In Vitro Shoot Regeneration of Limnophila aromatica (Lamk.) Merr. from Nodal and Internodal Explants

Abstract

Limnophila aromatica (Lamk.) Merr. is an important medicinal and aromatic plant widely used in

the traditional medicine. In this study, nodal and internodal explants of L. aromatica (Lamk.) Merr. were cultured

for multiple and rapid production on Murashige and Skoog (MS) medium containing combinations of 0.10 mg

L-1 Gibberellic acid (GA3) and 0.05, 0.10, 0.20, 0.40, 0.80 and 1.60 mg L-1 Thidiazuron (TDZ) for eight weeks.

In general, high shoot regeneration percentages were obtained for both types of explants. In nodal explants, the

maximum number of shoots per explant (21.44) and the highest shoot length (1.68 cm) were determined in MS

medium containing 0.10 mg L-1 GA3 + 0.40 mg L-1 TDZ and 0.10 mg L-1 GA3 + 0.10 mg L-1 TDZ, respectively.

In internodal explants, the maximum number of shoots per explant (17.46) and the highest shoot length (1.60 cm)

were obtained in MS medium supplemented with 0.10 mg L-1 GA3 + 0.20 mg L-1 TDZ and 0.10 mg L-1 GA3 + 0.05

mg L-1 TDZ, respectively. With the effect of plant growth regulators, the nodal explants gave more regenerated

shoots than the internodal explants. Regenerated shoots were successfully rooted on MS medium containing 0.25

mg L-1 indole butyric acid (IBA). Successful adaptation of the rooted shoots to the aquarium environment has been

achieved.

Keywords

• Nodal explant,propagation,in vitro,Limnophila aromatica,TDZ

Limnophila aromatica (Lamk.) Merr.’nın Boğum ve Boğum Arası Eksplantlarından In Vitro Sürgün Rejenerasyonu

Abstract

Limnophila aromatica (Lamk.) Merr. geneneksel tıpta yaygın şekilde kullanılan önemli tıbbi ve aromatik

bir bitkidir. Bu çalışmada, L. aromatica’nın çoklu ve hızlı üretimi için boğum ve boğum arası eksplantları 0.10 mg

L-1 Gibberellik asit (GA3) ve 0.05, 0.10, 0.20, 0.40, 0.80 ve 1.60 mg L-1 Tidiazuron (TDZ) kombinasyonlarını içeren

Murashige ve Skoog (MS) besin ortamında sekiz hafta boyunca kültüre alınmıştır. Genel olarak her iki tip eksplant

için de yüksek sürgün rejenerasyon yüzdeleri elde edilmiştir. Boğum eksplantlarında eksplant başına sürgün sayısı

(21.44 adet) ve en uzun sürgünler (1.68 cm) sırasıyla 0.10 mg L-1 GA3 + 0.40 mg L-1 TDZ ve 0.10 mg L-1 GA3 +

0.10 mg L-1 TDZ içeren MS ortamında tespit edilmiştir. Boğum arası eksplantlarında, maksimum eksplant başına

sürgün sayısı (17.46 adet) ve en uzun sürgünler (1.60 cm) sırasıyla 0.10 mg L-1 GA3 + 0.20 mg L-1 TDZ ve 0.10

mg L-1 GA3 + 0.05 mg L-1 TDZ eklenmiş MS ortamında elde edilmiştir. Bitki büyüme düzenleyicilerinin etkisi ile

boğum eksplantı, boğumarası eksplantından daha fazla rejenere sürgün vermiştir. Rejenere sürgünler 0.25 mg L-1

indol butirik asit (IBA) içeren MS ortamında başarılı bir şekilde köklendirilmiştir. Köklenen sürgünlerin akvaryum

ortamına alıştırılması başarıyla sağlanmıştır.

Keywords

• Boğum eksplant,çoğaltım,in vitro,Limnophila aromatica,TDZ

References

1. Anis M, Ahmad N, 2016. Plant Tissue Culture: A Journey from Research to Commercialization. In: Anis M., Ahmad N. (eds) Plant Tissue Culture: Propagation, Conservation and Crop Improvement. Springer, Singapore.
2. Babaoğlu M, Yorgancılar M, Akbudak MA, 2001. Doku Kültürleri: Tarihçe ve Temel Teknikleri. Bitki Biyoteknolojisi I-Doku Kültürü ve Uygulamaları, Editörler: Babaoğlu, M., E. Gürel, ve S. Özcan, Selçuk Üniversitesi Vakfı Yayınları, Konya. 1-35 p.
3. Brahmachari G, 2014. Limnophila (Scrophulariaceae): Chemical and Pharmaceutical Aspects - An Update. The Open Natural Products Journal, 7: 1-14.
4. Bui ML, Grayer RJ, Veitch NC, Hung Tran GC, Knguyen Q, 2004. Uncommon 8-oxygenated flavonoids from Limnophila aromatica (Scrophulariaceae). Biochemical Systematics and Ecology, 32: 943-947.
5. Cheruvathur MK, Abraham J, Mani B, Thomas TD, 2010. Adventitious shoot induction from cultured internodal explants of Malaxis acuminata D. Don, a valuable terrestrial medicinal orchid. Plant Cell, Tissue and Organ Culture, 101: 163-170.
6. Depnath SC, 2005. Micropropagation of lingonberry: influence of genotype, explant orientation, and overcoming TDZ induced inhibition of shoot elongation using zeatin. HortScience, 40: 185-188.
7. Do QD, Angkawijaya AE, Tran-Nguyen PL, Huyn LH, Soetaredjo FE, Ismadji S, Yi-Hsu J, 2014. Effect of extraction solvent on total phenol content, total flavonoid content, and antioxidant activity of Limnophila aromatica. Journal of Food and Drug Analysis, 22: 296-302.
8. Dogan M, Karatas M, Aasim M, 2015. An efficient in vitro plantlet regeneration of Ceratophyllum demersum L., an important medicinal aquatic plant. Fresenius Environmental Bulletin, 24(10b): 3499-3504.

9. Karataş M, Aasim M, 2015. In vitro whole plant regeneration of the medicial aquatic plant-Limnophilla aromatic. Fresenius Environmental Bulletin, 24: 1-4.
10. Kher MM, Joshi D, Nekkala S, Nataraj M, Raykundaliya DP, 2014. Micropropagation of Pluchea lanceolata (Oliver & Hiern.) using nodal explant. Journal of Horticultural Research, 22(1): 35-39.
11. Kukongviriyapan U, Luangaram S, Leekhaosoong S, Kukongviriyapan V, Preeprame S, 2007. Antioxidant and vascular protective activities of Cratoxylum formosum, Syzygium gratum and Limnophila aromatica, Biological and Pharmaceutical Bulletin, 30(4): 661-666.
12. Lata H, Chandra S, Wang YH, Raman V, Khan IA, 2013. TDZ-induced high frequency plant regeneration through direct shoot organogenesis in Stevia rebaudiana Bertoni: an important medicinal plant and a natural sweetener. American Journal of Plant Sciences, 4: 117-128.
13. Lincy A, Sasikumar B, 2010. Enhanced adventitious shoot regeneration from aerial stem explants of ginger using TDZ and its histological studies. Turkish Journal of Botany, 34: 21-29.
14. Murashige T, Skoog F, 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiological Plantarum, 15: 473-497.
15. Neumann KH, Kumar A, Imani J, 2009. Plant Cell and Tissue Culture - A Tool in Biotechnology, Principles and Practice, Springer-Verlag Berlin Heidelberg, Germany. 1 p.
16. Öztürk M, 2008. AKVARYUM bitkileri Hygrophila difformis ve Microsorium pteropus’un in vitro koşullarda çoğaltımı. Ankara Üniversitesi, Biyoteknoloji Enstitüsü, Doktora Tezi.
17. Prathanturarug S, Soonthornchareonnon N, Chuakul W, Phaidee Y, Saralamp P, 2005. Rapid micropropagation of Curcuma longa using bud explants pre-cultured in thidiazuron-supplemented liquid medium. Plant Cell, Tissue and Organ Culture 80: 347-351.
18. Raghuvanshi SS, Srivastava A, 1995. Plant regeneration of Mangifera indica using liquid shaker culture to reduce phenolic exudation. Plant Cell, Tissue and Organ Culture, 41(1): 83-85.
19. Sajid ZA, Aftab F, 2009. Effect of thidiazuron (TDZ) on in vitro micropropagation of Solanum tuberosum L. Cvs. desiree and cardinal. Pakistan Journal of Botany, 41: 1811-1815.
20. Siddique I, Anis M, 2007. Rapid micropropagation of Ocimum basilicum using shoot tip explants pre-cultured in thidiazuron supplemented liquid medium. Biologia Plantarum, 51(4): 787-790.
21. Wawrosch C, 2010. In Vitro Propagation of Medicinal Plants for Conservation and Quality Assurance, Medicinal Plant Biotechnology, Editörler: Arora, R., CAB International, Oxfordshire, UK. 94 p.