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Molecular characterization of the 14-3-3 gene family in rice and its expression studies under abiotic stress

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Main Conclusion

14-3-3 isoforms were relatively less conserved at the C-terminal region across plant groups. Both Os 14-3-3f and Os 14-3-3g were inducible with differential gene expression levels under different abiotic stress and developmental stages in sensitive and tolerant indica rice cultivars as confirmed both at transcript and protein level.

Plant 14-3-3s has been well characterized to function in several signaling pathways, biotic as well as abiotic stress and nutrient metabolism. We attempted comprehensive analysis of 14-3-3 genes in different plant lineages such as green algae (Chlamydomonas reinhardtii), moss (Physcomitrella patens) and lycophyte (Selaginella moellendorffii), dicot Arabidopsis thaliana and monocot Oryza sativa sub sp. japonica at the gene and protein level. Sequence alignment results revealed that 14-3-3 isoforms were evolutionarily conserved across all taxa with variable C-terminal end. Phylogenetic analysis indicated that the majority of 14-3-3 isoforms in rice belong to the non-epsilon group that clustered separately from the dicot group. Segmental duplication event played a significant role in the expansion of both, Arabidopsis and rice, 14-3-3 isoforms as revealed by synteny studies. In silico gene expression using Massive Parallel Signature Sequencing and microarray analysis revealed that 14-3-3 isoforms have variable expression in different tissue types and under different abiotic stress regime in Arabidopsis and japonica rice. Both, semi-quantitative and qPCR results, confirmed that Os14-3-3f and Os14-3-3g were inducible under abiotic stress in lamina and roots of indica rice and relatively higher under salinity and cold stress in Nonabokra, under dehydration stress in N-22 and under exogenous ABA in IR-29 usually after 3–6 h of treatment. Both, 14-3-3f and 14-3-3g, were highly expressed in flag leaves, stems and panicles and mature roots. These results were further confirmed by immunoblot analysis of rice cultivars using Os14-3-3f antibody generated from recombinant Os14-3-3f protein. The results provide the first comprehensive report of Os14-3-3 gene expression in indica rice cultivars which differ in tolerance to abiotic stress that might be useful for further research.

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Abbreviations

DAP:

Days after pollination

MPSS:

Massive parallel signature sequences

TPM:

Transcript per million

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Acknowledgements

We gratefully acknowledge the Director, Bose Institute, for providing fellowship to NY and for infrastructural support. We are also thankful to Mr. Jadab Ghosh, Mrs. Kaberi Ghosh and Mr. Mrinal Das, Bose Institute, for their technical help. We also thank Dr. Subarna Thakur (Bioinformatics Centre, Bose Institute) for helping in synteny map preparation.

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Correspondence to Dibyendu Narayan Sengupta.

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The authors declare that they have no competing interests.

Ethical statement regarding use of plant material

The authors state that rice seeds, leaves and other planting materials were procured as per local/national regulations. All experimental materials were grown in a contained facility without causing any harm to the natural resource and biodiversity.

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Yashvardhini, N., Bhattacharya, S., Chaudhuri, S. et al. Molecular characterization of the 14-3-3 gene family in rice and its expression studies under abiotic stress. Planta 247, 229–253 (2018). https://doi.org/10.1007/s00425-017-2779-4

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  • DOI: https://doi.org/10.1007/s00425-017-2779-4

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