NDLI: Reference gene identification for real-time PCR analyses in soybean leaves under fungus (Cercospora kikuchii) infection and treatments with salicylic and jasmonic acids

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Reference gene identification for real-time PCR analyses in soybean leaves under fungus (Cercospora kikuchii) infection and treatments with salicylic and jasmonic acids

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Australasian Plant Pathology : Volume 45, Issue 1, February 2016

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Reference gene identification for real-time PCR analyses in soybean leaves under fungus (Cercospora kikuchii) infection and treatments with salicylic and jasmonic acids

Content Provider	Springer Nature Link
Author	Costa, Jose H. Saraiva, Katia D. C. Morais, Vanessa D. Oliveira, Jose T. A. Sousa, Daniele O. B. Melo, Dirce Fernandes Morais, Janne K. S. Vasconcelos, Ilka M.
Copyright Year	2016
Abstract	Quantitative PCR (qPCR) is a robust technology for comparing the expression profiles of target genes, however the data produced need normalization with appropriate reference genes. The purpose of this study was to evaluate the expression stability of eight candidate reference genes (EF1A 1a1, EF1A 2a, EF1A 2b, EF1B, ACT11, UKN 1, ACT and SKIP 16) for qPCR assays in soybean leaves under fungus (Cercospora kikuchii) infection and treatments with salicylic and jasmonic acids. Four programs, GeNorm, NormFinder, BestKeeper, and RefFinder were used to evaluate the expression stability. The leaves were treated according the following conditions: 1) infected with Cercospora kikuchii (CK); (2) treated with salicylic acid (SA); (3) treated with SA and infected with C. kikuchii; (4) treated with jasmonic acid (JA); and (5) treated with JA and infected with C. kikuchii. For all studied conditions, GeNorm analyses revealed that combinations of six genes were always needed for gene expression normalization. Three EF1A genes (EF1A 2a, EF1A 1a1 and EF1A 2b) were the most stable in all tested conditions and then, they were always included in gene combinations. The other three genes varied according the different conditions. In analyses with other programs, at least two EF1A genes were often ranked among the three best stable genes. The expression of a PR3 (class I chitinase) gene was used to validate the reference genes across the total samples. Our results provide a shortlist of reference genes to normalize qPCR assays in soybean under CK infection and treatments with SA and JA.
Starting Page	191
Ending Page	199
Page Count	9
File Format	PDF
ISSN	08153191
Journal	Australasian Plant Pathology
Volume Number	45
Issue Number	2
e-ISSN	14486032
Language	English
Publisher	Springer Netherlands
Publisher Date	2016-02-11
Publisher Institution	Australasian Plant Pathology Society
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Glycine max Cercospora kikuchii Jasmonic acid Salicylic acid Gene expression qPCR Plant Pathology Plant Sciences Agriculture Entomology Ecology
Content Type	Text
Resource Type	Article
Subject	Plant Science

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Reference gene identification for real-time PCR analyses in soybean leaves under fungus (Cercospora kikuchii) infection and treatments with salicylic and jasmonic acids

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Reference gene identification for real-time PCR analyses in soybean leaves under fungus (Cercospora kikuchii) infection and treatments with salicylic and jasmonic acids