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In vitro Direct Regeneration and Agrobacterium Tumefaciens mediated in planta Transformation of Ocimum sanctum L.

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Title In vitro Direct Regeneration and Agrobacterium Tumefaciens mediated in planta Transformation of Ocimum sanctum L.
 
Creator Khan, Sana
Husain, Zakir
Rahman, Laiq ur
 
Subject Gus-A
Kanamycin
MAPs
Npt-II
O. tenuiflorum
Petiole
 
Description 1019-1028
An in vitro regeneration system for propagation has been successfully developed for a valuable medicinal and aromatic
plant ‘Ocimum sanctum L’. In the present study, petiole explants, from in-vitro grown cultures of O. sanctum, was used for
direct regeneration. The developed protocol employed 98% of regeneration frequency in addition to 9.6 shoots per explant
when cultured on Murashige and Skoog (MS) medium fortified with 3 mg/L benzylamino purine (BAP) and 1 mg/L
Naphthalene acetic acid (NAA). Furthermore, Agrobacterium tumefaciens Mediated genetic Transformation (ATMT)
protocol (transient and stable) was also developed using LBA4404 strain harboring pBI121 with uid-A and neomycin
phosphotransferase genes. The regenerated transformants were shifted on MS with kanamycin (50 mg/L) and afterwards
placed on the half-strength MS medium. The validation was done through polymerase chain reaction (PCR) with neomycin
phosphotransferase-II (npt-II) & β-glucoronidase (uid-A) gene primers. The maximum stable transformation frequency of
70% ± 0.35 was achieved. Hence, it is apparent that the established protocols i.e. in vitro direct regeneration and ATMT are
appropriate for integrating novel enzymes/genes through high throughput techniques such as gene tagging, and targeted
gene replacement to modulate the primary as well as secondary metabolic flux towards desired agronomic product
or trait in planta
 
Date 2022-10-28T09:56:00Z
2022-10-28T09:56:00Z
2022-10
 
Type Article
 
Identifier 0022-4456 (Print); 0975-1084 (Online)
http://nopr.niscpr.res.in/handle/123456789/60748
https://doi.org/10.56042/jsir.v81i10.59665
 
Language en
 
Publisher NIScPR-CSIR, India
 
Source JSIR Vol.81(10) [Oct 2022]