Drishti Mandal

St Xavier’s College

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Co-Authors

Ronita Nag Chaudhuri

St Xavier’s College

Senjuti Sinharoy

National Institute of Plant Genome Research

Saptarshi Datta

St Xavier’s College

Sicon Mitra

Sharda University

Pranab Kumar Mondal

Indian Institute of Technology Guwahati

G. N. Mukherjee

University of Calcutta

Nilesh Kumar Sharma

National Institute of Plant Genome Research

Anagh Ray

National Institutes of Health

Rahul Gupta

Indian Institute of Chemical Biology

Dhrubajyoti Chattopadhyay

University of Calcutta

Cooperative Institutions

University of Calcutta

National Institute of Plant Genome Research

St Xavier’s College

Bose Institute

National Institute of Pharmaceutical Education and Research

Indian Institute of Chemical Biology

Council of Scientific and Industrial Research

University of California, San Diego

Indian Institute of Science Bangalore

Saha Institute of Nuclear Physics

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ABI3 regulates ABI1 function to control cell length in primary root elongation zone

bioRxiv (Cold Spring Harbor Laboratory) (2023)

Saptarshi Datta Drishti Mandal Sicon Mitra Ronita Nag Chaudhuri

Abstract Post-embryonic primary root growth is effectively an interplay of several hormone signalling pathways. Here, we show that the ABA-responsive transcription factor ABI3, controls primary root growth through regulation of JA signalling molecule JAZ1 along with ABA responsive factor ABI1. In absence of ABI3, primary root elongation zone is shortened with significantly reduced cell length. Expression analyses and ChIP based assays indicate that ABI3 negatively regulates JAZ1 expression by occupying its upstream regulatory sequence and enriching repressive histone modification mark H3K27 trimethylation, thereby occluding RNAPII occupancy. Previous studies have shown that JAZ1 interacts with ABI1, the protein phosphatase 2C, that works during ABA signalling. Our results indicate that in absence of ABI3, when JAZ1 expression levels are high, ABI1 protein shows increased stability, compared to when JAZ1 is absent, or ABI3 is overexpressed. Consequently, in abi3 mutant, due to higher stability of ABI1, reduced phosphorylation of plasma membrane H + ATPase (AHA2) occurs. HPTS staining further indicated that, abi3 root cell apoplasts show reduced protonation, compared to wild type and ABI3 overexpressing seedlings. Such impeded proton extrusion, negatively affects cell length in primary root elongation zone. ABI3 therefore controls cell elongation in primary root by affecting ABI1-dependent protonation of root cell apoplasts. In summary, ABI3 controls expression of JAZ1 and in turn modulates function of ABI1 to regulate cell length in the elongation zone during primary root growth.

Wild type

10.1101/2023.10.16.562476

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Citations (2)

Root‐specific expression of chickpea cytokinin oxidase/dehydrogenase 6 leads to enhanced root growth, drought tolerance and yield without compromising nodulation

Plant Biotechnology Journal (2020)

Hitaishi Khandal Santosh Kumar Gupta Vikas Dwivedi Drishti Mandal Nilesh Kumar Sharma

Cytokinin group of phytohormones regulate root elongation and branching during post-embryonic development. Cytokinin-degrading enzymes cytokinin oxidases/dehydrogenases (CKXs) have been deployed to investigate biological activities of cytokinin and to engineer root growth. We expressed chickpea cytokinin oxidase 6 (CaCKX6) under the control of a chickpea root-specific promoter of CaWRKY31 in Arabidopsis thaliana and chickpea having determinate and indeterminate growth patterns, respectively, to study the effect of cytokinin depletion on root growth and drought tolerance. Root-specific expression of CaCKX6 led to a significant increase in lateral root number and root biomass in Arabidopsis and chickpea without any penalty to vegetative and reproductive growth of shoot. Transgenic chickpea lines showed increased CKX activity in root. Soil-grown advanced chickpea transgenic lines exhibited higher root-to-shoot biomass ratio and enhanced long-term drought tolerance. These chickpea lines were not compromised in root nodulation and nitrogen fixation. The seed yield in some lines was up to 25% higher with no penalty in protein content. Transgenic chickpea seeds possessed higher levels of zinc, iron, potassium and copper. Our results demonstrated the potential of cytokinin level manipulation in increasing lateral root number and root biomass for agronomic trait improvement in an edible legume crop with indeterminate growth habit.

Root (linguistics)

10.1111/pbi.13378

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Citations (62)

CRISPR-Cas9 system: A new-fangled dawn in gene editing

Life Sciences (2019)

Darshana Gupta Oindrila Bhattacharjee Drishti Mandal Madhab Kumar Sen Dhritiman Dey

10.1016/j.lfs.2019.116636

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Citations (228)

ABI3 regulates ABI1 function to control cell length in primary root elongation zone

The Plant Journal (2024)

Saptarshi Datta Drishti Mandal Sicon Mitra Swarnavo Chakraborty Ronita Nag Chaudhuri

SUMMARY Post‐embryonic primary root growth is effectively an interplay of several hormone signalling pathways. Here, we show that the ABA‐responsive transcription factor ABI3 controls primary root growth through the regulation of JA signalling molecule JAZ1 along with ABA‐responsive factor ABI1. In the absence of ABI3, the primary root elongation zone is shortened with significantly reduced cell length. Expression analyses and ChIP‐based assays indicate that ABI3 negatively regulates JAZ1 expression by occupying its upstream regulatory sequence and enriching repressive histone modification mark H3K27 trimethylation, thereby occluding RNAPII occupancy. Previous studies have shown that JAZ1 interacts with ABI1, the protein phosphatase 2C, that works during ABA signalling. Our results indicate that in the absence of ABI3, when JAZ1 expression levels are high, the ABI1 protein shows increased stability, compared to when JAZ1 is absent, or ABI3 is overexpressed. Consequently, in the abi3‐6 mutant, due to the higher stability of ABI1, reduced phosphorylation of plasma membrane H + ‐ATPase (AHA2) occurs. HPTS staining further indicated that abi3‐6 root cell apoplasts show reduced protonation, compared to wild‐type and ABI3 overexpressing seedlings. Such impeded proton extrusion negatively affects cell length in the primary root elongation zone. ABI3 therefore controls cell elongation in the primary root by affecting the ABI1‐dependent protonation of root cell apoplasts. In summary, ABI3 controls the expression of JAZ1 and in turn modulates the function of ABI1 to regulate cell length in the elongation zone during primary root growth.

Wild type

10.1111/tpj.17121

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Citations (0)

ABSCISIC ACID INSENSITIVE 3 promotes auxin signalling by regulating SHY2 expression to control primary root growth in response to dehydration stress

Journal of Experimental Botany (2024)

Drishti Mandal Saptarshi Datta Sicon Mitra Ronita Nag Chaudhuri

Plants combat dehydration stress through different strategies including root architectural changes. Here we show that when exposed to varying levels of dehydration stress, primary root growth in Arabidopsis is modulated by regulating root meristem activity. Abscisic acid (ABA) in concert with auxin signalling adjust primary root growth according to stress levels. ABSCISIC ACID INSENSITIVE 3 (ABI3), an ABA-responsive transcription factor, stands at the intersection of ABA and auxin signalling and fine-tunes primary root growth in response to dehydration stress. Under low ABA or dehydration stress, induction of ABI3 expression promotes auxin signalling by decreasing expression of SHY2, a negative regulator of auxin response. This further enhances the expression of auxin transporter gene PIN1 and cell cycle gene CYCB1;1, resulting in an increase in primary root meristem size and root length. Higher levels of dehydration stress or ABA repress ABI3 expression and promote ABSCISIC ACID INSENSITIVE 5 (ABI5) expression. This elevates SHY2 expression, thereby impairing primary root meristem activity and retarding root growth. Notably, ABI5 can promote SHY2 expression only in the absence of ABI3. Such ABA concentration-dependent expression of ABI3 therefore functions as a regulatory sensor of dehydration stress levels and orchestrates primary root growth by coordinating its downstream regulation.

Lateral root

Root hair

Ectopic expression

10.1093/jxb/erae237

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Citations (2)

RAV1 mediates cytokinin signalling for regulating primary root growth in Arabidopsis

bioRxiv (Cold Spring Harbor Laboratory) (2022)

Drishti Mandal Saptarshi Datta Giridhar Ravindra Pranab Kumar Mondal Ronita Nag Chaudhuri

Abstract Root growth dynamics is an outcome of complex hormonal crosstalk. The primary root meristem size for example, is determined by antagonizing actions of cytokinin and auxin. Here we show that RAV1, a member of the AP2/ERF family of transcription factors, mediates cytokinin signalling in roots to regulate meristem size. The rav1 mutants have prominently longer primary roots, with a meristem that is significantly enlarged and contain higher cell numbers, compared to wild type. The mutant phenotype could be restored on exogenous cytokinin application or by inhibiting auxin transport. At the transcript level, primary cytokinin-responsive genes like ARR1, ARR12 were significantly downregulated in the mutant root, indicating impaired cytokinin signalling. In concurrence, cytokinin induced regulation of SHY2 , an Aux/IAA gene, and auxin efflux carrier PIN1 was hindered in rav1 , leading to altered auxin transport and distribution. This effectively altered root meristem size in the mutant. Notably, CRF1 another member of the AP2/ERF family implicated in cytokinin signalling, is transcriptionally repressed by RAV1 to promote cytokinin response in roots. Further correlating RAV1 to cytokinin signalling, our results demonstrate that cytokinin upregulate RAV1 expression through ARR1, during post-embryonic root development. Regulation of RAV1 expression is a part of secondary cytokinin response that eventually represses CRF1 to augment cytokinin signalling. To conclude, in Arabidopsis, RAV1 functions in a branch pathway downstream to ARR1 that regulates CRF1 expression to enhance cytokinin action during primary root development.

Lateral root

Crosstalk

10.1101/2022.07.13.499994

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Citations (0)

Optimization of Hairy Root Transformation for the Functional Genomics in Chickpea: A Platform for Nodule Developmental Studies

Methods in molecular biology (2019)

Drishti Mandal Deevita Srivastava Senjuti Sinharoy

Root system

Nodule (geology)

Functional Genomics

Root hair

10.1007/978-1-0716-0235-5_18

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Citations (6)

A toolbox for nodule development studies in chickpea: a hairy-root transformation protocol and an efficient laboratory strain of Mesorhizobium sp

bioRxiv (Cold Spring Harbor Laboratory) (2018)

Drishti Mandal Senjuti Sinharoy

Abstract Mesorhizobium sp . produces root nodules in chickpea. Chickpea and model legume Medicago truncatula are members of inverted repeat lacking clade (IRLC). The rhizobia after internalization inside plant cell called ‘bacteroid’. Nodule Specific Cysteine-rich (NCR) peptides in IRLC legumes guide bacteroids to a ‘terminally differentiated swollen (TDS)’ form. Bacteroids in chickpea are less TDS than those in Medicago . Nodule development in chickpea indicates recent evolutionary diversification and merits further study. A hairy root transformation protocol and an efficient laboratory strain are prerequisites for performing any genetic study on nodulation. We have standardized a protocol for composite plant generation in chickpea with a transformation frequency above 50%, as shown by fluorescent markers. This protocol also works well in different ecotypes of chickpea. Localization of subcellular markers in these transformed roots is similar to Medicago . When checked inside transformed nodules, peroxisomes were concentrated along the periphery of the nodules, while ER and golgi bodies surrounded the symbiosomes. Different Mesorhizobium strains were evaluated for their ability to initiate nodule development, and efficiency of nitrogen fixation. Inoculation with different strains resulted in different shapes of TDS bacteroids with variable nitrogen fixation. Our study provides a toolbox to study nodule development in the crop legume chickpea.

Medicago truncatula

Leghemoglobin

Nodule (geology)

Mesorhizobium

Strain (injury)

10.1101/362947

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Citations (1)

A Tripartite Interaction among the Basidiomycete Rhodotorula mucilaginosa, N₂-Fixing Endobacteria, and Rice Improves Plant Nitrogen Nutrition

The Plant Cell (2019)

Karnelia Paul Chinmay Saha Mayurakshi Nag Drishti Mandal Haraprasad Naiya

Nitrogen (N) limits crop yield, and improvement of N nutrition remains a key goal for crop research; one approach to improve N nutrition is identifying plant-interacting, N2-fixing microbes. Rhodotorula mucilaginosa JGTA-S1 is a basidiomycetous yeast endophyte of narrowleaf cattail (Typha angustifolia). JGTA-S1 could not convert nitrate or nitrite to ammonium but harbors diazotrophic (N2-fixing) endobacteria (Pseudomonas stutzeri) that allow JGTA-S1 to fix N2 and grow in a N-free environment; moreover, P. stutzeri dinitrogen reductase was transcribed in JGTA-S1 even under adequate N. Endobacteria-deficient JGTA-S1 had reduced fitness, which was restored by reintroducing P. stutzeri. JGTA-S1 colonizes rice (Oryza sativa), significantly improving its growth, N content, and relative N-use efficiency. Endofungal P. stutzeri plays a significant role in increasing the biomass and ammonium content of rice treated with JGTA-S1; also, JGTA-S1 has better N2-fixing ability than free-living P. stutzeri and provides fixed N to the plant. Genes involved in N metabolism, N transporters, and NODULE INCEPTION-like transcription factors were upregulated in rice roots within 24 h of JGTA-S1 treatment. In association with rice, JGTA-S1 has a filamentous phase and P. stutzeri only penetrated filamentous JGTA-S1. Together, these results demonstrate an interkingdom interaction that improves rice N nutrition.

Pseudomonas stutzeri

Oryza

10.1105/tpc.19.00385

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Citations (37)

RAV1 mediates cytokinin signaling for regulating primary root growth in Arabidopsis

The Plant Journal (2022)

Drishti Mandal Saptarshi Datta Raveendar Giridhar Pranab Kumar Mondal Ronita Nag Chaudhuri

Root growth dynamics is an outcome of complex hormonal crosstalk. The primary root meristem size, for example, is determined by antagonizing actions of cytokinin and auxin. Here we show that RAV1, a member of the AP2/ERF family of transcription factors, mediates cytokinin signaling in roots to regulate meristem size. The rav1 mutants have prominently longer primary roots, with a meristem that is significantly enlarged and contains higher cell numbers, compared with wild-type. The mutant phenotype could be restored on exogenous cytokinin application or by inhibiting auxin transport. At the transcript level, primary cytokinin-responsive genes like ARR1, ARR12 were significantly downregulated in the mutant root, indicating impaired cytokinin signaling. In concurrence, cytokinin induced regulation of SHY2, an Aux/IAA gene, and auxin efflux carrier PIN1 was hindered in rav1, leading to altered auxin transport and distribution. This effectively altered root meristem size in the mutant. Notably, CRF1, another member of the AP2/ERF family implicated in cytokinin signaling, is transcriptionally repressed by RAV1 to promote cytokinin response in roots. Further associating RAV1 with cytokinin signaling, our results demonstrate that cytokinin upregulates RAV1 expression through ARR1, during post-embryonic root development. Regulation of RAV1 expression is a part of secondary cytokinin response that eventually represses CRF1 to augment cytokinin signaling. To conclude, RAV1 functions in a branch pathway downstream to ARR1 that regulates CRF1 expression to enhance cytokinin action during primary root development in Arabidopsis.

Crosstalk

Lateral root

10.1111/tpj.16039

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Citations (18)