CHEN Yun,GAO Yuanqing,WANG Jilian,et al.Effect of Different Preservation Methods on Genomic DNA Extraction of Pomegranate[J].Northern Horticulture,2016,40(08):100-103.[doi:10.11937/bfyy.201608028]
不同保存方法对石榴成熟叶片基因组DNA提取效果的影响
- Title:
- Effect of Different Preservation Methods on Genomic DNA Extraction of Pomegranate
- Keywords:
- Punica granatum L.; preserved methods; DNA extraction
- 文献标志码:
- A
- 摘要:
- 以石榴（Punica granatum L.）成熟叶片为试材，设置6种保存方法，采用改良的CTAB法提取石榴基因组DNA，研究了不同保存方法对石榴叶片基因组DNA提取效果的影响。结果表明：采用改良的CTAB法，新鲜叶片、-70℃保存6个月、4℃保存5 d的样品均能得到浓度较高的DNA；-20℃保存7 d和14 d的样品均能得到完整DNA，但浓度略低；硅胶干燥常温保存5个月的样品用同样的方法未能提取到DNA，在对提取方法进行优化后，可提取纯度较高的基因组DNA，但其浓度仅约为鲜叶的50%。上述方法保存的样品提取的基因组DNA均能得到清晰、稳定的SRAP-PCR 扩增图谱。
- Abstract:
- Taking leaves of Punica granatum L.as materials，six methods of preserving fresh leaves were designed，genomic DNA was extracted by improved CTAB method，the effect of different preserving methods on genomic DNA extraction was studied.The results showed that the DNA extracted from leaves stored about 6 months at the -70℃ and 5 days at 4℃ could get the same high quality comparing with the fresh leaves.Genomic DNA also could be extracted from sample stored 7 and 14 days at -20℃，but the concentration was slightly lower than that extracted from fresh leaves.However，DNA from silica gel-dried sample was not obtained by the same way.After optimization of the DNA extraction method，the high-purity DNA from silica gel-dried sample was got,but the concentration of DNA were about 50% comparing with fresh leaves.Genomic DNA extracted from samples preserved as the above-mentioned six methods could meet the demands for SRAP amplification.Clear and perfect SRAP amplification patterns were obtained.
参考文献/References:
[1]董周永,郭松年,赵国建,等.石榴果皮提取物抑菌活性研究[J].西北植物学报,2008,28(3):582-587. [2]郑菊映,卢成瑜,杨权生,等.番石榴叶治疗小儿轮状病毒肠炎的临床疗效观察[J].现代中西医结合杂志,2010,19(2):140-141. [3]杨建宇,陈韵,郑春兰,等.石榴的生物活性及其抗炎和抗肿瘤作用[J].云南大学学报 (自然科学版),2008,30(S1):430-438. [4]王婧茹,赵晶晶,叶春玲,等.番石榴叶总三萜对2型糖尿病大鼠的降血糖和血脂作用[J].中国病理生理杂志,2012,28(6)：1109-1113. [5]冯立娟,陶吉寒,尹燕雷,等.石榴功能物质鞣花酸研究进展[J].食品科学,2014，35(23):325-330. [6]王富荣,赵剑波,章镇,等.适于AFLP分析用的桃成熟叶片DNA提取方法[J].果树学报,2006（4）:638-641. [7]MOHAMMAD H S,MAJID T,BADRALDIN E S.Use of SRAP markers to assess genetic diversity and population structure of wild，cultivated，and ornamental pomegranates(Punica granatum L.) in different regions of Iran[J].Plant Syst Evol,2012,298:1141-1149. [8]LI G，QUIROS C F.Sequence-related amplified polymorphism(SRAP)，a new marker system based on a simple PCR reaction:its application to mapping and gene tagging in Brassica[J].Theor Appl Genet，2001,103:455-461. [9]胡翔宇,丁琼,宋希强,等.2种保存方法对五唇兰基因组DNA提取质量的影响[J].热带作物学报,2014,35(8):1546-1550. [10]李志真,谢一青,黄儒珠,等.不同保存方法对光皮桦总DNA提取效果的影响[J].分子植物育种,2006,4(1):131-134. [11]靖相密,褚云霞,汤庚国,等.不同保存方法对蜡梅总DNA提取效果的影响及ISSR-PCR验证[J].分子植物育种,2008,6(2):387-392. [12]吕杰,马媛,金湘,等.硅胶干燥胡杨叶片DNA提取与RAPD反应体系的建立[J].北方园艺,2011(14):131-134. [13]李莉,彭建营,白瑞霞.不同方法对枣叶片总DNA 提取效果的影响[J].果树学报,2007,24(3):389-392.
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备注/Memo
第一作者简介：陈芸（1980-），女，浙江天台人，硕士，讲师，现主要从事植物分子遗传等研究工作。E-mail：chenyun8111@126.com. 责任作者：马刘峰（1979-），女，安徽蚌埠人，博士，副教授，现主要从事植物基因工程等研究工作。E-mail:maliufeng@126.com. 基金项目：新疆维吾尔自治区高校科研计划资助项目（XJEDU2013S35）；喀什大学校内课题资助项目（（11）2411）。