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| Reference Gene Selection for Quantitative Real-time PCR(qRT-PCR) in Anthurium (Anthurium andraeanum Linden) Tissues Under Bacterial Blight Stress |
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Abstract Bacterial blight, caused by Xanthomonas axonopodis pv. dieffenbachiae (Xad), is the most destructive disease in anthurium (Anthurium andraeanum Linden). Appropriate reference gene selection is essential for accurate genes expression quantification about blight response by using quantitative Real-time PCR (qRT-PCR), and to elucidate the phytopathology mechanism of anthurium-bacteria interaction. In this study, 6 commonly used candidates including elongation factor1-α (EF1α), glyceraldehyde-3-phosphate dehydrogenase (GADPH), ubiquitin 7(UBQ7), histidine 3 (His3), α-tubulin (TUB) and β-Actin (ACTB), were employed to identify the most reliable reference genes. The genes amplification efficiency was assessed by qRT-PCR, and expression stability in the processes of foliage and systemic infection were analyzed on both resistant and susceptible cultivars, then generated dates were evaluated comprehensively using geNorm3.5, BestKeeper0.953 and NormFinder1.0 programs. As a result, the tested genes showed different expression characters in various samples, among which, the expression of UBQ7 was much more stable in different tissues and different varieties, could be recommended as reference gene for data normalization. The result will facilitate expression analysis of anthurium genes responding to blight.
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Received: 25 December 2014
Published: 09 July 2015
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