Identification and Validation of Reference Genes in the Adzuki Bean ( Vigna angularis ) Under Iron Deficiency Using Quan
- PDF / 2,233,729 Bytes
- 12 Pages / 595.276 x 790.866 pts Page_size
- 112 Downloads / 185 Views
ORIGINAL PAPER
Identification and Validation of Reference Genes in the Adzuki Bean (Vigna angularis) Under Iron Deficiency Using Quantitative Real-Time PCR Junshuai Wang 1 & Kai Yang 1 & Bo Zhao 1 & Yisong Li 2 & Ping Wan 1
# Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Adzuki bean (Vigna angularis var. angularis), an important food legume, is grown in more than 30 countries, especially in East Asia. Almost 25% of the world population is affected by iron deficiency. Iron is very important on adzuki bean yield and quality. The identification of suitable reference genes in adzuki bean is important for exploring gene expression and the molecular mechanisms of gene, yet few reference genes have been reported for adzuki bean. This research systematically evaluated the stability of 11 candidate reference genes (ACT, ACT1, CYP, CYP2, SKIP16, ELF1B, HDC, Lectin, TUA, PEPKR1, and 18SrRNA) using the geNorm, NormFinder, BestKeeper, and RefFinder protocols under iron-deficient stress. The best combined number of reference genes for all samples was two. ELF1B/Lectin was best for qRT-PCR analysis of roots and combined analysis of roots, stems, and leaves under iron deficiency, ELF1B/CYP was optimum for leaves, and SKIP16/PEPKR1 was optimum for stems. The identification of these reference genes will enable accurate quantification of gene expression under iron deficiency and enhance our comprehension of reference genes in adzuki bean. Keywords Adzuki bean (Vigna angularis var. angularis) . Reference gene . Quantitative real-time PCR . Iron deficiency
Introduction Adzuki bean (Vigna angularis), in the family Phaseoleae, was domesticated in China about 12,000 years ago (Liu et al. 2013). It is an important grain legume in East Asia (Yamada et al. 2001). The adzuki bean is rich in protein, starch, iron (7.4 mg/100 g), and vitamins (Lin et al. 2005; Xu et al. 2008). Iron is an essential element for plant growth and development.
Higher soil pH of the major adzuki bean-planting region in China results in iron uptake barrier, and further induces chlorosis and a reduction in the yield of adzuki bean. Transcriptional profile analysis is widely used for exploring the molecular mechanisms of responses to biotic and abiotic stresses in plants. Methods include real-time quantitative PCR (RT-qPCR), Northern blotting, ribonuclease protection assay, and gene chips. Of these, RT-qPCR has high sensitivity
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11105-019-01166-1) contains supplementary material, which is available to authorized users. * Yisong Li [email protected]
1
Key Laboratory of New Technology in Agricultural Application, College of Plant Science and Technology, Beijing University of Agriculture, Huilongguan Beinonglu 7, Changping District, Beijing 102206, China
2
Key Laboratory of Urban Agriculture (North) of Ministry of Agriculture, College of Biological Science and Engineering, Beijing University of Agriculture, Huilongguan Beinonglu 7, Changpin
Data Loading...
