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Selection and Validation of Reference Genes for Gene Expression Studies by RT-PCR in Dalbergia odorifera
Perennial tree Dalbergia odorifera T. Chen could form the precious heartwood used to produce chinese traditional medicine, rosewood furniture and fragrances. However the formation of heartwood is time-consuming and low efficient, leading to the severe destruction of its wild resources. Thus, it is u...
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Published in: | Scientific reports 2019-03, Vol.9 (1), p.3341-3341, Article 3341 |
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Main Authors: | , , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
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Summary: | Perennial tree
Dalbergia odorifera
T. Chen could form the precious heartwood used to produce chinese traditional medicine, rosewood furniture and fragrances. However the formation of heartwood is time-consuming and low efficient, leading to the severe destruction of its wild resources. Thus, it is urgent to study the molecular mechanism of heartwood formation in
D
.
odorifera
. But till now, there is no report about the reference gene selection in this species. In this study, the expression stability of nine candidate reference genes were evaluated across different tissues and stems treated by wound and chemical stimulators. Four algorithms were applied to obtain the robust genes. The results support
HIS2
,
GAPDH
, and
CYP
to be the most stable reference genes in samples under different wound treatments while
DNAj
was the least stable. In different tissues,
HIS2
,
UBQ
, and
RPL
were the most stable reference genes while
DNAj
was the least stable. The selected reference genes were validated through the normalization of the qRT-PCR data of six heartwood related genes in terpene biosynthesis pathway and ethylene signal pathway. The results showed that their expression levels were accurate when they were normalized by the most stable reference gene
HIS2
, or by the combination of the two or three most stable reference genes. These results demonstrated that these selected reference genes are reliable. |
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ISSN: | 2045-2322 2045-2322 |
DOI: | 10.1038/s41598-019-39088-3 |