بررسی اثر تنظیم کننده‌های رشد و انواع ریزنمونه بر پینه‌زایی و باززایی گونه کُنار Ziziphus spina christti (L.) Willd

نوع مقاله: مقاله پژوهشی

نویسندگان

1 کارشناس‌ارشد جنگلداری گروه جنگلداری دانشکده منابع طبیعی دانشگاه علوم کشاورزی و منابع طبیعی ساری

2 استادیار گروه جنگلداری دانشکده منابع طبیعی دانشگاه علوم کشاورزی و منابع طبیعی ساری

3 دانشیار گروه جنگلداری دانشکده منابع طبیعی دانشگاه علوم کشاورزی و منابع طبیعی ساری

چکیده

کُنار یک گونه چوبی دارویی مهم با خواص درمانی فراوان می­باشد که در طب سنتی ایران جایگاه بسیار ویژه‌ای دارد. امروزه استفاده از زیست‌فنآوری در اصلاح و تکثیر گونه کُنار مرسوم شده و لازم است تا مراحل ریخت‌زایی با استفاده از ریزنمونه و ترکیبات هورمونی بهینه، بهبود داده شود. این مطالعه جهت بررسی عکس‌العمل ریزنمونه­های گره، ریشه، برگ و تأثیر ترکیبات مختلف هورمونی بر روی آنها از نظر پینه­زایی و باززایی گونه کُنار در سال 1391 در آزمایشگاه بیوتکنولوژی دانشگاه علوم کشاورزی و منابع طبیعی ساری انجام گرفت. برای پینه­زایی، سه ریزنمونه از گیاه سترون روی محیط­کشت MS حاوی  هورمون­های BAP,NAA، TDZ و 2,4-D (5/0، 0/1 و 0/2 میلی‌گرم در لیتر) کشت شدند. سپس پینه‌های تولید شده در محیط MS حاوی هورمون­های BAP، TDZ و Kin (0/1، 0/3 و 0/5 میلی‌گرم در لیتر)  قرار داده و برای ریشه­دار کردن گیاهچه­ها از محیط 1/2MS با هورمون­های IBA و NAA (5/0، 1 و 2 میلی‌گرم در لیتر) استفاده شد. آزمایشات به‌صورت فاکتوریل در قالب طرح کاملاً تصادفی در 4 تکرار انجام شدند. نتایج نشان داد که مقدار پینه­زایی برای ریزنمونه­ برگ نسبت به دو ریزنمونه­ دیگر، در محیط‌کشت حاوی 2,4-D با غلظت 5/0 میلی­گرم در لیتر (0±10) از همه بیشتر می­باشد. در صورتی‌که میزان باززایی پینه با منشاء گره تحت تأثیر تیمارهای 0/5 میلی­گرم در لیتر Kin (03/1± 84/8) و 0/1 میلی­گرم در لیتر TDZ ( 95/8± 08/1) نسبت به دو نوع پینه حاصل از ریزنمونه برگ و ریشه از همه بیشتر  بود (95 درصد). مقدار ریشه­زایی توسط هورمون IBA (6/90 درصد) در همه غلظت‌ها نسبت به هورمون NAA (58 درصد) بیشتر است. با توجه به نتایج ذکر شده کنار نسبت به کشت درون‌شیشه‌ای و ریزافزایی واکنش خوبی نشان می­دهد. از این نتایج می­توان برای اهداف مختلف از جمله تکثیر و تولید ماده مؤثره بیشتر در شرایط کشت درون‌شیشه­ای استفاده نمود.

کلیدواژه‌ها


عنوان مقاله [English]

The effect of plant growth regulators and explant types on callus induction and regeneration of Christ’s thorn (Ziziphus spina christti (L.) Willd)

نویسندگان [English]

  • elahe ahmadi 1
  • seyed mohammad hoseini nasr 2
  • hamid jalilvand 3
چکیده [English]

Christ’s thorn is one of important medicinal trees with many valued medicinal properties that in folklore medicinally have a special position in Iran. Nowadays, usage of biotechnology in breeding and proliferation of Christ’s thorn is prevalent and it is necessary to be improved the regeneration using optimize explants and hormone combinations. In order to evaluation the performance of node, leaf and root explants and the effect of different hormonal combinations on three phases of callus induction, regeneration and rooting of Christ’s thorn this study has been carried out in 2013 at biotechnology laboratory of Sari Agricultural Sciences and Natural Resources University. For callus induction, explants of aseptic seedlings were cultured on MS medium containing 2, 4-D, NAA, TDZ and BAP in three levels (0.5, 1, 2 mg/l). Then, produced calluses were taken on MS medium containing different levels of  BAP, TDZ and Kin (1, 3, 5 mg/l) and for rooting of seedlings 1/2MS medium containing IBA and NAA (0.5, 1 ,2 mg/l) were used. The treatments were arranged in factorial based on completely randomized design with four replications. The results showed that the highest rate of callogenesis was observed in leaf explants on MS medium containing 2, 4-D at concentrations of 0.5 mg/l (0 ± 10). While, the shoot regeneration obtained from callus produced on node explant by 5 mg/l Kin (10.03± 8.84) and 1mg/l TDZ (1.08±8.95) better as regards to callus produced on leaf and root explants. The rooting rate produced by IBA (90.6%) better than NAA (58%) at all concentrations of the hormone. According to the mentioned results Christ’s thorn than in vitro culture and regeneration show good response. The results can be used for various purposes, including reproduction and production of active ingredient in vitro culture.

کلیدواژه‌ها [English]

  • Auxin
  • Christ’s thorn
  • Callogenesis
  • medium
  • Regeneration
Al-Sulaiman, M. A. and Barakat, M. N. 2010. In vitro shoot multiplication of Ziziphus spina-christi by shoot tip culture. African Journal Biotechenology, 9: 850-857.

Aea, M., Rha, E. W. and Ma, H. 1998. Batanouny atanouny, phenology, germination and propagation of some wild trees and shrubs in south sinai, Egypt. Egypt Journal Botany, 36: 91-107.

Abass, N. H. 2010. Rapid in vitro multiplication and ex vitro rooting of Ziziphus mauritiana. Journal Applied Science Research, 6(1): 63-69.

Al-Wasel, A. S. 2000. Micropropagation of Acacia seyal Del. in vitro. Journal of Arid Environment, 46: 425-431.

Ahmad, N., Faisal, M., Anis, M. and Aref, I. F. 2010. In vitro callus induction and plant regeneration from leaf explants of Ruta graveolens L. South African Journal Botany, 76: 597-600.

Arefi, H., Norouzi, M. and Bagheri, N. A. 2003. An investigation of callus induction and regeneration of rice genotypes (Oryza sativa L.) by anther culture. Iran Journal Agricalture Science, 35: 293-299.

Assareh, M.H. 2009. Biological characteristics of trees along in Iran and introduction of other species Ziziphus. Research Institute of Forests and Rangelands, publisher, Tehran, 571 p.

Assareh, M. H. and Sardabi, H. 2005. Macropropagation and micropropagation of Ziziphus spina-christi. Pesqua Agropec Brasalia, 5: 459-465.

Asnaashari, M., Zokaee khosrashahi, M. H. 2011. Comprehensive Guide for Plant Tissue Culture. Boalsina publisher, Haamedan, 469 p.

Castillo, A. M., Egana, B., Sanz, J. M. and Cistue, L. 1998. Somatic embryogenesis and plant regeneration from barley cultivars grown in Spain. Plant Cell Reports, 17: 902-906.

Chitra, S. and Anander Kumar, C. R. 2006. Plant regeneration from scutellum-derived callus of Assam rice collection (Oryza sativa L.). Madras Agricallture Journal, 93: 73-75.

Chen, J. T. and Chang, W. C. 2000. Efficient plant regeneration through somatic embryogenesis from callus cultures of Oncidium (Orchidaceae). Plant Science, 160: 87-93.

Chenghou, W., Yehua, H.,  BixiaL, X. and  Fangming, H. 2004. In vitro Culture of Jujube Stem Segment. Journal of Fruit Science DOI: cnki: ISSN: 1001-7364.0.2004-06-023.

Dalal, N. V. and Rai, V. R. 2004. In vitro propagation of Oroxylum indicum Vent. a medicinally important forest tree. Journal forest Research, 9: 61-65.

Gomez-Leyva, J. F., Martinez-Acosta, L. A., Lopez-Muraira, I.G., Silos-Espino, H., Ramirez-Cervantes, F. and Andrade-Gonzalez, I. 2008. Multiple shoot regeneration of roselle (Hibiscus sabdariffa L.) from a shoot apex culture system. International Journal Botany, 4: 326-330.

Gomes, F., Simoes, M., Lopes, M.L. and Canhoto, J.M. 2010. Effect of plant growth regulators and genotype on the micropropagation of adult trees of Arbutus unedo L. (strawberry). New Biotechnology, 27: 882-892.

Guo, D. P., Zhu, Z. J., Hu, X. X. and Zheng, S. J. 2005. Effect of cytokinins on shoot regeneration fromcotyledon and leaf segment of stemmustard (Brassica juncea var. tsatsai). Plant Cell Tissue Organ Culture, 83: 123-127.

Hare, P. D. and Van staden, J. 1994. Inhibitory effect of thidiazuron on the activity of cytokinin oxidase isolated fromsoybean callus. Plant cell physiology, 35: 1121-1125.

Hasandokht, M. and Ebrahimi, R. 2009. Basics of Plant Tissue Culture. Marze danesh publisher, Tehran, 328 p.

Kumar, N., Anand, K. G. V. and Reddy, M. P. 2011. In vitro regeneration from petiole explants of non-toxic Jatropha curcas. Industrial Crops Production, 33: 146-151.

Karami, O. 2008. Induction of Embryogenic Callus and Plant Regeneration in Carnation (Dianthus caryophyllus L.). On Line Journal Biology Science, (4): 68-72.

Khalafalla, M. M., Elaleem, K. G. and Modawi, R.S. 2010. Callus formation and organogenesis of potato (solanum tuberosum l.) Cultivar almera. Journal of Phytology, 2(5): 40-46.

Lu, M.C. 2002. Micropropagation of Morus latifolia Poilet using axillary buds from mature trees. Scienctia Horticulture, 96: 329-34.

Maliti, C. M., Basile, D. V. and Corpe, W. A. 2005. Effects of Methylobacterium spp. Strains on rice Oryza sativa L. callus induction, plantlet regeneration, and seedings growth. In vitro Journal Torrey Botany Society, 132: 355-367.

Marques, N. T., Nolasco, G. B. and Leitão, J. P. 2011. Factors affecting in vitro adventitious shoot formation on internode explants of Citrus aurantium L. cv.Brazilian. Science Horticalture, 129: 176-182.

Machur, N., Ramawat, K. G. and Sonie, K. C. 1993. In vitro propagation of Ziziphus nummularia. Annals of Arid Zone, 32(4): 219-222.

Mungole, A., Awati, R., Dey, S., Chaturvedi, A. and Zanwar, P. 2009. In-vitro callus induction and shoot regeneration in Ipomoea obscura (L.): potent Indian medicinal plant. Indian Journal of Science and Technology, 2: 24-26.  

Mamun, A. N. K., Islam, R., Reza, M. A. and Joadar, O. I. 1996. In vitrodifferentiation of plantlet of tissue culture of Samonea saman. Plant Tissue Cullture, 6: 1-5.

Mansor far, K. 2011. Advanced statistical methods with the computer program. Tehran University publisher, Tehran. 459 p.

Meiners, J., Schwab, M. and Szankowski, I. 2007. Efficient in vitro regeneration systems for Vaccinium species. Plant Cell Tissue Organ Cullture, 89:169-176.

Murthy, B. N. S., Murch, S. J. and Saxena, P. K. 1998. Thidiazuron: a potent regulator of in vitro plant morphogenesis. In vitro Cell development Biology-plant, 34: 267-275.

Naik, S. K., Pattnaik, S. and Chand, P. K. 1999. In vitro propagation of pomegranate (Punica granatum L. cv. Ganesh) through axillary shoot proliferation from nodal segments of mature tree. Scienctia Horticullture, 79: 175-183.

Nagori, R. and Purohit, S.D. 2004. In vitro plantlet regeneration in Annona squamosa through direct shoot bud differentiation on hypocotyl segments. Scientia Horticulturae, 99: 89-98.

Onofrio, C. D. and Morini, S. 2006. Somatic embryo, adventitious root and shoot regeneration in in vitro grown quince leaves as influenced by treatments of different length with growth regulators. Scientia Horticulturae, 107: 194-199.

Qu, L., James, P. and Nicholi, V. 2000. A highly efficient in vitro cranberry regeneration system using leaf explants. Horticullture Science, 35: 948-952.

Rathore, J. S., Rathore, M. S., Singh, M., Singh, R. P. and Shekhawat, N. S. 2007. Micropropagation of mature tree of Citrus limon. Indian journal of biotechnology, 6: 239-244.

Sadeghi, S. M. and Farar, N. 2002. Konar, value and user. Promote management and popular participation of Agriculture jahad Organization of the Bushehr. Journal promo 20 p.

Sanatombi, K. and Sharma, G. J. 2007. Micropropagation of Capsicum frutescens L. using axillary shoot explants. Scienctia Horticulture, 113: 96-99.

Singh, J. and Tiwari, K. N. 2010. High-frequency in vitro multiplication system for commercial propagation of pharmaceutically important Clitoria ternatea L.-A valuable medicinal plant. Industrial Crops and Products, 32: 534-538.

Sharma, S., Kumar, N. and Reddy, M. P. 2011. Regeneration in Jatropha curcas: Factors affecting the efficiency of in vitro regeneration. Industrial Crops Production, 34: 943-951.

Shrivastava, V. and Kant. T. 2010. Micropropagation of Pongamia pinnata (L.). Pierre- a Native Indian Biodiesel Tree from Cotyledonary Node. International Journal of Biotechnology and Biochemistry, 4: 555-560.

Sohail, M., Saied, A. S., Gebauer, J. and Buerkert, A. 2009. Effect of NaCl salinity on growth and mineral composition of Ziziphus spina-christi (L.) Willd. Journal Agriculture Rural Development Tropic Subtropic, 110:107-114.

Sudhersan, C. and Hussain, J. 2003. In vitro Clonal Propagation of a Multipurpose Tree, Ziziphus spina christi(L.) Desf. Turkish Journal of Botany 27:167-171.

Tian, D., Tilt, K. M., Dane, F., Woods, F. M. and Sibley, J. L. 2010. Comparison of shoot induction ability of different explants in herbaceous peony (Paeonia lactiflora Pall.) Daike. Scientia Horticullture, 123: 385-389.

Vijaya Chitra, D. S. and Padmaja, G. 1999. Clonal propagation of mulberry (Morus indica L. cultivar M-5) through in vitro culture of nodal explants. Scienctia Horticullture, 80: 289-298.

Xiaojia, G., Zhaohui, C., Yongjun, L. and Shiping, W. 2006. A tissue culture system for different germplasms of indica rice. Plant Cell Reports 25: 392-402.

Yan, M. M., Xu, C., Kim, C. H., Um, Y. C., Bah, A. A. and Guo, D. P. 2009. Effects of explant type, culture media and growth regulators on callus induction and plant regeneration of Chinese jiaotou (Allium chinense). Scientia Horticulturae, 123: 124-128

Zhang, J. R. and Gu, X. F. 2005. An efficient adventitious shoot regeneration system for Zhanhua winter jujube (Zizyphus jujuba Mill.) using leaf explants. Plant Cell Reports, 23:775-779.