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

Fatemeh Mahdavi, Maziah Mahmood, Normah Mohd. Noor

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)888-894
Number of pages7
JournalBiologia (Poland)
Volume69
Issue number7
DOIs
Publication statusPublished - 2014

Fingerprint

Musa
male flowers
bananas
flower
Green Fluorescent Proteins
green fluorescent protein
Gold
gold
DNA
Pressure
Genes
Vacuum
protein
gene
protein synthesis
Plant DNA
Musa paradisiaca
transgenic plant
Helium
helium

Keywords

  • banana
  • genetic engineering
  • green fluorescent protein
  • male inflorescence
  • particle bombardment
  • pCambia1304

ASJC Scopus subject areas

  • Plant Science
  • Animal Science and Zoology
  • Ecology, Evolution, Behavior and Systematics
  • Biochemistry
  • Cell Biology
  • Genetics
  • Molecular Biology

Cite this

Optimization of particle bombardment parameters for DNA delivery into the male flowers of banana. / Mahdavi, Fatemeh; Mahmood, Maziah; Mohd. Noor, Normah.

In: Biologia (Poland), Vol. 69, No. 7, 2014, p. 888-894.

Research output: Contribution to journalArticle

Mahdavi, Fatemeh ; Mahmood, Maziah ; Mohd. Noor, Normah. / Optimization of particle bombardment parameters for DNA delivery into the male flowers of banana. In: Biologia (Poland). 2014 ; Vol. 69, No. 7. pp. 888-894.
@article{4f21d814531446d3ac9c4756c9a3673e,
title = "Optimization of particle bombardment parameters for DNA delivery into the male flowers of banana",
abstract = "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.",
keywords = "banana, genetic engineering, green fluorescent protein, male inflorescence, particle bombardment, pCambia1304",
author = "Fatemeh Mahdavi and Maziah Mahmood and {Mohd. Noor}, Normah",
year = "2014",
doi = "10.2478/s11756-014-0391-7",
language = "English",
volume = "69",
pages = "888--894",
journal = "Biologia",
issn = "0006-3088",
publisher = "de Gruyter",
number = "7",

}

TY - JOUR

T1 - Optimization of particle bombardment parameters for DNA delivery into the male flowers of banana

AU - Mahdavi, Fatemeh

AU - Mahmood, Maziah

AU - Mohd. Noor, Normah

PY - 2014

Y1 - 2014

N2 - 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.

AB - 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.

KW - banana

KW - genetic engineering

KW - green fluorescent protein

KW - male inflorescence

KW - particle bombardment

KW - pCambia1304

UR - http://www.scopus.com/inward/record.url?scp=84906307499&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84906307499&partnerID=8YFLogxK

U2 - 10.2478/s11756-014-0391-7

DO - 10.2478/s11756-014-0391-7

M3 - Article

AN - SCOPUS:84906307499

VL - 69

SP - 888

EP - 894

JO - Biologia

JF - Biologia

SN - 0006-3088

IS - 7

ER -