摘要
以欧亚种酿酒葡萄品种赤霞珠（Vitis viniferaL.cv.Cabernet Sauvignon）的胚性愈伤组织（EC）和非胚性愈伤组织（NEC）为材料,进行了总RNA不同提取方法的比较.结果表明葡萄EC和NEC的总RNA的最佳提取方法是RNAp lant试剂法.通过此方法提取的总RNA完整性好,28S和18S条带清晰,且OD260/280 nm值都在1.8~2.0之间.以此RNA为模板,对actin,tubulin,CLV等基因进行了半定量RT-PCR对比,发现EC与NEC在不同基因表达上存在差异.以上结果说明,以RNAp lant试剂提取的RNA质量,可以满足cDNA合成和qRT-PCR等研究用途.这为深入研究葡萄花药愈伤组织胚性与非胚性细胞系的基因表达特点,以及葡萄体细胞胚发育过程中的基因时序表达奠定了基础.
The quality of total RNA extracted by different methods from embryogenic callus （EC） and non - embryogenic callus （NEC） of Vitis vinifera L. cv. Cabernet Sauvignon was compared. The result demonstrated that ＂RNAplant＂ was a suitable reagent for high quality total RNA extraction from grape EC and NEC. The OD260/280nm of total RNA was between 1.8 and 2.0, and the 28S and 18S bands were clear on the agarose gel after electrophoresis. The expression of a set of genes including actin, tubulin, CLA and other three genes in EC and NEC were successfully compared by semi - quantitative RT - PCR with the total RNA as templates. The results showed that the total RNA extracted with this protocol was qualified for cDNA and qRT - PCR use, and could be further applied in gene expression and regulation research of grapevine somatic embryogenesis.
出处
《云南大学学报（自然科学版）》
CAS
CSCD
北大核心
2009年第4期410-415,共6页
Journal of Yunnan University(Natural Sciences Edition)
基金
国家自然科学基金项目资助(30500347&39970469)
云南省省院省校科技合作项目资助(2005WX16)
