GUO Li-li,LI Jun,LI Jin-hang,et al.Extraction of Total RNA From Potted Tree Peony Leaves at Different Stages[J].Northern Horticulture,2014,38(12):99-102.
不同时期盆栽牡丹叶片RNA的提取研究
- Title:
- Extraction of Total RNA From Potted Tree Peony Leaves at Different Stages
- 文章编号:
- 1001-0009(2014)12-0099-04
- 关键词:
- 牡丹; CTAB-LiCl改良法; 总RNA提取
- 分类号:
- S 685. 11
- 文献标志码:
- A
- 摘要:
- 以牡丹品种‘洛阳红’为试材,采用CTAB-LiCl改良法,对小风铃期、大风铃期、圆桃期、平桃期、破绽期、初开期和谢花期的牡丹叶片进行总RNA提取。利用琼脂糖凝胶电泳检测RNA完整性,紫外分光光度计检测RNA纯度。结果表明:7个时期提取的总RNA经电泳均检测到28S、18S和5S rRNA条带,吸光度比值A260/A280均介于1.8~2.0之间,其中大风铃期、圆桃期、平桃期、破绽期和初开期的rRNA条带整齐清晰,并且28S rRNA亮度高于18S rRNA;上述表明CTAB-LiCl改良法适合从牡丹叶片中提取总RNA,且大风铃期、圆桃期、平桃期、破绽期和初开期是最适合提取总RNA的5个时期。
- Abstract:
- Taking Paeonia suffruticosa Andr.cv.‘Luoyanghong’ as test material,total RNA were extracted by CTAB-LiCl improved method from tree peony leaves at small bells stage,big bells stage,circular peach stage,flat peach stage,flaw stage,initial bloom stage and fading stage.RNA integrity was detected by agarose gel electrophoresis,RNA purity was detected by ultraviolet spectrophotometer.The results showed that after electrophoresis,the RNA extracted in seven stages all had three bands-28S,18S and 5S rRNA,the ratio of A260/A280 ranged between 1.8 and 2.0.The rRNA bands at big bells stage,circular peach stage,flat peach stage,flaw stage,and initial bloom stage were tidy and clear,and the lightness of 28S rRNA band was higher than 18S rRNA band.The above showed that CTAB-LiCl improved method was suitable for extraction of total RNA from tree peony leaves,and big bells stage,circular peach stage,flat peach stage,flaw stage and initial bloom stage were the optimal five stages for total RNA extraction.
参考文献/References:
[1] Chomczynski P,Sacchi N.Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction[J].Analytical Biochemistry,1987,162(1):156-159.
[2] Van Driessche E,Beeckmans S,Dejaegere R,et al.The antioxidant of choice for the purification of proteins from phenol-rich plant tissues[J].Analytical Biochemistry,1984,141(1):184-188.
[3] 侯小改,段春燕,刘素云,等.不同土壤水分条件下牡丹的生理特性研究[J].华北农学报,2007,22(3):80-83.
[4] 李永华,翟敏,李颖旭,等.干旱胁迫下牡丹叶片光合作用与抗氧化酶活性变化[J].河南农业科学,2007(5):91-93.
[5] 杜希华,陆海,高述民,等.文冠果花药总RNA提取方法研究[J].北京林业大学学报,2003,25(1):10-13.
[6] 张玉进,孟祥春,潘瑞炽,等.非洲菊花瓣总RNA提取方法的改进[J].植物学通报,2001,18(6):722-726.
[7] 王玉成,杨传平,姜静.木本植物组织总RNA提取的要点与原理[J].东北林业大学学报,2002,30(2):1-4.
[8] Chang S,Puryear J,Cairney J.A simple and efficient method for isolating RNA from pine trees[J].Plant Molecular Biology Reporter,1993,11(2):113-116.
[9] 张今今,王跃进,王西平,等.葡萄总RNA提取方法的研究[J].果树学报,2003,20(3):178-181.
[10] 赵双宜,吴耀荣,夏光敏.介绍一种简单高效的植物总RNA提取方法[J].遗传,2002,24(3):337-338.
[11] 李大力.一种从富含次生物质的植物中提取RNA的方法[J].南京理工大学学报,2001,25(5):547-549.
[12] 陈国梁,陈宗礼,孙萍,等.枣树幼叶总RNA提取方法的比较研究[J].北方园艺,2012(19):131-133.
[13] 高双成,施江,王世华,等.一种牡丹花瓣总RNA的提取方法[J].河南农业科学,2007(10):93-94.
[14] 郝福玲,刘雅莉,王跃进,等.百合花瓣总RNA提取方法的研究[J].西北植物学报,2005,25(6):1143-1147.
[15] 蒋建雄,张天真.利用CTAB/酸酚法提取棉花组织总RNA[J].棉花学报,2003,15(3):166-167.
[16] 黄凤兰,李长海,孙婷婷,等.芍药花瓣总RNA的提取[J].生物技术通讯,2005,16(3):282-284.
[17] 王玉成,薄海侠,杨传平.胡杨、柽柳总RNA提取方法的建立[J].东北林业大学学报,2003,31(5):99-100.
[18] 曾凡锁,南楠,詹亚光.富含多糖和次生代谢产物的白桦成熟叶中总RNA的提取[J].植物生理学通讯,2007,43(5):913-916.
[19] 夏海武,吕柳新,陈桂信,等.羊蹄甲果荚中RNA提取的新方法[J].分子生物育种,2006,4(1):147-149.
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备注/Memo
第一作者简介:郭丽丽(1982-),女,博士,讲师,研究方向为逆境生理与分子生物学。E-mail:guolili0928@126.com.
责任作者:孔祥生(1955-),男,硕士,教授,研究方向为植物生理生态。E-mail:kxsh55@163.com.
基金项目:国家自然科学基金资助项目(31200468)。
收稿日期:2014-01-21