Crc1,6RhaT is involved in the synthesis of hesperidin of the main bioactive substance in the Citrus reticulata ‘Chachi’ fruit

Hesperidin is a dihydroflavonoids, accounting for more than 50% of the total flavonoids in Citri Reticulatae Pericarpium (CRP) of traditional Chinese medicine. It is an effective antioxidant and free radical scavenger that has anti-inflammatory, antiviral and hypoglycemic properties. The latest stud...

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Main Authors: Na Shang, Panpan Tong, Peng Ye, Jianmu Su, Mei Bai, Hong Wu
Format: Article
Language:English
Published: KeAi Communications Co., Ltd. 2024-11-01
Series:Horticultural Plant Journal
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Online Access:http://www.sciencedirect.com/science/article/pii/S2468014123001164
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author Na Shang
Panpan Tong
Peng Ye
Jianmu Su
Mei Bai
Hong Wu
author_facet Na Shang
Panpan Tong
Peng Ye
Jianmu Su
Mei Bai
Hong Wu
author_sort Na Shang
collection DOAJ
description Hesperidin is a dihydroflavonoids, accounting for more than 50% of the total flavonoids in Citri Reticulatae Pericarpium (CRP) of traditional Chinese medicine. It is an effective antioxidant and free radical scavenger that has anti-inflammatory, antiviral and hypoglycemic properties. The latest studies reported that hesperidin has a potential for novel coronavirus resistance. However, little is known about the synthesis regulation and accumulation site of hesperidin in plants. In this study, hesperidin synthase gene Crc1,6RhaT was cloned, and the protein can be completely transformed flavanone-7-O-glucoside into hesperidin in vitro and in vivo. Studies on biological characteristics of ovary walls and exocarps showed that the relative expression levels of the Crc1,6RhaT gene and protein decreased gradually with the development of citrus fruits, and the relative content of hesperidin firstly increased, then sequentially decreased. In situ hybridization results further revealed that Crc1,6RhaT transcription was mainly concentrated in the secretory cavity cells, which are revealed to be the site of flavonoid synthesis. Immunocytochemistry localization results showed that the Crc1,6RhaT was mainly located in the endoplasmic reticulum, nucleus and vacuole of secretory cells. We inferred that the Crc1,6RhaT was synthesized in the endoplasmic reticulum, then was transported into the vacuoles through enlarged vesicles at the end of the endoplasmic reticulum. Our results not only revealed that Crc1,6RhaT may be involved in the synthesis of hesperidin of the main bioactive substance in the medicinal plant Citrus reticulata ‘Chachi’ fruit, but also provided novel insights into the main subcellular sites of hesperidin biosynthesis in vacuoles.
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spelling doaj-art-f861d50dc7024fa09442d30ad28fa62e2025-08-20T02:11:38ZengKeAi Communications Co., Ltd.Horticultural Plant Journal2468-01412024-11-011061321133510.1016/j.hpj.2022.10.012Crc1,6RhaT is involved in the synthesis of hesperidin of the main bioactive substance in the Citrus reticulata ‘Chachi’ fruitNa Shang0Panpan Tong1Peng Ye2Jianmu Su3Mei Bai4Hong Wu5State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangdong Laboratory for Lingnan Modern Agriculture, College of Life Sciences, South China Agricultural University, Guangzhou, Guangdong 510642, ChinaState Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangdong Laboratory for Lingnan Modern Agriculture, College of Life Sciences, South China Agricultural University, Guangzhou, Guangdong 510642, ChinaState Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangdong Laboratory for Lingnan Modern Agriculture, College of Life Sciences, South China Agricultural University, Guangzhou, Guangdong 510642, ChinaState Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangdong Laboratory for Lingnan Modern Agriculture, College of Life Sciences, South China Agricultural University, Guangzhou, Guangdong 510642, ChinaState Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangdong Laboratory for Lingnan Modern Agriculture, College of Life Sciences, South China Agricultural University, Guangzhou, Guangdong 510642, China; Corresponding authors.State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangdong Laboratory for Lingnan Modern Agriculture, College of Life Sciences, South China Agricultural University, Guangzhou, Guangdong 510642, China; Guangdong Technology Research Center for Traditional Chinese Veterinary Medicine and Natural Medicine, South China Agricultural University, Guangzhou, Guangdong 510642, China; Corresponding authors.Hesperidin is a dihydroflavonoids, accounting for more than 50% of the total flavonoids in Citri Reticulatae Pericarpium (CRP) of traditional Chinese medicine. It is an effective antioxidant and free radical scavenger that has anti-inflammatory, antiviral and hypoglycemic properties. The latest studies reported that hesperidin has a potential for novel coronavirus resistance. However, little is known about the synthesis regulation and accumulation site of hesperidin in plants. In this study, hesperidin synthase gene Crc1,6RhaT was cloned, and the protein can be completely transformed flavanone-7-O-glucoside into hesperidin in vitro and in vivo. Studies on biological characteristics of ovary walls and exocarps showed that the relative expression levels of the Crc1,6RhaT gene and protein decreased gradually with the development of citrus fruits, and the relative content of hesperidin firstly increased, then sequentially decreased. In situ hybridization results further revealed that Crc1,6RhaT transcription was mainly concentrated in the secretory cavity cells, which are revealed to be the site of flavonoid synthesis. Immunocytochemistry localization results showed that the Crc1,6RhaT was mainly located in the endoplasmic reticulum, nucleus and vacuole of secretory cells. We inferred that the Crc1,6RhaT was synthesized in the endoplasmic reticulum, then was transported into the vacuoles through enlarged vesicles at the end of the endoplasmic reticulum. Our results not only revealed that Crc1,6RhaT may be involved in the synthesis of hesperidin of the main bioactive substance in the medicinal plant Citrus reticulata ‘Chachi’ fruit, but also provided novel insights into the main subcellular sites of hesperidin biosynthesis in vacuoles.http://www.sciencedirect.com/science/article/pii/S2468014123001164Citri Reticulatae PericarpiumCitrusHesperidinFlavonoidsCrc1,6RhaTSecretory cavity
spellingShingle Na Shang
Panpan Tong
Peng Ye
Jianmu Su
Mei Bai
Hong Wu
Crc1,6RhaT is involved in the synthesis of hesperidin of the main bioactive substance in the Citrus reticulata ‘Chachi’ fruit
Horticultural Plant Journal
Citri Reticulatae Pericarpium
Citrus
Hesperidin
Flavonoids
Crc1,6RhaT
Secretory cavity
title Crc1,6RhaT is involved in the synthesis of hesperidin of the main bioactive substance in the Citrus reticulata ‘Chachi’ fruit
title_full Crc1,6RhaT is involved in the synthesis of hesperidin of the main bioactive substance in the Citrus reticulata ‘Chachi’ fruit
title_fullStr Crc1,6RhaT is involved in the synthesis of hesperidin of the main bioactive substance in the Citrus reticulata ‘Chachi’ fruit
title_full_unstemmed Crc1,6RhaT is involved in the synthesis of hesperidin of the main bioactive substance in the Citrus reticulata ‘Chachi’ fruit
title_short Crc1,6RhaT is involved in the synthesis of hesperidin of the main bioactive substance in the Citrus reticulata ‘Chachi’ fruit
title_sort crc1 6rhat is involved in the synthesis of hesperidin of the main bioactive substance in the citrus reticulata chachi fruit
topic Citri Reticulatae Pericarpium
Citrus
Hesperidin
Flavonoids
Crc1,6RhaT
Secretory cavity
url http://www.sciencedirect.com/science/article/pii/S2468014123001164
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