نوع مقاله : علمی - پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد علوم باغبانی، دانشکده کشاورزی، دانشگاه زنجان، زنجان، ایران

2 دانشیار، گروه علوم باغبانی، دانشکده کشاورزی، دانشگاه زنجان، زنجان، ایران

3 دانشیار، پژوهشکده بیوتکنولوژی کشاورزی ایران، کرج، ایران

4 کارشناس پژوهشکده بیوتکنولوژی کشاورزی ایران، کرج، ایران

چکیده

چکیده
این پژوهش در سال 1391 در آزمایشگاه کشت بافت دانشگاه زنجان و با هدف بهینه‌سازی ریز‌ازدیادی یکی از ژنوتیپ‌های ارزشمند و بومی سیب گوشت سرخ انجام شد. آزمایش‌ها شامل بررسی اثر ترکیبات متفاوت تنظیم‌کننده‌های رشد GA3 (در سه سطح صفر، 3 و 6 میکرومولار) وBAP  (در چهار سطح صفر، 2، 4 و 6 میکرومولار) بر جنبه‌های مختلف پرآوری شامل تعداد برگ و ریز‌شاخه تولید‌شده در هر ریزنمونه، میزان رشد طولی ریزشاخه‌هاو وزن تر و خشک ریزنمونه‌هابود. آزمایش‌ها به‌صورت فاکتوریل درقالب طرح کاملاً تصادفی با سه تکرار صورت گرفتند. همچنین، تأثیر غلظت‌های مختلف تنظیم‌کننده‌های رشدی IBA (در پنج سطح صفر، 1، 2، 3 و 4 میکرومولار) و NAA (در پنج سطح صفر، 1، 2، 3 و 4 میکرومولار) بر ریشه‌زایی ریزنمونه‌ها بررسی شد. بهترین نتیجه برای پرآوری (9/7 تعداد ریزشاخه/ ریزنمونه/ 8 هفته) در محیط کشت حاوی چهار میکرومولار BAp و سه میکرومولار GA3 به‌دست‌آمد. به‌علاوه، گیاهان تکثیر‌شده در این تیمار نسبت به تیمارهای دیگر پژوهش، نتایج مطلوب‌تری در کلیه صفات مورد بررسی شامل تعداد برگ، میزان رشد طولی ریزشاخه‌هاو وزن تر و خشک ریز‌نمونه‌ها نشان دادند. نتایج نشان داد، محیط کشت MS 2/1 حاوی سه میکرومولار IBA با 2/85 درصد ریشه‌زایی طی هشت هفته بهترین ترکیب برای ریشه‌زایی بود. نتایج سازگاری نشان داد گیاهان ریشه‌دار‌شده در تیمارهای حاوی IBA بسیار موفق‌تر (100درصد) از گیاهچه‌های ریشه‌دار‌شده در تیمارهای حاوی NAA (40 درصد) بودند، لذا محیط کشت حاوی سه میکرومولار IBA کاراترین ترکیب جهت ریشه‌دار شدن و بقای سیب گوشت سرخ در مرحله سازگاری بود.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Optimizing Micro Propagation of Red Flesh Apple Genotype

نویسندگان [English]

  • Nooshin Kazemi 1
  • Mohammad Esmaiil Amiri 2
  • Maryam Jaffarkhani Kermani 3
  • Zahra sadat Hosseini 4

1 M.Sc. Student of Horticultural Sciences, Faculty of Agriculture, University of Zanjan, Zanjan, Iran

2 Associate Professor, Department of Horticultural Sciences, Faculty of Agriculture, University of Zanjan, Zanjan, Iran

3 Associate Professor, Agriculture Biotechnology Research Institute of Iran (ABRII), Karaj, Iran

4 Agriculture Biotechnology Research Institute of Iran (ABRII), Karaj, Iran

چکیده [English]

Abstract
 
Background and Objectives
Red flesh apple belongs to Malus niedzwetzkyana of Rosaceae family. It has high levels of important phytochemicals like antioxidants, flavonoids and anthocyanins in its cortex, which, in addition to attractiveness, is a source of benefit compound. The aim of the present investigation was to optimize efficient protocols for micro propagation of red flesh apple in order to use the results in future breeding strategies.
 
Materials and Methods
During the early spring, stem cuttings of growing red flesh apple trees were surface-sterilized in 70% ethanol and 50% sodium hypochlorite. Then, sterilized samples were placed in the test tubes containing MS medium supplemented with 2 µM BAP. At the proliferation stage, interactive effects of BAP (0, 2, 4, 6 mM) and GA3 (0, 3, 6 mM) on various aspects of proliferation (multiplication rate and vegetative growth characteristics such as number of produced shoots per explant, shoots length, leaf numbers, and the fresh and dry weight of explants) were investigated. At rooting stage, the effect of different concentrations of IBA or NAA (0, 1, 2, 3,4 µM) on the percentage of rooting, number of roots per explant and length of roots were compared. At acclimatization stage, the plantlets were transferred into pots with a volume of 200 ml containing sterile compound at the greenhouse condition.
 
Results
The results showed that there was a significant difference between the effects of various concentrations of BAP and GA3 on micro propagation of red flesh apple. The best result in proliferation (7.9 of axillary shoots after 8 months) was obtained from explants that were cultured in the medium containing 4 mM BAP and 3 mM GA3. At rooting stage, the maximum rooting percentage (85.2%) was obtained in the medium containing 3 mM IBA. In addition, rooted explants from IBA treatments were successfully acclimatized (100%) in the greenhouse, but rooted explants in NAA treatments were weak in acclimatized stage (40%).
 
Discussion
Measuring the growth rate is the major economic parameter for successful plant production at in vitro conditions, which is usually characterized by the number of axillary shoots, number of regenerated leaves and increase in the length of shoots and fresh and dry weight. We obtained the highest growth rate of red flesh apple in the culture medium containing 4 μM of BAP. The efficiency of BAP as an important cytokinin in apple proliferation has been reported in other studies as well. The results of the present investigation showed that highest rooting percentage was obtained from culture medium containing 3 μM of IBA. In addition, morphology of roots varied in callus formation and longitudinal growth and the samples rooted in IBA treatments had stronger roots than the rooted samples in NAA treatments. It seems that exposure to NAA instead of IBA stimulates indirect organogenesis and produced undesirable side effects on roots, such as callus formation, which can cause plantlets weakness during the period of acclimatization.
 

کلیدواژه‌ها [English]

  • Compatibility
  • Multiplication
  • Plant growth regulator
  • Rooting
References
Ballester, A., Vidal, N., Vieitez, A. M., Niemi, K. and Scagel, C. (2009). Developmental stages during in vitro rooting of hardwood trees from material with juvenile and mature characteristics: Adventitious root formation of forest trees and horticultural plants from genes to applications. Research Singpost, Kerala,1, 277-299.
Boudabous, M., Mars, M., Marzougui, N. and Ferchichi, A. (2010). Micropropagation of apple (Malus domestica L. cultivar Douce de Djerba) through in vitro culture of axillary buds. Acta Botanica Gallica, 157(3), 513-524.
Brown, S. (2012). Fruit breeding, handbook of plant breeding. In M. L. Badenes and D. H. Byrne, (Eds), Fruit breeding (pp. 329-367). Berlin, Germany: Springer Science & Business Media.
Ciccotti, A. M., Bisognin, C., Battocletti, I., Salvadori, A., Herdemertens, M., Wallbraun, M. and Jarausch, W. (2009). Micropropagation of malus sieboldii hybrids resistant to apple proliferation disease. Acta Horticulturae, 839(1), 35-41.
Cornille, A., Gladieux, P., Smulders, M. J., Roldan-Ruiz, I., Laurens, F., Le Cam, B., Nersesyan, A., Clavel, J., Olonova, M., Feugey, L. and Gabrielyan, I. (2012). New insight into the history of domesticated apple: Secondary contribution of the European wild apple to the genome of cultivated varieties. PLoS genetics, 8(5), e1002703.
De Klerk, G. J. (2002). Rooting of microcuttings: Theory and practice. In Vitro Cellular & Developmental Biology-Plant, 38(5), 415-422.
Dobranszki, J. and Da Silva, J. A. T. (2010). Micropropagation of apple-a review. Biotechnology Advances, 28(4), 462-488.
Etherton, P. M., Hecker, K. D., Bonanome, A., Coval, S. M., Binkoski, A. E., Hilpert, K. F., Griel, A. E. and Etherton, T. D. (2002). Bioactive compounds in foods: Their role in the prevention of
cardiovascular disease and cancer. The American Journal of Medicine, 113(9), 71-88.
George, E. F., Hall, M. A. and De Klerk, G. J. (Eds.) (2008). Plant propagation by tissue culture. Dordrecht: Springer.
Gerdakaneh, M., Badakhshan, H., Mohamadi, M., and Arji, I. (2019). Effect of Different Media and Growth Regulators on Micropropagation of GF677. Plant Productions, 43(2), 241-254. [In Farsi]
Han, H., Zhang, S. and Sun, X. (2009). A review on the molecular mechanism of plants rooting modulated by auxin. African Journal of Biotechnology, 8(3), 348-353.
Hunt, M. A., Trueman, S. J. and Rasmussen, A. (2011). Indole-3-butyric acid accelerates adventitious root formation and impedes shoot growth of Pinus elliottii var. elliottii × P. caribaea var. hondurensis cuttings. New Forests, 41(3), 349-360.
Jafarkhani Kermani, M., Hosseini, Z. S. and Habashi, A. A. (2008). A refined tissue culture medium for in vitro proliferation of apple rootstocks. Acta Horticulturae, 829, 313-318.
Jamshidi, S., Yadollahi, A., Ahmadi, H., Arab, M. M. and Eftekhari, M. (2016). Predicting in vitro culture medium macro-nutrients composition for pear rootstocks using regression analysis and neural network models. Frontiers in Plant Science, 7, 1-12.
Luckman, G. A. and Menary, R. C. (2002). Increased root initiation in cuttings of Eucalyptus nitens by delayed auxin application. Plant Growth Regulation, 38(1), 31-35.
Magyar-Tabori, K., Dobranszky, J., Jambor-Benczur, E., Lazanyi, J., Szalai, J. and Ferenczy, A. (2002). Effects of indole-3-butyric acid levels and activated charcoal on rooting of in vitro shoots of apple rootstocks. International Journal of Horticultural Science, 8(3-4), 25-28.
Mazza, G. and Velioglu, Y. S. (1992). Anthocyanins and other phenolic compounds in fruits of red-flesh apples. Food Chemistry, 43(2), 113-117.
Modgil, M., Sharma, D. R. and Thakur, M. (2009). Commercially feasible protocol for rooting and acclimatization of micropropagated apple rootstocks. Acta Horticulturae, 839(1), 209-215.
Mohamadzadeh Moghadam, N. and Hamidi, H. (2017). Investigating the effects of medium, sterilization and hormonal treatment on micropropagation of some apple (Mallus Domestica Borkh.) rootstocks. Plant Productions, 40(1), 41-54. [In Farsi]
Murashige, T. and Skoog, F. (1962). A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum, 15(3), 473-497.
Naija, S., Elloumi, N., Jbir, N., Ammar, S. and Kevers, C. (2008). Anatomical and biochemical changes during adventitious rooting of apple rootstocks MM 106 cultured in vitro. Comptes Rendus Biologies, 331(7), 518-525.
Saito, A. and Suzuki, M. (1999). Plant regeneration from meristem-derived callus protoplasts of apple (Malus domestica cv. Fuji'). Plant Cell Reports, 18(7-8), 549-553.
Soni, M., Thakur, M. and Modgil, M. (2011). In vitro multiplication of Merton I. 793-An apple rootstock suitable for replantation. Indian Journal of Biotechnology, 10(3), 362-368.
Tang, H., Luo, Y. and Liu, C. (2008). Plant regeneration from in vitro leaves of four commercial Pyrus species. Plant Soil and Environment, 54(4), 140-148.
Xu, J., Wang, Y., Zhang, Y. and Chai, T. (2008). Rapid in vitro multiplication and ex vitro rooting of Malus zumi (Matsumura) rehd. Acta Physiologiae Plantarum, 30(1), 129-132.
Yepes, L. M. and Aldwinckle, H. S. (1994). Micropropagation of thirteen Malus cultivars and rootstocks, and effect of antibiotics on proliferation. Plant Growth Regulation, 15(1), 55-67.
 
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