Identification of suitable reference genes in leaves and roots of rapeseed (Brassica napus L.) under different nutrient deficiencies
Identification of suitable reference genes in leaves and roots of rapeseed(Brassica napus L.)under different nutrient deficiencies作者机构：Oil Crops Research Institute Chinese Academy of Agricultural Sciences/Key Laboratory of Biology and Genetics Improvement of Oil Crops Ministry of Agriculture Wuhan 430062 P.R. China
出 版 物：《Journal of Integrative Agriculture》 (农业科学学报（英文版）)
年 卷 期：2017年第16卷第4期
页 面：809-819页
核心收录：
基 金：supported by the grants from the Agricultural Science and Technology Innovation Program,Chinese Academy of Agricultural Sciences(CAAS-ASTIP-2013OCRI) the Excellent Young Scientist Fund of Chinese Academy of Agricultural Sciences(1610172015004) an open project funded by State Key Laboratory for the Conservation and Utilization of Subtropical Agro-bioresources,China(SKLCUSA-b201403)
主 题：reference genes rapeseed (Brassica napus L.) nutrient deficiency leaves roots
摘 要：Nutrient deficiency stresses often occur simultaneously in soil. Thus, it＇s necessary to investigate the mechanisms underlying plant responses to multiple stresses through identification of some key stress-responsive genes. Quantitative real-time PCR （qRT-PCR） is essential for detecting the expression of the interested genes, of which the selection of suitable reference genes is a crucial step before qRT-PCR. To date, reliable reference genes to normalize qRT-PCR data under different nutrient deficiencies have not been reported in plants. In this study, expression of ten candidate reference genes was detected in leaves and roots of rapeseed （Brassica napus L.） after implementing different nutrient deficiencies for 14 days. These candidate genes, included two traditionally used reference genes and eight genes selected from an RNA- Seq dataset. Two software packages （GeNorm, NormFinder） were employed to evaluate candidate gene stability. Results showed that VHA-E1 was the highest-ranked gene in leaves of nutrient-deficient rapeseed, while VHA-G1 and UBC21 were most stable in nutrient-deficient roots. When rapeseed leaves and roots were combined, UBC21, HTB1, VHA-G1 and A CT7 were most stable among all samples. To evaluate the stabilities of the highest-ranked genes, the relative expression of two target genes, BnTrxl;1 and BnPhtl;3 Were further determined. The results showed that the relative expression of BnTrxl;1 depended on reference gene selection, suggesting that it＇s necessary to evaluate the stability of reference gene prior to qRT-PCR. This study provides suitable reference genes for gene expression analysis of rapeseed responses to different nutrient deficiencies, which is essential for elucidation of mechanisms underlying rapeseed responses to multiple nutrient deficiency stresses
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