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Evaluation of somaclonal variation of Arnebia pulchra (Boraginaceae) calli versus seeds using ISSR markers | ||
Iranian Journal of Genetics and Plant Breeding | ||
دوره 10، شماره 1 - شماره پیاپی 19، تیر 2021، صفحه 39-46 اصل مقاله (596.95 K) | ||
نوع مقاله: Research paper | ||
شناسه دیجیتال (DOI): 10.30479/ijgpb.2022.16406.1306 | ||
نویسندگان | ||
Mahsa Tavakoli؛ Amir Mousavi* ؛ Kamahldin Haghbeen | ||
Department of Agricultural Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran. | ||
تاریخ دریافت: 09 آبان 1400، تاریخ بازنگری: 05 اردیبهشت 1401، تاریخ پذیرش: 16 خرداد 1401 | ||
چکیده | ||
This is the first report on the analysis ISSR markers for assessing the genetic variation in Arnebia pulchra. As the rate of cell division and propagation in A. pulchra is very low, the achievement of biomass in this valuable plant requires techniques and strategies that can guarantee the survival of the callus. The present study was performed to investigate genetic variation in two in vitro culture samples of the medicinal plant A. pulchra including new (one month old) and old (four years old) calli versus seeds by Inter Simple Sequence Repeat (ISSR) marker. In total, 79 bands were produced by 10 ISSR primers. The total expected heterozygosity (He) was calculated 0.028 and the greatest value was contributed to seeds samples (0.054) followed by old and new calli, respectively. The maximum values of different alleles (Na), effective alleles (Ne) and Shannon’s index were also observed in seeds. Cluster analysis was performed in the form of dendogram to indicate the genetic stability of the samples. UPGMA tree discriminated samples in two major groups and principal component analysis (PCoA) confirmed the clustering result. The first major cluster included seeds and the second involved new and old calli. Although it was thought that the morphological changes of A. pulchra could be due to the somaclonal variation caused by successive subcultures, however, the results indicated that the new and old calli were clustered in the same group. Therefore, it can be concluded that the morphological changes of the old and new calli could be due to environmental parameters or epigenetic changes but not directly due to somaclonal variation. | ||
کلیدواژهها | ||
Cluster analysis؛ ISSR markers؛ Somaclonal variation؛ Successive subculture؛ Tissue culture | ||
عنوان مقاله [English] | ||
ارزیابی تنوع سوماکلونی در کالوس آرنبیا پولچرا (تیره گاوزبان) در مقایسه با بذور آن با استفاده از نشانگر آی.اس.اس.آر | ||
نویسندگان [English] | ||
مهسا توکلی؛ امیر موسوی؛ کمال الدین حق بین | ||
گروه بیوتکنولوژی کشاورزی، پژوهشکده زیست فناوری کشاورزی، پژوهشگاه ملی مهندسی ژنتیک و زیست فناوری، تهران، ایران. | ||
چکیده [English] | ||
این مقاله نخستین گزارش کاربرد نشانگرهای آی.اس.اس.آر جهت ارزیابی تنوع ژنتیکی در گیاه آرنبیا پولچرا است. از آنجایی که سرعت تقسیم سلولی و تکثیر در این گیاه باارزش کم است، دستیابی به زیستتوده، نیازمند تامین فناوری و راکارهایی است که بتواند بقای کالوس را تضمین نماید. مطالعه حاضر به منظور بررسی تنوع ژنتیکی در دو نمونه کشت درون شیشهای گیاه دارویی آرنبیا پولچرا، شامل کالوسهای جدید (یک ماهه) و کالوس قدیمی ( چهار ساله) در مقابل بذر این گیاه با استفاده از نشانگر آی.اس.اس.آر انجام شده است. 79 باند توسط ده آغازگر آی.اس.اس.آر تولید شد. هتروزیگوتی کل مورد انتظار (He)، 028/0 محاسبه شد. بیشترین مقدار هتروزیگوتی مربوط به نمونه بذر بوده (054/0) که بیشترین تعداد آللهای مختلف و مؤثر و شاخص شانون را نیز به خود اختصاص داده است. تجزیه خوشهای به صورت دندوگرام برای نشان دادن پایداری ژنتیکی نمونهها انجام شد. درختچه UPGMA ، نمونهها را در دو گروه اصلی تفکیک کرد و تجزیه به مؤلفههای اصلی (PCoA) این خوشهبندی را تأیید کرد. اولین خوشه اصلی شامل بذور و خوشه دوم شامل کالوسهای جدید و قدیمی بود. گرچه تصور میشد تغییرات مورفولوژیکی کالوسهای آرنبیا پولچرا میتواند ناشی از تنوع سوماکلونال حاصل از واکشتهای متوالی باشد، اما نتایج نشان داد کالوسهای جدید و قدیمی در یک گروه قرار گرفتهاند. بنابراین میتوان نتیجه گرفت که تغییرات مورفولوژیکی کالوسهای قدیمی و جدید میتواند ناشی از عوامل محیطی یا تغییرات اپیژنتیک باشد اما مستقیماً به دلیل تنوع سوماکلونال نبوده است. | ||
کلیدواژهها [English] | ||
تجزیه خوشه ای, نشانگرهای آی.اس.اس.آر, تنوع سوماکلونی, واکشت های متوالی, کشت بافت | ||
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