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197590

FACTORS INFLUENCING SUCCESSFUL AGROBACTERIUMMEDIATED GENETIC TRANSFORMATION OF MUSKMELON

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Last updated: 05 Jan 2025

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Abstract

The development of a robust Agrobacterium-mediated
transformation protocol for muskmelon (Cucumis melo L.)
requires the identification and optimization of the factors affecting
DNA delivery and plant regeneration. We have used pieces of
mature cotyledon from cultivar Hales Best Jumbo and the
Agrobacterium tumefaciens strain LBA4404, to investigate and
optimize regeneration and transformation. Agrobacterium strain
harboring the binary vectors which contains phosphino-thricin
acetyl transferase gene (bar) as selectable marker for herbicide
resistance (glufosinat ammonium) and β-glucuronidase gene
(uidA) as reporter. Factors that produced differences in DNA
delivery and regeneration included bacterial concentration, length
of exposure, the time allowed for co-cultivation and antibiotic
concentration (claforan, cefotaximum). These factors combined as
a whole led to successful transformation. Bacterial concentration,
time of exposure of the explants to the bacteria, co-cultivation
period and antibiotic concentration were found to affect
regeneration and transformation efficiency. Analysis of these
parameters allowed the development of an optimized protocol for
Agrobacterium-mediated transformation of Cucumis melo cv.
Hales Best Jumbo cotyledon explants. We fully describe a
protocol that allowed efficient DNA delivery and gave rise stable
transgenic muskmelon plants. Selectable marker gene expression
and reporter gene assay demonstrated that transgenes were
integrated into the muskmelon genome. Genetic transformation of
calli and regenerated plantlets was confirmed by histochemical β-
glucuronidase assays.

DOI

10.21608/fjard.2006.197590

Keywords

transformation, Muskmelon, Agrobacterium-mediated, factors, herbicide, β-glucuronidase

Authors

First Name

Eissa Ahmed

Last Name

Eissa

MiddleName

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Affiliation

Department of Genetics, Faculty of Agriculture, Fayoum University, Egypt

Email

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City

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Orcid

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First Name

Velich

Last Name

I.

MiddleName

-

Affiliation

Department of Genetics and Plant Breeding, Faculty of Horticultural Sciences, Szent István University, Budapest, Hungary

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Orcid

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Volume

20

Article Issue

2

Related Issue

27952

Issue Date

2006-07-01

Receive Date

2021-10-03

Publish Date

2006-07-01

Page Start

111

Page End

120

Print ISSN

1110-7790

Online ISSN

2805-2528

Article File

FJARD_Volume 20_Issue 2_Pages 111-120.pdf

PDF . 654.4kB

Link

https://fjard.journals.ekb.eg/article_197590.html

Detail API

https://fjard.journals.ekb.eg/service?article_code=197590

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Type

Research articles.

Type Code

1,920

Publication Type

Journal

Publication Title

Fayoum Journal of Agricultural Research and Development

Publication Link

https://fjard.journals.ekb.eg/

MainTitle

FACTORS INFLUENCING SUCCESSFUL AGROBACTERIUMMEDIATED GENETIC TRANSFORMATION OF MUSKMELON

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Article

Created At

23 Jan 2023