تکثیر درون‌شیشه‌ای و عاری‌سازی ژنوتیپ‌های برتر محلب (‏Prunus mahaleb‏) از ویروس‌های بذرزاد ‏از طریق گرمادرمانی و کشت مریستم ‏

نوع مقاله: مقاله کامل

نویسندگان

1 دانش‌آموخته کارشناسی ارشد تولیدات گیاهی، دانشگاه آزاد اسلامی، واحد گرمسار

2 دانشیار، سازمان تحقیقات، آموزش و ترویج کشاورزی، مؤسسه تحقیقات باغبانی، پژوهشکده میوه‌های معتدله و سردسیری، کرج ‏

3 دانشیار، سازمان تحقیقات، آموزش و ترویج کشاورزی، مؤسسه تحقیقات ثبت و گواهی بذر و نهال، کرج

4 دانشیار، سازمان تحقیقات، آموزش و ترویج کشاورزی، پژوهشکده بیوتکنولوژی کشاورزی، کرج

چکیده

محلب (Prunus mahaleb) از مهم­ترین منابع تولید پایه بذری و کلونال و رایج­ترین پایه بذری گیلاس و آلبالو در ایران است. هدف از این تحقیق دست­یابی به پایه­های انتخابی محلب NB5176، NBVP1و NBVP2 عاری از ویروس­های بذرزاد آبله آلو (Plum pox virus, PPV)، کوتولگی آلو (Prune dwarf virus, PDV)، لکه حلقوی بافت مرده درختان هسته­دار (Prunus necrotic ringspot virus, PNRSV) بود.  برای این منظور، ابتدا آلودگی درختان مادری توسط آزمون سرولوژیک الیزا (ELISA) بررسی شد. مریستم­های انتهایی این ژنوتیپ­ها درمحیط MS مستقر و شش هفته بعد به محیط­های پرآوری MS (حاوی 5/0 میلی­گرم بر لیتر BAP، 1/0 میلی­گرم بر لیتر NAA و 3/0 میلی­گرم بر لیتر پکتین) و DKW (حاوی 5/0 میلی­گرم بر لیتر BAP)  و WPM (حاوی 3/0 میلی­گرم بر لیتر پکتین) منتقل شدند. پس از مرحله تکثیر اولیه، گیاهچه­ها درون شیشه گرمادرمانی شدند و سلامت گیاهان بازیابی شده از گرمادرمانی از سه ویروس فوق توسط RT-PCR بررسی و گیاهچه‌های سالم، تکثیر، ریشه­دار و به گلدان منتقل شدند. بر اساس نتایج آزمون الیزا، برخی درختان مادری به ویروس­های PDV و PNRSV آلوده بودند. بیشترین درصد استقرار مریستم 5/42% مربوط به ژنوتیپ NBVP1 بود. در مرحله پرآوری، هیچ ژنوتیپی در محیط­های MSوWPM  رشد نیافت و تنها محیط ­DKW مناسب بود. از نظر ریشه­زایی، محیط­های DKWوQL ‌ تغییریافته حاوی 5/1 میلی­گرم بر لیتر IBA مناسب بودند. ژنوتیپ NBVP1 در کلیه محیط‌های ریشه­زایی بهتر از دو ژنوتیپ دیگر ریشه‌دار شد. 40-25 درصد از گیاهچه­های درون شیشه­ای به شرایط گرمادرمانی تحمل داشته و براساس نتایج RT-PCR، عاری ازPDV ، PNRSV، PPV بودند. نهال­های گلدانی به­دست­آمده می­توانند به باغ ایزوله منتقل شده و برای تولید بذر سالم محلب به کار روند.

کلیدواژه‌ها


عنوان مقاله [English]

Obtaining seed-borne virus free mahaleb (Prunus mahaleb) genotypes through ‎meristem tip culture and in vitro thermotherapy

نویسندگان [English]

  • Maliheh Gamshidiha 1
  • Mansureh Keshavarzi 2
  • Masoud Naderpour 3
  • Naser Bouzari 2
  • Ali Mohammad Shakib 4
1 Former M.Sc. Student, Islamic Azad University, Garmsar, Iran‎
2 Associate Professor, Agricultural Research, Extension and Education Organisation (AREEO), Horticultural Research Institute, ‎Temperate Fruit Research Center, Karaj, Iran‎
3 Associate Professor, AREEO, Seed and Plant Registration Organization, Karaj, Iran
4 Associate Professor, AREEO, Agriciltural Biotechnology Research Institute, Karaj, Iran
چکیده [English]

Mahaleb (Prunus mahaleb) plant is one of the most important sources of cherry clonal and seed base rootstock production in Iran. Obtaining seed-borne viruse-free (Prune dwarf virus, Plum pox virus, Prunus necrotic ringspot virus) sources for selected mahaleb rootstocks (NB5176, NBVP1, NBVP2), For this purpose, the viral infection status of genotypes was initially studied in orchard using ELISA serological method. Then the meristem tips were cultured in vitro and 6 months later, transferred to MS medium containing 0.5mg/l BAP and 0.1 mg/l NAA, DKW contacting 0.5 mg/l BAP and WMP contacting 0.3 mg/l pectin. After the initial proliferation stage, the plantlets were heat-treated and then health status of the remained plantlets was studied using RT-PCR. Then the healthy plants were rooted and transferred to pots. Based on ELISA result, some genotypes were probably infected to PNRSV and PDV. The highest shoot tip establishment was recorded as 42.5% by NBPV1. No genotype was proliferated in MS and WPM, so only DKW was apropriate for proliferation. For plantlet rooting, 1/2DKW and modified QL containing 1.5 mg/l IBA were appropriate. NBVP2 was the best in all rooting media. Most of plantlets from three genotypes toleratedin-vitro thermotherapy condition and were free of PNRSV, PDV, PPV based on RT-PCR result. The healthy plants were transferred to pots and could be cultured in an isolated orchard to obtain healthy mahaleb seeds.

کلیدواژه‌ها [English]

  • ELISA
  • PNRSV
  • PPV
  • proliferation
  • RT-PCR
  • Rootstock
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