Assessment of genetic diversity in different types of tobacco using molecular markers

Document Type : Research Paper

Authors

1 Researcher, Plant Breeding, Faculty of Agricultural Sciences, University of Guilan, Rasht, Iran.

2 Associate Professor, Department of Agricultural Biotechnology, Faculty of Agricultural Sciences, University of Guilan, Rasht, Iran.

3 Professor, Department of Agricultural Biotechnology, Faculty of Agricultural Sciences, University of Guilan, Rasht, Iran.

Abstract

Information about the amount of genetic diversity in the germplasm and genetic relations of genotypes is essential for analyzing and designing breeding programs and could be used for assisting in genetic identification and improvement of the germplasm. In this research, the genetic diversity of 48 genotypes of the flue-cured, burley, and oriental types of tobacco was investigated by 12 ISSR, 10 SSR, and five IRAP primers. The maximum number of bands was observed in the UBC825, RTR-10, and mixed primer UBC817 + UBC826 with 17 bands, UBC817 with 16 bands, and RTR-8 with 15 bands. UBC824 with 10 and UBC823 with 11 bands showed the minimum number of bands among ISSR and IRAP primers, and PT30044 and PT30046 primers with two alleles showed the minimum number of alleles among SSR markers. In ISSR and IRAP, the observed percentage of polymorphism ranged from 76.92% for RTR-1 and RTR-7 to 94.11% for RTR-10, and in SSR markers, it was 100%. The average polymorphism percentage was 90.7%, which indicated suitable genetic diversity among the tobacco genotypes. The polymorphism information content ranged from 0.31 to 0.5 with an average of 0.42. The average diversity of Nei and Shannon indices were 0.39 and 0.58, respectively. Cluster analysis by the UPGMA method classified the 48 tobacco genotypes into five groups, containing 5, 12, 10, 6, and 15 genotypes, respectively. Principal coordinate analysis showed that the 14 first components could explain 51.49% of the total variance. The primers used in this study had high efficiency, which can be used to study the genetic diversity in tobacco.

Keywords

Main Subjects


Article Title [Persian]

بررسی تنوع ژنتیکی انواع مختلف توتون از طریق نشانگرهای مولکولی

Authors [Persian]

  • مصطفی مهدی زاده 1
  • محمد محسن زاده گلفزانی 2
  • حبیب ا... سمیع زاده 3
1 محقق اصلاح نباتات، دانشکده علوم کشاورزی، دانشگاه گیلان، رشت
2 دانشیار گروه بیوتکنولوژی کشاورزی، دانشکده علوم کشاورزی، دانشگاه گیلان، رشت
3 استاد گروه بیوتکنولوژی کشاورزی، دانشکده علوم کشاورزی، دانشگاه گیلان، رشت
Abstract [Persian]

ا
اطلاعات مربوط به میزان تنوع ژنتیکی در ژرم­ پلاسم و روابط ژنتیکی بین ژنوتیپ ­ها برای بررسی و طراحی برنامه­ های به‌نژادی مهم می­ باشد و می‌تواند برای کمک به شناسایی و بهبود ژنتیکی ژرم­ پلاسم به ­کار رود. در این تحقیق، تنوع ژنتیکی 48 ژنوتیپ توتون از تیپ‌های گرمخانه‌ای، بارلی و شرقی توسط 12 نشانگر ISSR، 10 نشانگر SSR و 5 نشانگر IRAP بررسی شد. آغازگرهای UBC825، RTR-10 و آغازگر ترکیبی UBC817 + UBC826 با تعداد 17 نوار و بعد از آن، آغازگر UBC817 با 16 نوار و RTR-8 با 15 نوار بیشترین تعداد نوار و آغازگر UBC824 با 10 و UBC823 با 11 نوار کمترین تعداد نوار در بین آغازگرهای ISSR و IRAP را داشتند. همچنین آغازگرهای PT30044 و PT30046 با 2 نوار از کمترین تعداد نوار در بین آغازگرهای SSR برخوردار بودند. درصد چندشکلی به دست آمده از 76.92 درصد برای RTR-1 و RTR-7 تا 94.11 درصد برای RTR-10 در آغازگرهای ISSR و IRAP متغیر بود و چندشکلی در آغازگرهای SSR صد درصد به دست آمد. میانگین چندشکلی 90.7 درصد، وجود تنوع ژنتیکی مناسب بین ژنوتیپ ­های توتون را نشان می ­دهد. محتوای اطلاعات چندشکلی در این تحقیق بین 0.31 تا 0.5 و میانگین محتوای اطلاعات چندشکلی 0.42 بود. میانگین تنوع ژنی نی و شاخص شانون به ترتیب 0.39 و 0.58 به دست آمد. تجزیه خوشه ­ای به روش UPGMA،  ژنوتیپ­ های توتون مورد بررسی را در پنج گروه قرار داد که به ترتیب شامل 5، 12، 10، 6 و 15 ژنوتیپ شدند. بر اساس تجزیه به بردارهای اصلی 14 مؤلفه اول 51.49 درصد از واریانس کل را توجیه کردند. پرایمرهای مورد استفاده در این مطالعه کارایی بالایی داشتند که می ­توان از آن­ ها برای بررسی تنوع ژنتیکی در توتون استفاده کرد.

Keywords [Persian]

  • بارلی
  • شرقی
  • گرمخانه ای
  • IRAP
  • ISSR
  • SSR
miraju J, Dholakia B, Santra D, Singh H, Lagu M, Tamhankar S, Dhaliwal H, Rao V, Gupta V, Ranjekar P. 2001. Identification of inter simple sequence repeat (ISSR) markers associated with seed size in wheat. Theor Appl Genet. 102(5): 726-732.
Azizi H, Bernousi I, Abdollahi MB, Darvishzadeh R. 2012. Study of genetic structure and diversity in cultivated alfalfa populations (Medicago sativa L.) using ISSR markers. Mod Gent J. 4(27): 61-69 (In Persian with English abstract).
Blair M, Panaud O, McCouch S. 1999. Inter-simple sequence repeat (ISSR) amplification for analysis of microsatellite motif frequency and fingerprinting in rice (Oryza sativa L.). Theor Appl Genet. 98(5): 780-792.
Brummer EC, 1999. Capturing heterosis in forage crop cultivar development. Crop Sci. 39(4): 943-954.
Brummer E, Bouton J, Kochert G. 1995. Analysis of annual Medicago species using RAPD markers. Genome 38(2): 362-367.
Dadras A, Sabouri H, Sabouri A. 2014. Association analysis of loci controlling some important morphological traits of tobacco using AFLP markers under osmotic stress conditions. Iran J Field Crops Res. 45(1): 73-84 (In Persian with English abstract).
Davalieva K, Maleva I, Filiposki K, Spiroski O, Efremov GD. 2010. Genetic variability of Macedonian tobacco varieties determined by microsatellite marker analysis. Diversity 2(4): 439-449.
Denduangboripant J, Setaphan S, Suwanprasart W, Panha S. 2010. Determination of local tobacco cultivars using ISSR molecular marker. Chiang Mai J Sci. 37(2): 293-303.
Doyle J, Doyle J. 1987. A rapid DNA isolation procedure for small amounts of fresh leaf material. Photochem Bul. l 19: 11-15.
Edrisi Maryan K, Samizadeh Lahiji H, Shoaei Deylami M. 2012. Assessing the genetic diversity of tobacco (Nicotiana tabacum L.) varieties. Crop Breed J. 2(2): 125-132.
Emel S. 2010. Evaluation of ISSR markers to assess genetic variability and relationship among winter triticale (Triticosecale Wittmack) cultivars. Pak J Bot. 42(4): 2755-2763.
Flavell AJ, Dunbar E, Anderson R, Pearce SR, Hartley R, Kumar A. 1992. Ty1–copia group retrotransposons are ubiquitous and heterogeneous in higher plants. Nucleic Acids Res. 20(14): 3639-3644.
Ghafari Azar A, Darvishzadeh R, Aghaali Z, Kahrizi D, Darvishi B. 2019. Assessment of genetic diversity and grouping of maize lines (Zea mays L. ) using ISSR markers. Cell Mol Res. 32(2): 230-241 (In Persian with English abstract).
Hassani TSF, Samizadeh LH, Shoaei DM. 2014. Study of genetic diversity among and within types of tobacco (Nicotiana tabacum l.) using ISSR markers. Mod Genet J. 9(1): 1-12. (In Persian with English abstract).
Hassani TSF, Samizadeh LH, and Shoaei DM, 2016. Assessment of genetic diversity among and within different types of tobacco (Nicotiana tabacum L.) using IRAP and REMAP markers. J Crop Breed. 7(16): 1-9. (In Persian with English abstract).
He B, Geng R, Cheng L, Yang X, Ge H, Ren M. 2020. Genetic diversity and fingerprinting of 33 standard flue-cured tobacco varieties for use in distinctness, uniformity, and stability testing. BMC Plant Biol. 20: 378.
Hoshyardel F, Darvishzadeh R, Ashkan B, Hatami Maleki H. 2016. Association mapping of agronomic traits in oriental tobacco. J Crop Breed. 8(18): 134-143 (In Persian with English abstract).
Kalendar R, Grob T, Regina M, Suoniemi A, Schulman A. 1999. IRAP and REMAP: two new retrotransposon-based DNA fingerprinting techniques. Theor Appl Genet. 98(5): 704-711.
Kantety RV, Zeng X, Bennetzen JL, Zehr BE. 1995. Assessment of genetic diversity in dent and popcorn (Zea mays L.) inbred lines using inter-simple sequence repeat (ISSR) amplification. Mol Breeding. 1(4): 365-373.
Liu ZQ, Pei Y, and Pu ZJ, 1999. Relationship between hybrid performance and genetic diversity based on RAPD markers in wheat, Triticum aestivum L. Plant breeding 118(2):119-123.
Mohsenzadeh Golfazani M, Samizade lahiji H, Alami A, shoayi deylami M, and Talesh Sasani S, 2012. Study of Genetic Diversity of Flue-Cured Tobacco (Nicotiana Tabacum L.) Genotypes using ISSR and Retrotransposon Markers. Iranian Journal of Field Crop Science 43(2):371-380 (In Persian with English abstract).
Mohsenzadeh Golfazani M, Samizadeh H, Olfati J, and Mostashari-rad F, 2021. Grouping of some bean genotypes in various drought stress treatments in Germination Stages and the expression investigating of some genes in resistant and susceptible. Iranian Journal of Seed Sciences and Research 8(3):227-243 (In Persian with English abstract).
Moon H, Nifong J, Nicholson J, Heineman A, Lion K, Van der Hoeven R, Hayes A, Lewis R. 2009. Microsatellite‐based analysis of tobacco (Nicotiana tabacum L.) genetic resources. Crop Sci. 49(6): 2149-2159.
Nagaoka T, Ogihara Y. 1997. Applicability of inter-simple sequence repeat polymorphisms in wheat for use as DNA markers in comparison to RFLP and RAPD markers. Theor Appl Genet. 94(5): 597-602.
Nazir M, Mahajan R, Hashim MJ, Iqbal J, Alyemeni MN, Ganai BA, Zargar SM. 2021. Deciphering allelic variability and population structure in buckwheat: an analogy between the efficiency of ISSR and SSR markers. Saudi J Biol Sci. 28(11): 6050-6056.
Nei M. 1972. Genetic distance between populations. Am Natur. 106(949): 283-292.
Nezamivand Chegini M, Samizadeh Lahiji H, Ramezani Malekroudi M, Mohsenzadeh Golfazani M, Seighalani R. 2016. Assessment of morphological and molecular variation in local olive (Olea europaea L.) in the northern part of Iran. Acta Agric Slov. 107(2): 397-408.
Piano Ld, Abet M, Sorrentino C, Acanfora F, Cozzolino E, Muro Ad. 2000. Genetic variability in Nicotiana tabacum and Nicotiana species as revealed by RAPD markers: 1. Development of the RAPD procedure. Beitr Tab Int. 19(1): 1-15.
Pradeep Reddy M, Sarla N, Siddiq EA. 2002. Inter simple sequence repeat (ISSR) polymorphism and its application in plant breeding. Euphytica 128(1): 9-17.
Raju KS, Madhav M, Sharma R, Murthy T, Mohapatra T. 2008. Genetic polymorphism of Indian tobacco types as revealed by amplified fragment length polymorphism. Curr Sci. 94(5): 633-639.
Sabouri H, Karim MR, Dadras AR. 2019. Association analysis of genes controlling morphological traits in rice using ISSR markers in flooding and drought conditions. J. Appl Biol. 32(3): 83-103 (In Persian with English abstract).
Salehian M, Darvishzadeh R, Rezazad Bari M. 2021. Assessment of genetic diversity and association analysis for agro-morphological traits in pepper (Capsicum spp.) using ISSR markers. J Plant Res. (Iran J Biol.) 34(1): 126-138 (In Persian with English abstract).
 Samizade Lahiji H, Mohsenzade M, Edrisi Maryan K, Shoayi deylami M, Alami A. 2013. Assessing the genetic diversity of 89 flue-cured tobacco varieties using morphological traits and inter-simple sequence repeat markers. Crop Breed J. 3(2): 79-85.
Serrote CML, Reiniger LRS, Silva KB, Rabaiolli SMDS, Stefanel CM. 2020. Determining the polymorphism information content of a molecular marker. Gene. 726: 144175.
Shannon CE. 1948. A mathematical theory of communication. Bell Syst Tech J. 27(3): 379-423.
Wicker T, Sabot F, Hua-Van A, Bennetzen JL, Capy P, Chalhoub B, Flavell A, Leroy P, Morgante M, Panaud O. 2007. A unified classification system for eukaryotic transposable elements. Nat Rev Genet. 8(12): 973-982.
Xiao B, Yang B. 2007. Assessment of genetic diversity among tobacco germplasms by ISSR markers. Sci Agric Sin. 40(10): 2153-2161.
Yang B-C, Xiao B-G, Chen X-J, Shi C-H. 2005. Genetic diversity of flue-cured tobacco varieties based on ISSR markers. Hereditas. 27(5): 753-758.
Yang B, Xiao B, Chen X, Shi C. 2007. Assessing the genetic diversity of tobacco germplasm using inter simple sequence repeat and inter‐retrotransposon amplification polymorphism markers. Ann Appl Biol. 150(3): 393-401.
Zhu J, Gale M, Quarrie S, Jackson M, Bryan G. 1998. AFLP markers for the study of rice biodiversity. Theor Appl Genet. 96(5): 602-611.