Modified TP-M13 Automated Fluorescent-labelled SSR Method and Its Application in Identification of Pecan Cultivars
-
摘要: 用TP-M13自动荧光法检测了常用10个美国山核桃品种中的8个SSR位点,发现原有直接使用三引物方法扩增的有效性和可靠性较低,而经过改进的间接三引物法则有较大的实用价值,在此基础上进一步研究后认为混合三引物法在适当控制混合引物的对数和提高特异性后,在提高间接三引物法的扩增效率方面是可行的。Abstract: Although TP-M13 automated fluorescent-labelled system has reduced the cost of fluorescent detection of SSR genotyping by introducing a universal fluorescent labelled primer, the complexity of PCR reaction brought by three primers limits its application on more species. TP-M13 method using three primers directly was implemented to detect eight SSR loci of regular 10 pecan (Carya illinoensis ) cultivars, which revealed the low availability and liability. A modified TP-M13 detection method called indirect three primer method was proved to be effective and valuable in solving this problem. Furthermore, the mixing of SSR primers based on this indirect three primer method could further improve the PCR efficiency
-
Keywords:
- automated fluorescent-labelled system /
- TP-M13 /
- Carya illinoensis /
- SSR
-
-
[1] Li Y C, Korol A B, Fahima T, et al. Microsatellites: genomic distribution, putative functions and mutational mechanisms: a review[J]. Mol Ecol, 2002, 11(12):2 453-2 465.
[2] Litt M, Luty J A. A hypervariable microsatellite revealed by in vitro amplification of a dinucleotide repeat within the cardiac muscle actin gene[J]. Am J Hum Genet, 1989, 44(3):397401.
[3] Tautz D. Hypervariability of simple sequences as a general source for polymorphic DNA markers[J]. Nucl Acids Res, 1989, 17(16):64 63-6 471.
[4] Kresovich S, Szewc-Mcfadden A K, et al. Abundance and characterization of simple-sequence repeats (SSRs) isolated from a size-fractionated genomic library of Brassica napus L. (rapeseed)[J]. Theor Appl Genet, 1995, 91(2):206211.
[5] 郝晨阳,王兰芬,贾继增,等. SSR荧光标记和银染技术的比较分析[J]. 作物学报, 2005, 31(2):144-149. [6] Oetting W S, Lee H K, Flanders D J, et al. Linkage analysis with multiplexed short tandem repeat polymorphisms using infrared fluorescence and M13 tailed primers[J]. Genomics, 1995, 30(3):450458.
[7] Schuelke M. An economic method for the fluorescent labeling of PCR fragments[J]. Nat Biotechnol, 2000, 18(2):233234.
[8] Jia H M, Shen Y T, Jiao Y, et al. Development of 107 SSR markers from whole genome shotgun sequences of Chinese bayberry (Myrica rubra) and their application in seedling identification[J]. J Zhejiang Univ Sci B, 2014, 15(11):9971005.
[9] Yue X Y, Liu G Q, Zong Y, et al. Development of genic SSR markers from transcriptome sequencing of pear buds[J]. J Zhejiang Univ Sci B, 2014, 15(4):303312.
[10] 高源,王昆,田路明,等. TP-M13-SSR技术及其在苹果种质资源遗传多样性研究中的应用[J]. 植物遗传资源学报,2011,12(2):228-233. [11] Liu Zhi-Zhai, Guo Rong-hua, Zhao jiu-ran, et al. Genetic Diversity of Two Important Groups of Maize Landraces with Same Name in China Revealed by M13 Tailed-Primer SSRs[J]. Agric Sci Chin, 2009, 8(1):15-23.
[12] Barkley N A, Dean R E, Pittman R N, et al. Genetic diversity of cultivated and wild-type peanuts evaluated with M13-tailed SSR markers and sequencing[J]. Genet Res, 2007, 89(2):93106.
[13] 刘志斋,王天宇,黎裕. TP—M13-SSR技术及其在玉米遗传多样性研究中的应用[J]. 玉米科学,2007,15(6):10-15. [14] 李会勇,王天宇,黎裕,等. TP-M13自动荧光检测法在高梁SSR基因型鉴定中的应用[J]. 植物遗传资源学报,2005,6(1):68-70. [15] 张日清,李江,吕芳德,等. 我国引种美国山核桃历程及资源现状研究[J]. 经济林研究,2003,21(4):107-109. [16] 彭华正. 竹子分生组织发育特点以及相关基因的克隆和功能研究[D]. 杭州:浙江大学,2005. [17] Grauke L J, Iqbal M J, Reddy A S, et al. Developing Microsatellite DNA Markers in Pecan[J]. J. Am. Soc. Hor. Sci, 2003, 128(3):374380.
[18] Grauke L J, Price H J, Johnston J S. Genome Size of Pecan as Determined by Flow Cytometry[J]. HortScience, 2001, 36(4):814.
计量
- 文章访问数: 535
- HTML全文浏览量: 0
- PDF下载量: 303