ارزیابی تنوع ژنتیکی برخی از ژنوتیپ‎های سیب ایران با استفاده از نشانگر RAPD

نوع مقاله: پژوهشی

نویسندگان

1 دانشجوی دکتری، گروه علوم باغبانی دانشکده کشاورزی دانشگاه گیلان

2 عضو هیأت علمی گروه علوم باغبانی، دانشگاه آزاد اسلامی واحد میانه، میانه، ایران

3 کارشناس ارشد موسسه تحقیقات اصلاح و تهیه نهال و بذر کرج

چکیده

      جمع­آوری و ارزیابی ژرم­پلاسم­ها در برنامه­های اصلاحی درختان میوه دارای نقش اساسی است. نشانگرهای مولکولی متفاوتی برای این نوع ارزیابی­ها به­کار برده شده­اند که مارکر RAPD جزء مارکرهای پرکاربرد در مورد شناسایی ژنوتیپ­ها و ارقام گوناگون سیب می­باشد. در این پژوهش تنوع ژنتیکی 23 ژنوتیپ بومی ایران (از سه منطقه آذربایجان، البرز مرکزی و زاگرس مرکزی) و رقم تجاری گرانی­اسمیت به عنوان شاهد توسط 11 آغازگر RAPD سری TIB MOLBIOL بررسی شد. تعداد متوسط  نوار به‎ازای هر آغازگر 27/10 بود. تجزیه و تحلیل داده­ها با نرم­افزار NTSYS با کاربرد ضریب تشابه DICE برای تعیین میزان تشابه انجام شد و دندروگرام به‎وسیله الگوریتم UPGMA رسم گردید. در حالت کلی، نتایج داده­های مولکولی، نمونه­های هر منطقه را به­طور جداگانه­ای از سایر مناطق  جدا کرد. البته برخی نمونه­های مناطق در خوشه­های منطقه­ای جز منطقه خود قرار گرفتند که این موضوع می­تواند به دلیل انتقال ژنوتیپ­ها بین مناطق در زمان گذشته باشد. میزان چند شکلی به­دست آمده در این پژوهش بالا بود (62/68 درصد). بر اساس داده­های مولکولی، دامنه شباهت بین نمونه­ها از 378/0 تا 723/0 متغیر بود. با توجه به نتایج به­دست آمده می­توان گفت که تنوع ژنتیکی بالایی در ژنوتیپ‏های سیب مناطق مختلف وجود دارد، ولی تفاوت درون منطقه‎ای جزئی بود. به­طور کلی، بررسی تنوع ژنتیکی نشان داد که مارکر RAPD می­تواند در شناسایی نواحی چند شکلی و تخمین فاصله ژنتیکی و مدیریت ژرم­پلاسم ژنوتیپ­ها و ارقام سیب مفید باشد.

کلیدواژه‌ها


عنوان مقاله [English]

Investigation of genetic variation among some Iranian apple genotypes using RAPD marker

نویسندگان [English]

  • Shahin Jahangirzadeh Khiavi 1
  • Hassan Noorafkan 2
  • Sima Damyar 3
1 Ph.D. Student, Hortculture Science, Agriculture Faculty, Gilan University, Gilan, Iran.
2 Faculty Member of Miyaneh Branch, Islamic Azad University, Miyaneh, Iran.
3 M.Sc. of Seed and Plant Improvement Research Institute, Karaj, Iran.
چکیده [English]

Collection and evaluation of germplasms play an important role in fruit tree breeding programs and different molecular markers have been used for this purpose. RAPD marker is one of the most common markers to identify different apple genotypes and varieties. In this study, the genetic diversity of 23 indigenous apple genotypes collected from Azerbaijan, Central Alborz and Central Zagros regions of Iran was evaluated by 11 RAPD primers serie TIB MOLBIOL and commercial cultivar of Granny Smith was consdered as the control. Data were analyzed using NTSYS software, dendrogram was drawn based on UPGMA results and DICE similarity index was used for measuring genetic similarities. Average number of bands was 10.27 for each primer. In general, samples of each region were separate from other regions and were classified in different clusters. However, some samples were placed in the cluster of other regions that it could be due to genotype transition among areas in the past tense. High polymorphism rates were obtained (68.62%). Based on the results, similarity of samples ranged from 0.378 to 0.723. There was high genetic difference among apple genotypes collected from the studied areas, but low difference was observed among samples of each region. In conclusion, investigation of genetic variation indicated that RAPD marker is suitable approach to determine polymorphic loci and to estimate the genetic distance among apple genotypes and cultivars.

کلیدواژه‌ها [English]

  • apple
  • germplasm
  • genetic diversity
  • similarity index
  • RAPD

Adebayo OL, Bola O, Opeyemi W, Gloria M, Temitope OO (2009) Phylogenetic and genomic relationships in the genus Malus based on RAPDs. African Journal of Biotechnology (15): 3387-3391.

Autio WR, Schupp JR, Ferree DC, Glavin R, Mulcahy DL (1998) Application of RAPDs to DNA extracted from apple rootstocks. Hort Science 33: 333-335.

Crosby JA, Janick J, Pecknold PC, Korban SS, O’Connor PA, Ries SM, Goffreda S, Voordeckers A (1992) Breeding apple for scab resistance. Acta Horticulturae 317: 43-70.

Deverno, LL, Charest PJ, Bonen L (1994) Mitochondrial DNA variation in somatic embryogenic cultures of Larix. Theoretical and Applied Genetics 88: 727-732.

Duneman F, Kahnau R, Schmidt H (1994) Genetic relationships in Malus evaluated by RAPD fingerprinting of cultivars and wild species. Plant Breeding 113: 150-159.

Erturk U, Akcay MA (2010) Genetic variability in accessions of ‘Amasya’ apple cultivar using RAPD markers. Notulae Botanicae Horti Agrobotanici Cluj-Napoca 38 (3): 239-245.

Gardiner SE, Bassett HCM, Madie C, Noition DAM (1996) Isozyme, randomly amplified polymorphic DNA (RAPD), restriction fragment length polymorphism (RFLP) markers to deduce a putative parent for the ‘Braeburn’ apple. Journal of the American Society for Horticultural Science 121: 996-1001.

Gygax, M, Gianfranceschi L, Liebhard R, Kellerhals M, Gessler C, Patocchi A (2004) Molecular markers linked to the apple scab resistance gene Vbj derived from Malus baccata Jackii. Theoretical and Applied Genetics 109: 1702-1709.

Harada T, Maksukawa K, Sato T, Ishikawa Niizeki R, Saito KM (1993) DNA-RAPD detect genetic variation and paternity in Malus. Euphytica 65: 87-91.

Hemmat, M, Weeden NF, Manganaris AG, Lawson DM (1994) Molecular marker linkage map for apple. Journal of Heredity 85: 4-11.

Jahangirzadeh khiavi SH, Zamani Z, Mardi M, Fatahi Moghaddam M (2013) Evaluation of chloroplast relationship between some apple genotype from Azerbaijan of Iran and their comparison with other local genotypes, cultivars and rootstocks. African Journal of Agricultural Research 8(1): 106-112.

Khadivi khob A, Zamani Z, Bouzari N, Fatahi Moghaddam MR (2009) Evaluation of genetic diversity in some Iranian sweet cherry cultivars using some morphological characteristics and RAPD markers. Seed and plant Improvement Journal 25(1): 195- 209. [In Persian with English Abstract]

Koller BA, Lehmann J, Modermott M, Gessier C (1993) Identification of apple cultivars using RAPD markers. Theoretical and Applied Genetics 85: 6-7.

Lavi U, Cregan P, Schaap T, Hillel J (1994) Application of DNA markers for identification and breeding of perennial fruit crops. In: Janick J (ED.), Plant breeding reviews. John Wiley and Sons, Inc, New York. pp. 195–226.

Liebhard R, Koller B, Gianfranceschi L, Gessler C (2003). Creating a saturated reference map for the apple (Malus domestica Borkh.) genome. Theoretical and Applied Genetics 106: 1497-1508.

Luis G, Cabrita L, Oliveira CM, Leitao JM (2001) Comparing RAPD and AFL analysis in discrimination and estimation of genetic similarities among apple (Malus domestica Borkh) cultivars. Euphytica 119: 250-270

Mohanty A, Martín JP, Aguinagalde I (2001) Chloroplast DNA study in wild populations and some cultivars of Prunus avium L. Theoretical and Applied Genetics 103: 112–117

Mulcahy DL, Cresti M, Sansavini S, Douglas GC, Linskens HF, Mulcahy GB, Vignani R, Pancaldi M (1993) The use of random amplified polymorphic DNAs to fingerprint apple genomes. Scientia Horticulturae 54: 89-96.

Nybon H, Schaal BA (1990) DNA ‘fingerprints’ applied to paternity analysis in apples (Malus x domestica). Theoretical and Applied Genetics 79: 763- 768.

Rohlf FJ (2008) NTSYS-PC numerical taxonomy and multivariate analysis system. Version 2.2. Exeter Software, Setauket, New York.

Royo JB, Itoiz R (2004) Evaluation of the discriminance capacity of RAPD, isoenzymes and morphologic markers in apple (Malus x domestica Borkh.) and the congruence among classifications. Genetic Resources and Crop Evolution 51: 153-160.

Watilon B, Druart P, Du Jardin P, kettmann R, Boxus P, Burny A (1991) Use of random cDNA probes to detect restriction fragment length polymorphisms among apple clones. Scientia Horticulturae 46: 235-243.

Weeden NF, Lamb RC (1985) Identification of apple cultivars by isoenzyme phenotypes. Journal of the American Society for Horticultural Science 110: 509–515

Williams JGK, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acid Research 18: 6531–6535.

Williams, JGK, Hanafey MK, Rafalski JA, Tingey SV (1993) Genetic analysis using random amplified polymorphic DNA markers. Methods in Enzymology 218: 704-740.

Zhou ZQ, Li YN (2000) The RAPD evidence for the phylogenetic relationship of the closely related species of cultivated apple. Genetic Resources and Crop Evolution 47: 353-357.

Zhu SL, Manfredi P, Monti LM, Rao R (1997) RAPD markers useful for the identification of the ‘Annurca’ apple variety and its sport ‘Rossa Del Sud’. Advances in Horticultural Science 11: 120-122.