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向日葵不是盐生作物,但具有较强的耐盐和耐旱能力。甜菜碱醛脱氢酶基因(BADH)是植物重要的抗逆基因之一。检测BADH基因表达有助于研究向日葵对逆境胁迫的响应机制。因此本文以真核延伸因子1A(eEF1A)为内参,采用SYBR Green I荧光染料法,建立向日葵BADH基因表达的定量PCR检测方法。BADH和eEF1A的扩增曲线基线平整、指数区明显,熔解曲线上仅显示单一特异峰,扩增效率分别为1.90和1.96,Cq值平均变异系数分别为0.51%和0.65%。这些数据表明检测方法特异性强、重复性好、结果可靠。应用建立的检测方法初步检测了盐和干旱胁迫对向日葵BADH基因表达的影响,结果显示BADH受盐和干旱胁迫的诱导表达。
Sunflower is not a salt crop, but has strong salt tolerance and drought tolerance. The betaine aldehyde dehydrogenase gene (BADH) is one of the important stress-resistant genes in plants. Detection of BADH gene expression helps to study the response mechanism of sunflower to stress. Therefore, eukaryotic elongation factor 1A (eEF1A) was used as an internal reference, and SYBR Green I fluorescent dye method was used to establish a quantitative PCR detection method for BADH gene expression in sunflower. The amplification curve of BADH and eEF1A showed a flat baseline, a significant exponential region, and a single specific peak on the melting curve. The amplification efficiencies were 1.90 and 1.96, respectively, and the average Cq values were 0.51% and 0.65%, respectively. These data indicate that the assay is specific, reproducible and reliable. The effects of salt and drought stress on the expression of BADH gene in sunflower were preliminarily detected by using the established detection method. The results showed that BADH was induced by salt and drought stress.