红砂正常和脱水组织中总RNA提取的改进CTAB法

摘要/Abstract

摘要： 红砂属于柽柳科超旱生小灌木，是荒漠半荒漠地区植被中的优势种。为了研究促分裂原激活的蛋白激酶(MAPK)信号传导通路在这类植物抗旱中的作用，我们尝试了很多RNA提取的方法都没有获得完整的RNA。在脱水组织中由于多糖、多酚以及其他与核酸能结合或共沉淀的化合物的含量升高而更不容易得到高质量的RNA。以CTAB为基础的改进的RNA提取方法，能成功地从少量的红砂植物的正常组织和脱水组织中获得高质量的RNA。这种方法经济且操作方便，提取的RNA纯度和完整性都很好，可用于RT-PCR反应并成功克隆了550 bp 左右的MAPK基因片段。

关键词: 红砂, 脱水组织, RNA提取

Abstract: Reaumuria soongorica belongs to the family of the Tamaricaceae, a perennial super-xerophytic semi-shrub. It is a predominant specces in arid and semi-arid areas. In attempts to evaluate the mechanism of mitogen-activated protein kinases (MAPKs) signaling transduction of drought tolerance in these plants, we experienced that many published protocols for nucleic acid isolation failed to yield RNA of sufficient quality for analysis. It is harder in dehydrated tissues because of higher levels of accumulated polysaccharides, RNases and other compounds that bind and/or coprecipitate with RNA. Therefore, we have modified CTAB (hexadecyltrimethylammonium bromide)-based extraction protocols for isolating intact purified RNA from dehydrated and non-dehydrated organs as a routine isolation of high-quality RNA from small amounts of R. soongorica tissues. The methods developed are simple and effective. The RNA was of sufficient quality for use in a RT-PCR reaction that amplified an about 550 bp fragment of the R. soongorica MAPK gene.

Key words: Reaumuria soongorica, dehydrated tissues, RNA isolation

中图分类号:

Q946

刘玉冰;张腾国;安黎哲;王刚*. 红砂正常和脱水组织中总RNA提取的改进CTAB法[J]. 中国沙漠, 2006, 26(4): 600-603.

LIU Yu-bing;ZHANG Teng-guo;AN Li-zhe;WANG Gang. Isolation of RNA Using Modified Method of CTAB from Dehydrated and Nondehydrated Reaumuria soongorica Tissues[J]. JOURNAL OF DESERT RESEARCH, 2006, 26(4): 600-603.

参考文献

[1]刘家琼,邱明新,蒲锦春,等. 我国荒漠典型超旱生植物——红砂[J]. 植物学报,1982,24(5):485-488.
[2]马茂华,孔令韶. 新疆呼图壁绿洲外缘的琵琶柴生物生态学特性研究[J]. 植物生态学报, 1998, 22(3): 237-244.
[3]王彦荣,曾彦军,付华,等. 过牧及封育对红砂荒漠植被演替的影响[J].中国沙漠,2002, 22(4):321-327.
[4]崔大方. 中国琵琶柴属分类、分布、生态和形态解剖学的初步研究[J].干旱区研究,1988, 5(1): 65-69.
[5]宋耀选,周茂先,张小由,等. 额济纳绿洲主要植物的水势与环境因子的关系[J].中国沙漠,2005,25(4):496-499.
[6]李骁,王迎春,征荣.西鄂尔多斯地区强旱生小灌木水分参数的研究(Ⅰ)[J].中国沙漠,2005,25(4):581-586.
[7]陈拓,冯虎元,徐世建,等.荒漠植物叶片碳同位素组成及其水分利用效率[J].中国沙漠,2002,22(3):288-291.
[8]Stokes S B, Rugh C L, Kamalay J C. RNA and DNA isolation from recalcitrant plant tissues[J]. Biotechniques, 1990, 9: 268-272.
[9]Camacho-Villasana Y M, Ochoa-Alejo N, Walling L, et al. An Improved Method for Isolating RNA From Dehydrated and Nondehydrated Chili Pepper (Capsicum annuum L.) Plant Tissues[J]. Plant Molecular Biology Reporter, 2002, 20: 407-414.
[10]刘志学,陈妍珂. 沙漠植物DNA 快速提取方法[J]. 中国沙漠,1997,17(3):274-279.
[11]陶玲,刘志学,何兴东,等. 用于RAPD 分析的沙拐枣DNA 提取方法[J].中国沙漠, 2001, 21(3): 300-302.
[12]Chang S, Pryear J, Cairney J. A simple and efficient method for isolating RNA from pine trees[J]. Plant Molecular Biology Reporter, 1993, 11: 113-117.
[13]Bugos R C, Chiang V L, Zhang X H, et al. RNA isolation from plant tissues recalcitrant to extraction in guanidine[J]. Biotechniques, 1995, 19: 734-737.
[14]Claros M G, Canovas F M. Rapid high quality RNA preparation from Pine seedlings[J]. Plant Molecular Biology Reporter, 1998, 16: 9-18.
[15]李宏,王新力. 植物组织RNA 提取的难点及对策[J]. 生物技术通报, 1999, (1):36-39.