Sains Malaysiana 45(3)(2016): 373–381

In vitro Regeneration and Comparison of Phenolic Content, Antioxidant and Antityrosinase Activity of in vivo and in vitro Grown Asparagus officinalis

(Penjanaan Semula in vitro dan Perbandingan Kandungan Fenolik, Antioksida danAktiviti Antitirosinase Asparagus officinalis Ditanam Secara in vivo dan in vitro)

ARASH KHORASANI ESMAEILI*, ROSNA MAT TAHA, SADEGH MOHAJER & BEHROOZ BANISALAM

Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur

Malaysia

Received: 7 August 2014/Accepted: 7 September 2015

ABSTRACT

Asparagus officinalis as a valuable medicinal plant has a low multiplication rate using the conventional methods. This study was carried out to establish an efficient in vitro propagation protocol and also to compare some biological activities of in vivo and in vitro grown Asparagus. The nodal explants were cultured on MS medium supplemented with different concentrations of 6-benzylaminopurine (BAP) and 1-Naphthaleneacetic acid (NAA) or kinetin (Kn) and Indolebutyric acid (IBA), under light and dark conditions. After 6 weeks of culture, the highest percentage (100%) of callus formation was found in 17 of treatments under dark condition and 3 treatments under light condition. Also between the two groups of hormones, Kn +IBA showed better results in promoting callus formation. The highest average number of shoots (4.25) of size 4 mm or more per explant, formed under dark condition using 1.5 mg/L BAP mixed with 0.05 mg/L NAA. Rooting was best induced in shoots excised from shoot cultures which were proliferated on MS medium supplemented with an optimal concentration of 0.4 mg/L IBA (2 roots per explant). In the second part of the study, the extracts of in vivo and in vitro grown plants as well as callus tissue were tested for their total phenolic and flavonoid content, antioxidant and antityrosinase activities, using two different extraction solvents (methanol and hexane). The methanol extract of in vivo grown plants showed a significantly higher amount of total phenolic and flavonoid content. The antioxidant activity of tested samples followed this order; in vivo plant > callus > in vitro plant.

Keywords: Antioxidant; antityrosinase; flavonoid; phenolic; propagation

ABSTRAK

Asparagus officinalis sebagai tumbuhan ubatan yang bernilai mempunyai kadar pembiakan yang rendah apabila dibiakkan secara konvensional. Kajian ini bertujuan untuk menghasilkan kaedah pembiakan secara in vitro yang cekap dan untuk membandingkan aktiviti biologi daripada Asparagus officinalis yang ditanam secara in vivo (biasa) dan in vitro (kaedah kultur tisu). Eksplan nodal dikultur menggunakan media MS yang ditambah kepekatan hormon 6-benzilaminopurin ( BAP) dan asid 1- naftalena (NAA) atau kinetin (Kn) dan asid indolbutrik (IBA) di bawah keadaan cahaya dan gelap. Selepas 6 minggu, peratus tertinggi (100%) pembentukan kalus didapati daripada 17 rawatan yang diletakkan di bawah keadaan gelap dan 3 rawatan di bawah cahaya. Didapati daripada 2 kumpulan hormon, Kn dan IBA telah menunjukkan keputusan yang lebih baik dalam pembentukan kalus. Purata pembentukan pucuk tertinggi (4.25) bersaiz 4 mm atau lebih bagi setiap eksplan, terbentuk di bawah keadaan gelap menggunakan 1.5 mg/L BAP beserta 0.05 mg/L NAA. Pertumbuhan akar didapati terbaik apabila pucuk diambil daripada kultur yang dibiakkan dalam media MS yang ditambah dengan 0.4 mg/L IBA (2 akar setiap pucuk). Dalam bahagian kedua eksperimen, ekstrak daripada tumbuhan yang ditanam secara in vivo, in vitro dan juga tisu kalus telah dikaji untuk mengetahui jumlah fenolik dan kandungan flavonoid, aktiviti antioksidan serta antitirosinase menggunakan 2 pelarut (metanol dan heksan). Ekstrak metanol daripada tumbuhan in vivo menunjukkan jumlah fenolik dan kandungan flavonoid yang ketara dan signifikan. Aktiviti antioksidan bagi sampel yang telah dikaji adalah dalam susunan berikut: tumbuhan in vivo> kalus > tumbuhan in vitro.

Kata kunci: Antioksidan; antitirosinase; fenolik; flavonoid; propagasi

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*Corresponding author; email: arash_khorasani@yahoo.com