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Validation of reference genes for normalization of gene expression by qRT-PCR in a resveratrol-producing entophytic fungus (Alternaria sp. MG1)

Overview of attention for article published in AMB Express, November 2016
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Title
Validation of reference genes for normalization of gene expression by qRT-PCR in a resveratrol-producing entophytic fungus (Alternaria sp. MG1)
Published in
AMB Express, November 2016
DOI 10.1186/s13568-016-0283-z
Pubmed ID
Authors

Jin-xin Che, Jin-xin Che, Jun-ling Shi, Yao Lu, Yan-lin Liu

Abstract

Alternaria sp. MG1, an endophytic fungus isolated from Vitis vinifera, can independently produce resveratrol, indicating that this species contains the key genes for resveratrol biosynthesis. Identification of these key genes is essential to understand the resveratrol biosynthesis pathway in this strain, which is currently unknown in microorganisms. qRT-PCR is an efficient and widely used method to identify the key genes related to unknown pathways at the level of gene expression. Verification of stable reference genes in this strain is essential for qRT-PCR data normalization, although results have been reported for other Alternaria sp. strains. In this study, nine candidate reference genes including TUBA, EF1, EF2, UBC, UFD, RPS5, RPS24, ACTB and 18S were evaluated for expression stability in a diverse set of six samples representing different growth periods. We compared cell culture conditions and an optimized condition for resveratrol production. The comparison of the results was performed using four statistical softwares. A combination of TUBA and EF1 was found to be suitable for normalization of Alternaria sp. MG1 in different developmental stages, and 18S was found to be the least stable. The reference genes verified in this study will facilitate further research to explore gene expression and molecular mechanisms as well as the improvement of secondary metabolite yields in Alternaria sp. MG1. To our knowledge, this is the first validation of reference genes in Alternaria with the capability to produce resveratrol. Additionally, these results provide useful guidelines for the selection of reference genes in other Alternaria species.

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Geographical breakdown

Country Count As %
Unknown 14 100%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 4 29%
Student > Master 2 14%
Student > Doctoral Student 2 14%
Professor 1 7%
Lecturer 1 7%
Other 1 7%
Unknown 3 21%
Readers by discipline Count As %
Agricultural and Biological Sciences 9 64%
Biochemistry, Genetics and Molecular Biology 3 21%
Unknown 2 14%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 09 November 2016.
All research outputs
#15,392,529
of 22,899,952 outputs
Outputs from AMB Express
#446
of 1,236 outputs
Outputs of similar age
#197,221
of 312,900 outputs
Outputs of similar age from AMB Express
#17
of 46 outputs
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