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Item Differential expression of genes during recovery of Nicotiana tabacum from tomato leaf curl Gujarat virus infection(Springer Nature Publishing AG, 2023) Namgial, T.; Singh, A. K.; Singh, N. P.; Francis, A.; Chattopadhyay, Debasis; Voloudakis, A.; Chakraborty, S.Elucidating the role of host factors in response to viral infection is crucial in understanding the plant host–virus interaction. Begomovirus, a genus in the family Geminiviridae, is reported throughout the globe and is known to cause serious crop diseases. Tomato leaf curl Gujarat virus (ToLCGV) infection in Nicotiana tabacum resulted in initial symptom expression followed by a quick recovery in the systemic leaves. Transcriptome analysis using next-generation sequencing (NGS) revealed a large number of diferentially expressed genes both in symptomatic as well as recovered leaves when compared to mock-inoculated plants. The virus infected N. tabacum results in alteration of various metabolic pathways, phytohormone signaling pathway, defense related protein, protease inhibitor, and DNA repair pathway. RT-qPCR results indicated that Germin-like protein subfamily T member 2 (NtGLPST), Cysteine protease inhibitor 1-like (NtCPI), Thaumatin-like protein (NtTLP), Kirola-like (NtKL), and Ethylene-responsive transcription factor ERF109-like (NtERTFL) were down-regulated in symptomatic leaves when compared to recovered leaves of ToLCGV-infected plants. In contrast, the Auxin-responsive protein SAUR71-like (NtARPSL) was found to be diferentially down-regulated in recovered leaves when compared to symptomatic leaves and the mock-inoculated plants. Lastly, Histone 2X protein like (NtHH2L) gene was found to be down-regulated, whereas Uncharacterized (NtUNCD) was up-regulated in both symptomatic as well as recovered leaves compared to the mock-inoculated plants. Taken together, the present study suggests potential roles of the diferentially expressed genes that might govern tobacco’s susceptibility and/or recovery response towards ToLCGV infection.Item Characterization of a pollen-preferential gene OSIAGP from rice (Oryza sativa L. subspecies indica) coding for an arabinogalactan-protein homolog and analysis of its promoter activity during pollen development and pollen tube growth(Springer, 2010) Anand, Saurabh; Tyagi, Akhilesh K.During differential screening of inflorescence-specific cDNA libraries from Oryza sativa indica, an arabinogalactan protein (OSIAGP) cDNA (586 bp) expressing preferentially in the inflorescence has been isolated. It encodes an arabinogalactan protein of 59 amino acids (6.4 kDa) with a transmembrane domain and a secretory domain at the N terminus. The protein shows homology with AGP23 from Arabidopsis, and its homologue in japonica rice is located on chromosome 6. OSIAGP transcripts also accumulate in shoots and roots of rice seedling grown in the dark, but light represses expression of the gene. Analysis of a genomic clone of OSIAGP revealed that its promoter contains several pollen-specificity and light-regulatory elements. The promoter confers pollen-preferential activity on gus, starting from the release of microspores to anther dehiscence in transgenic tobacco, and is also active during pollen tube growth. Analysis of pollen preferential activity of the promoter in the transgenic rice system revealed that even the approximately 300 bp fragment has activity in pollen and the anther wall and further deletion down to approximately 100 bp completely abolishes this activity, which is consistent with in-silico analysis of the promoter. Arabinogalactan proteins have been shown to be involved in the cell elongation process. The homology of OSIAGP with AGP23 and the fact that seedling growth in the dark and pollen tube growth are events based on cell elongation strengthen the possibility of OSIAGP performing a similar function.Item Expression of CAP2, an AP2-family transcription factor from chickpea enhances growth and tolerance to dehydration and salt tress in transgenic tobacco(American Society of Plant Biologists, 2006) Shukla, Rakesh K.; Raha, Sumita; Tripathi, Vineeta; Chattopadhyay, DebasisThe APETALA2 (AP2) domain defines a large family of DNA-binding proteins that play important roles in plant morphology, development, and stress response. We describe isolation and characterization of a gene (CAP2) from chickpea (Cicer arietinum) encoding a novel AP2-family transcription factor. Recombinant CAP2 protein bound specifically to C-repeat/dehydration- responsive element in gel-shift assay and transactivated reporter genes in yeast (Saccharomyces cerevisiae) one-hybrid assay. CAP2 appeared to be a single/low copy intronless gene, and the protein product localized in the nucleus. Transcript level of CAP2 increased by dehydration and by treatment with sodium chloride, abscisic acid, and auxin, but not by treatment with low temperature, salicylic acid, and jasmonic acid. The 35S promoter-driven expression of CAP2 in tobacco (Nicotiana tabacum) caused drastic increase in the leaf cell size, and, thereby, in leaf surface area and number of lateral roots. Transgenic plants demonstrated more tolerance to dehydration and salt stress than the wild-type plants. Transgenic plants expressed higher steady-state transcript levels of abiotic stress-response genes NtERD10B and NtERD10C and auxin-response genes IAA4.2 and IAA2.5. Taken together, our results indicated a mutual interrelation between plant growth-development and abiotic stress- response pathways and a probable involvement of CAP2 in both the signaling pathways.Item Regulation of TOP2 by various abiotic stresses including cold and salinity in pea and transgenic tobacco plants(Oxford University Press, 2005) Hettiarachchi, Gardhi H. C. M.; Reddy, Malireddy K.; Sopory, Sudhir K.; Chattopadhyay, SudipAlthough abiotic stress affects plant growth and development, demonstration of its direct effect on regulation of the components of the DNA replication machinery is largely unknown. In this work, we demonstrate that the expression of TOP2 (encoding topoisomerase II) is up-regulated by various abiotic stresses including salinity and low temperature, and phytohormones such as ABA (abscisic acid) and SA (salicylic acid). Transgenic studies with different deletion versions of the TOP2 promoter in tobacco define several promoter determinants responsible for specific abiotic stress responsiveness. Taken together, these results demonstrate a direct involvement of stress in the transcriptional regulation of TOP2.
