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The sprawling 852-acre IIT Jodhpur campus is a majestic tribute to the deep-rooted diverse cultural heritage of the region. The campus is an architectural masterpiece, which is slated to become an international exemplar of sustainability with net-zero energy, and advanced water and waste management strategies.&rdquo;

IIT Jodhpur

Hydrogen peroxide (H2O2) is a reactive oxygen species that affects cell signaling in various plant defense responses and induces programmed cell death. To identify the new components associated with H2O2 signaling and tolerance, we conducted a genome-wide association study (GWAS) on the root growth of 133 Arabidopsis thaliana accessions grown in the presence of toxic H2O2 levels. The most significant SNPs were associated with a cluster of chromosome 4 genes encoding an aquaporin NODULIN 26-LIKE INTRINSIC PROTEIN 1; 1 (NIP1;1), an NB-ARC domain-containing disease resistance protein (AT4G19050), and a putative membrane lipoprotein (AT4G19070). The expression level of NIP1;1 was relatively high in A. thaliana accessions sensitive to H2O2. Additionally, overexpression of NIP1;1 in a tolerant accession (e.g., Col-0) increased the sensitivity of transgenic plants to H2O2. An in planta β-glucuronidase reporter assay revealed that variations in the NIP1;1 promoter were responsible for the differences of its expression level in H2O2-tolerant and -sensitive accessions. Cell death was extensive and H2O2 levels were high in the roots of H2O2-sensitive and NIP1;1-overexpressing accessions. Together, our results indicate that the aquaporin NIP1;1 is a key determinant of the sensitivity of A. thaliana to H2O2, and contributes to the phenotypic variations detected by our GWAS. © 2017 The Author

Department of Bioscience & Bioengineering

World University root node

Molecular plant

Genome-wide association study reveals that the aquaporin NIP1;1 contributes to variation in hydrogen peroxide sensitivity in Arabidopsis thaliana

Plants have evolved a series of tolerance mechanisms to saline stress, which perturbs physiological processes throughout the plant. To identify genetic mechanisms associated with salinity tolerance, we performed linkage analysis and genome-wide association study (GWAS) on maintenance of root growth of Arabidopsis thaliana in hydroponic culture with weak and severe NaCl toxicity. The top 200 single-nucleotide polymorphisms (SNPs) determined by GWAS could cumulatively explain approximately 70% of the variation observed at each stress level. The most significant SNPs were linked to the genes of ATP-binding cassette B10 and vacuolar proton ATPase A2. Several known salinity tolerance genes such as potassium channel KAT1 and calcium sensor SOS3 were also linked to SNPs in the top 200. In parallel, we constructed a gene co-expression network to independently verify that particular groups of genes work together to a common purpose. We identify molecular mechanisms to confer salt tolerance from both predictable and novel physiological sources and validate the utility of combined genetic and network analysis. Additionally, our study indicates that the genetic architecture of salt tolerance is responsive to the severity of stress. These gene datasets are a significant information resource for a following exploration of gene function. © 2016 John Wiley &amp; Sons Ltd.

Plant, cell \& environment

Joint genetic and network analyses identify loci associated with root growth under NaCl stress in Arabidopsis thaliana

Agrobacterium tumefaciens-mediated transformation is an efficient method for incorporating genes and recovering stable transgenic plants in cowpea because this method offers several advantages such as the defined integration of transgenes, potentially low copy number, and preferential integration into transcriptional active regions of the chromosome. Cotyledonary node explants of cowpea present an attractive target for T-DNA delivery followed by regeneration of shoots via axillary proliferation without involvement of a de novo regeneration pathway. In this chapter, we describe a detailed protocol for Agrobacterium-mediated transformation of the cowpea variety Pusa Komal. The seedling cotyledonary node explants are used for cocultivation with an Agrobacterium strain EHA105 harboring standard binary vector, pCAMBIA2301 or pNOV2819, and putative transformed plants are selected using aminoglycoside antibiotic or mannose as sole carbon source, respectively. The entire process includes explant infection to transgenic seed generation in greenhouse. © Springer Science+Business Media New York 2015.

Agrobacterium Protocols

Cowpea [Vigna unguiculata (L.) Walp.]

Cowpea (Vigna unguiculata L. Walp) is an important grain legume cultivated in drought-prone parts of the world, having higher tolerance to heat and drought than many other crops. The transcription factor, Dehydration-Responsive Element-Binding protein 2A (DREB2A), controls expression of many genes involved in osmotic and heat stress responses of plants. In Arabidopsis, DREB2A-interacting proteins (DRIPs), which function as E3 ubiquitin ligases (EC 6.3.2.19), regulate the stability of DREB2A by targeting it for proteasome-mediated degradation. In this study, we cloned the cowpea ortholog of DRIP (VuDRIP) using PCR based methods. The 1614bp long VuDRIP mRNA encoded a protein of 433 amino acids having a C3HC4-type Really Interesting New Gene (RING) domain in the N-terminus and a C-terminal conserved region, similar to Arabidopsis DRIP1 and DRIP2. We found VuDRIP up-regulation in response to various abiotic stresses and phytohormones. Using yeast (Saccharomyces cerevisae) two-hybrid analysis, VuDRIP was identified as a VuDREB2A-interacting protein. The results indicate negative regulation of VuDREB2A by ubiquitin ligases in cowpea similar to Arabidopsis along with their other unknown roles in stress and hormone signaling pathways. © 2014 Elsevier Masson SAS.

Plant physiology and biochemistry

The cowpea RING ubiquitin ligase VuDRIP interacts with transcription factor VuDREB2A for regulating abiotic stress responses

Isolation and functional characterization of a stress-inducible transcription factor VuDREB2A from cowpea

Main conclusion: VuDREB2A exists in cowpea as a canonical DREB2-type transcription factor, having the ability to bind dehydration-responsive elements in vitro and confer enhanced drought resistance in transgenic Arabidopsis. Cowpea (Vigna unguiculata L. Walp) is an important cultivated legume that can survive better in arid conditions than other crops. But the molecular mechanisms involved in the drought tolerance of this species remain elusive with very few reported candidate genes. The Dehydration-Responsive Element-Binding Protein2 (DREB2) group of transcription factors plays key roles in plant responses to drought. However, no DREB2 ortholog has been reported from cowpea so far. In this study, we isolated and characterized a gene from cowpea, namely VuDREB2A, encoding a protein of 377 amino acids exhibiting features of reported DREB2-type proteins. In cowpea, VuDREB2A transcript accumulation was highly induced by desiccation, heat and salt, but slightly by exogenous abscisic acid (ABA) treatment. We also isolated the VuDREB2A promoter and predicted stress-responsive cis-elements in it using Arabidopsis microarray data. The E. coli-expressed VuDREB2A protein showed binding to synthetic oligonucleotides with Dehydration-Responsive Elements (DREs) from Arabidopsis, in electrophoretic mobility shift assays. Heterologous expression of VuDREB2A in Arabidopsis significantly improved plant survival under drought. In addition, overexpression of a truncated version of VuDREB2A, after removal of a putative negative regulatory domain (between amino acids 132-182) led to a dwarf phenotype in the transgenic plants. Microarray and quantitative PCR analyses of VuDREB2A overexpressing Arabidopsis revealed up-regulation of stress-responsive genes having DRE overrepresented in their promoters. In summary, our results indicate that VuDREB2A conserves the basic functionality and mode of regulation of DREB2A in Arabidopsis and could be a potent candidate gene for the genetic improvement of drought resistance in cowpea. © 2014 Springer-Verlag Berlin Heidelberg.

Planta

VuDREB2A, a novel DREB2-type transcription factor in the drought-tolerant legume cowpea, mediates DRE-dependent expression of stress-responsive genes and confers enhanced drought resistance in transgenic Arabidopsis

Plants being sessile are susceptible to heavy metals (HMs) toxicity and respond differentially to hostile environments. The toxicity of HM is governed by the type of ion and its concentration, plant physiology and stage of plant growth. Plants counteract the HMs stress by overexpressing numerous stress related proteins, glutathione mediated tolerance pathways and signaling proteins involving networks of various stress regulations. Though the response may vary and be specific in its stress networks regulation for each HM. The intricacy of HM tolerance response involves the set of molecular regulation, which demands to be understood to yield HM tolerant plant. Topical advancements in molecular biology and genomics have facilitated studies in transcriptomics and proteomics to identify regulatory genes implied in HM tolerance in plants. The integration of resources obtained through these studies will be of extreme significance, combining the diverse fields of plant biology to dissect the actual HM stress response network. In this review, we put an endeavor to describe the specific aspects of the molecular mechanisms of a plant response to HMs which may contribute to better understanding of the mode of HMs action and overlaps in metal sensing and signaling/crosstalk to other stresses. © 2012 Springer Science+Business Media, LLC.

Biometals

Molecular mechanistic model of plant heavy metal tolerance

Chemical genomics is a newly emerged and rapidly progressing field in biology, where small chemical molecules bind specifically and reversibly to protein(s) to modulate their function(s), leading to the delineation and subsequent unravelling of biological processes. This approach overcomes problems like lethality and redundancy of classical genetics. Armed with the powerful techniques of combinatorial synthesis, high-throughput screening and target discovery chemical genomics expands its scope to diverse areas in biology. The well-established genetic system of Arabidopsis model allows chemical genomics to enter into the realm of plant biology exploring signaling pathways of growth regulators, endomembrane signaling cascades, plant defense mechanisms and many more events.

Indian Journal of Biochemistry and Biophysics

Chemical genomics in plant biology

An improved method of Agrobacterium-mediated transformation of cowpea was developed employing both sonication and vacuum infiltration treatments. 4 day-old cotyledonary nodes were used as explants for co-cultivation with Agrobacterium tumefaciens strain EHA105 harbouring the binary vector pSouv-cry1Ac. Among the different injury treatments, vacuum infiltration and their combination treatments tested, sonication for 20 s followed by vacuum infiltration for 5 min with A. tumefaciens resulted in highest transient GUS expression efficiency (93% explants expressing GUS at regenerating sites). After 3 days of co-cultivation, the explants were cultured in 150 mg/l kanamycin-containing selection medium and putative transformed plants were recovered. The presence, integration and expression of nptII and cry1Ac genes in T 0 transgenic plants were confirmed by polymerase chain reaction (PCR), genomic Southern and qualitative reverse transcription (RT)-PCR analysis. Western blot hybridization and enzyme-linked immunosorbent assay (ELISA) detected and demonstrated the accumulation of Cry1Ac protein in transgenic plants. The cry1Ac gene transmitted in a Mendelian fashion. The stable transformation efficiency increased by 88. 4% using both sonication-assisted Agrobacterium-mediated transformation (SAAT) and vacuum infiltration than simple Agrobacterium-mediated transformation in cowpea. © 2011 Springer-Verlag.

Plant cell reports

Improved Agrobacterium-mediated transformation of cowpea via sonication and vacuum infiltration

Rice (Oryza sativa) is a significant agricultural crop consumed by more than half of the global population. Its demand is expected to increase due to rising consumption and a growing global population. Moreover, the rice plant is frequently exposed to disease-causing pathogens, such as bacteria, fungi, viruses, and nematodes. Thus, cultivating disease-resistant varieties is an efficient way of disease control compared to pesticide applications. However, the rice plant has a well-defined defense system to prevent the onset of disease, including Pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) and effector-triggered immunity (ETI). The defense system is controlled by various disease-resistance proteins, such as resistance (R) proteins and pathogen recognition receptors (PRRs). Therefore, the identification of disease-resistance proteins not only reduces the amount of pesticides used in rice fields but also increases their yield. Though some resistant proteins have been characterized, their rapid identification, precise diagnosis, and appropriate management are still lacking. However, few methods based on sequence-similarity and de novo prediction, such as Machine Learning (ML), usually have low prediction power. In this study, we built a state-of-the-art classifier based on Deep Learning (DL) for the early detection of disease-resistance proteins in rice and related species. We compared the DL-based Multi-layer Perceptron (MLP) model with the five well-established ML-based methods using a protein dataset of rice and its related species. The DL-based MLP model outperformed all of the five classifiers on 10-fold cross-validation. The accuracy, Area Under Receiving Operating Characteristic (ROC) curve (AUC), F1-score, precision, and recall were superior in the DL-based MLP model. In conclusion, the MLP model is an effective DL model for predicting disease-resistance proteins with high scores in performance metrics. This study will provide insight to the breeders in developing disease-resistant rice varieties and assist in transforming traditional rice farming practices into a new age of smart rice farming.