An efficient protocol for DNA isolation from the genus Pyrus

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Research Paper 01/04/2013
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An efficient protocol for DNA isolation from the genus Pyrus

Mohammad Islam, Habib Ahmad, Imtiaz Ahmad Khan
Int. J. Biosci.3( 4), 122-127, April 2013.
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Abstract

This paper communicates an economical, handy, quicker and reliable protocol of DNA isolation from different parts e.g. herbarium specimens, bark, wood, leaves etc. of pears. The protocol is recommended under the normal lab conditions without using any sophisticated equipment and costly kits/chemicals. Best quality of DNA was extracted from any part of the plant. The PCR product was amplified for 18S rRNA genes of various Pyrus genotypes. The genes were cleaned with gel elution Kit (K0513, Fermentas) and sequenced accordingly. The sequence data obtained was BLAST against the entire available nucleotide database using online tool at NCBI access and the land races were successfully discriminated. The protocol presented here is recommended for reliable extraction of best quality and high yield of DNA from woody plants.

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Aganga AA, Tshwenyane SO.  2003. Feeding values –anti nutritive factors of forage tree legumes. Pakistan Journal of Nutrition 2, 170-173. http://dx.doi.org/jn.2003.170

Barzegari A, Vahed SZ, Atashpaz S, Khani S, Omidi Y. 2010. Rapid and simple methodololgy for isolation of high quality  genomic  DNA  from coniferous tissues( Taxus baccata). Molecular Biology Reporter 833-837. http://dx.doi.org/10.1007/s1133-009-9634-z

Bell RL. 1990. Pears (Pyrus). In: J N Moore and J R Ballington Jr. (Eds.), Genetics Resources of Temperate Fruit and Nut Crops I. Pp. 655-697. International Society for Horticultural Science, Wageningen, The Netherlands.

Bell RL, Quamme HA, Layne REC, Skirvin RM. 1996. Pears. In: J.Janick and J.N. Moore (Eds.), Fruits Breeding, Vol. I. Tree and Tropical Fruits p. 441-514. John Wiley and Sons Inc.

Chevereau E, Leuliette S, Gallet M.  1997. Inheritance  and  Linkage  of  isozyme  Loci  in  Pear (Pyrus communis L.).Theoritical & Applied Genetics 94, 498-506. http://dx.doi.org/10.1007/s00122-012-2016-2

Do N, Adams RP. 1991. A simple technique of removing plant polysaccharides contaminants from DNA. Biotechnique 10, 162-166.

Doyle JJ. 1991.  DNA Protocols for plants. Pp. 283-293 in: G. Hewitt, A.W.B. Johnson, and J. P. W. Young (eds.), Molecular Techniques in taxonomy. NATO ASI Series H, Cell Biology 57. http://dx.doi.org/10.1083/jcb

Frankhan R, Ballou GD, Briscoe DA and Michlnnes KH. 2004. A primer of Conservation Genetics.Cambridge University Press Publisher. ISN. 978-0-511-18661-5.

Hussain F, Islam M, Zaman A. 2006. Ethnobotanical Profile of plants of Shawar valley, District Swat, Pakistan. International Journal of Biology & Biotechnology 3, 301-307.

Islam M, Ahmad H. 2012. Morphological Evaluation of Pyrus genotypes of Kaghan Valley, Pakistan through quantitative parameters. International Journal of Biosciences 2, 1-6.

Jang JT, Tanab K, Tamura F, Banno K. 1991. Identification of Pyrus species by peroxidase isozyme phenotypes of flower buds. Journal of Japan Societies’ for Horticulture Sciences 60, 513-519.

John, ME. 1992. Nucleic Acids Res. 20: 3281. MEDLINE Abstract.

Kim CS, Lee CH, Han GP, Ryu DH, Lee KH. 2000. SCARS Markers derived from RAPD for cultivar identification in Pyrus pyrifolia. J. Korcoci Horticulture Sciences 41, 125-128. http://dx.doi.org/10.1007/s00122-003-1582-8

Kim SC, Lee CH, Shin  JS, Chung YS,  Hyung NI. 1990. A  simple and rapid method for isolation of high  quality genomic DNA from fruit trees and conifers using PVP. Nuclic Acid Research 25, 1085-1086. http://dx.doi.org/10.1093/nar/25.5.1085

Kumar A, Pushpangadan P, Mehrotra S. 2003. Extraction of high molecular DNA from dry root tissues of Berberis lyceum suitable for RAPD. Plant Molecular Reporter 21, 309a-309d. http://dx.doi.org/.1007/BF02772807

Li JF, Li L,    Sheen J. 2010. Protocol: a rapid and economical procedure for purification of plasmid or plant DNA with diverse applications in plant biology. Plant Methods, 6, p.1. http://dx.doi.org/10.1186/1746-4811-6-1

Peng S, Iwahori S. 2000. Variety, distribution and major cultivars of domesticated Pyrus spp. in China. Agriculture Horticulture 75, 763-772.

Saghai-Maroof M, Solman A, Jorgensen KM, Allard RW. 1984. Ribosomal DNA apacer-length polymorphisms in Barley: Mendelian inheritance, chromosomal location and population dynamics. Proceeding of National A c a d e m y o f Sciences. USA., 81, 8014-8018.

Scott KD, Playford J. 1996. DNA extraction technique for PCR in rain forest plant species. Biotechniques 20, 974- 978.

Shepherd MC, McLay TGB. 2011. Two microscale protocols for the isolation of DNA from polysaccharide- rich plant tissues. Journal of Plant Researc, 124, 311-314. http://dx.doi.org/10.1007/s10265-010-0379-5

Shepherd MC, Stockoe LR,  Scott LJ, Jones ME. 2002. High throughput DNA extraction from forest trees. Plant Molecular Biology Reporter 20, 425a-524j. http://dx.doi.org/10.1007/BF02772134

Smyth, RP. 2010. Reducing chimera formation during PCR amplification to ensure accurate genotyping. Genetics, 469, 45-51. http://dx.doi.org/10.1016/j.gene.2010.08.009

Teng Y, Tanaba K. 2004. Reconsideration on the origin of cultivated pears native to East Asia. Acta Horticulture, 634, 175-182.

Verma A, Padh H, Shrivastav N. 2007. Plant genomic DNA isolation: an art or a science. Biotechnology Journal 2, 386-392. http://dx.doi.org/10.1002/biot.200600195

Yamamoto T. 2003. Parentage analysis in pear cultivars characterized by SSR markers. Journal of Japan Society of Horticultural Sciences, 72, 182-189.