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204911

Agrobacterium-Mediated Genetic Transformation and Regeneration of Salt Tolerant Transgenic of Sour Orange Rootstock (Citrus au-rantium L.)

Article

Last updated: 04 Jan 2025

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Abstract

Traditional breeding techniques of citrus are complicated and time intensive. In this study, an effective and simple method for the production of sour orange transgenic plants incorporating salt tolerance gene (Pr10a) has been established. Transgenic plants were produced by Agrobacterium tumefaciens -mediated gene transfer using hypocotyl explants under the selection pressure of phosphinothericin and NaCl as selective agents. Hypocotyl segments were excised from in vitro seedlings and incubated with Agrobacterium tumefaciens EHA105 strain harboring the transgene (Pr10a) under the control of mannopine synthase promoter (p-MAS). In addition, the used dicistronic binary vector contained luciferase (Luc) gene linked to internal ribosome entry site elements in the T-DNA region. MS medium containing 1.5 mg/l 6-benzylaminopurine (BA) and 0.2 mg/l indole butyric acid (IBA) proved optimal for shoot regeneration (97%) from sour orange hypocotyl. The number of marker gene (Luc) expressing plants increased as the inoculation time and bacterial density (OD600) increased up to20 min and 0.4, respectively. Under this condition, the transformation efficiency of hypocotyl segments was 24.2%. PCR analysis using Pr10a and Luc primers confirmed the existence of the transgenes in transgenic plants where the expected band size of 480 and 837 bp were obtained for both transgenes, respectively. The integration of T-DNA containing Pr10a and Luc genes in the transgenic sour orange was revealed by Southern blot analysis and verifiedthe results of luciferase enzyme activity and PCR analysis of Pr10a and Luc genes. A simple integration pattern for all the transgenes including the single copy integration was predominantly observed.

DOI

10.21608/jpp.2021.204911

Keywords

genetic transformation, Shoot organogenesis, Sour orange, Pr10a, salinity

Authors

First Name

Doaa

Last Name

Abou Elyazid

MiddleName

Mahmoud

Affiliation

Department of Horticulture , Faculty of Agriculture, Kafrelsheikh university

Email

delgiar@agr.kfs.edu.eg

City

Kafr El Sheikh

Orcid

0000-0001-7269-2833

First Name

A. N.

Last Name

El-Banna

MiddleName

-

Affiliation

Genetics Department, Faculty of Agriculture, Kafrelsheikh University, Kafr El-Sheikh 33516, Egypt

Email

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City

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Orcid

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Volume

12

Article Issue

11

Related Issue

28871

Issue Date

2021-11-01

Receive Date

2021-11-15

Publish Date

2021-11-01

Page Start

1,145

Page End

1,150

Print ISSN

2090-3669

Online ISSN

2090-374X

Link

https://jpp.journals.ekb.eg/article_204911.html

Detail API

https://jpp.journals.ekb.eg/service?article_code=204911

Order

2

Type

Original Article

Type Code

887

Publication Type

Journal

Publication Title

Journal of Plant Production

Publication Link

https://jpp.journals.ekb.eg/

MainTitle

Agrobacterium-Mediated Genetic Transformation and Regeneration of Salt Tolerant Transgenic of Sour Orange Rootstock (Citrus au-rantium L.)

Details

Type

Article

Created At

22 Jan 2023