A peep into the Plant miRNA world

microRNAs constitute a major class of the small regulatory molecules that are involved in regulating the intrinsic normal growth of cells and development of organisms as well as in maintaining the integrity of genomes. The plant miRNA research gained momentum, 2002 onwards with identification of new miRNA molecules and their targets. This was accompanied by the discovery of plant homologs of proteins involved in miRNA biogenesis, including a new member SERRATE. The identification of several diverging and converging functions of miRNAs indicate that they play versatile roles in regulating cell differentiation and tissue development. This article provides an update on the conservation and identification of plant miRNAs. The classical miRNA biogenesis pathway and the associated proteins are discussed along with the emerging concept on the processing of miRNA-encoding introns (mirtrons). It also contains a concise account of plant miRNA targets and functions with focus on the recent successful attempt on engineering synthetic miRNAs to study gene function as well as to impart virus resistance in plants

Identification of genes differentially expressed in onion infected with Iris yellow spot virus

Iris yellow spot virus (IYSV) causes severe damage and economic losses in onion production. Differential display-PCR was used to study changes in the gene expression of IYSV-infected onion plants. Representative up-regulated and down-regulated genes were selected for further study. Based on sequence analysis, the up-regulated genes were identified as retrotransposon protein, disease resistance-like proteins, chitinase, pathogenesis-related protein, cytochrome oxidase, cytochrome c, pentatricopeptide repeat-containing protein and pectin methylesterase. A DNA-binding transcriptional repressor protein gene was greatly down-regulated. . Most of the identified genes are known to play essential roles in plant defence systems, and are newly identified in onion sequences

Identification of mirtrons in rice using MirtronPred: a tool for predicting plant mirtrons.

article i nfo Studies from flies and insects have reported the existence of a special class of miRNA, called mirtrons that are produced from spliced-out introns in a DROSHA-independent manner. The spliced-out lariat is debranched and refolded into a stem-loop structure resembling the pre-miRNA, which can then be processed by DICER into mature ~21 nt species. The mirtrons have not been reported from plants. In this study, we present Mir- tronPred, a web based server to predict mirtrons from intronic sequences. We have used the server to predict 70 mirtrons in rice introns that were put through a stringent selection filter to shortlist 16 best sequences. The prediction accuracy was subsequently validated by northern analysis and RT-PCR of a predicted Os- mirtron-109. The target sequences for this mirtron were also found in the rice degradome database. The pos- sible role of the mirtron in rice regulon is discussed. The MirtronPred web server is available at http://bioinfo. icgeb.res.in/mirtronPred

When an Intruder Comes Home: GM and GE Strategies to Combat Virus Infection in Plants

Viruses are silent enemies that intrude and take control of the plant cell’s machinery for their own multiplication. Infection by viruses and the resulting damage is still a major challenge in the agriculture sector. Plants have the capability to fight back, but the ability of viruses to mutate at a fast rate helps them to evade the host’s response. Therefore, classical approaches for introgressing resistance genes by breeding have obtained limited success in counteracting the virus menace. Genetic modification (GM)-based strategies have been successful in engineering artificial resistance in plants. Several different approaches based on pathogen-derived resistance, antisense constructs, hairpin RNAs, double-stranded RNA, etc., have been used to enhance plants’ resistance to viruses. Recently, genome editing (GE) strategies mainly involving the CRISPR/Cas-mediated modifications are being used for virus control. In this review, we discuss the developments and advancements in GM- and GE-based methods for tackling viral infection in plants

Effect of antibiotics on callus regeneration during transformation of IR 64 rice

We report here the effect of antibiotics on the regeneration potential of recalcitrant indica rice cultivar, IR64. Different protocols reporting high-efficiency agro-bacterium-mediated transformation of mature seed-derived regenerative calli were used and compared. The putative transgenic (T0) plants were analyzed for integration of the transgene through polymerase chain reaction and Southern blotting analyses. It was observed that the high-efficiency transformation of scutellar-derived regenerative calli could be obtained by using maltose as a carbon source and increased quantity of 2,4-D on a medium containing a higher concentration of gelling agent. The percentage of regeneration is greatly affected by the presence of antibiotics

Comparative miRomics of Salt-Tolerant and Salt-Sensitive Rice

Increase in soil salt causes osmotic and ionic stress to plants, which inhibits their growth and productivity. Rice production is also hampered by salinity and the effect of salt is most severe at the seedling and reproductive stages. Salainity tolerance is a quantitative property controlled by multiple genes coding for signaling molecules, ion transporters, metabolic enzymes and transcription regulators. MicroRNAs are key modulators of gene-expression that act at the post-transcriptional level by translation repression or transcript cleavage. They also play an important role in regulating plant’s response to salt-stress. In this work we adopted the approach of comparative and integrated data-mining to understand the miRNA-mediated regulation of salt-stress in rice. We profiled and compared the miRNA regulations using natural varieties and transgenic lines with contrasting behaviors in response to salt-stress. The information obtained from sRNAseq, RNAseq and degradome datasets was integrated to identify the salt-deregulated miRNAs, their targets and the associated metabolic pathways. The analysis revealed the modulation of many biological pathways, which are involved in salt-tolerance and play an important role in plant phenotype and physiology. The end modifications of the miRNAs were also studied in our analysis and isomiRs having a dynamic role in salt-tolerance mechanism were identified

Signaling through MAP kinase networks in plants

Protein phosphorylation is the most important mechanism for controlling many fundamental cellular processes in all living organisms including plants. A specific class of serine/threonine protein kinases, the mitogen-activated protein kinases (MAP kinases) play a central role in the transduction of various extra- and intracellular signals and are conserved throughout eukaryotes. These generally function via a cascade of networks, where MAP kinase (MAPK) is phosphorylated and activated by MAPK kinase (MAPKK), which itself is activated by MAPKK kinase (MAPKKK). Signaling through MAP kinase cascade can lead to cellular responses including cell division, differentiation as well as response to various stresses. In plants, MAP kinases are represented by multigene families and are organized into a complex network for efficient transmission of specific stimuli. Putative plant MAP kinase cascades have been postulated based on experimental analysis of in vitro interactions between specific MAP kinase components. These cascades have been tested in planta following expression of epitope-tagged kinases in protoplasts. It is known that signaling for cell division and stress responses in plants are mediated through MAP kinases and even auxin, ABA and possibly ethylene and cytokinin also utilize a MAP kinase pathway. Most of the biotic (pathogens and pathogen-derived elicitors) including wounding and abiotic stresses (salinity, cold, drought, and oxidative) can induce defense responses in plants through MAP kinase pathways. In this article we have covered the historical background, biochemical assay, activation/inactivation, and targets of MAP kinases with emphasis on plant MAP kinases and the responses regulated by them. The cross-talk between plant MAP kinases is also discussed to bring out the complexity within this three-component module

Fine tuning of auxin signaling by miRNAs

microRNAs (miRNAs) constitute a major class of endogenous non-coding regulatory small RNAs. They are present in a variety of organisms from algae to plants and play an important role in gene regulation. The miRNAs are involved in various biological processes, including differentiation, organ development, phase change, signaling, disease resistance and response to environmental stresses. This review provides a general background on the discovery, history, biogenesis and function of miRNAs. However, the focus is on the role for miRNA in controlling auxin signaling to regulate plant growth and development

Salinity- and ABA-induced up-regulation and light-mediated modulation of mRNA encoding glycine-rich RNA-binding protein from Sorghum bicolor

Glycine-rich RNA-binding proteins play an important role in post-transcriptional regulation of gene expression, including RNA processing, and are known to be up-regulated in response to a number of external stimuli. However, their regulation in response to salinity stress has not been reported. We have isolated a light- and salt-regulated, full-length cDNA clone encoding a putative glycine-rich protein containing conserved ribonucleoprotein motif from Sorghum bicolor designated as sbGR-RNP. Sequence analysis of the 701 bp insert revealed that the open reading frame of 513 bp encodes a 170 amino acid protein, with an apparent molecular mass of 16.68 kDa and calculated pI of 6.59. The deduced amino acid sequence also revealed that protein is hydrophilic in nature and contains 38% glycine residues. Northern blot analysis revealed a transcript size of 630 nucleotides, which shows regulation by blue and red light. The transcript is initially up- and down-regulated rapidly within 5 min of irradiation with blue and red light, respectively. This kind of rapid and opposite regulation by different light wavelengths could be a novel behavior of this photo-regulated gene. Furthermore, NaCl (500 mM) and abscisic acid (10 μM) also stimulated the transcript levels of sbGR-RNP to fourfold and sevenfold, respectively. These novel regulations of sbGR-RNP in response to light and salinity are important phenomena, which will be helpful in understanding the molecular mechanisms of cross-talk between abiotic stress and light signaling in plants

Pea DNA helicase 45 overexpression in tobacco confers high salinity tolerance without affecting yield

Salt tolerance is an important trait that is required to overcome salinity-induced reduction in plant productivity. We have reported previously the isolation of a pea DNA helicase 45 (PDH45) that exhibits striking homology with the eukaryotic translation initiation factor eIF-4A. Here, we report that PDH45 mRNA is induced in pea seedlings in response to high salt, and its overexpression driven by a constitutive cauliflower mosaic virus-35S promoter in tobacco plants confers salinity tolerance, thus suggesting a previously undescribed pathway for manipulating stress tolerance in crop plants. The T0 transgenic plants showed high levels of PDH45 protein in normal and stress conditions, as compared with WT plants. The T0 transgenics also showed tolerance to high salinity as tested by a leaf disk senescence assay. The T1 transgenics were able to grow to maturity and set normal viable seeds under continuous salinity stress without any reduction in plant yield in terms of seed weight. Measurement of Na+ ions in different parts of the plant showed higher accumulation in the old leaves and negligible accumulation in seeds of T1 transgenic lines as compared with the WT plants. The possible mechanism of salinity tolerance is discussed. Overexpression of PDH45 provides a possible example of the exploitation of DNA/RNA unwinding pathways for engineering salinity tolerance without affecting yield in crop plants