Identification of Differentially Expressed Genes in Sweetpotato Storage Roots Between Kokei No. 14 and Its Mutant Nongdafu 14 Using PCR-Based cDNA Subtraction

doi:10.1016/S2095-3119(13)60276-2

Journal of Integrative Agriculture ›› 2013, Vol. 12 ›› Issue (4): 589-595.DOI: 10.1016/S2095-3119(13)60276-2

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

收稿日期:2011-12-15 出版日期:2013-04-01 发布日期:2013-04-07
通讯作者: Correspondence LIU Qing-chang, Tel/Fax: +86-10-62733710, E-mail: liuqc@cau.edu.cn
基金资助:
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).

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

Received:2011-12-15 Online:2013-04-01 Published:2013-04-07
Contact: Correspondence LIU Qing-chang, Tel/Fax: +86-10-62733710, E-mail: liuqc@cau.edu.cn
Supported by:
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).

摘要/Abstract

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.

关键词: Ipomoea batatas (L.) Lam. , carotenoids , suppression subtractive hybridization , differentially expressed ESTs

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.

Key words: Ipomoea batatas (L.) Lam. , carotenoids , suppression subtractive hybridization , differentially expressed ESTs

CHEN Wei, ZHAI Hong, YANG Yuan-jun, HE Shao-zhen, LIU De-gao , LIU Qing-chang. Identification of Differentially Expressed Genes in Sweetpotato Storage Roots Between Kokei No. 14 and Its Mutant Nongdafu 14 Using PCR-Based cDNA Subtraction[J]. Journal of Integrative Agriculture, 2013, 12(4): 589-595.

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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

Identification of Differentially Expressed Genes in Sweetpotato Storage Roots Between Kokei No. 14 and Its Mutant Nongdafu 14 Using PCR-Based cDNA Subtraction

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