Establishment of Gypsophila paniculata root culture for biomass, saponin, and flavonoid production

Authors

  • Ahmed A. ELATEEQ Northeast Forestry University, Key Laboratory of Forest Plant Ecology, Ministry of Education, Harbin 150040; Al-Azhar University, Faculty of Agriculture, Horticulture Department, Nasr City, Cairo (EG) https://orcid.org/0000-0002-9417-2684
  • Ahmed M.M. GABR Department of Plant Biotechnology, Biotechnology Research Institute, National Research Centre (NRC), Cairo, 12622 (EG)
  • Aisha M. ABDELKAWY Al-Azhar University, Faculty of Science (Girls Branch), Botany and Microbiology Department, Nasr City, Cairo (EG)
  • Nabil M. TOAIMA Al-Azhar University, Faculty of Agriculture, Horticulture Department, Nasr City, Cairo (EG)
  • Hussein A. BOSILA Al-Azhar University, Faculty of Agriculture, Horticulture Department, Nasr City, Cairo (EG)
  • Mostafa M. ZARAD Al-Azhar University, Faculty of Agriculture, Horticulture Department, Nasr City, Cairo (EG)
  • Hanan S. EBRAHIM Department of Plant Biotechnology, Biotechnology Research Institute, National Research Centre (NRC), Cairo, 12622 (EG)
  • Jiao JIAO Northeast Forestry University, Key Laboratory of Forest Plant Ecology, Ministry of Education, Harbin 150040 (CN)
  • Hongyi PAN Northeast Forestry University, Key Laboratory of Forest Plant Ecology, Ministry of Education, Harbin 150040 (CN)
  • Shakir ULLAH Northeast Forestry University, Key Laboratory of Forest Plant Ecology, Ministry of Education, Harbin 150040 (CN)
  • Yujie FU Northeast Forestry University, Key Laboratory of Forest Plant Ecology, Ministry of Education, Harbin 150040; Beijing Forestry University, The College of Forestry, Beijing 100083 (CN)

DOI:

https://doi.org/10.15835/nbha50312886

Keywords:

adventitious roots, baby's breath, DPPH, flavonoid, hairy root, saponin

Abstract

Baby’s breath (Gypsophila paniculata L.) roots are valuable as pharmaceuticals due to the content of triterpenoid saponins and other bioactive phytochemicals. However, the long root harvest period and fluctuation of these constituent’s content are among the constraints to traditional agricultural production. The present study shows an efficient and reliable liquid root culture of G. paniculata cv. ‘Perfecta’ established in a shake flask system using different auxins, media, and sugars. Hairy root (HR) induction in G. paniculata through Rhizobium rhizogenes-mediated transformation was also investigated. α-Naphthaleneacetic acid (NAA) was optimized for adventitious roots (AR) biomass, saponin, and flavonoid production at 1 or 2 mg/L compared to IBA. Full strength Gamborg’s medium (B5) recorded higher saponin content; however, the highest yield of total saponin and total flavonoids was achieved by full strength Murashige and Skoog's (MS) medium. Sucrose was more essential for root growth and accumulated total saponins and flavonoids rather than fructose and glucose. Polymerase chain reaction (PCR) analysis showed that G. paniculata HR carried rolC gene of R. rhizogenes A4 strain in its genome but not virD2 gene. Compared to non-transformed root, saponin content of leaf and stem-derived HR was 2.7 and 2.3-fold, while total flavonoid content was 2.1 and 2.0-fold, respectively. The DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity was also higher in HR extracts than in non-transgenic roots. This study established an efficient protocol for G. paniculata root cultures for sustainable production of important natural saponins and flavonoids.

References

Ataslar E, Guloren OT, Filik-Iscen C, Ilhan S (2019). In vitro antimicrobial and antioxidant activities of four Gypsophila L. species plant extracts. Fresenius Environmental Bulletin 28(3):1841-1851.

Bulgakov VP, Shkryl YN, Veremeichik GN, Gorpenchenko TY, Vereshchagina YV (2013). Recent advances in the understanding of Agrobacterium rhizogenes-derived genes and their effects on stress resistance and plant metabolism. In: Doran P (Ed). Biotechnology of Hairy Root Systems. Advances in Biochemical Engineering/Biotechnology. vol 134. Springer, Berlin, Heidelberg, pp 1-22. https://doi.org/10.1007/10_2013_179

Chang C, Yang M, Wen H, Chern J (2002). Estimation of total flavonoid content in Propolis by two complementary colorimetric methods. Journal of Food and Drug Analysis 10(3):178-182. https://doi.org/10.38212/2224-6614.2748

Cordeiro LS, Simões-Gurgel C, Albarello N (2015). Multiplication and cryopreservation of adventitious roots of Cleome rosea Vahl. In Vitro Cellular & Developmental Biology-Plant 51:249-257. https://doi.org/10.1007/s11627-015-9688-7

Cui XH, Murthy HN, Zhang JD, Song HL, Jiang YJ, Qi WW, Li YY, Paek KY, Park SY (2020). Effect of nutritional factors on the accretion of secondary metabolites in Malaysian ginseng adventitious root cultures. Plant Biotechnology Reports 14:381-386. https://doi.org/10.1007/s11816-019-00592-7

Dowom AS, Abrishamchi P, Radjabian T, Salami SA (2022). Elicitor-induced phenolic acids accumulation in Salvia virgata Jacq. hairy root cultures. Plant Cell, Tissue and Organ Culture 148:107-117. https://doi.org/10.1007/s11240-021-02170-8

Doyle JJ, Doyle JL (1987). A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin 19(1):11-15. https://worldveg.tind.io/record/33886/

Duncan DB (1995). Multiple range and multiple F tests. Biometrics 11:1-42. https://doi.org/10.2307/3001478

Elateeq AA, Saad Z, Eissa M, Ullah S (2021). Effect of chitosan and light conditions on the production of callus biomass, total flavonoids and total phenolics in Ginkgo biloba L. Al-Azhar Journal of Agricultural Research 46(1):28-42. https://doi.org/10.21608/AJAR.2021.218197

Elateeq AA, Sun Y, Nxumalo W, Gabr AM (2020). Biotechnological production of silymarin in Silybum marianum L.: A review. Biocatalysis and Agricultural Biotechnology 29:101775. https://doi.org/10.1016/j.bcab.2020.101775

Elshahawy OAM, Zeawail ME-F, Hamza MA, Elateeq AA, Omar MA (2022). Improving the production of total phenolics and flavonoids and the antioxidant capacity of Echinacea purpurea callus through biotic elicitation. Egyptian Journal of Chemistry 65(12):137-149. https://doi.org/10.21608/EJCHEM.2022.145210.6328

Folkes LK, Wardman P (2001). Oxidative activation of indole-3-acetic acids to cytotoxic species–a potential new role for plant auxins in cancer therapy. Biochemical Pharmacology 61(2):129-136. https://doi.org/10.1016/S0006-2952(00)00498-6

Fulcheri C, Morard P, Henry M (1998). Stimulation of the growth and the triterpenoid saponin accumulation of Saponaria officinalis cell and Gypsophila paniculata root suspension cultures by improvement of the mineral composition of the media. Journal of Agricultural and Food Chemistry 46(5):2055-2061. https://doi.org/10.1021/jf970627+

Gabr AMM, Mabrok HB, Abdel-Rahimd EA, El-Bahra MK, Smetanskab I (2018). Determination of lignans, phenolic acids and antioxidant capacity in transformed hairy root culture of Linum usitatissimum. Natural Product Research 32(15):1867-1871. https://doi.org/10.1080/14786419.2017.1405405

Gabr AMM, Mabrok HB, Ghanem KZ, Blaut M, Smetanska I (2016). Lignan accumulation in callus and Agrobacterium rhizogenes-mediated hairy root cultures of flax (Linum usitatissimum). Plant Cell, Tissue and Organ Culture 126(2):255-267. https://doi.org/10.1007/s11240-016-0995-4

Gabr AMM, Sytar O, Ghareeb H, Brestic M (2019). Accumulation of amino acids and flavonoids in hairy root cultures of common buckwheat (Fagopyrum esculentum). Physiology and Molecular Biology of Plants 25(3):787-797. https://doi.org/10.1007/s12298-019-00669-1

Gai QY, Jiao J, Luo M, Wei ZF, Zu YG, Ma W, Fu YJ (2015). Establishment of hairy root cultures by Agrobacterium rhizogenes mediated transformation of Isatis tinctoria L. for the efficient production of flavonoids and evaluation of antioxidant activities. Plos One 10(3):e0119022. https://doi.org/10.1371/journal.pone.0119022

Gamborg O, Miller R, Ojima K (1968). Nutrient requirements of suspension cultures of soybean root cells. Experimental Cell Research 50(1):151-158. https://doi.org/10.1016/0014-4827(68)90403-5

Gehlot A, Chaudhary N, Devi J, Joshi R, Kumar D, Bhushan S (2022). Induction and submerged cultivation of Valeriana jatamansi adventitious root cultures for production of valerenic acids and its derivatives. Plant Cell, Tissue and Organ Culture 148:347-361. https://doi.org/10.1007/s11240-021-02193-1

Gevrenova R, Joubert O, Mandova T, Zaiou M, Chapleur Y, Henry M (2014). Cytotoxic effects of four Caryophyllaceae species extracts on macrophage cell lines. Pharmaceutical Biology 52(7):919-925. https://doi.org/10.3109/13880209.2013.868492

Gevrenova R, Stancheva T, Voynikov Y, Laurain-Mattar D, Henry M (2010). Root in vitro cultures of six Gypsophila species and their saponin contents. Enzyme and Microbial Technology 47(3):97-104. https://doi.org/10.1016/j.enzmictec.2010.05.007

Hanafy MS, Abou-Setta LM (2007). Saponins production in shoot and callus cultures of Gypsophila paniculata. Journal of Applied Sciences Research 3(10):1045-1049.

Henry M (1993). Gypsophila paniculata L. (Baby’s Breath): in vitro culture and the production of gypsogenin saponins. In: Bajaj YPS (Ed). Biotechnology in Agriculture and Forestry. Vol. 21. Medicinal and Aromatic Plants IV. Springer-Verlag Berlin Heidelberg, pp 187-206. https://doi.org/10.1007/978-3-642-77004-3_13

Henry M, Rochd M, Bennini B (1991). Biosynthesis and accumulation of saponins in Gypsophila paniculata. Phytochemistry 30(6):1819-1821. https://doi.org/10.1016/0031-9422(91)85020-Z

Herold MC, Henry M (2001). UDP-glucuronosyltransferase activity is correlated to saponin production in Gypsophila paniculata root in vitro cultures. Biotechnology Letters 23:335-337. https://doi.org/10.1023/A:1005689104107

Ho TT, Lee JD, Jeong CS, Paek KY, Park SY (2018). Improvement of biosynthesis and accumulation of bioactive compounds by elicitation in adventitious root cultures of Polygonum multiflorum. Applied Microbiology and Biotechnology 102(1):199-209. https://doi.org/10.1007/s00253-017-8629-2

Hossain MS, Hossain MM, Hossain T, Haque MM, Zakaria M, Sarkar MD (2017). Varietal performance of potato on induction and development of microtuber in response to sucrose. Annals of Agricultural Sciences 62(1):75-81. https://doi.org/10.1016/j.aoas.2017.05.002

Jiang YJ, Piao XC, Liu JS, Jiang J, Lian ZX, Kim MJ, Lian ML (2015). Bioactive compound production by adventitious root culture of Oplopanax elatus in balloon-type airlift bioreactor systems and bioactivity property. Plant Cell, Tissue and Organ Culture 123:413-425. https://doi.org/10.1007/s11240-015-0845-9

Jiao J, Gai QY, Fu YJ, Ma W, Peng X, Tan SN, Efferth T (2014). Efficient production of isoflavonoids by Astragalus membranaceus hairy root cultures and evaluation of antioxidant activities of extracts. Journal of Agricultural and Food Chemistry (62):12649-12658. https://doi.org/10.1021/acs.jafc.0c02600

Jiao J, Gai QY, Wang X, Liu J, Lu Y, Wang ZY, Xu XJ, Fu YJ (2020). Effective production of phenolic compounds with health benefits in pigeon pea [Cajanus cajan (L.) Millsp.] hairy root cultures. Journal of Agricultural and Food Chemistry 68(31):8350-8361. https://doi.org/10.1021/acs.jafc.0c02600

Kołodziej B, Sęczyk Ł, Sugier D, Kędzia B, Chernetskyy M, Gevrenova R, Henry M (2019). Determination of the yield, saponin content and profile, antimicrobial and antioxidant activities of three Gypsophila species. Industrial Crops and Products 138:111422. https://doi.org/10.1016/j.indcrop.2019.05.071

Korchowiec B, Korchowiec J, Kwiecińska K, Gevrenova R, Bouguet-Bonnet S, Deng C, Henry M, Rogalska E (2022). The molecular bases of the interaction between a saponin from the roots of Gypsophila paniculata L. and model lipid membranes. International Journal of Molecular Sciences 23(6):3397. https://doi.org/10.3390/ijms23063397

Krstić-Milošević D, Janković T, Vinterhalter B, Menković N, Aljančić I, Vinterhalter D (2013). Influence of carbohydrate source on xanthone content in root cultures of Gentiana dinarica Beck. Plant Growth Regullation 71:147-155. https://doi.org/10.1007/s10725-013-9815-6

Kumar SS, Manoj P, Giridhar P (2015). Optimization of an in vitro protocol for the production of ascorbic acid in Hibiscus cannabinus leaf-derived normal root cultures. EurAsian Journal of BioSciences 9:38-45. http://dx.doi.org/10.5053/ejobios.2015.9.0.5

Lee YS, Yang TJ, Park SU, Baek JH, Wu SQ, Lim KB (2011). Induction and proliferation of adventitious roots from Aloe vera leaf tissues for in vitro production of aloe-emodin. Plant Omics Journal 4(4):190-194. https://search.informit.org/doi/10.3316/informit.307923778436899

Li F, Gao Y, Jin C, Wen X, Geng H, Cheng Y, Qu H, Liu X, Feng S, Zhang F, Ruan J, Yang C, Zhang L, Wang J (2022). The chromosome-level genome of Gypsophila paniculata reveals the molecular mechanism of floral development and ethylene insensitivity. Horticulture Research uhac176. https://doi.org/10.1093/hr/uhac176

Li F, Mo X, Wu L, Yang C (2020). A novel double-flowered cultivar of Gypsophila paniculata mutagenized by 60Co γ-Ray. HortScience 55(9):1531-1532. https://doi.org/10.21273/HORTSCI15137-20

Li J, Wang J, Li J, Li J, Liu S, Gao W (2016). Salicylic acid induces the change in the adventitious root of Glycyrrhiza uralensis Fisch.: bioactive compounds and antioxidant enzymes. Research on Chemical Intermediates 42:1503-1519. https://doi.org/10.1007/s11164-015-2099-x

Liao Y, Li Z, Zhou Q, Sheng M, Qu Q, Shi Y, Yang J, Lv L, Dai X, Shi X (2021). Saponin surfactants used in drug delivery systems: A new application for natural medicine components. International Journal of Pharmaceutics 603:120709. https://doi.org/10.1016/j.ijpharm.2021.120709

Madaan R, Banasal G, Kumar S, Sharma A, (2011). Estimation of total phenols and flavonoids in extracts of Actaea spicata roots and antioxidant activity studies. Indian Journal of Pharmaceutical Sciences 73(6):666-669. http://dx.doi.org/10.4103/0250-474X.100242

Matvieieva N, Shakhovsky A, Tashyreva H, Ratushnyak Y, Duplij V, Bohdanovych T, Kuchuk M (2022). Study of superoxide dismutase activity in long-term cultivated Artemisia and Althaea “hairy” roots. Current Microbiology, 79:14. https://doi.org/10.1007/s00284-021-02709-0

Medina-Bolivar F, Condori J, Rimando AM, Hubstenberger J, Shelton K, O’Keefe SF, Bennett S, Dolan MC (2007). Production and secretion of resveratrol in hairy root cultures of peanut. Phytochemistry 68(14):1992-2003. https://doi.org/10.1016/j.phytochem.2007.04.039

Murashige T, Skoog F (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15(3):473-497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x

Murthy HN, Dalawai D, Bhat MA, Dandin VS, Paek KY, Park SY (2021). Biotechnological production of useful phytochemicals from adventitious root cultures. In: Ramawat KG, Ekiert HM, Goyal S (Eds). Plant Cell and Tissue Differentiation and Secondary Metabolites. Reference Series in Phytochemistry. Springer, Cham. pp 469-485. https://doi.org/10.1007/978-3-030-30185-9_19

Nagella P, Murthy HN (2010). Establishment of cell suspension cultures of Withania somnifera for the production of withanolide A. Bioresource Technology 101(17):6735-6739. https://doi.org/10.1016/j.biortech.2010.03.078

Nasiri A, Rashidi-Monfared S, Ebrahimi A, Charkhabi NF, Moieni A (2022). Metabolic engineering of the diosgenin biosynthesis pathway in Trigonella foenum-graceum hairy root cultures. Plant Science 323:111410. https://doi.org/10.1016/j.plantsci.2022.111410

Pakdin Parizi A, Farsi M, Nematzadeh GA, Mirshamsi A (2015). Impact of different culture media on hairy roots growth of Valeriana officinalis L. Acta Agriculturae Slovenica 103(2):299-305. http://dx.doi.org/10.14720/aas.2014.103.2.14

Pandey P, Singh S, Negi AS, Banerjee S (2022a). Harnessing the versatility of diverse pentacyclic triterpenoid synthesis through hairy root cultures of various Ocimum species: An unprecedented account with molecular probing and up-scaling access. Industrial Crops and Products 177:114465. https://doi.org/10.1016/j.indcrop.2021.114465

Pandey S, Sundararajan S, Ramalingam S, Pant B (2022b). Elicitation and plant growth hormone-mediated adventitious root cultures for enhanced valepotriates accumulation in commercially important medicinal plant Valeriana jatamansi Jones. Acta Physiologiae Plantarum 44:4. https://doi.org/10.1007/s11738-021-03319-w

Pauthe-Dayde D, Rochd M, Henry M (1990). Triterpenoid saponin production in callus and multiple shoot cultures of Gypsophila spp. Phytochemistry 29(2):483-487. https://doi.org/10.1016/0031-9422(90)85101-K

Popova P, Zarev Y, Shkondrov A, Krasteva I, Ionkova I (2022). In vitro production of flavonoids in cultures of Gypsophila glomerata. Pharmacia, 69(1):107-111. https://doi.org/10.3897/pharmacia.69.e77769

Qian Y, Zhang S, Yao S, Xia J, Li Y, Dai X, Wang W, Jiang X, Liu Y, Li M, Gao L, Xia T, (2018). Effects of vitro sucrose on quality components of tea plants (Camellia sinensis) based on transcriptomic and metabolic analysis. BMC Plant Biology 18:121. https://doi.org/10.1186/s12870-018-1335-0

Reis RV, Borges APPL, Chierrito TPC, de Souto ER, de Souza LM, Iacomini M, de Oliveira AJB, Gonçalves RAC (2011). Establishment of adventitious root culture of Stevia rebaudiana Bertoni in a roller bottle system. Plant Cell, Tissue and Organ Culture 106:329-335. https://doi.org/10.1007/s11240-011-9925-7

Sahayarayan JJ, Udayakumar R, Arun M, Ganapathi A, Alwahibi MS, Aldosari NS, Morgan AM (2020). Effect of different Agrobacterium rhizogenes strains for in-vitro hairy root induction, total phenolic, flavonoids contents, antibacterial and antioxidant activity of (Cucumis anguria L.). Saudi Journal of Biological Sciences 27(11):2972-2979. https://doi.org/10.1016/j.sjbs.2020.08.050

Sarkar S, Ghosh I, Roychowdhury D, Jha S (2018). The effects of rol genes of Agrobacterium rhizogenes on morphogenesis and secondary metabolite accumulation in medicinal plants. In: Kumar N (Ed). Biotechnological Approaches for Medicinal and Aromatic Plants. Springer, Singapore, pp 27-51. https://doi.org/10.1007/978-981-13-0535-1_2

Sivanesan I, Jeong BR. (2009). Induction and establishment of adventitious and hairy root cultures of Plumbago zeylanica L. African Journal of Biotechnology 8(20):5294-5300. https://www.ajol.info/index.php/ajb/article/view/65963

Teale WD, Paponov IA, Palme K (2006). Auxin in action: signaling, transport and the control of plant growth and development. Nature Reviews Molecular Cell Biology 7:847-859. https://doi.org/10.1038/nrm2020

Toaima N, Bosila H, El-Ateeq AA (2016). In vitro growth regulators, gelling agents and sucrose levels affect micropropagation of Gypsophila paniculata L. Middle East Journal of Agriculture Research 5(3):313-323. http://www.curresweb.com/mejar/mejar_July-sep-2016.html

Tusevski O, Stanoeva JP, Stefova M, Spasenoski M, Simic SG (2019). State of antioxidant systems and phenolic compounds’ production in Hypericum perforatum L. hairy roots. Acta Physiologiae Plantarum 41:132. https://doi.org/10.1007/s11738-019-2919-5

Vettori L, Schiff S, Tani C, Pasqualetto P, Bennici A (2015) Morphological and cytological observations of wild species and hybrids of Gypsophila. Plant Biosystems 149(2):322-328. https://doi.org/10.1080/11263504.2013.824041

Wu CH, Tang J, Jin ZX, Wang M, Liu ZQ, Huang T, Lian ML (2018). Optimizing co-culture conditions of adventitious roots of Echinacea pallida and Echinacea purpurea in air-lift bioreactor systems. Biochemical Engineering Journal 132:206-216. https://doi.org/10.1016/j.bej.2018.01.024

Yin S, Gao W, Liang Y, Wang J, Liu H, Wei C, Zuo B (2013a). Influence of sucrose concentration and phosphate source on biomass and metabolite accumulation in adventitious roots of Pseudostellaria heterophylla. Acta Physiologiae Plantarum 35:1579-1585. https://doi.org/10.1007/s11738-012-1199-0

Yin S, Liang Y, Gao W, Wang J, Jing S, Zhang Y, Liu H (2013b). Influence of medium salt strength and nitrogen source on biomass and metabolite accumulation in adventitious root cultures of Pseudostellaria heterophylla. Acta Physiologiae Plantarum 35:2623-2628. https://doi.org/10.1007/s11738-013-1270-5

Zdraveva P, Pencheva I, Popova P, Ionkova I, Krasteva I (2015). Production of saponarin in in vitro cultures of Gypsophila species. Journal of Chemical and Pharmaceutical Research 7(1):829-832.

Zhang Z, Yu Z, Jin Z, Liu J, Li Y (2013). Liquid culture of adventitious roots is a potential alternative to field cultivation for Psammosilene tunicoides, a rare and endangered endemic medicinal plant. Advance Journal of Food Science and Technology 5(2):127-131. http://dx.doi.org/10.19026/ajfst.5.3231

Zhang ZS, Wang ZQ (2009). Effects of culture conditions on the saponin accumulation in Astragalus membranaceus callus. Journal of Dalian Polytechnic University 28:174-177.

Zheleznichenko T, Banaev E, Asbaganov S, Voronkova M, Kukushkina T, Filippova E, Mazurkova N, Shishkina L, Novikova T, (2018). Nitraria schoberi L. hairy root culture as a source of compounds with antiviral activity against influenza virus subtypes А(H5N1) and А(H3N2). 3 Biotech 8:260. https://doi.org/10.1007/s13205-018-1280-5

Zolfaghari F, Rashidi-Monfared S, Moieni A, Abedini D, Ebrahimi A (2020). Improving diosgenin production and its biosynthesis in Trigonella foenum-graecum L. hairy root cultures. Industrial Crops and Products 145:112075. https://doi.org/10.1016/j.indcrop.2019.112075

Downloads

Published

2022-12-06

How to Cite

ELATEEQ, A. A., GABR, A. M., ABDELKAWY, A. M., TOAIMA, N. M., BOSILA, H. A., ZARAD, M. M., EBRAHIM, H. S., JIAO, J., PAN, H., ULLAH, S., & FU , Y. (2022). Establishment of Gypsophila paniculata root culture for biomass, saponin, and flavonoid production. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 50(4), 12886. https://doi.org/10.15835/nbha50312886

Issue

Vol. 50 No. 4 (2022)

Section

Research Articles
CITATION
DOI: 10.15835/nbha50312886

License

Copyright (c) 2022 Ahmed A. ELATEEQ, Ahmed M.M. GABR, Aisha M. ABDELKAWY, Nabil M. TOAIMA, Hussein A. BOSILA, Mostafa M. ZARAD, Hanan S. EBRAHIM, Jiao JIAO, Hongyi PAN, Shakir ULLAH, Yujie FU

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

License:

© Articles by the authors; licensee UASVM and SHST, Cluj-Napoca, Romania. Papers published in the journal Notulae Botanicae Horti Agrobotanici Cluj-Napoca are open access distributed under the terms and conditions of the Creative Commons Attribution License (http://creativecommons.org/licenses).

Open Access Journal:

The journal allows the author(s) to retain publishing rights without restriction. Users are allowed to read, download, copy, distribute, print, search, or link to the full texts of the articles, or use them for any other lawful purpose, without asking prior permission from the publisher or the author.