Browsing by Author "Yadav, Sandeep"

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Auxin signaling modulates LATERAL ROOT PRIMORDIUM1 (LRP1) expression during lateral root development in Arabidopsis
(John Wiley & Sons, 2020) Singh, Sharmila; Yadav, Sandeep; Singh, Alka; Mahima; Singh, Archita; Gautam, Vibhav; Sarkar, Ananda K.
Auxin signaling mediated by various Auxin/Indole-3-Acetic Acid (Aux/IAAs) and AUXIN RESPONSE FACTORs (ARFs) regulate lateral root (LR) development by controlling the expression of downstream genes. LATERAL ROOT PRIMORDIUM1 (LRP1), a member of SHORT INTERNODES/STYLISH (SHI/STY) family, was identified as an auxin inducible gene. Precise developmental role and molecular regulation of LRP1 in root development remains to be understood. Here we show that LRP1 is expressed in all stages of LR development, besides primary root. The expression of LRP1 is regulated by histone deacetylation in an auxin dependent manner. Our genetic interaction studies showed that LRP1 acts downstream of auxin responsive Aux/IAAs-ARFs modules during LR development. We showed that auxin mediated induction of LRP1 is lost in slr-1 and arf7arf19 mutants roots. NPA treatment studies showed that LRP1 acts after LR founder cell specification and asymmetric division. Overexpression of LRP1 (LRP1 OE) showed increased number of LRPs at stage I, IV and V, resulting in reduced emerged LR density, which suggests that it is involved in LRP development. Interestingly, LRP1 induced expression of YUC4, which is involved in auxin biosynthesis, contributes to increased accumulation of endogenous auxin in LRP1 OE roots. LRP1 interacts with SHI, STY1, SRS3 and SRS7 proteins of SHI/STY family indicating their possible redundant role during root development. Our results suggest that auxin and histone deacetylation affect LRP1 expression, and it acts downstream of LR forming auxin response modules to negatively regulate LRP development by modulating auxin homeostasis in Arabidopsis thaliana.
Coevolution pattern and functional conservation or divergence of miR167s and their targets across diverse plant species
(Nature Publishing Group, 2015) Barik, Suvakanta; Kumar, Ashutosh; Das, Shabari Sarkar; Yadav, Sandeep; Gautam, Vibhav; Singh, Archita; Singh, Sharmila; Sarkar, Ananda K.
microRNAs (miRNAs), a class of endogenously produced small non-coding RNAs of 20-21 nt length, processed from precursor miRNAs, regulate many developmental processes by negatively regulating the target genes in both animals and plants. The coevolutionary pattern of a miRNA family and their targets underscores its functional conservation or diversification. The miR167 regulates various aspects of plant development in Arabidopsis by targeting ARF6 and ARF8. The evolutionary conservation or divergence of miR167s and their target genes are poorly understood till now. Here we show the evolutionary relationship among 153 MIR167 genes obtained from 33 diverse plant species. We found that out of the 153 of miR167 sequences retrieved from the "miRBase", 27 have been annotated to be processed from the 3' end, and have diverged distinctively from the other miR167s produced from 5' end. Our analysis reveals that gma-miR167h/i and mdm-miR167a are processed from 3' end and have evolved separately, diverged most resulting in novel targets other than their known ones, and thus led to functional diversification, especially in apple and soybean. We also show that mostly conserved miR167 sequences and their target AUXIN RESPONSE FACTORS (ARFs) have gone through parallel evolution leading to functional diversification among diverse plant species.
Conserved LBL1-ta-siRNA and miR165/166-RLD1/2 modules regulate root development in maize
(The Company of Biologists, 2021) Gautam, Vibhav; Singh, Archita; Yadav, Sandeep; Singh, Sharmila; Kumar, Pramod; Das, Shabari Sarkar; Sarkar, Ananda K.
Root system architecture and anatomy of monocotyledonous maize is significantly different from dicotyledonous model Arabidopsis. The molecular role of non-coding RNA (ncRNA) is poorly understood in maize root development. Here we address the role of LEAFBLADELESS1 (LBL1), a component of maize trans-acting short-interfering RNA (ta-siRNA), in maize root development. We report that the root growth, anatomical patterning, number of lateral roots (LRs) and monocot-specific crown roots (CRs) and seminal roots (SRs) are significantly affected in lbl1-rgd1 mutant, which is defective in production of ta-siRNA, including tasiR-ARF that targets AUXIN RESPONSE FACTOR3 (ARF3) in maize. Altered accumulation and distribution of auxin, due to differential expression of auxin biosynthesis and transporter genes, created an imbalance in auxin signaling. Altered expression of microRNA165/166 (miR165/166) and its targets ROLLED1/2 (RLD1/2) contributed to the changes in lbl1-rgd1 root growth and vascular patterning, as was evident by altered root phenotype of Rld1-O semi-dominant mutant. Thus, LBL1/ta-siRNA module regulates root development, possibly by affecting auxin distribution and signaling, in crosstalk with miR165/166-RLD1/2 module. We further showed that ZmLBL1 and its Arabidopsis homolog AtSGS3 proteins are functionally conserved.
CRISPR-Cas9 directed genome engineering for enhancing salt stress tolerance in rice
(Elsevier B.V., 2019) Farhat, Sufia; Jain, Neha; Singh, Nisha; Sreevathsa, Rohini; Dash, Prasanta K.; Rai, Rhitu; Yadav, Sandeep; Kumar, Pramod; Sarkar, Ananda K.; Jain, Ajay; Singh, Nagendra K.; Rai, Vandna
Crop productivity in rice is harshly limited due to high concentration of salt in the soil. To understand the intricacies of the mechanism it is important to unravel the key pathways operating inside the plant cell. Emerging state-of-the art technologies have provided the tools to discover the key components inside the plant cell for salt tolerance. Among the molecular entities, transcription factors and/or other important components of sensing and signaling cascades have been the attractive targets and the role of NHX and SOS1 transporters amply described. Not only marker assisted programs but also transgenic approaches by using reverse genetic strategies (knockout or knockdown) or overexpression have been extensively used to engineer rice crop. CRISPR/Cas is an attractive paradigm and provides the feasibility for manipulating several genes simultaneously. Here, in this review we highlight some of the molecular entities that could be potentially targeted for generating rice amenable to sustain growth under high salinity conditions by employing CRISPR/Cas. We also try to address key questions for rice salt stress tolerance other than what is already known.
Expression dynamics of miRNAs and their targets in seed germination conditions reveals miRNA-ta-siRNA crosstalk as regulator of seed germination
(Nature Publishing Group, 2018) Das, Shabari Sarkar; Yadav, Sandeep; Singh, Archita; Gautam, Vibhav; Sarkar, Ananda K.; Nandi, Asis K.; Karmakar, Prakash; Majee, Manoj; Sanan-Mishra, Neeti
Seed germination paves the way for the dormant embryo to establish itself as a new plant marking the first critical step in postembryonic plant growth and development. Germination starts with the uptake of water (imbibition), followed by induction of transcription, translation, energy metabolism, and cell division processes. Although small RNAs have been implicated in many developmental processes, their role during seed germination stages and conditions remained elusive. Here we show that seed germination conditions, like imbibition and temperature, dynamically regulate the expression of many developmentally important miRNAs and their targets. We have identified 58 miRNAs belonging to 30 different families at different seed germination conditions. Amongst these, 15 miRNAs and their targets were significantly differentially expressed in Arabidopsis seeds in dry and 12 h, 24 h and 48 h of imbibition. Interestingly, differential expression of miR390, which targets trans-acting siRNA locus (TAS3) derived transcripts, resulted in alteration of tasiR-ARF mediated regulation of expression of target AUXIN RESPONSE FACTORs (ARF2/3/4). Our results suggest that the dynamic expression of several miRNAs, their targets, and a crosstalk between miRNA and ta-siRNA pathways contribute to the regulation of seed germination in Arabidopsis thaliana.
Iron availability affects phosphate deficiency-mediated responses, and evidences of cross talk with auxin and zinc in Arabidopsis
(Oxford University Press, 2015) Rai, Vandna; Sanagala, Raghavendrarao; Sinilal, Bhaskaran; Yadav, Sandeep; Sarkar, Ananda K.; Dantu, Prem Kumar; Jain, Ajay
Phosphate (Pi) is pivotal for plant growth and development. Pi deficiency triggers local and systemically regulated adaptive responses in Arabidopsis thaliana. Inhibition of primary root growth (PRG) and retarded development of lateral roots (LRs) are typical local Pi deficiency-mediated responses of the root system. Expression of Pi starvation-responsive (PSR) genes is regulated systemically. Here, we report the differential influence of iron (Fe) availability on local and systemic sensing of Pi by Arabidopsis. P-Fe- condition disrupted local Pi sensing, resulting in an elongated primary root (PR). Altered Fe homeostasis in the lpsi mutant with aberration in local Pi sensing provided circumstantial evidence towards the role of Fe in the maintenance of Pi homeostasis. Reporter gene assays, expression analysis of auxin-responsive genes (ARGs) and root phenotyping of the arf7arf19 mutant demonstrated the role of Fe availability on local Pi deficiency-mediated LR development. In addition, Fe availability also exerted a significant influence on PSR genes belonging to different functional categories. Together, these results demonstrated a substantial influence of Fe availability on Pi deficiency-mediated responses of ontogenetically distinct traits of the root system and PSR genes. The study also provided evidence of cross-talk between Pi, Fe and Zn, highlighting a complex tripartite interaction amongst them for maintaining Pi homeostasis.
Phytohormonal crosstalk modulates the expression of miR166/165s, target Class III HD-ZIPs, and KANADI genes during root growth in Arabidopsis thaliana
(Nature Publishing Group, 2017) Singh, Archita; Roy, Shradha; Singh, Sharmila; Das, Shabari Sarkar; Gautam, Vibhav; Yadav, Sandeep; Kumar, Ashutosh; Singh, Alka; Samantha, Sukanya; Sarkar, Ananda K.
Both phytohormones and non-coding microRNAs (miRNAs) play important role in root development in Arabidopsis thaliana. Mature miR166/165 s, which are derived from precursor transcripts of concerned genes, regulate developmental processes, including leaf and root patterning, by targeting Class III HOMEODOMAIN LEUCINE-ZIPPER (HD-ZIP III) transcription factors (TFs). However, their regulation through hormones remained poorly understood. Here, we show that several phytohormones dynamically regulate the spatio-temporal expression pattern of miR166/165 and target HD-ZIP IIIs in developing roots. Hormone signaling pathway mutants show differential expression pattern of miR166/165, providing further genetic evidence for multilayered regulation of these genes through phytohormones. We further show that a crosstalk of at least six different phytohormones regulate the miR166/165, their target HD-ZIP IIIs, and KANADI (KANs). Our results suggest that HD-ZIP IIIs mediated root development is modulated both transcriptionally through phytohormones and KANs, and post-transcriptionally by miR166/165 that in turn are also regulated by the phytohormonal crosstalk.
Potassium chloroaurate-mediated in vitro synthesis of gold nanoparticles improved root growth by crosstalk with sucrose and nutrient-dependent auxin homeostasis in Arabidopsis thaliana
(MDPI AG, 2022) Yadav, Sandeep; Yugandhar, Poli; Alavilli, Hemasundar; Raliya, Ramesh; Singh, Archita; Sahi, Shivendra V.; Sarkar, Ananda K.; Jain, Ajay
In a hydroponic system, potassium chloroaurate (KAuCl4 ) triggers the in vitro sucrose (Suc)-dependent formation of gold nanoparticles (AuNPs). AuNPs stimulate the growth of the root system, but their molecular mechanism has not been deciphered. The root system of Arabidopsis (Arabidopsis thaliana) exhibits developmental plasticity in response to the availability of various nutrients, Suc, and auxin. Here, we showed the roles of Suc, phosphorus (P), and nitrogen (N) in facilitating a AuNPs-mediated increase in root growth. Furthermore, the recuperating effects of KAuCl4 on the natural (IAA) auxin-mediated perturbation of the root system were demonstrated. Arabidopsis seedlings harboring the cell division marker CycB1;1::CDB-GUS provided evidence of the restoration efficacy of KAuCl4 on the IAA-mediated inhibitory effect on meristematic cell proliferation of the primary and lateral roots. Arabidopsis harboring synthetic auxin DR5rev::GFP exhibited a reinstating effect of KAuCl4 on IAA-mediated aberration in auxin subcellular localization in the root. KAuCl4 also exerted significant and differential recuperating effects on the IAA-mediated altered expression of the genes involved in auxin signaling and biosynthetic pathways in roots. Our results highlight the crosstalk between KAuCl4 -mediated improved root growth and Suc and nutrient-dependent auxin homeostasis in Arabidopsis.
Role of abiotic stress responsive miRNAs in Arabidopsis root development
(Springer Nature Publishing AG, 2020) Singh, Archita; Gandhi, Nidhi; Mishra, Vishnu; Yadav, Sandeep; Rai, Vandna; Sarkar, Ananda K.
Abiotic stresses like drought, extreme temperature, and lack of sufficient water or nutrients adversely affect plant growth and productivity. The physiological responses of higher plants to the environmental stresses are largely influenced by the root system, which can quickly modulate its developmental pattern under changing water, nutrient, and temperature, as an adaptive response. Protein coding genes, phytohormones and microRNAs (miRNAs) are among the key players which imparts crucial intrinsic role in shaping the root development and its environment adaptive growth pattern. Among these factors, miRNAs belong to a class of small non-coding RNAs of 21–24 nucleotides in length, which regulates various aspects of plant growth and development by negatively regulating their target genes through either transcriptional cleavage or translational inhibition. Although many miRNAs have been identified to be differentially regulated under various abiotic stress conditions, only a limited number of them have been characterized, due to the complex nature of its regulation. However, some of the miRNAs, such as miR156, miR165/166, miR169 etc., have recently been shown to be involved in both abiotic stress response and root development, indicating the diverse role of miRNA mediated gene regulation. The field of miRNA mediated gene regulation is dynamically expanding and more miRNAs are being characterized for their function. Current review focuses on miRNAs that are differentially regulated by major abiotic stresses as well as are involved in root development in Arabidopsis thaliana. We highlight their role in regulation of multiple and diverse aspects of developmental and physiological processes in Arabidopsis.
Role of chromatin modification and remodeling in stem cell regulation and meristem maintenance in Arabidopsis
(Oxford University Press, 2020) Singh, Sharmila; Singh, Alka; Singh, Archita; Mahima; Yadav, Sandeep; Bajaj, Ishita; Kumar, Shailendra; Jain, Ajay; Sarkar, Ananda K.
In higher plants, pluripotent stem cells reside in the specialized microenvironment called stem cell niches (SCNs) harbored at the shoot apical meristem (SAM) and root apical meristem (RAM), which give rise to the aerial and underground parts of a plant, respectively. The model plant Arabidopsis thaliana (Arabidopsis) has been extensively studied to decipher the intricate regulatory mechanisms involving some key transcriptions factors and phytohormones that play pivotal roles in stem cell homeostasis, meristem maintenance, and organ formation. However, there is increasing evidence to show the epigenetic regulation of the chromatin architecture, gene expression exerting an influence on an innate balance between the self-renewal of stem cells, and differentiation of the progeny cells to a specific tissue type or organ. Post-translational histone modifications, ATP-dependent chromatin remodeling, and chromatin assembly/ disassembly are some of the key features involved in the modulation of chromatin architecture. Here, we discuss the major epigenetic regulators and illustrate their roles in the regulation of stem cell activity, meristem maintenance, and related organ patterning in Arabidopsis.
Sirtinol, a Sir2 protein inhibitor, affects stem cell maintenance and root development in Arabidopsis thaliana by modulating auxin-cytokinin signaling components
(Nature Publishing Group, 2017) Singh, Sharmila; Singh, Alka; Yadav, Sandeep; Gautam, Vibhav; Singh, Archita; Sarkar, Ananda K.
In Arabidopsis thaliana, besides several key transcription factors and chromatin modifiers, phytohormones auxin and cytokinin play pivotal role in shoot and root meristem maintenance, and lateral root (LR) development. Sirtinol, a chemical inhibitor of Sir2 proteins, is known to promote some auxin induced phenotypes in Arabidopsis. However, its effect on plant stem cell maintenance or organ formation remained unaddressed. Here we show that sirtinol affects meristem maintenance by altering the expression of key stem cell regulators, cell division and differentiation by modulating both auxin and cytokinin signaling in Arabidopsis thaliana. The expression of shoot stem cell niche related genes WUSCHEL (WUS) and CLAVATA3 (CLV3) was upregulated, whereas SHOOT MERISTEMLESS (STM) was downregulated in sirtinol treated seedlings. The expression level and domain of key root stem cell regulators PLETHORA (PLTs) and WUS-Related Homeobox 5 (WOX5) were altered in sirtinol treated roots. Sirtinol affects LR development by disturbing proper auxin transport and maxima formation, similar to 2,4-dichlorophenoxyacetic acid (2,4-D). Sirtinol also affects LR formation by altering cytokinin biosynthesis and signaling genes in roots. Therefore, sirtinol affects shoot and root growth, meristem maintenance and LR development by altering the expression of cytokinin-auxin signaling components, and regulators of stem cells, meristems, and LRs.
Tweaking microRNA-mediated gene regulation for crop improvement
(Elsevier B.V., 2020) Yadav, Sandeep; Das, Shabari Sarkar; Kumar, Pramod; Mishra, Vishnu; Sarkar, Ananda K.
Many beneficial agronomic traits of crops have been lost in the course of domestication and selective introgression processes. One of the best strategies to prevent the exclusion of beneficial traits during the introgression of novel traits is to selectively fine-tune the expression of some candidate genes or microRNAs (miRNAs). Noncoding regulatory miRNAs of 20–24 nucleotide (nt) length have evolved as specific posttranscriptional regulators that negatively regulate the transcript abundance of their target genes via either cleavage of mRNAs or translational inhibition. In this chapter, we summarize the current knowledge on the role of miRNAs and elaborate their use in miRNA-mediated gene regulation for the improvement of agronomic traits in crop plants. Recent CRISPR-Cas9-based genome engineering technology can be applied to selectively tweak the specific miRNA-target regulation, and thus, the function of the candidate genes to improve crop plants by conferring better productivity or resistance to abiotic or biotic stresses.
Uncovering the molecular signature underlying the light intensity-dependent root development in Arabidopsis thaliana
(BioMed Central Ltd, 2019) Kumari, Sony; Yadav, Sandeep; Patra, Debadutta; Singh, Sharmila; Sarkar, Ananda K.; Panigrahi, Kishore C. S.
Background Root morphology is known to be affected by light quality, quantity and direction. Light signal is perceived at the shoot, translocated to roots through vasculature and further modulates the root development. Photoreceptors are differentially expressed in both shoot and root cells. The light irradiation to the root affects shoot morphology as well as whole plant development. The current work aims to understand the white light intensity dependent changes in root patterning and correlate that with the global gene expression profile. Results Different fluence of white light (WL) regulate overall root development via modulating the expression of a specific set of genes. Phytochrome A deficient Arabidopsis thaliana (phyA-211) showed shorter primary root compared to phytochrome B deficient (phyB-9) and wild type (WT) seedlings at a lower light intensity. However, at higher intensity, both mutants showed shorter primary root in comparison to WT. The lateral root number was observed to be lowest in phyA-211 at intensities of 38 and 75 μmol m − 2 s − 1. The number of adventitious roots was significantly lower in phyA-211 as compared to WT and phyB-9 under all light intensities tested. With the root phenotypic data, microarray was performed for four different intensities of WL light in WT. Here, we identified ~ 5243 differentially expressed genes (DEGs) under all light intensities. Gene ontology-based analysis indicated that different intensities of WL predominantly affect a subset of genes having catalytic activity and localized to the cytoplasm and membrane. Furthermore, when root is irradiated with different intensities of WL, several key genes involved in hormone, light signaling and clock-regulated pathways are differentially expressed. Conclusion Using genome wide microarray-based approach, we have identified candidate genes in Arabidopsis root that responded to the changes in light intensities. Alteration in expression of genes such as PIF4, COL9, EPR1, CIP1, ARF18, ARR6, SAUR9, TOC1 etc. which are involved in light, hormone and clock pathway was validated by qRT-PCR. This indicates their potential role in light intensity mediated root development.
A unique miR775-GALT9 module regulates leaf senescence in Arabidopsis during post-submergence recovery by modulating ethylene and the abscisic acid pathway
(The Company of Biologists, 2022) Mishra, Vishnu; Singh, Archita; Gandhi, Nidhi; Das, Shabari Sarkar; Yadav, Sandeep; Kumar, Ashutosh; Sarkar, Ananda K.
Submergence-induced hypoxic condition negatively affects the plant growth and development, and causes early onset of senescence. Hypoxia alters the expression of a number of microRNAs (miRNAs). However, the molecular function of submergence stress-induced miRNAs in physiological or developmental changes and recovery remains poorly understood. Here we show that miR775 is an Arabidopsis thaliana-specific young and unique miRNA that possibly evolved non-canonically. miR775 post-transcriptionally regulates Galactosyltransferase (GALT9) and their expression is inversely affected at 24 hours of complete submergence stress. The overexpression of miR775 (miR775-Oe) confers enhanced recovery from submergence stress and reduced accumulation of RBOHD and ROS, in contrast to wild type and MIM775 Arabidopsis shoot. A similar recovery phenotype of galt9 mutant indicates the role of miR775-GALT9 module in post-submergence recovery. We predicted Golgi-localized GALT9 to be potentially involved in protein glycosylation. The altered expression of senescence-associated genes (SAG12, SAG29, and ORE1), ethylene signalling (EIN2 and EIN3) and ABA biosynthesis (NCED3) pathway genes in miR775-Oe, galt9 and MIM775 plants. Thus, our results indicate the role of miR775-GALT9 module in post-submergence recovery through a crosstalk with ethylene and ABA pathway.