Molecular Characterization and Expression Profiles of Myrosinase Gene (RsMyr2) in Radish (Raphanus sativus L.)
Myrosinase is a defense-related enzyme and is capable of hydrolyzing glucosinolates into a variety of compounds, some of which are toxic to pathogens and herbivores. Many studies revealed that a number of important vegetables or oil crops contain the myrosinase-glucosinolate system. However, the rel...
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KeAi Communications Co., Ltd.
2014-09-01
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| Series: | Journal of Integrative Agriculture |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2095311913606449 |
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| author | Yan PAN Yuan-yuan XU Xian-wen ZHU Zhe LIU Yi-qin GONG Liang XU Mao-yong GONG Li-wang LIU |
| author_facet | Yan PAN Yuan-yuan XU Xian-wen ZHU Zhe LIU Yi-qin GONG Liang XU Mao-yong GONG Li-wang LIU |
| author_sort | Yan PAN |
| collection | DOAJ |
| description | Myrosinase is a defense-related enzyme and is capable of hydrolyzing glucosinolates into a variety of compounds, some of which are toxic to pathogens and herbivores. Many studies revealed that a number of important vegetables or oil crops contain the myrosinase-glucosinolate system. However, the related promoter and genomic DNA sequences as well as expression profiles of myrosinase gene remain largely unexplored in radish (Raphanus sativus). In this study, the 2 798 bp genomic DNA sequence, designated as RsMyr2, was isolated and analyzed in radish. The RsMyr2 consisting of 12 exons and 11 introns reflected the common gene structure of myrosinases. Using the genomic DNA walking approach, the 5′-flanking region upstream of RsMyr2 with length of 1 711 bp was successfully isolated. PLACE and PlantCARE analyses revealed that this upstream region could be the promoter of RsMyr2, which contained several basic cis-regulatory elements including TATA-box, CAAT-box and regulatory motifs responsive to defense and stresses. Furthermore, recombinant pET-RsMyr2 protein separated by SDS-PAGE was identified as myrosinase with mass spectrometry. Real-time PCR analysis showed differential expression profiles of RsMyr2 in leaf, stem and root at different developmental stages (e.g., higher expression in leaf at cotyledon stage and lower in flesh root at mature stage). Additionally, the RsMyr2 gene exhibited up-regulated expression when treated with abscisic acid (ABA), methyl jasmonate (MeJA) and hydrogen peroxide (H2O2), whereas it was down-regulated by wounding (WO) treatment. The findings indicated that the expression of RsMyr2 gene was differentially regulated by these stress treatments. These results could provide new insight into elucidating the molecular characterization and biological function of myrosinase in radish. |
| format | Article |
| id | doaj-art-5d469209ea9948b6a4658edf431fe6e2 |
| institution | Kabale University |
| issn | 2095-3119 |
| language | English |
| publishDate | 2014-09-01 |
| publisher | KeAi Communications Co., Ltd. |
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| series | Journal of Integrative Agriculture |
| spelling | doaj-art-5d469209ea9948b6a4658edf431fe6e22025-08-20T03:56:18ZengKeAi Communications Co., Ltd.Journal of Integrative Agriculture2095-31192014-09-011391877188810.1016/S2095-3119(13)60644-9Molecular Characterization and Expression Profiles of Myrosinase Gene (RsMyr2) in Radish (Raphanus sativus L.)Yan PAN0Yuan-yuan XU1Xian-wen ZHU2Zhe LIU3Yi-qin GONG4Liang XU5Mao-yong GONG6Li-wang LIU7National Key Laboratory of Crop Genetics and Germplasm Enhancement/Engineering Research Center of Horticultural Crop Germplasm Enhancement and Utilization, Ministry of Education/Key Laboratory of Horticultural Crop Biology and Genetic Improvement (East China), Ministry of Agriculture/College of Horticulture, Nanjing Agricultural University, Nanjing 210095, P.R. China; PAN YanNational Key Laboratory of Crop Genetics and Germplasm Enhancement/Engineering Research Center of Horticultural Crop Germplasm Enhancement and Utilization, Ministry of Education/Key Laboratory of Horticultural Crop Biology and Genetic Improvement (East China), Ministry of Agriculture/College of Horticulture, Nanjing Agricultural University, Nanjing 210095, P.R. ChinaDepartment of Plant Sciences, North Dakota State University, Fargo, ND 58105, USANational Key Laboratory of Crop Genetics and Germplasm Enhancement/Engineering Research Center of Horticultural Crop Germplasm Enhancement and Utilization, Ministry of Education/Key Laboratory of Horticultural Crop Biology and Genetic Improvement (East China), Ministry of Agriculture/College of Horticulture, Nanjing Agricultural University, Nanjing 210095, P.R. ChinaNational Key Laboratory of Crop Genetics and Germplasm Enhancement/Engineering Research Center of Horticultural Crop Germplasm Enhancement and Utilization, Ministry of Education/Key Laboratory of Horticultural Crop Biology and Genetic Improvement (East China), Ministry of Agriculture/College of Horticulture, Nanjing Agricultural University, Nanjing 210095, P.R. ChinaNational Key Laboratory of Crop Genetics and Germplasm Enhancement/Engineering Research Center of Horticultural Crop Germplasm Enhancement and Utilization, Ministry of Education/Key Laboratory of Horticultural Crop Biology and Genetic Improvement (East China), Ministry of Agriculture/College of Horticulture, Nanjing Agricultural University, Nanjing 210095, P.R. ChinaNational Key Laboratory of Crop Genetics and Germplasm Enhancement/Engineering Research Center of Horticultural Crop Germplasm Enhancement and Utilization, Ministry of Education/Key Laboratory of Horticultural Crop Biology and Genetic Improvement (East China), Ministry of Agriculture/College of Horticulture, Nanjing Agricultural University, Nanjing 210095, P.R. ChinaNational Key Laboratory of Crop Genetics and Germplasm Enhancement/Engineering Research Center of Horticultural Crop Germplasm Enhancement and Utilization, Ministry of Education/Key Laboratory of Horticultural Crop Biology and Genetic Improvement (East China), Ministry of Agriculture/College of Horticulture, Nanjing Agricultural University, Nanjing 210095, P.R. China; Correspondence LIU Li-wang, Tel: +86-25-84395563, Fax: +86-25-84395266Myrosinase is a defense-related enzyme and is capable of hydrolyzing glucosinolates into a variety of compounds, some of which are toxic to pathogens and herbivores. Many studies revealed that a number of important vegetables or oil crops contain the myrosinase-glucosinolate system. However, the related promoter and genomic DNA sequences as well as expression profiles of myrosinase gene remain largely unexplored in radish (Raphanus sativus). In this study, the 2 798 bp genomic DNA sequence, designated as RsMyr2, was isolated and analyzed in radish. The RsMyr2 consisting of 12 exons and 11 introns reflected the common gene structure of myrosinases. Using the genomic DNA walking approach, the 5′-flanking region upstream of RsMyr2 with length of 1 711 bp was successfully isolated. PLACE and PlantCARE analyses revealed that this upstream region could be the promoter of RsMyr2, which contained several basic cis-regulatory elements including TATA-box, CAAT-box and regulatory motifs responsive to defense and stresses. Furthermore, recombinant pET-RsMyr2 protein separated by SDS-PAGE was identified as myrosinase with mass spectrometry. Real-time PCR analysis showed differential expression profiles of RsMyr2 in leaf, stem and root at different developmental stages (e.g., higher expression in leaf at cotyledon stage and lower in flesh root at mature stage). Additionally, the RsMyr2 gene exhibited up-regulated expression when treated with abscisic acid (ABA), methyl jasmonate (MeJA) and hydrogen peroxide (H2O2), whereas it was down-regulated by wounding (WO) treatment. The findings indicated that the expression of RsMyr2 gene was differentially regulated by these stress treatments. These results could provide new insight into elucidating the molecular characterization and biological function of myrosinase in radish.http://www.sciencedirect.com/science/article/pii/S2095311913606449radishRsMyr2promoterheterologous expressionqRT-RCR |
| spellingShingle | Yan PAN Yuan-yuan XU Xian-wen ZHU Zhe LIU Yi-qin GONG Liang XU Mao-yong GONG Li-wang LIU Molecular Characterization and Expression Profiles of Myrosinase Gene (RsMyr2) in Radish (Raphanus sativus L.) Journal of Integrative Agriculture radish RsMyr2 promoter heterologous expression qRT-RCR |
| title | Molecular Characterization and Expression Profiles of Myrosinase Gene (RsMyr2) in Radish (Raphanus sativus L.) |
| title_full | Molecular Characterization and Expression Profiles of Myrosinase Gene (RsMyr2) in Radish (Raphanus sativus L.) |
| title_fullStr | Molecular Characterization and Expression Profiles of Myrosinase Gene (RsMyr2) in Radish (Raphanus sativus L.) |
| title_full_unstemmed | Molecular Characterization and Expression Profiles of Myrosinase Gene (RsMyr2) in Radish (Raphanus sativus L.) |
| title_short | Molecular Characterization and Expression Profiles of Myrosinase Gene (RsMyr2) in Radish (Raphanus sativus L.) |
| title_sort | molecular characterization and expression profiles of myrosinase gene rsmyr2 in radish raphanus sativus l |
| topic | radish RsMyr2 promoter heterologous expression qRT-RCR |
| url | http://www.sciencedirect.com/science/article/pii/S2095311913606449 |
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