Structural analysis of hairy and non transgenic roots of Luffa cylindrica

Subject Areas : Developmental biology of plants and animals , development and differentiation in microorganisms

leila Ghazizadeh ¹ , Parastoo Ehsani ² , alireza iranbakhsh ³

1 - cell bank,pasteur institute of IRAN,Tehran
2 - Molecular Biology, Pasteur Institute of Iran
3 - Department of Biology and Science and Research Branch, Islamic Azad University, Tehran, IRAN

Received: 2018-02-27 Accepted : 2018-07-01 Published : 2018-05-22

Keywords: Agrobactrium rhizogenes, Luffin, RIP,

Abstract :

In this study, the hairy roots of the Luffacylindrica, a medicinal plant, were obtained following co cultivation of plant tissue with three different strains of Agrobacterium rhizogenes named 1583, 9534 and A4. The results indicated an increase in biomass in the hairy roots induced by agrobacteria strains 15834 and 9534. After transplantation of transgenic and non-transgenic roots, the surface area of the layers of the epidermis, cortex and vascular tissues, as well as the size of the parenchymal cells were analyzed using ImageJ software. Results showed that vascular system in transgenic roots, is more compact than in the control plant. So that the size of the xylems of hairy roots are higher than the control plant sample and is as following: 15834(393.5µm)>A4(324.6 µm)>9534(238.8 µm)>ctrl(175 µm). Moreover, pheloem region in the control plant is 2.6 times larger than 15834 produced hairy root which contain the highest surface of pheloem among transgenic species. The surface of cortex layer, the thickness of the epidermis and the parenchymal cells were also 1.67, 1.154 and 1.8 times larger in the 15834 hairy roots than control roots respectively. The results of this study confirmed the significant effect of Agrobactrium rhizogenes strains on increasing biomass. Therefore, by selecting the suitable strain, the amount of biomass due to enlargement of reservoir compartments such as parenchyma could be modified. Thus the amount of metabolites in the transgenic plant such as Luffacylindrica hairy roots could be optimized.

References:

[1]     افسانه صمدی، مراد جعفری^*، ناصر عباسپور، محمدرضا دینی ترکمانی، 1393، القا و
بهینه­سازی شرایط رشد ریشه‌های موئین گیاه سنبل الطیب(ValerianaofficinalisL). حاصل از تلقیح آگروباکتریوم رایزوژنز (Agrobacteriumrhizogenes)، مجله سلول و بافت، 23-30.

[2]     پورخسروانی محسن، ولی عباسعلی، مسعود معیری. 1388. بررسی ارتباط مورفولوژی گیاهی با خصوصیات مورفومتری گونه (نبکاهای روماریا تورسستانیکا). پژوهش­های جغرافیای طبیعی. 99-113.

[3]     حسینی.1391. کاربرد بیوتکنولوژی در افزایش بهره وری گیاهان دارویی.

[4]     زمانزاده زهرا، احسانپور علی­اکبر، امینی فریبا، 1392، بررسی میزان اکسین در گیاهان باززایی شده از ریشه­های تراریخت تنباکو (Nicotianatabacum)، مجله علمی پژوهشی سلول و بافت، جلد 1، شماره 2، 1-7.

[5]     سارا کبیرنتاج، الناز قطبی­راوندی^*، فرخنده رضانژاد و بهزاد شاهین کلیبر. 1392. تأثیر سویه­های مختلف اگروباکتریوم رایزوژنزبر میزان بیوسنتز ترکیبات فنولی و کلروژنیک اسید در ریشه­های موئین گیاه کاسنی. مجله علمی پژوهشی زیست فناوری گیاهان زراعی، 69-79.

[6]     سید طباطبایی ابراهیم، 1394، کشت بافت و سلول گیاهی، انتشارات دانشگاه تهران.

[7]     مظفریان، ولی ا....1391. کتاب شناخت گیاهان دارویی و معطرایران. انتشارات فرهنگ معاصر.

[8]     مظفریان، ولی ا... . 1373. کتاب فرهنگ نام­های گیاهان دارویی ایران. چاپ چهارم. انتشارات فرهنگ معاصر.

[9]     Baiza AMet al, 2000, Geneticstability of hairy root culturs of Daturastramonium. Plant cell tissue and org culture. 59: 9-17.

[10] Bevan MW, Chilton MD, 1982, T-DNA of the Agrobacterium Ti and Ri plasmids, Annu Rev Genet., 16: 357-84.

[11] BłażejewskaK et al, 2017, Mature Luffa Leaves (Luffacylindrica L.) as a Tool for Gene Expression Analysis by Agroinfiltration, Front Plant Sc, 21(8): 228.

[12] Gururaja HB et al, 2006, Agrobactriumrhizagenes mediated genetic transformation resulting in hairy root formation is inhanced by ultrasonication and acetosyringonetreatment.Electronic J. of Biotech. 9 (4): 349-357.

[13] Gurusamy PD, Schäfer H, Ramamoorthy S, 2o17, Biologically active recombinant humanerythropoietinexpressed in hairy root cultures and regenerated plantlets of Nicotianatabacum L. PLoS One, 12 (8): e018236.

[14] Iranbakhsh A.R, Oshaghi M. A, Ebadi M, 2007, Growth and Production Optimization of Tropane Alkaloids in Daturastramonium

 

Cell Suspension Culture,Pakistan Journal of Biological Science, 10 (8): 1236-1242.

[15] Liling Liu1, Rupeng Wang, Wei He1, Fengtian He2, and Gang Huang ,2010, Cloning and soluble expression of mature a-luffin from Luffacylindrica and its antitumor activities in vitro, ActaBiochimBiophys, 42: 585–592.

[16] Liu. L,. Wang. R, He. W, F. He, and G. Huang, 2010, "Cloning and soluble expression of mature α-luffin from Luffacylindrica and its antitumor activities in vitro," ActaBiochimBiophys Sin, vol. 42, pp. 585-592.

[17] Luigi sanitadiToppi, Paola Gorini, et al, 1996, Production of ribosome-inactivating protein from hairy root culture of  Luffacylindrica (L.) Roem.Plant cell reports. 15: 910-913.

[18] Oboh, O. and Aluyor, E. O, 2009, Luffa Cylindrica an emerging cash crop. African J of Agricultrual.Res 4(8), 684-688.

[19] Omosum G et al, 2008, Growthandanatomy of Amarantthushybridus as affected by different crude oil concentrations. Am-Eurasian J Sci, 3(1):70-74.

[20] Skoog F, Murashig T, 1962, A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures, Physiologia Plantarum Volume 15, Issue 3, Pages 473–497

[21] Srivastava S1, Srivastava AK, 2007, Hairy root culture for mass-production of high-value secondary metabolites, Crit Rev Biotechnol, Jan-Mar;27(1):29-43.

[22] Sutharshana, 2013, Protectiv Role of Luffa Cylindrica. J.Pharm, Sci. & Res. Vol. 5(9), 184-186.

[23] Vivanca JM, Guimaraes RL, Flores HE, 2002, The biosynthetic potential of roots, Underground plant metabolism, pp 1045-1064