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Rapid regeneration and ploidy stability of 'cv IR36' indica rice (Oryza Sativa. L) confers efficient protocol for in vitro callus organogenesis and Agrobacterium tumefaciens mediated transformation.

ABSTRACT: Cereal crops are the major targets for transformation mediated crop improvement and IR36 is an early maturing, high yielding, insect and disease resistant rice variety however, it is abiotic stress sensitive. Hence, development of an efficient and reproducible micropropagation system via somatic embryogenesis and Agrobacterium tumefaciens mediated transformation is prerequisite to develop abiotic stress tolerant IR36. Further, Genetic stability of analysis of plantlets through RAPD and ISSR and Ploidy level through Flow cytometry (FCM) measurement of 2C DNA content is necessary for future application of transformed IR36.In this study, Mature seeds inoculated on (Murashige and Skoog) MS medium with 11.31 ?M 2, 4-dichlorophenoxyacetic acid (2, 4-D) and 0.3 ?M Kinetin (Kn) had highest callus induction frequency (98%). The highest regeneration frequency (80%) was observed in MS?+?13.28 ?M Benzyladenine (BA) with 8.06 ?M ?-naphthalene acetic acid (NAA). Randomly Amplified Polymorphic DNA (RAPD), Inter Simple Sequence Repeat (ISSR) and Flow Cytometry (FCM) analysis showed no significant variation in the 2C DNA (0.81 pg/2C) content and Ploidy level between wild type IR36 and in vitro maintained rice lines. Of the various OD bacterial culture, an optimum OD of 0.4 and inoculation duration of 10 min resulted in efficient Agrobacterium-mediated transformation. ?-glucuronidase activity was maximum in callus (99.05%).These results described here confirm the reliability of this protocol for micropropagation and delivery of desirable gene using A. tumefaciens into indica rice.

SUBMITTER: Krishnan SR

PROVIDER: S-EPMC5430341 | biostudies-literature | 2013 Dec

REPOSITORIES: biostudies-literature

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Rapid regeneration and ploidy stability of 'cv IR36' indica rice (Oryza Sativa. L) confers efficient protocol for in vitro callus organogenesis and Agrobacterium tumefaciens mediated transformation.

Krishnan Subramanian Radhesh SR Priya Arumugam Mohana AM Ramesh Manikandan M

Botanical studies 20131021 1

<h4>Background</h4>Cereal crops are the major targets for transformation mediated crop improvement and IR36 is an early maturing, high yielding, insect and disease resistant rice variety however, it is abiotic stress sensitive. Hence, development of an efficient and reproducible micropropagation system via somatic embryogenesis and Agrobacterium tumefaciens mediated transformation is prerequisite to develop abiotic stress tolerant IR36. Further, Genetic stability of analysis of plantlets through ...[more]

PMID: 28510899

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Project description:Arbuscular mycorrhizal fungi (AMF) symbionts not only promote the growth of host plant but also alleviate abiotic stresses. This study aimed to investigate the putative role of AMF in salt stress regulation of upland pigmented rice cv. Leum Pua (LP) comparing with Pokkali salt tolerant (positive check). In general, LP is a variety of glutinous rice that contains anthocyanin pigment in the black pericarp, due to which it possesses high antioxidant activities compared to non-pigmented rice. Pot experiment was conducted to evaluate the impact of inoculated AMF, Glomus etunicatum (GE), Glomus geosporum (GG), and Glomus mosseae (GM) strains, in the LP plantlets subjected to 0 (control) or 150 mM NaCl (salt stress) for 2 weeks in comparison with Pokkali (a salt tolerant rice cultivar), which was maintained as a positive check. Root colonization percentage under NaCl conditions ranged from 23 to 30%. Na+ content in the flag leaf tissues was increased to 18-35 mg g-1 DW after exposure to 150 mM NaCl for 14 days in both inoculated and un-inoculated LP plants, whereas Na:K ratio was very low in cv. Pokkali. Interestingly, sucrose content in the flag leaf tissues of un-inoculated LP plants under salt stress was increased significantly by 50 folds over the control as an indicator of salt stress response, whereas it was unchanged in all AMF treatments. Fructose and free proline in GE inoculated plants under salt stress were accumulated over control by 5.75 and 13.59 folds, respectively, for osmotic adjustment of the cell, thereby maintaining the structure and functions of chlorophyll pigments, Fv/Fm, ?PSII, and stomatal function. Shoot height, flag leaf length, number of panicles, panicle length, panicle weight, and 100-grain weight in GE inoculated plants of cv. LP under salt stress were maintained similar to cv. Pokkali. Interestingly, cyanidin-3-glucoside (C3G) and peonidin-3-glucoside (P3G) in the pericarp of cv. LP were regulated by GE inoculation under salt stress conditions. In summary, AMF-inoculation in rice crop is a successful alternative approach to reduce salt toxicity, maintain the yield attributes, and regulate anthocyanins enrichment in the pericarp of grains.

Dataset Information

Rapid regeneration and ploidy stability of 'cv IR36' indica rice (Oryza Sativa. L) confers efficient protocol for in vitro callus organogenesis and Agrobacterium tumefaciens mediated transformation.

Publications

Rapid regeneration and ploidy stability of 'cv IR36' indica rice (Oryza Sativa. L) confers efficient protocol for in vitro callus organogenesis and Agrobacterium tumefaciens mediated transformation.

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