliable alternatives that can contribute to sustainable agricultural practices to manage abiotic stress. Nanotechnology and its implementation in agriculture have emerged as a promising option to cater to the problem of abiotic stress. Induction of reactive oxygen species (ROS) is an inevitable phenomenon linked to stress. Nanoparticles (NPs) perform dual actions in regulating ROS biology. The bidirectional roles of NPs in modulating ROS generation and/or ROS detoxification is tightly coupled within the hormetic boundaries. Nonetheless, how these NPs control the ROS metabolism within hormetic limits demands extensive investigation. This review focuses on the details of ROS metabolism under normal versus stressed conditions. It shall elaborate on the types, modes and process of uptake and translocation of NPs. The molecular dissection of the role of NPs in controlling transcriptomic expressions and modulating molecular crosstalks with other growth regulators, ions, reactive nitrogen species and other signalling molecules shall also be detailed. Throughout, this review aims to summarise the potential roles and regulation of NPs and consider how they can be used for green synthesis within a sustainable agricultural industry.

Phylogenetic, structural, functional characterisation and effect of exogenous spermidine on rice (Oryza sativa) HAK transporters under salt stress.

The HAK (High-affinity K+ ) family members mediate K+ transport that confers normal plant growth and resistance against unfavourable environmental conditions. Rice (Oryza sativa L.) HAK transporters have been extensively investigated for phylogenetic analyses with other plants species with very few of them functionally characterised. But very little information is known about their evolutionary aspects, overall structural, functional characterisation, and global expression pattern of the complete HAK family members in response to salt stress. In this study, 27 rice transporters were phylogenetically clustered with different dicot and monocot family members. Subsequently, the exon-intron structural patterns, conserved motif analyses, evolutionary divergence based different substitution matrix, orthologous-paralogous relationships were studied elaborately. Structural characterisations included a comparative study of secondary and tertiary structure, post-translational modifications, correspondence analyses, normal mode analyses, K+ /Na+ binding affinities of each of the OsHAK gene members. Global expression profile under salt stress showed clade-specific expression pattern of the proteins. Additionally, five OsHAK genes were chosen for further expression analyses in root and shoot tissues of two rice varieties during short-term salinity in the presence and absence of exogenous spermidine. All the information can be used as first-hand data for dissecting the administrative role of rice HAK transporters under various abiotic stresses.

Molecular characterization, evolutionary and phylogenetic analyses of rice ACT/BAT-type amino acid transporters.

The Amino Acid-Polyamine-Organocation (APC) family transporters containing BAT/ACT (amino acid/choline transporters), PHS/LAT (polyamine H+-symporters) and CAT (cationic amino acid transporters) type transporters are ubiquitously reported in plants. ACT (containing BAT genes) which mediates bidirectional amino acid transport has been poorly characterized till date. In the present study, bioinformatics-based analytical studies have been performed to characterize the structural, functional, and evolutionary features of seven ACT/BAT transporters in the model crop Oryza sativa. Sequence homologies with allied transporters suggest that these seven transporter proteins can also have important roles in regulating amino acid transportation. Evolutionary and phylogenetic study of OsBATs proteins using different statistical models based on CDS, Protein, and UTR regions have brought forward some valuable information regarding their way and patterns of diversification over the time, possibly due to the influence of their prevailing growth conditions. Computational analyses based on promoter regions, exon-intron structural patterns, conserved motifs, secondary and tertiary structures and ligand binding sites with probable ligand names provided clues on their evolutionary structural diversification indicating changes in their functional specification probably needed to combat with the changing environment.

Molecular phylogenomic study and the role of exogenous spermidine in the metabolic adjustment of endogenous polyamine in two rice cultivars under salt stress.

Compelling evidences anticipated the well acclamation of involvement of exogenous and endogenous polyamines (PAs) in conferring salt tolerance in plants. Intracellular PA's anabolism and catabolism should have contributed to maintain endogenous PAs homeostasis to induce stress signal networks. In this report, the evolutionary study has been conducted to reveal the phylogenetic relationship of genes encoding enzymes of the anabolic and catabolic pathway of PAs among the five plant lineages including green algae, moss, lycophyte, dicot and monocot along with their respective exon-intron structural patterns. Our results indicated that natural selection pressure had considerable influence on the ancestral PA metabolic pathway coding genes of land plants. PA metabolic genes have undergone gradual evolution by duplication and diversification process leading to subsequent structural modification through exon-intron gain and loss events to acquire specific function under environmental stress conditions. We have illuminated on the potential regulation of both the pathways by investigating the real-time expression analyses of PA metabolic pathway related enzyme coding genes at the transcriptional level in root and shoot tissues of two indica rice varieties, namely IR 36 (salt sensitive) and Nonabokra (salt-tolerant) in response to salinity in presence or absence of exogenous spermidine (Spd) treatment. Additionally, we have performed tissue specific quantification of the intracellular PAs and tried to draw probable connection between the PA metabolic pathway activation and endogenous PAs accumulation. Our results successfully enlighten the fact that how exogenous Spd in presence or absence of salt stress adjust the intracellular PA pathways to equilibrate the cellular PAs that would have been attributed to plant salt tolerance.

Molecular phylogenetic study and expression analysis of ATP-binding cassette transporter gene family in Oryza sativa in response to salt stress.

ATP-binding cassette (ABC) transporter is a large gene superfamily that utilizes the energy released from ATP hydrolysis for transporting myriad of substrates across the biological membranes. Although many investigations have been done on the structural and functional analysis of the ABC transporters in Oryza sativa, much less is known about molecular phylogenetic and global expression pattern of the complete ABC family in rice. In this study, we have carried out a comprehensive phylogenetic analysis constructing neighbor-joining and maximum-likelihood trees based on various statistical methods of different ABC protein subfamily of five plant lineages including Chlamydomonas reinhardtii (green algae), Physcomitrella patens (moss), Selaginella moellendorffii (lycophyte), Arabidopsis thaliana (dicot) and O. sativa (monocot) to explore the origin and evolutionary patterns of these ABC genes. We have identified several conserved motifs in nucleotide binding domain (NBD) of ABC proteins among all plant lineages during evolution. Amongst the different ABC protein subfamilies, 'ABCE' has not yet been identified in lower plant genomes (algae, moss and lycophytes). The result indicated that gene duplication and diversification process acted upon these genes as a major operative force creating new groups and subgroups and functional divergence during evolution. We have demonstrated that rice ABCI subfamily consists of only half size transporters that represented highly dynamic members showing maximum sequence variations among the other rice ABC subfamilies. The evolutionary and the expression analysis contribute to a deep insight into the evolution and diversity of rice ABC proteins and their roles in response to salt stress that facilitate our further understanding on rice ABC transporters.

Genome-wide analysis and evolutionary study of sucrose non-fermenting 1-related protein kinase 2 (SnRK2) gene family members in Arabidopsis and Oryza.

The over-expression of plant specific SnRK2 gene family members by hyperosmotic stress and some by abscisic acid is well established. In this report, we have analyzed the evolution of SnRK2 gene family in different plant lineages including green algae, moss, lycophyte, dicot and monocot. Our results provide some evidences to indicate that the natural selection pressure had considerable influence on cis-regulatory promoter region and coding region of SnRK2 members in Arabidopsis and Oryza independently through time. Observed degree of sequence/motif conservation amongst SnRK2 homolog in all the analyzed plant lineages strongly supported their inclusion as members of this family. The chromosomal distributions of duplicated SnRK2 members have also been analyzed in Arabidopsis and Oryza. Massively Parallel Signature Sequencing (MPSS) database derived expression data and the presence of abiotic stress related promoter elements within the 1 kb upstream promoter region of these SnRK2 family members further strengthen the observations of previous workers. Additionally, the phylogenetic relationships of SnRK2 have been studied in all plant lineages along with their respective exon-intron structural patterns. Our results indicate that the ancestral SnRK2 gene of land plants gradually evolved by duplication and diversification and modified itself through exon-intron loss events to survive under environmental stress conditions.

In silico characterization and evolutionary analyses of CCAAT binding proteins in the lycophyte plant Selaginella moellendorffii genome: a growing comparative genomics resource.

NF-Y transcription factors encoded by HAP gene family, composed of three subunits (HAP2/NF-YA, HAP3/NF-YB and HAP5/NF-YC), are capable of transcriptional regulation of target genes with high specificity by binding to the CCAAT-containing promoter sequences. Here, we have characterized duplicated HAP genes in Selaginella moellendorffii and explored some features that might be involved in the regulation of gene expression and their function. Subsequently, the evolutionary relationships of LEC1-type of HAP3 genes have been studied starting from lycophytes to angiosperm to reveal the details of conservation and diversification of these genes during plant evolution. Computational analyses demonstrated the variation in length of cis-regulatory region of HAP3 duplicates in S. moellendorffii containing three thermodynamically stable and evolutionarily conserved RNA secondary structures. The homology modeling of NF-Y proteins, secondary structural details, DNA binding large positive patches, binding affinity of H2A-H2B interactive residues of NF-YC subunits on the duplicated NF-YB subunits, conserved domain analyses and protein structural alignments indicated that gene duplication process of HAP genes in S. moellendorffii, followed by structural diversification, provide specific hints about their functional specificity under various circumstances for the survival of this lycophytic plant. We have identified several conserved motifs in LEC1 proteins among all plant lineages during evolution.

Human GMDS gene fragment hypermethylation in chronic high level of arsenic exposure with and without arsenic induced cancer.

Arsenic, though a poor mutagen, is an accepted environmental carcinogen. Perturbation of DNA methylation pattern leading to aberrant gene expression has been hypothesized as the mechanism for arsenic induced carcinogenesis. We had earlier demonstrated the hypermethylation of promoter region of p53 and p16 genes in persons exposed to different doses of arsenic. Till now no genomic hot spot has been identified which is frequently hypermethylated or hypomethylated in persons chronically exposed to environmental arsenic. In the present work, we have identified one hypermethylated sequence by methyl-sensitive arbitrarily primed polymerase chain reaction in the peripheral blood leukocyte DNA of chronically arsenic exposed persons with and without arsenic induced skin cancer. The sequence is from GMDS gene responsible for fucose metabolism. Southern hybridization of the sequence to the amplification products of methyl sensitive restriction enzyme digested genome of persons exposed to different doses of arsenic indicated that methylation increased in a dose dependent manner.