با همکاری مشترک دانشگاه پیام نور و انجمن بیوتکنولوژی جمهوری اسلامی ایران

نوع مقاله : علمی پژوهشی

نویسندگان

1 دانشجوی دکتری اصلاح نباتات، گروه به‌نژادی و بیوتکنولوژی، دانشکده کشاورزی دانشگاه تبریز، تبریز، ایران

2 استاد گروه زراعت و اصلاح نباتات، دانشکده کشاورزی دانشگاه تهران، پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج، ایران

3 دانشیار گروه زراعت و اصلاح نباتات، دانشکده کشاورزی دانشگاه تهران، پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج، ایران

چکیده

به منظور شناسایی نشانگرهای پیوسته با صفات مرفوفنولوژیک در ژنوتیپ های لوبیا چشم بلبلی، از 22 جفت نشانگرهای ریزماهواره (SSR) استفاده شد. آغازگرها، 186 مکان در 32 ژنوتیپ لوبیا چشم بلبلی تولید کردند. میانگین تعداد مکان ها 45/8 مکان برای هر آغازگر و میزان اطلاعات چندشکلی (PIC) برای آغازگرها 445/0 از 25/0 (Vm25) تا 625/0 (Vm5) متغیر بود. به منظور بررسی فاصله ژنتیکی بین ژنوتیپ ها تجزیه خوشه ای برای داده های حاصل از نشانگرهای SSR براساس ضرایب تشابه دایس و جاکارد ژنوتیپ ها را در دو گروه اصلی قرار داد. به منظور شناسایی نشانگرهای مولکولی مثبت، تجزیه رگرسیون گام به گام بین داده های مولکولی به عنوان متغیرهای مستقل و صفات مورفولوژیک به عنوان متغیرهای وابسته در هر دو شرایط خشکی و نرمال انجام گرفت. نشانگرهای مرتبط با صفات عملکرد اقتصادی و بیولوژیک به ترتیب در شرایط نرمال Vm70 و Vm33، در خشکی Vm3 و Vm26 بودند. در تنش خشکی بیشترین تغییرات مربوط به صفات تعداد روز تا 50% گلدهی، تعداد روز تا 50% رسیدگی غلاف و طول غلاف (9/99درصد) به ترتیب توسط نشانگرهای Vm22، Vm31 و Vm26تبین شد و در شرایط نرمال بیشترین تغییرات مربوط به صفات ضخامت غلاف، عرض غلاف و تعداد روز تا 50 درصد گلدهی (9/99 درصد) توسط نشانگرهای Vm14، Vm34 و Vm34 تبین شدند. در هر دو شرایط تنش خشکی و نرمال اکثر آغازگرهای مورد استفاده روی صفات ارتباط نشان دادند، بنابراین می‌توان از این آغازگرها همراه با اطلاعات صفات مرفوفنولوژیک در اصلاح لوبیا چشم‌بلبلی جهت شناسایی ژنوتیپ‌های متحمل به تنش خشکی، مناسب برای تهیه جمعیت‌های نقشه‌یابی و تولید ارقام متحمل استفاده کرد.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Identification of SSR markers associated with morphological traits in cowpea (Vigna unguiculata L. Walp) genotypes under drought stress condition

نویسندگان [English]

  • Khosro Mafakheri 1
  • Mohammad Reza Bihamta 2
  • Ali Reza Abbasi 3

1 Ph.D Student, Department of Biotechnology and Plant Breeding, University of Tabriz, Tabriz, Iran.

2 Professor, Department of Agronomy and Plant Breeding, University of Tehran, Tehran, Iran

3 Associate Professor, Department of Agronomy and Plant Breeding, University of Tehran, Tehran, Iran.

چکیده [English]

In the current investigation, 22 SSR primer pairs were used to identify molecular markers associated with morphological traits in cowpea (Vigna unguiculata L.) based association analysis, markers produced 186 locations in 32 cowpea genotypes. 22 SSR primers pair amplified 8.45 for each primer location and the average polymorphism information content (PIC) ranged from 0.445 to 0.25 (Vm25) and 0.625 (Vm5) were varied, respectively. Cluster analysis based on Dice, Jaccard and Simple Maching similarity coefficients based on molecular traits revealed primarily two major groups. Stepwise regression analysis between molecular data as independent variables, and morphological data as dependent variables was performed to identify informative markers associated with the studied traits. SSR loci associated with economic and biological yield, in normal conditions and in drought stress condition were Vm70 and Vm33, and Vm3 and Vm26 respectively. In drought stress condition the maximum variation of number of days to 50% flowering, days to 50% maturity of pods and pod length (99.9%, respectively) was accounted by Vm22, Vm31 and Vm26 markers. While in the normal condition the maximum variation of traits sheath thickness, width of pod and days to 50% flowering (99.9%) was accounted by Vm14, Vm34 and Vm34 markers. In both normal and drought conditions most of the used SSR primers showed significant association with the studied traits, so we can use these markers along with morphological traits in breeding cowpea genotypes to identify drought tolerant and hardy cultivars suitable for the preparation and use mapping populations.

کلیدواژه‌ها [English]

  • Cowpea
  • Positive Markers
  • Morphological Traits
  • Molecular markers
Agriculture statistics, crops, (2014) First volume. Ministry of Agriculture. (In Persian).
Abdollahi Mandoulakani B, and Azizi H (2010) Identification of ISSR markers associated with morphological traits in cultivated alfalfa (Medicago sativa L.) populations. Journal of Cellular and Molecular 2: 260-268.
Abdollahi Mandoulakani B, Alami A, and Esfahani M (2010) Association analysis for morphological traits in peanut (Arachis hypogea L.) using microsatellite markers. Iranian Journal of Crop Sciences 12: 510-519.
Agarma HA, Tuinstra MR (2003) Phylogenetic diversity and relationship among sorghum accessiond using SSRs and RAPDs. Afr. J. Biotechnol. 2: 334-340.
Abdolshahi R, Omidi M, Talei A R, Yazdi Samadi, B (2010) Evaluation of bread wheat genotypes for drought tolerance. Esci. J. Crop Prod. 3: 159-171.
Agrama HA, Eizenga GC, Yan W (2007) Association mapping of yield and its components in rice cultivars. Mol. Breeding. 19: 341-356.
Brummer EC (1999) Capturing hetrosis in forage crop cultivar development. Crop Science. 32: 939-943.
Brummer E C, Bouton J H and Kochert G (1995) Analysis of annual Medicaogo species using RAPD markers. Genome 38: 362-367.13.
Belhassen E (Ed) (1996) Drought in higher plants: Genetical, Physiological and Molecular biological analysis. ENSAINRA SGAP, Montpellier, France.
Diouf D, Hilu KW (2005) Microsatellites and RAPD markers to study genetic relationships among cowpea breeding lines and local varieties in Senegal. Genetic Resources and Crop Evolution 52:1057-1067.
Fathi M (2005) Study of genetic diversity cowpea (Vigna unguiculater L.) with molecular and morphological markers. MSc. Thesis, College of Agriculture, University of Tehran, Karaj, Iran.
Gebhardt C, Ballvora A, Walkemeier B, Oberhagemann P, Schuler K (2004) Assessing genetic potential in germplasm collections of crop plants by marker-trait association: a case study for potatoes with Quantitative variation of resistance to late blight and maturity type. Molecular Breeding 13: 93–102.
Jun TH, Van K, Kim MY, Lee SH, Walker DR (2008) Association analysis using SSR markers to find QTL for seed protein content in soybean. Euphytica 62: 179–191.
Julier B, Huyghe CH, Ecall CH (2000) within and among cultivar genetic variation in alfalfa: forage quality, morphology and yield. Crop Science 4: 362-365.
Jolaei M (2010) The effect of mild water stress on some physiological characteristics and the level of enzyme activity of anti-oxidants of different cultivars of wheat. M.Sc. Thesis, College of Agriculture, University of Tehran, Karaj, Iran (in Persian).
Li X, Wei Y, Moore KJ, Michaud R, Viands DR, Hansen JL, Acharya A, and Brummer EC (2011) Association Mapping of Biomass Yield and Stem Composition in a tetraploid Alfalfa Breeding Population. The plant genome 4: 24-35
Li Q, Liu QC, Zhai H, Ma DF, Wang X, Li XQ, Wang YP (2008) Genetic diversity in main parents of sweet potato in china as revealed by ISSR Markers. Acta Agronomica Sinicia 34: 972-977.
Litt M, Luty JA (1989) A hyper variable microsatellite revealed by in vitro amplification of a dinucleotide repeat within the cardiac muscle actin gene. Am. J. Hum. Genet 44: 397–401.
Ivandic V, Hackett CA, Nevo E, Keith R, Thomas WTB, Forster BP (2002) Analysis of simple sequence repeats (SSRs) in wild barley from the Fertile Crescent: associations with ecology, geography and flowering time. Plant Molecular Biology. 48: 511–527.
International Board for Plant Genetic Resources (1983) Descriptors for cowpea. IBPGR Secretariat, Rome.
Mafakheri Kh, Bihamta M R, Abbasi A R, and Rasollnia A R (2015) Genetic Diversity of Cowpea (Vigna unguiculata (L.) Walp) Germplasm Based on Agronomic and Morphological Traits. Seed and Plant Improvement Journal 31: 135-162.
Majnoun Hosseinei N (2008) Legumes agriculture and production. The university press center, Tehran, Iran 103129.
Mohammad Ali Pour Yamchi H, Bihamta MR, Peighambari SA, Naghavi MR (2012) Identify informative markers microsatellite in Kabuli chickpea genotypes under drought stress .12th Iranian Genetic Congress, Tehran, Iran.
Mohammadi R, Naghavi M R, Maali Amiri R, Rezaii M (2010) Identify informative markers Microsatellite in cultivated alfalfa (Medicago sativa L.) Iran. Modern Genetics Journal 2: 57-66.
Mengoni A, Gori A and Bazzicalupo M (2000) Use of RAPD and microsatellite (SSR) variation to assess genetic relationships among populations of tetraploid alfalfa, Medicago sativa. Plant Breeding, 119: 113-117.
Naghavi M R, Mardi M, Ramshini H A, Fazelinasab B (2004) Comparative analysis of the genetic diversity among bread wheat genotypes based on RAPD and SSR markers. Iran. J. Biotech. 2: 195–202. (In Persian).
Naghavi M R, Ghareyazie B, and Hosseini Salak Deh Gh (1384) Molecular markers. The university press center, Tehran, Iran. (In Persian).
Nei M (1973) Analysis of gene diversity in subdivided populations. Proc Natl Acad Sci USA 70:3321-3323. Doi: 10.1073/pnas/70.12.3321.
Naghavi M, Jahansouz M R (2005) Variation in the Agronomic and Morphological Traits of Iranian Chickpea Accessions. J. of Integrative Plant Biology 47: 375-379.
Kashi Y, Soller M (1999) Functional roles of microsatellites and minisatellites. In: Goldstein D.B. and Schlِtterer C (Eds). Microsatellites: Evolution and Application. 10–23.Oxford University Press, Oxford.
Rashidi Monfared S, Mardi M, Hosseinzadeh A H, Naghavi M (2008) Association analysis between important Agronomic traits and SSAP markers in durum wheat. Modern Genetics Journal 3: 29-35. (In Persian).
Roy JK, Bandopadhyay R, Rustgi S, Balyan H S and Gupta P K (2006) Association analysis of agronomically important traits using SSR, SAMPL and AFLP markers in bread wheat. Current Sci. 5: 683-689.
Saeed A, Hovsepyan H, Darvishzadeh R, Imtiaz M, Panguluri S K and Nazaryan R (2011) Genetic diversity of Iranian accessions, improved lines of chickpea (Cicer arietinum L.) and their wild relatives using simple sequence repeats. Plant Molecular Biology Reporter DOI 10.1007/s11105-011-0294-5.
Sefera T, Abebie B, Gaur PM, Assefa K (2011) Characterization and genetic diversity analysis of selected chickpea cultivars of nine countries using simple sequence repeat (SSR) markers. Crop Pasture Sci 62: 177-187.
Skot L, Humphreys MO, Armstead IP, Heywood S, Skot KP, Sanderson R, Thomas ID, Chorlton KH, Sackville Hamilton NR (2005) An association mapping approach to identify flowering time genes in natural populations of Lolium perenne L. Molecular Breeding. 15: 233-245.
Shafii Khorshidi M, Bihamta MR, Khialparast F, Naghavi M (2012) Identify informative markers Microsatellite in beans (Phaseolus vulgaris.) genotypes 12th Iranian Genetic Congress, Tehran, Iran.
Takeda S, Matsuoka M (2008) Genetic approaches to crop improvement: responding to environmental and population change. Nature. 9: 444-457.
Takeda S, Matsuoka M (2008) Genetic approaches to crop improvement: responding to environmental and population change. Nature 9: 444-457.
Thudi M, Upadhyaya HD, Rathore A, Mal Gaur P, Krishnamurthy L, Roorkiwal M, Nayak SN, Kumar Chaturvedi S, Sarathi Basu P, Gangarao NVPR, Fikre A, Kimurto P, Sharma PC, Sheshashayee MS, Tobita S, Kashiwagi J, Ito O, Killian A, Kumar Varshney R (2014) Genetic Dissection of drought and heat tolerance in Chickpea through genome-wide and candidate gene based association mapping approaches. PloS ONE 9:e96758.
Yazdi Samadi B, Mohammadi V, Abd Mishani S (2010) Breeding crops. University Publication Center. Tehran. Iran.
Yeh FC, Boyle T, Rongcai Y, Ye Z, Xian JM (1999) POPGENE version 3.1. http://www.ualberta.ca/- fyeh/fyeh.
Zaccardelli M, Gnocchi S, Carelli M, Scotti C (2003) Variation among and within Italian alfalfa ecotypes by means of bioagronomic characters and amplified fragment length polymorphism analyses. Plant Breeding 122: 61-65.
Veronesi F, Charles B, Huyghe C (2010) Alfalfa. Springer Sci. 395-436.
Virk PS, Ford-Lioyd BV, Jackson MT, Pooni HS, Clemeno TP, Newbury HJ (1996) Marker-assisted prediction of agronomic traits using diverse rice germplasm. Third International Rice Genetics Symposium, Manilla, Philippines. 307-316.
Upadhyaya HD, Dwivedi SL, Baum M, Varshney RK (2008) Genetic structure, diversity and allelic richness in composite collection and reference set in chickpea (Cicer arietinum L.). BMC Plant Biol. 8: 106-112.