Optimization of particle bombardment parameters for DNA delivery into the male flowers of banana

The possibility of increasing the efficiency of banana transformation was investigated by particle bombardment of the male flowers of banana plants for constitutive expression of gfp gene. The effects of particle bombardment parameters, such as acceleration pressure, bombardment distance, chamber va...

詳細記述

保存先:
書誌詳細
主要な著者: Mahdavi, Fatemeh, Mahmood, Maziah, Mohd Noor, Normah
フォーマット: 論文
出版事項: Versita 2014
オンライン・アクセス:http://psasir.upm.edu.my/id/eprint/34780/
http://link.springer.com/article/10.2478%2Fs11756-014-0391-7
タグ: タグ追加
タグなし, このレコードへの初めてのタグを付けませんか!
id my.upm.eprints.34780
record_format eprints
spelling my.upm.eprints.347802015-12-23T01:14:55Z http://psasir.upm.edu.my/id/eprint/34780/ Optimization of particle bombardment parameters for DNA delivery into the male flowers of banana Mahdavi, Fatemeh Mahmood, Maziah Mohd Noor, Normah The possibility of increasing the efficiency of banana transformation was investigated by particle bombardment of the male flowers of banana plants for constitutive expression of gfp gene. The effects of particle bombardment parameters, such as acceleration pressure, bombardment distance, chamber vacuum pressure, gold microcarrier size, gold quantity, DNA quantity, number of bombardments and pre-culture were examined. Single cauliflower-like bodies (CLBs) clusters, induced from meristemic parts of Musa sapientum cv. Nangka (AAB) male flowers, were bombarded by pCambia1304 plasmid carrying gfp gene driven by the CaMV 35S promoter. Optimal transient expression of green-fluorescent protein (GFP) was obtained when the three-day old cultured tissues were bombarded two times at 1100 psi helium pressure. However, the highest GFP expression was observed when 9 cm was applied as bombardment distance with 28 mmHg chamber vacuum pressure. Gold particle with 1 μm diameter at 60 μg/μL concentrations coated with 1.5 μg/μL of DNA have been used as the optimum bombardment parameter since GFP expression was significantly different compared to other conditions. Application of optimized condition proved effective for the generation of stable transgenic banana plants. PCR and southern blot analyses confirmed the presence and integration of gfp gene in genomic DNA of transformed plants. Transformation frequency achieved with the optimized protocol was 7.5% which was significantly higher than the conventional protocol. Versita 2014 Article PeerReviewed Mahdavi, Fatemeh and Mahmood, Maziah and Mohd Noor, Normah (2014) Optimization of particle bombardment parameters for DNA delivery into the male flowers of banana. Biologia, 69 (7). pp. 888-894. ISSN 0006-3088; ESSN: 1336-9563 http://link.springer.com/article/10.2478%2Fs11756-014-0391-7 10.2478/s11756-014-0391-7
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
description The possibility of increasing the efficiency of banana transformation was investigated by particle bombardment of the male flowers of banana plants for constitutive expression of gfp gene. The effects of particle bombardment parameters, such as acceleration pressure, bombardment distance, chamber vacuum pressure, gold microcarrier size, gold quantity, DNA quantity, number of bombardments and pre-culture were examined. Single cauliflower-like bodies (CLBs) clusters, induced from meristemic parts of Musa sapientum cv. Nangka (AAB) male flowers, were bombarded by pCambia1304 plasmid carrying gfp gene driven by the CaMV 35S promoter. Optimal transient expression of green-fluorescent protein (GFP) was obtained when the three-day old cultured tissues were bombarded two times at 1100 psi helium pressure. However, the highest GFP expression was observed when 9 cm was applied as bombardment distance with 28 mmHg chamber vacuum pressure. Gold particle with 1 μm diameter at 60 μg/μL concentrations coated with 1.5 μg/μL of DNA have been used as the optimum bombardment parameter since GFP expression was significantly different compared to other conditions. Application of optimized condition proved effective for the generation of stable transgenic banana plants. PCR and southern blot analyses confirmed the presence and integration of gfp gene in genomic DNA of transformed plants. Transformation frequency achieved with the optimized protocol was 7.5% which was significantly higher than the conventional protocol.
format Article
author Mahdavi, Fatemeh
Mahmood, Maziah
Mohd Noor, Normah
spellingShingle Mahdavi, Fatemeh
Mahmood, Maziah
Mohd Noor, Normah
Optimization of particle bombardment parameters for DNA delivery into the male flowers of banana
author_facet Mahdavi, Fatemeh
Mahmood, Maziah
Mohd Noor, Normah
author_sort Mahdavi, Fatemeh
title Optimization of particle bombardment parameters for DNA delivery into the male flowers of banana
title_short Optimization of particle bombardment parameters for DNA delivery into the male flowers of banana
title_full Optimization of particle bombardment parameters for DNA delivery into the male flowers of banana
title_fullStr Optimization of particle bombardment parameters for DNA delivery into the male flowers of banana
title_full_unstemmed Optimization of particle bombardment parameters for DNA delivery into the male flowers of banana
title_sort optimization of particle bombardment parameters for dna delivery into the male flowers of banana
publisher Versita
publishDate 2014
url http://psasir.upm.edu.my/id/eprint/34780/
http://link.springer.com/article/10.2478%2Fs11756-014-0391-7
_version_ 1643831256863997952
score 13.251813