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Optimization of hairy root transformation for the functional genomics in chickpea: A platform for nodule developmental studies

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Title Optimization of hairy root transformation for the functional genomics in chickpea: A platform for nodule developmental studies
 
Creator Mandal, Drishti
Srivastava, Deevita
Sinharoy, Senjuti
 
Subject A. rhizogenes
Chickpea
Composite plants
Co-transformation
Hairy root transformation
Root nodule symbiosis
 
Description Accepted date: 1 Jan 2020
Chickpea is a major protein source in low socio-economic classes and cultivated in marginal soil without fertilizer or irrigation. As a result of its root nodule formation capacity chickpea can directly use atmospheric nitrogen. Chickpea is recalcitrant to stable transformation, particularly root regeneration efficiency of chickpea is low. The composite plant-based system with a non-transformed shoot and transformed root is particularly important for root biologist and this approach has already been used successfully for root nodule symbiosis, arbuscular mycorrhizal symbiosis, and other root-related studies. Use of fluorescent marker-based approach can accurately identify the transformed root from its non-transgenic counterpart. RNAi-based gene knockout, overexpression of genes, promoter GUS analysis to understand tissue specific expression and localization of protein can be achieved using the hairy root-based system. We have already published a hairy root-based transformation and composite plant regeneration protocol of chickpea. Here we are describing the recent modification that we have made to increase the transformation frequency and nodule morphology. Further, we have developed a pouch based artificial system, large number of plants can be scored for its nodule developmental phenotype, by using this system.
We thank J. Terpolilli, Murdoch University, Australia for providing
M. ciceri CC1192; Senthil-Kumar Muthappa, NIPGR, New Delhi
for providing chickpea seeds (BGD 112, PUSA 372, GNG 1958,
GNG 1488, GNG 1581, RSG 931); Michael Udvardi, Nobel
Research Institute, Oklahoma, USA for A. rhizogenes ARqua1
strain; Maria J. Harrison, Boyce Thompson Institute for Plant
Research Ithaca, USA for providing pAtUb driven subcellular
marker construct; M. DasGupta, Department of Biochemistry,
University of Calcutta, for providing pCAMBIA2301 vector;
NIPGR for their confocal facilities; and CIF-NIPGR; NIPGRDELCON
for their support. This work is supported by core
research grant from National Institute of Plant Genome Research,
Ramalingwaswami Re-entry grant, DBT (BT/RLF/Re-entry/41/
2013).
 
Date 2020-01-07T06:31:54Z
2020-01-07T06:31:54Z
2020
 
Type Article
 
Identifier Methods in Molecular Biology, 2107: 335-348
978-1-0716-0235-5
https://doi.org/10.1007/978-1-0716-0235-5_18
https://link.springer.com/protocol/10.1007%2F978-1-0716-0235-5_18
http://223.31.159.10:8080/jspui/handle/123456789/1029
 
Language en_US
 
Format application/pdf
 
Publisher Springer Nature Publishing AG