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Identification of Differentially Expressed Genes in Sweetpotato Storage Roots Between Kokei No. 14 and Its Mutant Nongdafu 14 Using PCR-Based cDNA Subtraction |
CHEN Wei, ZHAI Hong, YANG Yuan-jun, HE Shao-zhen, LIU De-gao , LIU Qing-chang |
Beijing Key Laboratory of Crop Genetic Improvement/Laboratory of Crop Heterosis and Utilization, Ministry of Education/China Agricultural University, Beijing 100193, P.R.China |
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摘要 The contents of carotenoids in the storage root of sweetpotato, Ipomoea batatas (L.) Lam. vary dramatically among different cultivars. However, so far little is known about the regulation of carotenoids synthesis in sweetpotato. In our laboratory, we identified a novel sweetpotato mutant, Nongdafu 14, which is a homogenous mutant derived from the wild type Kokei No. 14. The contents of carotenoids in the storage root of Nongdafu 14 were analyzed using high performance liquid chromatography (HPLC), and it was found that the amount of carotenoids, b-carotene, lutein and zeaxantion, three major types of carotenoids in sweetpotato storage roots, increased 2-26 folds in Nongdafu 14 compared to Kokei No. 14. Suppression subtractive hybridization (SSH) was used to identify genes that were differentially expressed in Nongdafu 14, and a differentially expressed cDNA library was constructed using the cDNA of Nongdafu 14 storage roots as tester and that of Kokei No. 14 storage roots as driver. Out of the 1 530 clones sequenced, we identified 292 nonredundant ESTs. GO and KEGG analyses of these differentially expressed ESTs indicated that diverse metabolism pathways were affected and candidate genes involved in regulation of carotenoids synthesis are suggested.
Abstract The contents of carotenoids in the storage root of sweetpotato, Ipomoea batatas (L.) Lam. vary dramatically among different cultivars. However, so far little is known about the regulation of carotenoids synthesis in sweetpotato. In our laboratory, we identified a novel sweetpotato mutant, Nongdafu 14, which is a homogenous mutant derived from the wild type Kokei No. 14. The contents of carotenoids in the storage root of Nongdafu 14 were analyzed using high performance liquid chromatography (HPLC), and it was found that the amount of carotenoids, b-carotene, lutein and zeaxantion, three major types of carotenoids in sweetpotato storage roots, increased 2-26 folds in Nongdafu 14 compared to Kokei No. 14. Suppression subtractive hybridization (SSH) was used to identify genes that were differentially expressed in Nongdafu 14, and a differentially expressed cDNA library was constructed using the cDNA of Nongdafu 14 storage roots as tester and that of Kokei No. 14 storage roots as driver. Out of the 1 530 clones sequenced, we identified 292 nonredundant ESTs. GO and KEGG analyses of these differentially expressed ESTs indicated that diverse metabolism pathways were affected and candidate genes involved in regulation of carotenoids synthesis are suggested.
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Received: 15 December 2011
Accepted:
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Fund: This work was supported by the China Agriculture Research System (CARS-11), the HarvestPlus Challenge Program, and the National High-Tech Research and Development Project of China (2011AA100607). |
Corresponding Authors:
Correspondence LIU Qing-chang, Tel/Fax: +86-10-62733710, E-mail: liuqc@cau.edu.cn
E-mail: liuqc@cau.edu.cn
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Cite this article:
CHEN Wei, ZHAI Hong, YANG Yuan-jun, HE Shao-zhen, LIU De-gao , LIU Qing-chang.
2013.
Identification of Differentially Expressed Genes in Sweetpotato Storage Roots Between Kokei No. 14 and Its Mutant Nongdafu 14 Using PCR-Based cDNA Subtraction. Journal of Integrative Agriculture, 12(4): 589-595.
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[1]Bartley G E, Scolnik P A. 1995. Plant carotenoids: pigments for photoprotection, visual attraction, and humanhealth. The Plant Cell, 7, 1027-1038[2]Bateman A. 2002. The SGS3 protein involved in PTGS findsa family. BMC Bioinformatics, 3, 21.Bouvier F, Suire C, d’Harlingue A, Backhaus R A, CamaraB. 2000. Molecular cloning of geranyl diphosphatesynthase and compartmentation of monoterpenesynthesis in plant cells. The Plant Journal, 24, 241-252[3]Cao W X, Epstein C, Liu H, DeLoughery C, Ge N X, Lin J Y,Diao R, Cao H, Long F, Zhang X, et al. 2004. Comparinggene discovery from Affymetrix GeneChip microarraysand Clontech PCR-select cDNA subtraction: a casestudy. BMC Genomics, 5, 26.[4]Christopher I, Cazzonelli C I, Pogson B J. 2010. Source tosink: regulation of carotenoid biosynthesis in plants.Trends of Plant Science, 15, 266-274[5]Cunningham F X, Gantt E. 1998. Genes and enzymes ofcarotenoid biosynthesis in plants. Annual Review ofPlant Physiology, 49, 557-583[6]Fraser P D, Bramley P M. 2004. The biosynthesis andnutritional uses of carotenoids. Progress in LipidResearch, 43, 228-265[7]Giuliano G, Tavazza R, Diretto G, Beyer P, Taylor M A.2008. Metabolic engineering of carotenoid biosynthesisin plants. Trends in Biotechnology, 26, 139-145[8]He S Z, Han Y F, Wang Y P, Zhai H, Liu Q C. 2009. In vitroselection and identification of sweetpotato (Ipomoeabatatas (L.) Lam.) plants tolerant to NaCl. Plant CellTissue and Organ Culture, 96, 69-74[9]Hirschberg J. 2001. Carotenoid biosynthesis in floweringplants. Current Opinion in Plant Biology, 4, 210-218[10]Kumakura N, Takeda A, Fujioka Y, Motose H, Takano R,Watanabe Y. 2009. SGS3 and RDR6 interact andcolocalize in cytoplasmic SGS3/RDR6-bodies. FEBSLetters, 583, 1261-1266[11]Liu Q C. 2011. Sweet potato omics and biotechnology inChina. Plant Omics Journal, 4, 295-301[12]Stigliani AL, Giorio G, D’Ambrosio C. 2011. Characterizationof P450 carotenoid beta- and epsilon-hydroxylases oftomato and transcriptional regulation of xanthophyllbiosynthesis in root, leaf, petal and fruit. Plant CellPhysiology, 52, 851-865[13]Tian L, Magallanes-Lundback M, Musetti V, DellaPennaD. 2003. Functional analysis of beta- and epsilon-ringcarotenoid hydroxylases in Arabidopsis. The PlantCell, 15, 1320-1332[14]Wang Y P, Wang F, Zhai H, Liu Q C. 2007. Production of auseful mutant by chronic irradiation in sweetpotato.Scientia Horticulturae, 111, 173-178[15]Zhang J Z, Li Z M, Liu L, Mei L, Yao J L, Hu C G. 2008.Identification of early-flower-related ESTs in an earlyfloweringmutant of trifoliate orange (Poncirustrifoliata) by suppression subtractive hybridization andmacroarray analysis. Tree Physiology, 28, 1449-1457 |
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