Isolation and Activity Analysis of Phytoene Synthase (<i>ClPsy1</i>) Gene Promoter of Canary-Yellow and Golden Flesh-Color Watermelon
Watermelon (<i>Citrullus lanatus</i>) is an economically important cucurbit crop. Its pulp is rich in antioxidant carotenoids, which confer a variety of flesh colors. <i>ClPsy1</i> (Phytoene Synthase) is the rate-limiting enzyme for carotenoid synthesis; however, the promoter...
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MDPI AG
2023-04-01
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| Series: | Agronomy |
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| Online Access: | https://www.mdpi.com/2073-4395/13/4/1080 |
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| author | Yue Cao Xufeng Fang Shi Liu Feishi Luan |
| author_facet | Yue Cao Xufeng Fang Shi Liu Feishi Luan |
| author_sort | Yue Cao |
| collection | DOAJ |
| description | Watermelon (<i>Citrullus lanatus</i>) is an economically important cucurbit crop. Its pulp is rich in antioxidant carotenoids, which confer a variety of flesh colors. <i>ClPsy1</i> (Phytoene Synthase) is the rate-limiting enzyme for carotenoid synthesis; however, the promoter activity of <i>ClPsy1</i> is still unknown. In the present study, promoter sequences were isolated from four watermelon accessions: Cream of Saskatchewan pale yellow (COS), canary yellow flesh (PI 635597), golden flesh (PI 192938), and red flesh (LSW-177), all of which express <i>ClPsy1</i> at extremely high levels. Sequence alignment and cis-element analysis disclosed six SNPs between the four lines all in COS, two of which (at the 598th and 1257th positions) caused MYC and MYB cis-element binding sequence variations, respectively. To confirm <i>ClPsy1</i> gene promoter activity, full-length and deletion fragments of the promoter were constructed and connected to a β-D-glucosidase (GUS) vector and transferred into tomato fruits. GUS staining was performed to analyze the key segment of the promoter. The activity of the PI 192938 <i>ClPsy1</i> full-length promoter exceeded that of COS. The deletion fragment from −1521 bp to −1043 bp exhibited strong promoter activity, and contained a MYB transcription factor-binding site mutation. We combined RNA-seq with qRT-PCR to analyze the gene expression pattern between the MYB transcription factor <i>Cla97C10G196920</i> and <i>ClPsy1</i> gene and found that <i>Cla97C10G196920</i> (ClMYB21) showed the same expression trend with <i>ClPsy1</i>, which positively regulates carotenoid synthesis and metabolism. |
| first_indexed | 2024-03-11T05:19:55Z |
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| last_indexed | 2024-03-11T05:19:55Z |
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| series | Agronomy |
| spelling | doaj.art-6777a5a3edf444aaa3c7ec2b967777042023-11-17T17:57:18ZengMDPI AGAgronomy2073-43952023-04-01134108010.3390/agronomy13041080Isolation and Activity Analysis of Phytoene Synthase (<i>ClPsy1</i>) Gene Promoter of Canary-Yellow and Golden Flesh-Color WatermelonYue Cao0Xufeng Fang1Shi Liu2Feishi Luan3Key Laboratory of Biology and Genetic Improvement of Horticulture Crops (Northeast Region), Ministry of Agriculture and Rural Affairs, Harbin 150030, ChinaKey Laboratory of Biology and Genetic Improvement of Horticulture Crops (Northeast Region), Ministry of Agriculture and Rural Affairs, Harbin 150030, ChinaKey Laboratory of Biology and Genetic Improvement of Horticulture Crops (Northeast Region), Ministry of Agriculture and Rural Affairs, Harbin 150030, ChinaKey Laboratory of Biology and Genetic Improvement of Horticulture Crops (Northeast Region), Ministry of Agriculture and Rural Affairs, Harbin 150030, ChinaWatermelon (<i>Citrullus lanatus</i>) is an economically important cucurbit crop. Its pulp is rich in antioxidant carotenoids, which confer a variety of flesh colors. <i>ClPsy1</i> (Phytoene Synthase) is the rate-limiting enzyme for carotenoid synthesis; however, the promoter activity of <i>ClPsy1</i> is still unknown. In the present study, promoter sequences were isolated from four watermelon accessions: Cream of Saskatchewan pale yellow (COS), canary yellow flesh (PI 635597), golden flesh (PI 192938), and red flesh (LSW-177), all of which express <i>ClPsy1</i> at extremely high levels. Sequence alignment and cis-element analysis disclosed six SNPs between the four lines all in COS, two of which (at the 598th and 1257th positions) caused MYC and MYB cis-element binding sequence variations, respectively. To confirm <i>ClPsy1</i> gene promoter activity, full-length and deletion fragments of the promoter were constructed and connected to a β-D-glucosidase (GUS) vector and transferred into tomato fruits. GUS staining was performed to analyze the key segment of the promoter. The activity of the PI 192938 <i>ClPsy1</i> full-length promoter exceeded that of COS. The deletion fragment from −1521 bp to −1043 bp exhibited strong promoter activity, and contained a MYB transcription factor-binding site mutation. We combined RNA-seq with qRT-PCR to analyze the gene expression pattern between the MYB transcription factor <i>Cla97C10G196920</i> and <i>ClPsy1</i> gene and found that <i>Cla97C10G196920</i> (ClMYB21) showed the same expression trend with <i>ClPsy1</i>, which positively regulates carotenoid synthesis and metabolism.https://www.mdpi.com/2073-4395/13/4/1080cis-elementMYB transcription factorphytoene synthasepromoterwatermelon |
| spellingShingle | Yue Cao Xufeng Fang Shi Liu Feishi Luan Isolation and Activity Analysis of Phytoene Synthase (<i>ClPsy1</i>) Gene Promoter of Canary-Yellow and Golden Flesh-Color Watermelon Agronomy cis-element MYB transcription factor phytoene synthase promoter watermelon |
| title | Isolation and Activity Analysis of Phytoene Synthase (<i>ClPsy1</i>) Gene Promoter of Canary-Yellow and Golden Flesh-Color Watermelon |
| title_full | Isolation and Activity Analysis of Phytoene Synthase (<i>ClPsy1</i>) Gene Promoter of Canary-Yellow and Golden Flesh-Color Watermelon |
| title_fullStr | Isolation and Activity Analysis of Phytoene Synthase (<i>ClPsy1</i>) Gene Promoter of Canary-Yellow and Golden Flesh-Color Watermelon |
| title_full_unstemmed | Isolation and Activity Analysis of Phytoene Synthase (<i>ClPsy1</i>) Gene Promoter of Canary-Yellow and Golden Flesh-Color Watermelon |
| title_short | Isolation and Activity Analysis of Phytoene Synthase (<i>ClPsy1</i>) Gene Promoter of Canary-Yellow and Golden Flesh-Color Watermelon |
| title_sort | isolation and activity analysis of phytoene synthase i clpsy1 i gene promoter of canary yellow and golden flesh color watermelon |
| topic | cis-element MYB transcription factor phytoene synthase promoter watermelon |
| url | https://www.mdpi.com/2073-4395/13/4/1080 |
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