Construction of Overexpression Vector with <i>TYR7523</i> Gene and Its Effect on Browning in <i>Macrocybe gigantea</i>

<i>Macrocybe gigantea</i> is a rare high-temperature edible fungus known for its resistance to browning. Previous studies suggested that the anti-browning property of the SCAU4 strain might be associated with low expression levels of the <i>TYR7523</i> gene. In this study, an...

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Main Authors: Jinyun Gao, Shuqing Song, Xinqian Liu, Zhuanlin Mo, Meihua Mo
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Journal of Fungi
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Online Access:https://www.mdpi.com/2309-608X/11/3/216
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author Jinyun Gao
Shuqing Song
Xinqian Liu
Zhuanlin Mo
Meihua Mo
author_facet Jinyun Gao
Shuqing Song
Xinqian Liu
Zhuanlin Mo
Meihua Mo
author_sort Jinyun Gao
collection DOAJ
description <i>Macrocybe gigantea</i> is a rare high-temperature edible fungus known for its resistance to browning. Previous studies suggested that the anti-browning property of the SCAU4 strain might be associated with low expression levels of the <i>TYR7523</i> gene. In this study, an overexpression vector for the <i>TYR7523</i> gene was constructed and introduced into SCAU4 mycelium using an Agrobacterium-mediated transformation method. After three rounds of hygromycin resistance screening, successful transformants were identified through PCR amplification and validated by qRT-PCR analysis, confirming a 3.47-fold upregulation of <i>TYR7523</i> expression. The overexpression strain <i>OE7523</i> was compared with the wild-type SCAU4 strain in terms of growth rate, browning degree, and tyrosinase activity. Although there was no significant difference in growth rate on the mother culture medium, <i>OE7523</i> showed faster growth on the stock culture and mycelium culture medium. In the late storage period, <i>OE7523</i> exhibited a higher browning degree and tyrosinase activity than SCAU4, suggesting a potential role of <i>TYR7523</i> in fruiting body browning. Physiological analyses indicated that low <i>TYR7523</i> expression may contribute to storage tolerance, while high expression influenced postharvest browning and preservation duration. The results provide data support for further study on the function of <i>TYR7523</i> gene of <i>Macrocybe gigantea</i>.
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spelling doaj-art-a520dcfb537748e68c8ba67c90c4d9cd2025-08-20T03:43:16ZengMDPI AGJournal of Fungi2309-608X2025-03-0111321610.3390/jof11030216Construction of Overexpression Vector with <i>TYR7523</i> Gene and Its Effect on Browning in <i>Macrocybe gigantea</i>Jinyun Gao0Shuqing Song1Xinqian Liu2Zhuanlin Mo3Meihua Mo4College of Food Science, South China Agricultural University, Guangzhou 510642, ChinaCollege of Food Science, South China Agricultural University, Guangzhou 510642, ChinaCollege of Food Science, South China Agricultural University, Guangzhou 510642, ChinaCenter for Basic Experiment and Practical Training, South China Agricultural University, Guangzhou 510642, ChinaCollege of Food Science, South China Agricultural University, Guangzhou 510642, China<i>Macrocybe gigantea</i> is a rare high-temperature edible fungus known for its resistance to browning. Previous studies suggested that the anti-browning property of the SCAU4 strain might be associated with low expression levels of the <i>TYR7523</i> gene. In this study, an overexpression vector for the <i>TYR7523</i> gene was constructed and introduced into SCAU4 mycelium using an Agrobacterium-mediated transformation method. After three rounds of hygromycin resistance screening, successful transformants were identified through PCR amplification and validated by qRT-PCR analysis, confirming a 3.47-fold upregulation of <i>TYR7523</i> expression. The overexpression strain <i>OE7523</i> was compared with the wild-type SCAU4 strain in terms of growth rate, browning degree, and tyrosinase activity. Although there was no significant difference in growth rate on the mother culture medium, <i>OE7523</i> showed faster growth on the stock culture and mycelium culture medium. In the late storage period, <i>OE7523</i> exhibited a higher browning degree and tyrosinase activity than SCAU4, suggesting a potential role of <i>TYR7523</i> in fruiting body browning. Physiological analyses indicated that low <i>TYR7523</i> expression may contribute to storage tolerance, while high expression influenced postharvest browning and preservation duration. The results provide data support for further study on the function of <i>TYR7523</i> gene of <i>Macrocybe gigantea</i>.https://www.mdpi.com/2309-608X/11/3/216<i>Macrocybe gigantea</i>tyrosinase geneoverexpressionbrowning
spellingShingle Jinyun Gao
Shuqing Song
Xinqian Liu
Zhuanlin Mo
Meihua Mo
Construction of Overexpression Vector with <i>TYR7523</i> Gene and Its Effect on Browning in <i>Macrocybe gigantea</i>
Journal of Fungi
<i>Macrocybe gigantea</i>
tyrosinase gene
overexpression
browning
title Construction of Overexpression Vector with <i>TYR7523</i> Gene and Its Effect on Browning in <i>Macrocybe gigantea</i>
title_full Construction of Overexpression Vector with <i>TYR7523</i> Gene and Its Effect on Browning in <i>Macrocybe gigantea</i>
title_fullStr Construction of Overexpression Vector with <i>TYR7523</i> Gene and Its Effect on Browning in <i>Macrocybe gigantea</i>
title_full_unstemmed Construction of Overexpression Vector with <i>TYR7523</i> Gene and Its Effect on Browning in <i>Macrocybe gigantea</i>
title_short Construction of Overexpression Vector with <i>TYR7523</i> Gene and Its Effect on Browning in <i>Macrocybe gigantea</i>
title_sort construction of overexpression vector with i tyr7523 i gene and its effect on browning in i macrocybe gigantea i
topic <i>Macrocybe gigantea</i>
tyrosinase gene
overexpression
browning
url https://www.mdpi.com/2309-608X/11/3/216
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