بررسی کشت در شیشه (invitro) گیاه هندوانه ابوجهل Citrullus colocynthis (L.) Schrad
محورهای موضوعی : ژنتیکسورمه قرهماتروسیان 1 , مهلقا قربانلی 2
1 - گروه زیست شناسی، دانشگاه آ زاد اسلامی واحد گرگان
2 - گروه زیست شناسی، دانشگاه آ زاد اسلامی واحد گرگان
کلید واژه: کینتین, هندوانه ابوجهل, بنزیل آمینوپورین, IAA, اندامزایی, نفتالن استیک اسید,
چکیده مقاله :
در این پژوهش تعیین بهترین محیط کشت برای اندامزایی ساقهای و انتخاب مناسبترین جدا کشت از دانه رستهای هندوانه ابو جهل مورد بررسی قرار گرفته است. برای به دست آوردن دانه رستهای استریل بازدارندگی پوسته ای دانه در اسید سولفوریک غلیظ به مدت 20 دقیقه بر طرف شد. بعد از شستشو با آب مقطر استریل دانهها به محیط کشت استریل منتقل شدند. 20 روز بعد، از دانه رستها قطعات کوچک (لپه، جوانه انتهایی، محور زیر لپه و ریشه) تهیه و به محیط کشت پایه MS با تیمارهای هورمونی مختلف منتقل شدند. تمامی جدا کشتها تولید کالوس کردند و فقط جوانه انتهایی با تیمار IAA و کینتین برابر با 1 میلیگرم در رلیترتولید کالوس کرده سپس به جوانه و شاخه تمایز یافت. برای تعیین مناسبترین محیط کشت ؛ دو محیط کشت در نظر گرفته شد. محیط کشت پایه MS با تیمار IAA و کینتین برابر با 1 میلی گرم و محیط کشت پایه MS، ویتامین 2 برابر محیط پایه MS و تیمار بنزیل آمینو پورین 2 میلیگرم در لیتر به همراه نفتالن استیک اسید 1/0 میلیگرم در لیتر. جوانه انتهایی از محیط کشت استریل جدا شد و در این دو محیط کشت قرار گرفت. طرح بر پایه بلوک کاملاً تصادفی انجام شد. جمع آوری دادهها درچهار مرحله انجام شد. قابلیت تولید کالوس در محیط کشت با تیمار بنزیل آمینو پورین به همراه نفتالن استیک اسید نسبت به محیط کشت IAA و کینتین بیشتر بود. در محیط کشت با تیمار IAA و کینتین با مقادیر برابر شاخه زایی و افزایش طول شاخهها معنی دار بود. در محیط کشت با تیمار بنزیل آمینو پورین به همراه نفتالن استیک اسید سبب تمایز کالوس به شاخه شد ولی شاخهزایی و افزایش طول شاخهها معنیدار نبود.
In this research, the determination of the best cultivation environment for stem organogenesis and the selection of the most suitable cultivar from Abu Jahl watermelon seeds have been studied. In order to obtain sterile roasted seeds, the inhibition of the seed shell was removed in concentrated sulfuric acid for 20 minutes. After washing with sterile distilled water, the seeds were transferred to a sterile culture medium. 20 days later, small pieces (cotyledon, terminal bud, cotyledon axis and root) were prepared from the roasted seeds and transferred to MS medium with different hormone treatments. All isolated cultures produced callus, and only the terminal bud treated with IAA and quintine equal to 1 mg in relitertolide produced callus, then it was differentiated into bud and branch. To determine the most suitable cultivation environment; Two cultivation environments were considered. MS basic culture medium with IAA and quintine treatment equal to 1 mg and MS basic culture medium, vitamin 2 equal to MS basic medium and benzyl aminopurine treatment 2 mg/liter along with naphthalene acetic acid 0.1 mg/liter. The terminal bud was separated from the sterile culture medium and placed in these two culture mediums. The design was based on a completely random block. Data collection was done in four stages. The ability to produce callus in the culture medium treated with benzyl aminopurine along with naphthalene acetic acid was higher than in the culture medium of IAA and quintine. In the cultivation environment with IAA and quintine treatment with equal amounts, branching and increase in the length of the branches were significant. In the culture medium, treatment with benzyl aminopurine along with naphthalene acetic acid caused the differentiation of callus into branches, but branching and increasing the length of branches were not significant.
Chaturvedi.M., Mali.P.C., Ansari.A.S (2003) Induction of reversible antifertility with crude ethanol extract of Citrullus colocynthis Schard fruit in male rats. International Journal of experimental and clinical pharmacology; Vol. 68:38-48
Culbreth. D.M. R., Ph.G., M.D (1927) Colocynthis, Colocynthe, U.S.P. A manual of material medica and pharmacology.
Compton. M.E., Gray.D.J (1993) Shoot organogenesis and plant regeneration from cotyledons of diploid, triploid, and tetraploid watermelon. Journal of the American Society for Horticultural Science, V.118, P.151-157.
Diwan. F.H., Abdel-Hassan. I.A.,Mohammed. S.T (2000) Effect of saponin on mortality and histopathological changes in mic. Vol. 6, P.345-351.
Dong, J.Z.: JIA, S.R (1991). High efficiency plant regeneration from cotyledons of watermelon (Citrullus vulgaris Shrad.). Plant Cell Reports, v.9, p.559-562,
Duke.J.A (1983) Citrullus colocyntis (L.) Schrad. Hand book of Energy crops.
Grieve. M (2005) Apple (Bitter). htt//www. Botanical.com.
Krug. M.G.Z., Stipp. L.C.L., Rodriguez. A.P.M., Mendes. B.M.J (2005) Invitro organogenesis in watermelon cotyledons. Pesq.agropec.Brasilia., Vol.40,n.9,P.861-865.
Lee. C.W., Thomas. J.C. (1985) Tissue culture propagation of buffalo gourd. Hortscience 20:218-219.
Lorenz, P.R., Lippmann, F.L., Durrling. K., Solf. M., Geissler, J. (2005) Pharmaco-toxicological and clinical studies with colocyth pulp extracts (Extr. Colocynthidis fructus)., Arzneimittel-Forschung. 55(11): 621-63
Nmila. R., Gross. R., Rchid. H., Roye. M., Manteghetti. M., Petit. P., Tijane. M., Ribes. G., Sauvaire. Y (2000). Insulinotropic effect of Citrullus colocyntis fruit extracts.Planta med., Jun.,66(5):418-23.
Sarowar. S., Oh. H. Y., Hyung. N.I., Min.B.W., Harn. C.H., Yang. S.K.,Ok. S.H., Shin. J.S (2003) Invitro micropropagation of a cucurbita interspecific hybrid cultivar-a root stock plant cell,tissue,organ culture 75:179-182.
Sawaya. W.N., Daghir. N.J., Khalil, J.K (1986) Citrullus colocyntis seeds as a potential source of protein for food and feed.J.Aric.Foodchem.34.285-288.
Schlising. R.L (1993) Cucurbitaceae gourd family. Treatment from the Jepson manual.university of California press.
Souza. F.V.D., Garcia-Sogo. B., Souza. A.S., San-Juan. A.P., Moreno.V. (2006) Morphogenetic Response of cotyledon and leaf explants of melon (Cucumis melo L.) cv.Amarillo Oro. Brazilian archives of biology and technology. Vol.49. n. 1:PP.21-27.
Srivastava.D.R.,Andrianov.V.M.,Piruzian.E.S(1989) Tissue culture and plant regeneration of watermelon (Citrullus vulgaris Schrad.cv. Melitoplski).Plant cell reports.V. 8,P. 300-302.
Unnoun (2000) Cultivating support of medical plant stakehoiders. Traffic bulletin Vol. 18No.2
Unnoun (2005) Citrullus colocynthis (L.)Schrad. The western Australian flora. htt://flora base. calm. wa. gov.au/
Wehner. T.C., R.M.Cade and Locy. R.D. (1990) Cell,tissue,and organ culture techniques for genetic improvement of Cucurbits. Biol. Util.Cucurbitaceae.Cornell Univ. Press, Ithaca, NY, P. 367-381.
Yaniv. Z., Shabelsky. E and Schafferman. D (1999) Colocynth: Potential arid land oilseed from an ancient Cucurbit.P.257-261.
_||_
Chaturvedi.M., Mali.P.C., Ansari.A.S (2003) Induction of reversible antifertility with crude ethanol extract of Citrullus colocynthis Schard fruit in male rats. International Journal of experimental and clinical pharmacology; Vol. 68:38-48
Culbreth. D.M. R., Ph.G., M.D (1927) Colocynthis, Colocynthe, U.S.P. A manual of material medica and pharmacology.
Compton. M.E., Gray.D.J (1993) Shoot organogenesis and plant regeneration from cotyledons of diploid, triploid, and tetraploid watermelon. Journal of the American Society for Horticultural Science, V.118, P.151-157.
Diwan. F.H., Abdel-Hassan. I.A.,Mohammed. S.T (2000) Effect of saponin on mortality and histopathological changes in mic. Vol. 6, P.345-351.
Dong, J.Z.: JIA, S.R (1991). High efficiency plant regeneration from cotyledons of watermelon (Citrullus vulgaris Shrad.). Plant Cell Reports, v.9, p.559-562,
Duke.J.A (1983) Citrullus colocyntis (L.) Schrad. Hand book of Energy crops.
Grieve. M (2005) Apple (Bitter). htt//www. Botanical.com.
Krug. M.G.Z., Stipp. L.C.L., Rodriguez. A.P.M., Mendes. B.M.J (2005) Invitro organogenesis in watermelon cotyledons. Pesq.agropec.Brasilia., Vol.40,n.9,P.861-865.
Lee. C.W., Thomas. J.C. (1985) Tissue culture propagation of buffalo gourd. Hortscience 20:218-219.
Lorenz, P.R., Lippmann, F.L., Durrling. K., Solf. M., Geissler, J. (2005) Pharmaco-toxicological and clinical studies with colocyth pulp extracts (Extr. Colocynthidis fructus)., Arzneimittel-Forschung. 55(11): 621-63
Nmila. R., Gross. R., Rchid. H., Roye. M., Manteghetti. M., Petit. P., Tijane. M., Ribes. G., Sauvaire. Y (2000). Insulinotropic effect of Citrullus colocyntis fruit extracts.Planta med., Jun.,66(5):418-23.
Sarowar. S., Oh. H. Y., Hyung. N.I., Min.B.W., Harn. C.H., Yang. S.K.,Ok. S.H., Shin. J.S (2003) Invitro micropropagation of a cucurbita interspecific hybrid cultivar-a root stock plant cell,tissue,organ culture 75:179-182.
Sawaya. W.N., Daghir. N.J., Khalil, J.K (1986) Citrullus colocyntis seeds as a potential source of protein for food and feed.J.Aric.Foodchem.34.285-288.
Schlising. R.L (1993) Cucurbitaceae gourd family. Treatment from the Jepson manual.university of California press.
Souza. F.V.D., Garcia-Sogo. B., Souza. A.S., San-Juan. A.P., Moreno.V. (2006) Morphogenetic Response of cotyledon and leaf explants of melon (Cucumis melo L.) cv.Amarillo Oro. Brazilian archives of biology and technology. Vol.49. n. 1:PP.21-27.
Srivastava.D.R.,Andrianov.V.M.,Piruzian.E.S(1989) Tissue culture and plant regeneration of watermelon (Citrullus vulgaris Schrad.cv. Melitoplski).Plant cell reports.V. 8,P. 300-302.
Unnoun (2000) Cultivating support of medical plant stakehoiders. Traffic bulletin Vol. 18No.2
Unnoun (2005) Citrullus colocynthis (L.)Schrad. The western Australian flora. htt://flora base. calm. wa. gov.au/
Wehner. T.C., R.M.Cade and Locy. R.D. (1990) Cell,tissue,and organ culture techniques for genetic improvement of Cucurbits. Biol. Util.Cucurbitaceae.Cornell Univ. Press, Ithaca, NY, P. 367-381.
Yaniv. Z., Shabelsky. E and Schafferman. D (1999) Colocynth: Potential arid land oilseed from an ancient Cucurbit.P.257-261.