ارزیابی مولکولی کارایی گرمادرمانی و کشت مریستم برای حذف شماری از بیماری‌های ویروسی در ‏رقم‌های مهم تجاری گلابی (‏Pyrus communis L.‎‏) ‏

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی سابق دکتری، گروه علوم باغبانی، دانشکده کشاوزری، دانشگاه تبریز

2 دانشیار، گروه علوم باغبانی، دانشکده کشاوزری، دانشگاه تبریز

3 دانشیار، پژوهشکده بیوتکنولوژی ایران (ابری)، کرج

چکیده

این پژوهش با هدف تولید نهال عاری از سه ویروس ACLSV (Apple chlorotic leafspot virus)، ASGV (Apple stem grooving virus) و ASPV (Apple stem pitting virus) در هفت رقم گلابی شامل ابته فتل، بیروتی، درگزی، کوشیا، لوئیزبون، ملینا و اسپادونا انجام شد. آزمایش­ها با ارزیابی اثربخشی دوره‌های گرمادرمانی شامل صفر، 7، 14 و 21 روز در دمای 38 درجه سانتی‌گراد و کشت مریستم انتهایی با اندازه کمتر از 2/0 میلی‌متر، بر میزان حذف ویروس از ریزنمونه­ها انجام شد. در ابتدا حضور ویروس‌های مورد مطالعه در ریزنمونه‌های مادری با روش RT-PCR مورد ارزیابی قرار گرفت و به جز، نمونه­های مادری رقم­های ابته­فتل و بیروتی که عاری از ویروس  ASPVبودند، سایر ریزنمونه‌ها به هر سه ویروس آلودگی داشتند. گرمادرمانی و کشت مریستم در شرایط درون شیشه‌ای انجام شد. ریزشاخه­های حاصل از گرمادرمانی و کشت مریستم توسط RT-PCR برای هر سه ویروس مورد بررسی قرار گرفتند و نتایج نشان داد میزان حذف سه ویروس ACLSV، ASGV و ASPV به‏‌‏ترتیب با درصدهای 63/26، 5/35 و 46/78 در رقم‌های مختلف با یکدیگر متفاوت بود. بیشترین میزان عاری شدن از ویروس در رقم کوشیا و کمترین آن در رقم اسپادونا مشاهده شد. افزایش طول مدت گرمادرمانی رابطه مستقیم با افزایش درصد عاری سازی ریزنمونه­ها از هر سه ویروس مورد مطالعه داشت، اما از طرفی این افزایش دوره زمانی گرمادرمانی در 21 روز باعث کاهش رشد و تکثیر و حتی از بین رفتن ریزنمونه­ها شد. بنابراین 14 روز گرمادرمانی مؤثرترین تیمار جهت حذف آلودگی ویروس­هایASGV ،ASPV  و ACLSV به‏‌‏ترتیب با 4/61، 100 و 5/45 درصد از ریزنمونه­های مورد مطالعه بود. در پایان آزمایش نمونه‌هایی که توسط RT-PCR سالم تشخیص داده شدند، تکثیر و ریشه‌دار شدند و در شرایط گلخانه­ای سازگار شدند.

کلیدواژه‌ها


عنوان مقاله [English]

Molecular assessment of thermotherapy and meristem culture efficiency on some ‎virus’s eradication from important commercial pear cultivars (Pyrus communis L.)‎

نویسندگان [English]

  • Nooshin Kazemi 1
  • Fariborz Zaare-Nahandi 2
  • Ali Akbar Habashi 3
  • Mohammad Reza Dadpour 2
1 Former Ph.D. Student and Associate Professor, Department of Horticultural Science, Faculty of Agriculture, University of Tabriz, ‎Tabriz, Iran
2 Associate Professor, Department of Horticultural Science, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
3 Associate Professor, Department of Agriculture Biotechnology Research Institute of Iran (ABRII), Karaj, Iran‎
چکیده [English]

This study was aimed to eradicate three viruses, Apple chlorotic leaf spot virus (ACLSV),Apple stem grooving virus (ASGV) and Apple stem pitting virus (ASPV) from seven pear cultivars including Abate Fetel, Beiruti, Dargazi, Coscia, Louise Bonne, Mellina, Spadona. Experiments were performed by evaluation of the effectiveness of thermotherapy duration including 0, 7, 14 and 21 days at 38 °C and meristem culturing in size of less than 0.2 mm on virus eradication rates of explants. At first mother samples were tested for ACLSV, ASGV and ASPV by and RT-PCR and all samples were infected by all three viruses, except Abate Fetel and Beiruti samples, that were free of ASP virus. Thermotherapy and meristem culturing were performed in vitro. Explants from thermotherapy and meristem culturing were tested by RT-PCR for all three viruses and the results showed that eradication rates of ACLSV, ASGV and ASPV, respectively 78.46, 26.63 and 35.5 percent were different in the different pear cultivars. The highest rate of virus eradication was related to Coscia and the lowest to Spadona cultivars. There was a direct relationship between increasing the duration of thermotherapy and virus elimination of explants, but this increased period of time up to 21 days reduced the growth and multiplication and even destroyed the explants. Therefore, 14 days of thermotherapy was the most effective treatment for elimination of ASGV, ASPV and ACLSV, respectively 61.4, 100 and 45.5 percent, from pear explants. At the end of the experiment, samples that were diagnosed virus free using RT-PCR, were proliferated, rooted and transferred to greenhouse condition for acclimation stage.

کلیدواژه‌ها [English]

  • Apple chlorotic leaf spot virus
  • Apple stem pitting virus
  • Apple stem grooving virus
  • RT-PCR
  • thermotherapy
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