بررسی اثر اندازه مریستم و منشاء پیوندک در ریزپیوندی گیلاس در شرایط درون شیشه‌ای

نوع مقاله : مقالات پژوهشی

نویسندگان

• محمد اسماعیل نداف ¹
• ابراهیم گنجی مقدم ²
• غلامرضا ربیعی ¹
• عبدالرحمان محمدخانی ¹

¹ گروه علوم باغبانی، دانشگاه شهرکرد، شهرکرد، ایران

² مرکز تحقیقــات و آمــوزش کشــاورزی و منــابع طبیعــی خراســان رضــوی، سازمان تحقیقات، آموزش و ترویج کشاورزی، مشهد، ایران

https://doi.org/10.22067/jhs.2021.70383.1050

چکیده

این مطالعه، با هدف بررسی تاثیر اندازه و منشاء پیوندک روی ریزپیوندی گیلاس در مرکز تحقیقات و آموزش کشاورزی و منابع طبیعی خراسان رضوی در طی سال‌های 1396-1394 انجام شد. این پژوهش در قالب آزمایش فاکتوریل بر پایه طرح کاملاَ تصادفی انجام شد. فاکتور اول: رقم در هفت سطح (’زرد‘، ’سیاه مشهد‘، ’دوم رس‘، ’بینگ‘، ’پیش رس‘، ’تکدانه ‘ و ’حاج یوسفی‘) و فاکتور دوم: نوع پیوندک در چهار سطح (ریزپیوندک با اندازه‌های دو و پنج میلی‌متری با منشاء درون شیشه­ای و برون شیشه­ای) بود. عملیات ریزپیوندی با ریزپیوندک‌هایی از نوک شاخه ارقام گیلاس روی پایه‌های ’گیزلا 6 ‘ به روش اسکنه انجام شد. گیاهان ریزپیوندی شده، ابتدا برروی محیط کشت MS حاوی یک میلیگرم برلیتر بنزیل آمینو پورین (BAP ) و بعد از سه هفته، در محیط پرلیت با محلول غذاییMS  حاوی یک میلی‌گرم بر لیتر ایندول بوتیریک اسید (IBA) قرار گرفتند و سپس به بستر پرلیت:پیت موس (به نسبت یک :یک) منتقل شدند. نتایج نشان داد که در همه شاخص‌ها بین تیمارهای نوع پیوندک و اثرات متقابل رقم در نوع پیوندک، به جز زمان گیرایی پیوند، اختلاف معنی‌دار بود، اما تفاوت معنی‌داری بین ارقام (به جز رشد طولی پیوندک) وجود نداشت. در نوع پیوندک، ریزپیوندک‌های پنج میلی‌متری با منشاء درون شیشه­ای دارای بیشترین درصد موفقیت پیوند (42 درصد)، تعداد برگ (7/3)، رشد طولی (3/6 سانتی‌متر) و زمان گیرایی پیوند (دو روز) بودند و در نهایت در بین تمام ریزپیوندهای موفق، درصد ریشه زایی، 41 درصد و درصد سازگاری، 3/25 درصد بدست آمد. در مجموع استفاده از ریز پیوندک‌های 5 میلی‌متری درون شیشه­ای در همه ارقام گیلاس، برای ریزپیوندی روی پایه ’گیزلا‘ کارایی بهتری داشت.

کلیدواژه‌ها

• آغازه‌ی برگی
• سازگاری
• عاری از ویروس
• کشت بافت
• نوک شاخه

20.1001.1.20084730.1401.36.4.7.0

موضوعات

• بیوتکنولوژی و اصلاح درختان و گل های زینتی

عنوان مقاله [English]

Effect of Meristem Size and Scion Origin on Micrografting of some Sweet Cherry Cultivars in vitro

نویسندگان [English]

• Mohammad Esmaeil Naddaf ¹
• Ebrahim Ganji Moghadam ²
• Gholmreza Rabiei ¹
• Abdolrahman Mohammadkhani ¹

¹ Department of Horticultural Science, Faculty of Agriculture, University of Shahrekord, Shahrekord, Iran

² Crop and Horticultural Science Research Department, Khorasan Razavi Agricultural and Natural Resources Research and Education Center, AREEO, Mashhad, I.R. Iran

چکیده [English]

Introduction
 Sweet Cherry (Prunus avium) belongs to the Rosaceae family, which due to vegetative propagation problems, in vitro propagation is recommended to increase mass and disease-free production. Micropropagation has many advantages over other vegetative methods. Although the most suitable organ that preserves the genetic characteristics of the cultivar is bud meristem, plant regeneration from meristem culture is difficult in many species of woody plants, so micro-grafting is a suitable technique to overcome these problems. The aim of this study was to investigate the effect of scion size and origin of commercial sweet cherry cultivars interact with micrografting on the vegetative rootstocks.
Materials and Methods
 In this study, factorial experiment was used as a test unit in a completely randomized design (CRD) with two factors in five replications and ten seedlings per replication. The first factor was cultivar in seven levels (B: Bing, D: Dovomres, H: Haj Yousefi, P: Pishres, S: Siah- Mashhad, T: Takdaneh, Z:  Zard) and the second factor was scion type in four levels (M1R1: 2 mm in vivo explant, M2R1: 5 mm in vivo explant , M1R2: 2 mm in vitro explant and  M2R2: 5 mm in vitro explant). To prepare the scion, 1.5 to 2 cm long explants were isolated from shoot tips and then disinfected with 75% ethanol and 20% Sodium hypochlorite. After rinsing with distilled water, the shoot tips with 2 and 5 mm length were extracted for in vivo explants. In vitro explants were obtained from shoo tips that was previously established in MS culture medium with supplement of 1 mg.l^-1 of BAP. The meristems were prepared in 2 and 5 mm and used as in vitro explant.  5 cm length in vitro shoots of sweet cherry ‘Gisella 6’ was used as rootstock. Micro-grafting was performed according to the standard method for sweet cherries. Micro-grafted plantlets were transferred to MS medium supplemented with 1 mg.l^{- l} BAP, and kept under low light (100 lux) condition for one week, then  transferred to growth chamber at 27.1 °C photoperiods 16/8 hrs light/darkness (1500 lux). In order to induce root, grafted plantlets were transferred to Perlite: MS medium supplement with 1 mg.l^{- l} IBA. After rooting, plants were placed in polyethylene pots containing perlite: peatmoss (1:1) for acclimation. Micro-grafting success indices were recorded in each of the micro-grafted plantlets. The data were analyzed by SAS statistical software (9.1) and the means were compared by Duncan's multiple range test (1 and 5 % of probability levels).
Results and Discussion
 The results showed that in all indices there was a significant difference between scion types and cultivar scion type interactions except grafting time, but there was no difference between cultivars (except longitudinal growth of scion). Among the scion types, the 5 mm in vitro scion (M2R2) had the highest micro-grafting success rate (42%), number of leaves (3.7), longitudinal growth (6.3 cm) and taken grafting time (two days). Finally, in successful micro-grafted plants, ‘Pishres’ cultivar had better results in rooting (32.8%) and ‘Zard’ cultivar in acclimation (3.4%) traits. Probably the presence of leaves led to better nutrient supply and surface contact, so it mostly improved the success of micrografting technique. In this study, micro-grafting success indices were lower than previous reports using seedling rootstocks. This might be due to difficult grafting operations, poor rootstock-scion communication, low physiological activity, and high in vitro oxidase activity. In the type of scion, micro-grafting success rate of 5 mm in vitro scions (include leaf primordia), was better than 2 mm scions (without leaf primordia). These results were consistent with most reports in sweet cherries and other stone fruit that were more successful in micro-grafting using larger in vitro explant.
Conclusion
 Based on our results, it can be concluded that the micro-grafting method in sweet cherry micro-propagation is a fast practical method with high potential for production and regeneration of healthy orchards, which is also possible for other cultivars. In micro-grafting success, in vitro explants are preferable to explants taken directly from in vivo mother trees, and the use of larger explants for scion is recommended due to the presence of leaf primordia in micro-grafting success. However, smaller-size explants are more likely to produce healthy plants.

کلیدواژه‌ها [English]

• Acclimation
• Leafy primordia
• Shoot tips
• Tissue culture
• Virus-free

مراجع

1. Aazami, M., & Bagher, M. (2010). In vitro micro-grafting of some Iranian grapevine cultivars. Romanian Biotechnology 15: 5576-5580.
2. Abbas, M., Khan, M., Fatima, B., Iftikhar, Y., Mughal, S., Jaskani, M., Khan, I., & Abbas, H. (2008). Elimination of Citrus tristeza closterovirus (CTV) and production of certified citrus plants through shoot-tip micrografting. Pakistan Journal of Botany 40: 1301-1312.
3. Amiri, M. (2006). In vitro technique to study the shoot tip grafting of Sweet cherry (Prunus avium var Seeyahe Mashad. Journal of Food, Agriculture & Environment 41: 151-154.
4. Daneshvar Hossini, A., Ganji Moghadam, E., & Anahid, S. (2010). Effects of media cultures and plant growth regulators in micro propagation of ‘Gisela 6’ rootstock. Annals of Biological Research 1(2): 135-141.
5. Deogratias, J., Lutz, A., & Dosba, F. (1986). In vitro micrografting of shoot tips from juvenile and adult Prunus avium and Prunus persica L. Batsch to produce virus-free plants. Acta Horticulturae 193: 139–45.
6. Estrada-Luna, A., López-Peralta, C., & Cárdenas-Soriano, E. (2002). In vitro micrografting and histology of the graft union for­mation of selected species of prickly pear cactus (Opuntia spp.). Science Horticulture 92:317-327.
7. (2015). FAOSTAT. Food and Agriculture Organization of the United Nations, Rome, Italy. Http: //faostat.fao.org/faostat.
8. Ganji Moghadam, E., & Buzari, N. (2007). Practical and practical guide to Cherry. Agricultural Extension Education and Extensions Press, Iran, 344p. (In Persian)
9. Hussain, G., Wani, M., Mir, M., Rather, Z., & Bhat, K. (2014). Micrografting for fruit crop improvement. African Journal of Biotechnology 13(25): 2474-2483.
10. Isikalan, C., Namli, S., Akbas, F., & Erol, A. (2011). Micrografting of almond (Amygdalus communis) cultivar ‘Nonpariel’. Australian Journal of Crop Science 5: 61-65.
11. Jonard, R., Lukman, D., Schall, F., & Villemur, P. (1990). Early testing of graft incompatibilities in apricot and lemon trees using in vitro Science Horticulture 43: 117-128.
12. Lauri, P., Caboni, E., & Damno, C. (2001). In vitro adventitious shoot regeneration from vegetative apices of almond and other prunus Acta Horticulturae 560: 403-406.
13. Martinez, J., Hugard, J., & Jonard, R. (1979). The different grafting of shoot tips realized in vitro between Peach (Prunus persica Batsch), Apricot (Prunus armeniaca) and Myrobolan (Prunus cerasifera Ehrh). Comala Ren Academic Science 288: 759-762.
14. Martinez-Gomez, P., & Gradziel, T.M. (2001). In vitro micrografts in almond and their application in breeding programs. Journal of Horticulture Science and Biotechnology 11: 313-
15. Murashige, T., & Skoog, F. (1962). A revised medium for rapid growth and bioassays with tobacco cultures. Physiolgy of Plant 15: 473-497.
16. Murashige, T., Bitters, W., Rangan, T., Nauer, E., Roistacher, C., & Holliday, P. (1972). A technique of shoot apex grafting and its utilization towards recovering virus-free Citrus clones. Horticulture Science 7: 118–119.
17. Nas, M., & Read, P. (2003). Simultaneous micrografting rooting and acclimatization of micropropagated American chestnut grapevine and hybrid hazelnut. European Journal of Horticultural Science 68: 234-237.
18. Navarro, L. (1988). Application of shoot-tip grafting in vitro to woody species. Acta Horticulturae 227: 43–55.
19. Navarro, L., Roistacher, C., & Murashige, T. (1975). Improvement of shoot-tip grafting in vitro for virusfree citrus. Journal of the American Society for Horticultural Sciences 100: 471–479.
20. Pathirana, R., & McKenzie, M. (2005). Early detection of grapevine leafroll virus in Vitis vinifera using in vitro Plant Cell Tissue and Organ Culture 81: 11-18.
21. Piagnani, M., Prinsi, B., & Bassi, D. (2006). Experimental approaches to in vitro grafting in Prunus armeniaca and spinosa. Horticulture Science 20(3): 224-230.
22. Rafail, S., & Mosleh, M. (2010). Factors involved in micropropagation and shoot tip grafting of apple Malus domestica) and pear (Pyrus sp. L.). Tropentag World Food System – A Contribution from Europe September 14 -16, 2010 in Zurich.
23. Ramanayake, S., & Kovoor, T. (1999). In vitro micrografting of cashew (Anacardium occidentale). Journal of Horticulture Science and Biotechnology 74: 265-68.
24. Richardson, F., Saoir, S., & Harvey, B. (1996). A study of the graft union in in vitro micrografted apple. Plant growth regulation 20: 17-23.
25. Wang, G., Li, X., Chen, Q., & Tian, J. (2010). Studies on factors affecting the micro-shoot grafting survival of walnut. Acta Horticulture 861: 327-331.