Reference gene selection for RT-qPCR analysis of Chrysanthemum lavandulifolium during its flowering stages

被引：39

作者：

Fu, Jianxin ^{[1
]}

Wang, Yi ^{[1
]}

Huang, He ^{[1
]}

Zhang, Chao ^{[1
]}

Dai, Silan ^{[1
]}

机构：

[1] Beijing Forestry Univ, Coll Landscape Architecture, Beijing 100083, Peoples R China

来源：

MOLECULAR BREEDING | 2013年 / 31卷 / 01期

基金：

中国国家自然科学基金;

关键词：

Reference genes; RT-qPCR; Chrysanthemum lavandulifolium; Flowering; GeNorm; REAL-TIME PCR; GENOME-WIDE IDENTIFICATION; INTERNAL CONTROL GENES; VALID REFERENCE GENES; EXPRESSION ANALYSIS; HOUSEKEEPING GENES; NORMALIZATION; LEAF;

D O I：

10.1007/s11032-012-9784-x

中图分类号：

S3 [农学（农艺学）];

学科分类号：

0901 ;

摘要：

Quantitative real-time RT-PCR (RT-qPCR) is a technology that can be used to analyze the abundance of gene expression. Reference genes, which are assumed to remain at constant levels in different tissues at various developmental stages and photoperiodic treatments, were selected to analyze the expression levels of flowering time genes and floral development genes. Using digital gene expression technology, nine reference genes with moderate expression in the leaves of Chrysanthemum lavandulifolium at the juvenile phase (CK1) and the squaring stage (W1) were selected as the candidate reference genes for further study. A total of 115 biological samples of C. lavandulifolium were analyzed, including different tissues under various developmental stages and leaves with varied photoperiodic treatments. The stability of the nine reference genes was slightly variable across the samples, but MTP, SKIP16 and PGK were the most stable genes overall. In addition, the relative expression level of ClFT in different tissues of plants with the competence to flower was analyzed to verify the reference genes selected in this study. These studies provide a guide for selecting reference genes for analyzing the expression pattern of flowering time genes and floral development genes in C. lavandulifolium.

引用

页码：205 / 215

页数：11

共 46 条

[1] Normalization of real-time quantitative reverse transcription-PCR data: A model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets [J].

Andersen, CL ;

Jensen, JL ;

Orntoft, TF .

CANCER RESEARCH, 2004, 64 (15) :5245-5250

[2] Enzyme co-localization in the pea leaf cytosol: 3-P-glycerate kinase, glyceraldehyde-3-P dehydrogenase, triose-P isomerase and aldolase [J].

Anderson, LE ;

Carol, AA .

PLANT SCIENCE, 2005, 169 (03) :620-628

[3] Identification and evaluation of new reference genes in Gossypium hirsutum for accurate normalization of real-time quantitative RT-PCR data [J].

Artico, Sinara ;

Nardeli, Sarah M. ;

Neto, Osmundo B. Oliveira ;

Grossi-de-Sa, Maria Fatima ;

Alves-Ferreira, Marcio .

BMC PLANT BIOLOGY, 2010, 10

[4] Validating internal controls for quantitative plant gene expression studies [J].

Brunner A.M. ;

Yakovlev I.A. ;

Strauss S.H. .

BMC Plant Biology, 4 (1)

[5] Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays [J].

Bustin, SA .

JOURNAL OF MOLECULAR ENDOCRINOLOGY, 2000, 25 (02) :169-193

[6]

Chen FH, 1957, CHRY DAHLIA, V7, P264

[7]

Chen JY, 1984, ACTA HORTIC, V167, P349

[8] Selection of candidate housekeeping controls in tomato plants using EST data [J].

Coker, JS ;

Davies, E .

BIOTECHNIQUES, 2003, 35 (04) :740-+

[9] Evaluation of coffee reference genes for relative expression studies by quantitative real-time RT-PCR [J].

Cruz, Fernanda ;

Kalaoun, Samara ;

Nobile, Paula ;

Colombo, Carlos ;

Almeida, Juliana ;

Barros, Leila M. G. ;

Romano, Eduardo ;

Grossi-de-Sa, Maria Fatima ;

Vaslin, Maite ;

Alves-Ferreira, Marcio .

MOLECULAR BREEDING, 2009, 23 (04) :607-616

[10] Genome-wide identification and testing of superior reference genes for transcript normalization in Arabidopsis [J].

Czechowski, T ;

Stitt, M ;

Altmann, T ;

Udvardi, MK ;

Scheible, WR .

PLANT PHYSIOLOGY, 2005, 139 (01) :5-17

← 1 2 3 4 5 →