Development of a direct transformation method by GFP screening and in vitro whole plant regeneration of Capsicum frutescens L.

Background Capsicum is a genus of important spice crop that belongs to the chili lineage. However, many Capsicum species (family Solanaceae) are known to be recalcitrant to genetic transformation and in vitro regeneration, thus hampering the effort in using Capsicum species for detailed biologica...

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Main Authors: Chee, Marcus Jenn Yang, Lycett, Grantley W., Chin, Chiew Foan
Format: Article
Published: Elsevier 2018
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Online Access:https://eprints.nottingham.ac.uk/51963/
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author Chee, Marcus Jenn Yang
Lycett, Grantley W.
Chin, Chiew Foan
author_facet Chee, Marcus Jenn Yang
Lycett, Grantley W.
Chin, Chiew Foan
author_sort Chee, Marcus Jenn Yang
building Nottingham Research Data Repository
collection Online Access
description Background Capsicum is a genus of important spice crop that belongs to the chili lineage. However, many Capsicum species (family Solanaceae) are known to be recalcitrant to genetic transformation and in vitro regeneration, thus hampering the effort in using Capsicum species for detailed biological investigation. In this study, we have developed an optimized protocol for the direct transformation of Capsicum frutescens L. cv. Hot Lava via a biolistic particle delivery system. In addition, in vitro whole plant regeneration from the hypocotyl explants of C. frutescens was established. Results In this biolistic system study, explant target distance, bombardment helium (He) pressure and the size of microcarrier were the key parameters to be investigated. The optimized parameters based on screening of GFP expression were determined to be 6 cm target distance, 1350 psi of helium pressure and 1.6 μm of gold particle (microcarrier) size. The greatest number of shoots were obtained from hypocotyl as explant using Murashige and Skoog medium supplemented with 5.0 mg/L BAP and 0.1 mg/L NAA. An average of 5 shoots per explant were formed. Out of which, one shoot managed to form root and developed into whole plant. Conclusions We have obtained an optimized protocol for the biolistic transformation of chili and in vitro regeneration of chili plantlets. The establishment of the protocols will provide a platform for molecular breeding and biological studies of the chili plants.
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spelling nottingham-519632025-09-09T06:00:11Z https://eprints.nottingham.ac.uk/51963/ Development of a direct transformation method by GFP screening and in vitro whole plant regeneration of Capsicum frutescens L. Chee, Marcus Jenn Yang Lycett, Grantley W. Chin, Chiew Foan Background Capsicum is a genus of important spice crop that belongs to the chili lineage. However, many Capsicum species (family Solanaceae) are known to be recalcitrant to genetic transformation and in vitro regeneration, thus hampering the effort in using Capsicum species for detailed biological investigation. In this study, we have developed an optimized protocol for the direct transformation of Capsicum frutescens L. cv. Hot Lava via a biolistic particle delivery system. In addition, in vitro whole plant regeneration from the hypocotyl explants of C. frutescens was established. Results In this biolistic system study, explant target distance, bombardment helium (He) pressure and the size of microcarrier were the key parameters to be investigated. The optimized parameters based on screening of GFP expression were determined to be 6 cm target distance, 1350 psi of helium pressure and 1.6 μm of gold particle (microcarrier) size. The greatest number of shoots were obtained from hypocotyl as explant using Murashige and Skoog medium supplemented with 5.0 mg/L BAP and 0.1 mg/L NAA. An average of 5 shoots per explant were formed. Out of which, one shoot managed to form root and developed into whole plant. Conclusions We have obtained an optimized protocol for the biolistic transformation of chili and in vitro regeneration of chili plantlets. The establishment of the protocols will provide a platform for molecular breeding and biological studies of the chili plants. Elsevier 2018-05-18 Article PeerReviewed Chee, Marcus Jenn Yang, Lycett, Grantley W. and Chin, Chiew Foan (2018) Development of a direct transformation method by GFP screening and in vitro whole plant regeneration of Capsicum frutescens L. Electronic Journal of Biotechnology, 34 . pp. 51-58. ISSN 0717-3458 Biolistics; Capsaicin; Capsicum frutescens; Direct transformation; GFP; Gold particle; Metabolic engineering; Particle delivery system; Shoots; Solanaceae; Tissue culture https://www.sciencedirect.com/science/article/pii/S0717345818300228 doi:10.1016/j.ejbt.2018.05.005 doi:10.1016/j.ejbt.2018.05.005
spellingShingle Biolistics; Capsaicin; Capsicum frutescens; Direct transformation; GFP; Gold particle; Metabolic engineering; Particle delivery system; Shoots; Solanaceae; Tissue culture
Chee, Marcus Jenn Yang
Lycett, Grantley W.
Chin, Chiew Foan
Development of a direct transformation method by GFP screening and in vitro whole plant regeneration of Capsicum frutescens L.
title Development of a direct transformation method by GFP screening and in vitro whole plant regeneration of Capsicum frutescens L.
title_full Development of a direct transformation method by GFP screening and in vitro whole plant regeneration of Capsicum frutescens L.
title_fullStr Development of a direct transformation method by GFP screening and in vitro whole plant regeneration of Capsicum frutescens L.
title_full_unstemmed Development of a direct transformation method by GFP screening and in vitro whole plant regeneration of Capsicum frutescens L.
title_short Development of a direct transformation method by GFP screening and in vitro whole plant regeneration of Capsicum frutescens L.
title_sort development of a direct transformation method by gfp screening and in vitro whole plant regeneration of capsicum frutescens l.
topic Biolistics; Capsaicin; Capsicum frutescens; Direct transformation; GFP; Gold particle; Metabolic engineering; Particle delivery system; Shoots; Solanaceae; Tissue culture
url https://eprints.nottingham.ac.uk/51963/
https://eprints.nottingham.ac.uk/51963/
https://eprints.nottingham.ac.uk/51963/