| Stipa grandis is a typical grassland species in Central Asia. With the drought stress treatment to Stipa grandis seedlings in greenhouse, the study as follows:According to the published sequences of nine candidate reference genes:the Stipa grandis actin (Act2), microtubule associated protein a (TUB-a), microtubule associated protein B (TUB-b),18S ribosomal RNA (18S rRNA), glyceraldehyde-3phosphate dehydrogenase (GAPDH), eukaryotic elongation factor (EF-1a), RNA polymerase II (RNA POL Ⅱ), adenine phosphoribosyl converting enzyme (APRT), translation factor (TLF), primers were designed for qRT-PCR. The stable value of nine candidate genes were determined by comparing the Ct value with using geNorm and NormFinder software. The results showed that the most stable reference gene expressed in Stipa grandis roots under drought stress is18S rRNA,followed by EF-1a and TLF, the most unstable reference gene is RNA POL Ⅱ; the most stable combination of reference genes is18S rRNA and EF-la. The order of reference gene stability sequence expressed in the leaves was18S rRNA,TLF and EF-la, the most unstable reference gene is TUB-b;the most stable combination of reference genes is18S rRNA and TLF.The expression of water channel protein gene PIP2;1and PIP2;2in Stipa grandis under drought stress was researched by using qRT-PCR method, and the design of primers referred to conserved region of PIP2;1and PIP2;2gene previous cloned sequences. The result showed that under drought stress, the expression of PIP2;1and PIP2;2of Stipa grandis in roots were much higher than in leaves, and expression of two genes increased with increasing. The PIP2;1gene expression was up to the highest level on the20d after drought stress, was about2.2times of the control, and then decreased; PIP2;2had the highest expression level on the15d after drought stress, is about2.5times of the control, then decreased. It implied that the PIP2;1and PIP2;2gene were involved in the physiological process of water transport in Stipa grandis. |
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