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Poster pdf file for Austin Winker, program number 594C at TAGC 2020 Online.

A cDNA clone, encoding 75-amino acids metallothionein type 1 protein (CcMT1), has been isolated from the cDNA library of pigeonpea. Northern blot analysis of pigeonpea plants subjected to heavy metal stress revealed increased levels of CcMT1 transcripts. Escherichia coli cells expressing CcMT1 exhibited enhanced metal tolerance and higher accumulation of metal ions than that of control cells. Over-expression of CcMT1 in transgenic Arabidopsis plants imparted increased copper and cadmium tolerance as evidenced by higher plant biomass and chlorophyll content besides profuse root growth when compared to the control plants. Transgenic plants expressing CcMT1 disclosed higher accumulations of Cu and Cd ions in roots as compared to the shoots. No worthwhile differences were observed in the expression levels of copper (AtZIP2) and cadmium (AtNRAMP) transporter genes of CcMT1-transgenics and control plants both under stressed and unstressed conditions. The overall results amply indicate that CcMT1 is contributing to the detoxification of metal ions, thereby providing marked tolerance against metal stresses. This study, first report of its kind, deals with the isolation, characterization and functional validation of CcMT1 in heterologous systems. As such, CcMT1 may be deployed as a potential candidate for enhancement of metal tolerance in diverse crop plants.

Pigeonpea, a major grain legume crop with remarkable drought tolerance traits, has been used for the isolation of stress-responsive genes. Herein, we report generation of ESTs, transcript profiles of selected genes and validation of candidate genes obtained from the subtracted cDNA libraries of pigeonpea plants subjected to PEG/water-deficit stress conditions. Cluster analysis of 124 selected ESTs yielded 75 high-quality ESTs. Homology searches disclosed that 55 ESTs share significant similarity with the known/putative proteins or ESTs available in the databases. These ESTs were characterized and genes relevant to the specific physiological processes were identified. Of the 75 ESTs obtained from the cDNA libraries of drought-stressed plants, 20 ESTs proved to be unique to the pigeonpea. These sequences are envisaged to serve as a potential source of stress-inducible genes of the drought stress-response transcriptome, and hence may be used for deciphering the mechanism of drought tolerance of the pigeonpea. Expression profiles of selected genes revealed increased levels of m-RNA transcripts in pigeonpea plants subjected to different abiotic stresses. Transgenic Arabidopsis lines, expressing Cajanus cajan hybrid-proline-rich protein (CcHyPRP), C. cajan cyclophilin (CcCYP) and C. cajan cold and drought regulatory (CcCDR) genes, exhibited marked tolerance, increased plant biomass and enhanced photosynthetic rates under PEG/NaCl/cold/heat stress conditions. This study represents the first report dealing with the isolation of drought-specific ESTs, transcriptome analysis and functional validation of drought-responsive genes of the pigeonpea. These genes, as such, hold promise for engineering crop plants bestowed with tolerance to major abiotic stresses.

A hybrid-proline-rich protein encoding gene (CcHyPRP) has been isolated and characterized, for the first time, from the subtracted cDNA library of pigeonpea (Cajanus cajan L.) plants subjected to drought stress. Functionality of CcHyPRP has been validated for abiotic stress tolerance using the heterologous yeast and Arabidopsis systems. The CcHyPRP contained a repetitive proline-rich (PR) N-terminal domain and a conserved eight cysteine motif (8CM) at the C-terminus. Southern analysis disclosed single-copy nature of CcHyPRP gene in the pigeonpea genome. Northern analysis revealed higher levels of CcHyPRP transcripts in PEG, NaCl, heat (42 degrees C), cold and ABA-treated plants compared with the weak signals observed in the untreated plants, suggesting stress-responsive nature of the CcHyPRP gene. In yeast, expression of CcHyPRP imparted marked tolerance against abiotic stresses exerted by PEG, high temperature, NaCl and LiCl. Transgenic Arabidopsis lines, expressing CcHyPRP under the control of CaMV35S and rd29A promoters, when subjected to PEG, mannitol, NaCl, LiCl and heat (42 degrees C) stress, developed into healthy plants with profuse root system and increased biomass in contrast to the weak-stunted wild-type plants. The CcHyPRP-transgenics driven by stress-inducible rd29A exhibited similar stress-tolerance as that of CaMV35S-lines without any negative effects on plant morphology, implying that stress-inducible promoters are preferable for production of stress tolerant transgenics. The overall results amply demonstrate the profound effect of CcHyPRP in bestowing multiple abiotic stress tolerance at cellular and whole plant levels. Accordingly, the multipotent CcHyPRP seems promising as a prime candidate gene to fortify crop plants with abiotic stress tolerance.

This study examines the process of divorce; especially the pre-divorce stage, using data from a family court in the city of Visakhapatnam, India. It examines the process of divorce along with the correlates and other determinants. Basing on the information provided by court records, only women divorcees were interviewed. Some of the findings like shorter marriages resulted in divorce and majority of the divorcees were young, comply with the universally acknowledged research findings. However, absence of association between wife’s education and divorce are some of the crucial findings that do not corroborate the established theory. This cross-cultural study calls for evolving a socially approved ritual to mark the transition from marriage to divorce; in order to facilitate the separation in a cordial atmosphere. In view of the linguistic and socio-cultural diversity of the Indian society the study conducted in this part of the country (state of Andhra Pradesh) which is first of its kind, assumes significance. It is also expected to contribute substantially to the discussion on gender related issues.

A full-length cDNA clone of pigeonpea (Cajanus cajan L.) encoding cyclophilin (CcCYP) has been isolated from the cDNA library of plants subjected to drought stress. Amino acid sequence of CcCYP disclosed similarity with that of single-domain cytosolic cyclophilins of various organisms. Expression profile of CcCYP in pigeonpea plants is strongly induced by different abiotic stresses, indicating its stress-responsive nature. Compared to the control plants, the transgenic Arabidopsis lines expressing CcCYP exhibited high-level tolerance against major abiotic stresses, viz., drought, salinity and extreme temperatures as evidenced by increased plant survival, biomass, chlorophyll content and profuse root growth. The CcCYP transgenics, compared to the controls, revealed enhanced peptidyl-propyl cis-trans isomerase (PPIase) activity under stressed conditions, owing to transcriptional activation of stress-related genes besides intrinsic chaperonic activity of the cyclophilin. The transgenic plants subjected to salt stress exhibited higher Na(+) ion accumulation in roots as compared to shoots, while a reverse trend was observed in the salt-stressed control plants, implicating the involvement of CcCYP in the maintenance of ion homeostasis. Expression pattern of CcCYP:GFP fusion protein confirmed the localization of CcCYP predominantly in the nucleus as revealed by intense green fluorescence. The overall results amply demonstrate the implicit role of CcCYP in conferring multiple abiotic stress tolerance at whole-plant level.

A full-length cDNA clone of pigeonpea (Cajanus cajan L.) encoding cyclophilin (CcCYP) has been isolated from the cDNA library of plants subjected to drought stress. Amino acid sequence of CcCYP disclosed similarity with that of single-domain cytosolic cyclophilins of various organisms. Expression profile of CcCYP in pigeonpea plants is strongly induced by different abiotic stresses, indicating its stress-responsive nature. Compared to the control plants, the transgenic Arabidopsis lines expressing CcCYP exhibited high-level tolerance against major abiotic stresses, viz., drought, salinity and extreme temperatures as evidenced by increased plant survival, biomass, chlorophyll content and profuse root growth. The CcCYP transgenics, compared to the controls, revealed enhanced peptidyl-propyl cis-trans isomerase (PPIase) activity under stressed conditions, owing to transcriptional activation of stress-related genes besides intrinsic chaperonic activity of the cyclophilin. The transgenic plants subjected to salt stress exhibited higher Na+ ion accumulation in roots as compared to shoots, while a reverse trend was observed in the salt-stressed control plants, implicating the involvement of CcCYP in the maintenance of ion homeostasis. Expression pattern of CcCYP:GFP fusion protein confirmed the localization of CcCYP predominantly in the nucleus as revealed by intense green fluorescence. The overall results amply demonstrate the implicit role of CcCYP in conferring multiple abiotic stress tolerance at whole-plant level.

A hybrid-proline-rich protein encoding gene (CcHyPRP) has been isolated and characterized, for the first time, from the subtracted cDNA library of pigeonpea (Cajanus cajan L.) plants subjected to drought stress. Functionality of CcHyPRP has been validated for abiotic stress tolerance using the heterologous yeast and Arabidopsis systems. The CcHyPRP contained a repetitive proline-rich (PR) N-terminal domain and a conserved eight cysteine motif (8CM) at the C-terminus. Southern analysis disclosed single-copy nature of CcHyPRP gene in the pigeonpea genome. Northern analysis revealed higher levels of CcHyPRP transcripts in PEG, NaCl, heat (42 degrees C), cold and ABA-treated plants compared with the weak signals observed in the untreated plants, suggesting stress-responsive nature of the CcHyPRP gene. In yeast, expression of CcHyPRP imparted marked tolerance against abiotic stresses exerted by PEG, high temperature, NaCl and LiCl. Transgenic Arabidopsis lines, expressing CcHyPRP under the control of CaMV35S and rd29A promoters, when subjected to PEG, mannitol, NaCl, LiCl and heat (42 degrees C) stress, developed into healthy plants with profuse root system and increased biomass in contrast to the weak-stunted wild-type plants. The CcHyPRP-transgenics driven by stress-inducible rd29A exhibited similar stress-tolerance as that of CaMV35S-lines without any negative effects on plant morphology, implying that stress-inducible promoters are preferable for production of stress tolerant transgenics. The overall results amply demonstrate the profound effect of CcHyPRP in bestowing multiple abiotic stress tolerance at cellular and whole plant levels. Accordingly, the multipotent CcHyPRP seems promising as a prime candidate gene to fortify crop plants with abiotic stress tolerance.