تنوع سوماکلونال در گیاهان باززایی‌شده لیلکی (.Gleditschia caspica Desf) با استفاده از نشانگرهای بین ریزماهواره ژنومی (ISSR-PCR)

نوع مقاله : علمی- پژوهشی

نویسندگان

1 نویسنده مسئول، دانش‌آموخته دکتری، گروه علوم و مهندسی جنگل، دانشکده منابع طبیعی، دانشگاه علوم کشاورزی و منابع طبیعی ساری، ساری، ایران

2 دانشیار، گروه علوم و مهندسی جنگل، دانشکده منابع طبیعی، دانشگاه علوم کشاورزی و منابع طبیعی ساری، ساری، ایران

3 دانشیار، گروه اصلاح نباتات، دانشکده علوم زراعی، دانشگاه علوم کشاورزی و منابع طبیعی ساری، ساری، ایران

4 استادیار، گروه علوم باغبانی، دانشکده تولیدات گیاهی، دانشگاه علوم کشاورزی و منابع طبیعی گرگان، گرگان، ایران

چکیده

تنوع سوماکلونال، آزمایش ژنتیکی است که با استفاده از آن می‌توان تغییرات ژنی و ثبات ژنتیکی گیاه تکثیرشده را نسبت به گیاه مادری ارزیابی کرد. هدف این پژوهش، بررسی تنوع سوماکلونال در گیاهان باززایی‌شده لیلکی (Gleditsia caspica Desf.) تحت تأثیر تنظیم‌کننده رشد BAP و بازدارنده رشد CPZ بود. پس از بذرگیری از یک درخت لیلکی در باغ گیاه‌شناسی دانشکده منابع طبیعی در دانشگاه علوم کشاورزی و منابع طبیعی ساری، ژنوتیپ‌های پایه مادری و گیاهان باززایی‌شده در محیط‌های کشت حاوی BAP (20 میکرومول) و CPZ (3/0، سه و 15 میکرومول) با استفاده از نشانگر مولکولی ISSR مقایسه شدند. بیشترین درصد شاخه‌زایی و تعداد شاخساره در تیمار BAP با غلظت 20 میکرومول مشاهده شد. وجود بازدارنده رشد CPZ در محیط ‌کشت به‌شدت سبب کاهش درصد شاخه‌زایی ریزنمونه‌ها شد. همچنین، روند کاهشی مشخصه‌های مورد مطالعه با افزایش غلظت CPZ تشدید یافت. استفاده از 10 نشانگر مولکولی ISSR درمجموع، 107 نوار DNA تولید کرد که 9/87 درصد آن‌ها، چندشکلی بودند. دارنگاره تجزیه خوشه‌‌ای براساس ماتریس تشابه جاکارد به‌روش UPGMA نشان داد که ژنوتیپ‌های مورد مطالعه با ضریب تشابه 27/0 در دو گروه قرار گرفتند. به‌نظر می‌رسد که وجود بازدارنده رشد CPZ به‌‌رغم اثر منفی بر رویش گیاه سبب افزایش تنوع و کاهش شباهت ژنتیکی بین گیاهان باززایی‌شده و پایه مادری می‌شود.

کلیدواژه‌ها

عنوان مقاله [English]

Somaclonal diversity in Caspian honey locust (Gleditsia caspica Desf.) regenerated plants using genomic microsatellite markers (ISSR-PCR)

نویسندگان [English]

  • M. Imani Rastabi 1
  • S.M. Hosseini-Nasr 2
  • Gh. Ranjbar 3
  • M. Khoshhal Sarmast 4
1 Corresponding author, Ph.D. Graduated, Department of Forestry Science and Engineering, Faculty of Natural Resources, Sari Agricultural Sciences and Natural Resources University, Sari, Iran
2 Associate Prof., Department of Forestry Science and Engineering, Faculty of Natural Resources, Sari Agricultural Sciences and Natural Resources University, Sari, Iran
3 Associate Prof., Department of Plant Breeding and Biotechnology, Sari Agricultural sciences and Natural Resources University, Sari, Iran
4 Assistant Prof., Department of Horticultural Sciences, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran
چکیده [English]

    Somaclonal diversity is a genetical test that can be used to study the genetic variation and genetic stability of a propagated plant relative to the mother plant. The aim of this study was to investigate the genetic stability of Caspian honey locust (Gleditsia caspica Desf.) regenerated plants under the influence of BAP growth regulator and CPZ growth inhibitor. For this purpose, native and regenerated plants genotypes were compared in culture media containing growth regulator BAP (20 μmol) and CPZ (0.3, 3 and 15 μmol) used by molecular marker of ISSR. The highest percentage of shoot proliferation and number of shoots was observed in 20 μmol BAP treatment. The presence of CPZ in the culture medium greatly reduced the shoot proliferation percentage and with increasing CPZ concentration, the decreasing trend of the studied characteristics increased. The results showed that the use of 10 ISSR molecular markers produced a total of 107 DNA bands that had 87.9% polymorphism. Cluster analysis dendrogram based on Jaccard similarity matrix by UPGMA method showed that the studied genotypes were divided into two groups with similarity coefficient of 0.27. It seems that the presence of CPZ, despite the growth inhibitory effect on the plant, will increase diversity and reduce the genetic affinity between the regenerated plants and the mother plant.

کلیدواژه‌ها [English]

  • Genetic diversity
  • growth inhibitor
  • growth regulator
  • tissue culture

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