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

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Title A toolbox for nodule development studies in chickpea: a hairy-root transformation protocol and an efficient laboratory strain of Mesorhizobium sp.
 
Creator Mandal, Drishti
Sinharoy, Senjuti
 
Subject chickpea
Mesorhizobium sp.
toolbox
 
Description Accepted date: 01 November 2018
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 into the plant cell are 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 the localization observed in transformed Medicago roots. 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.
We thank R. Varshney and S. Gopalakrishnan, ICRISAT, India; S. Bhatia, NIPGR-New Delhi, J. Terpolilli, Murdoch University, Australia for providing M. ciceri IC59, M. ciceri TAL 620, and M. ciceri CC1192 respectively; H.D. Upadhyaya and D. Sastry, ICRISAT, India for providing chickpea seeds (ICC4958, ICC17258, ICC1882, ICC8261); S. Bhatia, NIPGR for providing BDG256. D. J. Chattopadhyay, Amity University; M. DasGupta of Department of Biochemistry, University of Calcutta, and A. Seal, Department of Biotechnology, University of Calcutta, for their enormous support and allowing us to use their facility. A. Seal for providing pm-ck CFP CD3 1001 construct, M. DasGupta for providing pCAMBIA2301 vector and A. rhizogenes R1000, Michael Udvardi, Nobel Research Institute, Oklahoma, USA for A. rhizogenes ARqua1 and MSU440 strains, Maria J. Harrison, Boyce Thompson Institute for Plant Research Ithaca, USA for providing pAtUb driven subcellular marker constructs and Suzaki Takuya, University of Tsukuba, Japan for providing DR5-GFP-NLS construct. DBT-CU-IPLS and NIPGR for their confocal facilities; CIF-NIPGR; NIPGR-DELCON for their support. T Khanna, Jamia Millia Islamia, New Delhi for technical support and Dipanjan Ghosh, NIPER Kolkata, Dhritiman Dey and Arghya Mukherjee for valuable discussion. This work wassupported by core research grant from National Institute of Plant Genome Research and Ramalingwaswami Re-entry grant, DBT (BT/RLF/Re-entry/41/2013).
 
Date 2018-11-15T09:36:01Z
2018-11-15T09:36:01Z
2019
 
Type Article
 
Identifier Molecular Plant-Microbe Interactions, 32(4): 367-378
1943-7706
http://223.31.159.10:8080/jspui/handle/123456789/897
https://apsjournals.apsnet.org/doi/abs/10.1094/MPMI-09-18-0264-TA
https://doi.org/10.1094/MPMI-09-18-0264-TA
 
Language en_US
 
Format application/pdf
 
Publisher American Phytopathological Society