Shuang YANG, Muhammad IRFAN, Xue Lİ, Yan Bin LİU, Xing Jia SUN

Mısırda Bt geninin ve tohum saflığının tayini

Sunulan bu çalışmada, mısırda Bt geninin ve tohum saflığının PCR temelli yöntemle tayini sunulmuştur. Elli dört farklı mısır türü, 1950 bazçifti ve 301 bazçifti boyutuna sahip BT1 ve 35S primerleri kullanılarak Bt geninin tayini gerçekleştirilmiştir. Örnekler arasındaki onbeş örnek Bt geni içermektedir. Bt geni içermesi daha sonraki üreme işlemlerinde kullanılamayacağı anlamına gelmektedir. Doksanaltı mısır tohumu, bnIg 161 SSR primeri ile saflık kontrolu için kullanılmıştır. Sonuçlar, bu tohumların % 5 safsızlık ile % 95 saflıkta olduğunu göstermiştir. Ayrıca, bu sonuçlar PCR temelli işaretçilerin kültür tanımlanmasında çok faydalı olduğunu ve mısır yetiştirme programlarında iyileştirmeye olanak sağlayacağını göstermektedir

Anahtar Kelimeler:

Bt geni, tohum saflığı, PCR, mısır

Detection of Bt gene and seed purity in maize

In this study we have described the detection of Bt gene and seed purity in maize by using PCR based method. Fifty four maize varieties were used for the detection of Bt gene by using BT1 and 35S primer having size of 195bp and 301bp respectively. Of these, fifteen varieties contained Bt gene which means that these varieties can not be used for further breeding. Ninety six seeds of maize were used for purity check by bnlg 161 SSR primer. Results showed that these seeds were 95% pure with 5% impurity. So these results showed that PCR based markers were very helpful in cultivar identification which leads to the improvement in maize breeding programs.

Keywords:

Bt gene, seed purity, PCR, maize,

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M. Querci, M. Jeremini, V. G. den Eede. The analysis of food samples for the presence of genetically modified organisms. User Manual. JRC. Available from: URL: http://gmotraining.jrc.it/. (2006)
Z. Sieradzki, M. Mazur, K. Kwiatek. Validation of procedures based on PCR reactions for detection and identification of genetically modified maize & soybean. Bull. Vet. Inst. Pula., 52 (2008) 611-614.
C. Tengel, P. Schusler, E. Setzke, J. Balles, M. Sprenges- Hausels. PCR- based detection of genetically modified soy bean & maize in raw & highly processed food stuffs. Biotechnology 31(2001) 426-429.
ISAAA 2011. International Service for the Acquisition of Agri- biotech Application. Available from: URL: http://www.isaaa.org.
N. Bohorova, W. Zhang, P. Julstrum, S. Mclean, B. Luna, R.M. Brito, L. Diaz, M. E. Ramos, P. Estanol, M. Pacheco, M. Salgado, D. Hoisington. Production of transgenic tropical maize with cry1Ab and cry1Ac genes via microprojectile bombardment of immature embryos. Theor. Appl. Genet., 99 (1999) 437-444.
D. Bradley, M. A. Harkey, M. K. Kim, K. D. Biever, L. S. Bauer. The insecticidal Cry IB crystal protein of Bacillus thuringiensis ssp. thuringiensis has dual specificity to coleopteran and lepidopteran larvae. J Invert Path., 65 (1995) 162–173.
A. Germini, A. Zanetti, C. Salati, S. Rossi, C. Forre, S. Schmid, R. Marchelli. Development of a seven- target multiplex PCR for the simultaneous detection of transgenic soybean and maize in feeds and foods. J Agr Food Chem., 52 (2004) 3275-3280.
M. Rabiei, M. Mehdizadeh, M. Alebouyeh, H. Rastegar. 2011. Screening of genetically modified organisms and specific detection of Bt-11 maize in foodstuffs. Proceedings of the 2nd International Congress of Food Hygiene, April 30th- May 1st, Tehran, Iran.
R.R. Zillman, W. Bushuk. Wheat cultivar identification by gliadin electrophore-grams. III. Catalogue of electrophoregram formulas of Canadian wheat cultivars. Canadian J. Plant Sci., 59 (1979) 287–298.
R. Tkachuk, V.J. Mellish. Wheat cultivar identification by high voltage electrophoresis. Annals Technol. Agr., 29 (1980) 207–212.
B.A. Marchylo, D.W. Hatcher, J.E. Kruger. Identifi- cation of wheat cultivars by reversed-phase high- performance liquid chromatography of storage proteins. Cereal Chem., 65 (1988) 28–40.
M.G. Scanlon, H.D. Sapirstein, W. Bushuk. Computeri- zed wheat varietal identification by high-performance liquid chromatography. Cereal Chem., 66 (1989) 439– 443.
D.J. Perry. Identification of Canadian durum wheat varieties using a single PCR. Theor. App. Gen. 109 (2001) 55-61.
R.M. Devarumath, S. Nandy, V. Rani, S. Marimuthu, N. Muraleedharan, S.N. Raina. RAPD, ISSR and RFLP fingerprints as a useful markers to evaluate genetic integrity of micropropagated plants of three diploid and tetraploid elite tea clones representing Camellia sinensis (China type) and C. assamica sp. assamica (Assam-India type). Plant Cell Rep., 21 (2002) 166-173.
M.H. Rahman, O.P. Rajora. Microsatellite DNA somaclonal variation in micropropagated trembling aspen (Populus tremuloides). Plant Cell Reports 20 (2001) 531-536.
T. Bastia, N. Scotti, T. Cardi. Organelle DNA analysis of Solanum and Brassica somatic hybrids by PCR with universal primers. Theo. Appl. Genet., 102 (2001) 1265-1272.
A. Mohanty, J.P. Martin, I. Aguinagalde. Chloroplast DNA study in wild populations and some cultivars of Prunus avium L. Theo. Appl. Genet., 103(2001) 112-117.
M. Rahman, D. Hussain, Y. Zafar. Estimation of genetic divergence among elite cotton cultivars-genotypes by DNA fingerprinting technology. Crop Science 42 (2002) 2137-2144.
M. Rahman, M. Asif, I. Ullah, K.A. Malik, Y. Zafar. 2005. Overview of cotton genomic studies in Pakistan. Plant & Animal Genome Conference XIII. San Diego, CA. USA. PP81.
M. Asif, M. Rahman, Y. Zafar. DNA fingerprinting studies of some wheat (Triticum aestivum L.) genotypes using random amplified polymorphic DNA (RAPD) analysis. Pak J Bot., 37(2005) 271-277.
A.H. Paterson, Y. Saranga, M. Menz, C.X. Jiang, R.J. Wright. QTL analysis of genotype x environment interactions affecting cotton fiber quality. Theor. Appl. Genet., 106 (2003) 384-396.
R.V. Kantety, X. Zeng, J.L. Bennetzen, B. Zehr. Assessment of genetic diversity in dent and popcorn (Zea mays L.) inbred lines using Inter-Simple Sequence Repeat (ISSR) amplification. Mol. Breed., 1(1995) 365-373.
T. Lubberstedt. A.E. Melchinger, C. Dussle, M. Vuyls- teke, M. Kuiper. Relationships among early European maize inbreds: IV. Genetic diversity revealed with AFLP markers and comparison with RFLP, RAPD and pedigree data. Crop Sci., 40 (2000) 783-791.
M. Irfan, T.T. Zhang, Y. Wang, C. Zhang, Q. Miao, L. Zhang, F. Lin. Modification of CTAB protocol for maize genomic DNA extraction. Res. J. Biotechnol., 8 (2013) 41-45.
M. Rabieia, M. Mehdizadeh, H. Rastegar, H. Vahidic, M. Alebouyeh. Detection of Genetically Modified Maize in Processed Foods Sold Commercially in Iran by Qualitative PCR. Iranian J. Pharma. Res., 12 (2013) 25- 30.
J.W. Danson, M. Kimani, M. Mbogori. Detection of Bacillus thuringiensis genes in transgenic maize by the PCR method and FTA paper technology. Afr. J. Biotechnol., 5 (2006) 2345-2349.
E. Margarit, M.I. Reggiardo, R.H. Vallejos, H.R. Permingeat. Detection of BT transgenic maize in foodstuffs. Food Res. Int., 39 (2006) 250–255.
T.C. Lee, J. Zeng, M. Bailey, S. R. Sims, P. R. Sanders, R. L. Fuchs. Assessment of equivalence of insect protected corn and E.coli produced B.T.K. HD-1 protein. Plant Physiol. Suppl., 108 (1995) 151.
M. Wu, X. Jia , L. Tian, B. Lv. Rapid and Reliable Purity Identification of F1 Hybrids of Maize (Zea mays L.) Using SSR Markers. Mol. Plant Breed., 4(2006): 381- 384.
Q. Lin, H. Yang , K. Lei, S. Zhang, F. Hao, Z. Cai, H. Wu. Genetic purity identification of ‘ Yunuo’ serial fresh 2 table maize hybrids by RAPD technique. Southwest China J. Agr. Sci., 19(2006) 182-184.
M. Asif, M. Rahman, Y. Zafar. Genotyping analysis OF six maize (Zea mays L.) hybrids using DNA fingerprinting technology. Pak J. Bot., 38 (2006) 1425-1430.
K.C.P.C. Salgado, M.D.G.G.C. VIEIRA, É. V. D.V. Pinho, C.T.Guimarães, R.G. V. Pinho, L. V. Sousa. Genetic purity certificate in seeds of hybrid maize using molecular markers. Revista Brasil. de Sem., 28(2006): 169-175.