Functional Analysis of Pheophorbide a Oxygenase Gene and Its Promoter from Mango (Mangifera indica) Fruit

Pheophorbide a oxygenase (PAO) is a key enzyme in the process of chlorophyll degradation. It can metabolize a produced in the process of chlorophyll degradation into red chlorophyll metabolite. The full-length cDNA sequence of PAO gene was cloned from “Gui fei” mango peels, the relative expression o...

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Main Authors: Beibei Bai, Kuanliang Liu, Zhichang Zhao, Ruixiong Luo, Aiping Gao, Huangtuan, Wang Zisong
Format: Article
Language:English
Published: Taylor & Francis Group 2025-12-01
Series:International Journal of Fruit Science
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Online Access:https://www.tandfonline.com/doi/10.1080/15538362.2025.2519120
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author Beibei Bai
Kuanliang Liu
Zhichang Zhao
Ruixiong Luo
Aiping Gao
Huangtuan
Wang Zisong
author_facet Beibei Bai
Kuanliang Liu
Zhichang Zhao
Ruixiong Luo
Aiping Gao
Huangtuan
Wang Zisong
author_sort Beibei Bai
collection DOAJ
description Pheophorbide a oxygenase (PAO) is a key enzyme in the process of chlorophyll degradation. It can metabolize a produced in the process of chlorophyll degradation into red chlorophyll metabolite. The full-length cDNA sequence of PAO gene was cloned from “Gui fei” mango peels, the relative expression of three cultivars by RT-PCR was the highest in the green “Gui qi,” followed by the red “Gui fei” and the lowest in the yellow “Jin huang.” The analysis of carotenoid content in transgenic tomato fruit shows that overexpression of PAO can increase the content of β-carotene and inhibition of PAO gene expression can reduce the content of β-carotenein. Fragments of the 2.2 kb upstream region of the PAO gene were fused to the pGreenII 0800-Luc vector. We examined tobacco (Nicotiana tabacum) leaves carrying a series of nested PAO promoter – FLuc fusion for FLuc/RLuc activity. The region of −1218 to −3 in PAO promoter showed high activities in the three parts. Three 5’deletion analysis of the PAO promoter identified regions related to IAA-inducible and GA3-inducible expression as well as regions containing positive and negative elements. At low concentration, IAA was mainly negative regulation, GA3 was mainly positive regulation, and at high concentration, the results were opposite by measuring FLuc/RLuc activity. The study of PAO gene and promoter can provide some reference for us to regulate fruit color, especially the effect of the application of growth regulators on fruit color in mango production and cultivation.
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issn 1553-8362
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language English
publishDate 2025-12-01
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series International Journal of Fruit Science
spelling doaj-art-e2e8ec83e2ce4854a2f391a56b5b6f762025-08-20T03:23:11ZengTaylor & Francis GroupInternational Journal of Fruit Science1553-83621553-86212025-12-01251526110.1080/15538362.2025.2519120Functional Analysis of Pheophorbide a Oxygenase Gene and Its Promoter from Mango (Mangifera indica) FruitBeibei Bai0Kuanliang Liu1Zhichang Zhao2Ruixiong Luo3Aiping Gao4Huangtuan5Wang Zisong6Guangxi Key Laboratory of Biology for Mango, Baise University, Baise, Guangxi, ChinaTropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan, ChinaGuangxi Key Laboratory of Biology for Mango, Baise University, Baise, Guangxi, ChinaTropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan, ChinaGuangxi Key Laboratory of Biology for Mango, Baise University, Baise, Guangxi, ChinaGuangxi Key Laboratory of Biology for Mango, Baise University, Baise, Guangxi, ChinaGuangxi Key Laboratory of Biology for Mango, Baise University, Baise, Guangxi, ChinaPheophorbide a oxygenase (PAO) is a key enzyme in the process of chlorophyll degradation. It can metabolize a produced in the process of chlorophyll degradation into red chlorophyll metabolite. The full-length cDNA sequence of PAO gene was cloned from “Gui fei” mango peels, the relative expression of three cultivars by RT-PCR was the highest in the green “Gui qi,” followed by the red “Gui fei” and the lowest in the yellow “Jin huang.” The analysis of carotenoid content in transgenic tomato fruit shows that overexpression of PAO can increase the content of β-carotene and inhibition of PAO gene expression can reduce the content of β-carotenein. Fragments of the 2.2 kb upstream region of the PAO gene were fused to the pGreenII 0800-Luc vector. We examined tobacco (Nicotiana tabacum) leaves carrying a series of nested PAO promoter – FLuc fusion for FLuc/RLuc activity. The region of −1218 to −3 in PAO promoter showed high activities in the three parts. Three 5’deletion analysis of the PAO promoter identified regions related to IAA-inducible and GA3-inducible expression as well as regions containing positive and negative elements. At low concentration, IAA was mainly negative regulation, GA3 was mainly positive regulation, and at high concentration, the results were opposite by measuring FLuc/RLuc activity. The study of PAO gene and promoter can provide some reference for us to regulate fruit color, especially the effect of the application of growth regulators on fruit color in mango production and cultivation.https://www.tandfonline.com/doi/10.1080/15538362.2025.2519120MangoPAOchlorophyll degradationpromotercontent of β-carotene
spellingShingle Beibei Bai
Kuanliang Liu
Zhichang Zhao
Ruixiong Luo
Aiping Gao
Huangtuan
Wang Zisong
Functional Analysis of Pheophorbide a Oxygenase Gene and Its Promoter from Mango (Mangifera indica) Fruit
International Journal of Fruit Science
Mango
PAO
chlorophyll degradation
promoter
content of β-carotene
title Functional Analysis of Pheophorbide a Oxygenase Gene and Its Promoter from Mango (Mangifera indica) Fruit
title_full Functional Analysis of Pheophorbide a Oxygenase Gene and Its Promoter from Mango (Mangifera indica) Fruit
title_fullStr Functional Analysis of Pheophorbide a Oxygenase Gene and Its Promoter from Mango (Mangifera indica) Fruit
title_full_unstemmed Functional Analysis of Pheophorbide a Oxygenase Gene and Its Promoter from Mango (Mangifera indica) Fruit
title_short Functional Analysis of Pheophorbide a Oxygenase Gene and Its Promoter from Mango (Mangifera indica) Fruit
title_sort functional analysis of pheophorbide a oxygenase gene and its promoter from mango mangifera indica fruit
topic Mango
PAO
chlorophyll degradation
promoter
content of β-carotene
url https://www.tandfonline.com/doi/10.1080/15538362.2025.2519120
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