Contact us
 |
[email protected] |
 |
3275638434 |
 |
 |
| Paper Publishing WeChat |
Useful Links
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License
Article
Transgenic Date Palm Containing Endotoxin Cry3Aa Gene
Author(s)
Awatef Mahmoud Badr-Elden1, Ibrahim Abd El-Maksoud Ibrahim1, Hamdy Ahmed Emara1, Mahdia Farid Gabr2, Ahmed Abbas Nower1 and Tamer Mahfouz Abd Elaziem2
Full-Text PDF
XML 939 Views
DOI:10.17265/2161-6256/2017.04.003
Affiliation(s)
1. Department of Plant Biotechnology, Genetic Engineering and Biotechnology Research Institute (GEBRI), University of Sadat City, Sadat City 32897, Egypt
2. Department of Plant Genetic Resources, Desert Research Center, Cairo 11753, Egypt
ABSTRACT
Date palm, like all other
crops, is very sensitive to the injury by many insect pests,
which may lead to the death of the affected plant and causes decrease in yield. In the present study,
an efficient Agrobacterium for genetic
transformation was successfully achieved for well known date palm (Phoenix
dactylifera L.) cv. Medjool and Khalas using callus as explant. Embryogenic callus
were recorded 100% mortality when cultured on MS medium containing 100 mg/L kanamycin
with different cultivars, thus it was chosen for the selection of transformed
explants. Embryogenic callus of Medjool and Khalas were incubated with Agrobacterium
tumefaciens strain LBA 4404 for
0.5, 1, 2, 4 and 24 h on LB medium. After the incubation
periods, embryogenic callus was transferred to MS medium with 0.1 mg/L NAA,
0.05 mg/L BA, 250 mg/L carbenicillin and 100 mg/L kanamycin for detection of transgenic embryogenic callus.
Polymerase chain reaction (PCR) was used
for the rapid screening of Cry3Aa gene. For screening, total genomic DNA was isolated from transformants.
Using primer specific to Cry3Aa gene
(forward and reverse), a PCR product with a
size of about 2,000 bp was amplified when
all nucleic acid from
the transformants were utilized as templates. PCR analysis confirmed the
appearance of the transgene of 2,000 bp in one individual plantlet. Presence and
integration of foreign Cry3Aa gene in
regenerated kanamycin resistant embryogenic callus was also confirmed by Southern blot
hybridization. It was found that one transgenic embryogenic callus
for both Medjool and Khalas showed a single copy of gene integration. These
results signify the successful transfer of Cry3Aa gene into date palm plant.
KEYWORDS
Date palm, Agrobacterium tumefaciens, Cry3Aa gene, Southern blot analysis, transformation.
Cite this paper
References