Sains Malaysiana 42(10)(2013): 1505–1510

In Vitro Shoot Regeneration from Leaf Explants of Kenaf (Hibiscus cannabinus L.)

(Penjanaan Semula Pucuk Kenaf (Hibiscus cannabinus L.) Secara In Vitro Menggunakan

Eksplan Daun)

P.W. Samanthi¹, A. Mohd Puad¹, N. Suhaimi¹, S.M. Kumar² & A.S. Nor Aini^{2 3}*

¹Department of Cell and Molecular Biology, Faculty of Biotechnology and Bio Molecular Sciences

Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia

²Faculty of Forestry, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia

³Institute of Tropical Forest and Forest Products, Universiti Putra Malaysia, 43400 Serdang,

 Selangor, Malaysia

Diserahkan: 13 April 2013/Diterima: 12 Mei 2013

ABSTRACT

Kenaf (Hibiscus cannabinus L.) is a versatile plant with multiuse ranging from animal feed to a wide variety of biocomposite products such as pulp and paper and fibre reinforce plastic. Therefore genetically improved planting materials are needed to tailor made requirement of the industry. Thus, development of plant regeneration through callus is important for in vitro genetic manipulation of kenaf. Currently development of successful genetic transformation of kenaf is through in planta transformation means. In vitro shoot regeneration was conducted using leaf explants from varieties V36 and G4 treated to three different combinations of N⁶ Benzyl adenine (BA) and Indole-3-butyric acid (IBA). High percentage of healthy callus induction was produced in MS medium supplemented with combination of 1.5 mgL^-1 BA and 0.5 mgL^-1 IBA. In addition 68.7% plant regeneration was obtained in MS medium supplemented with 0.3 mgL^-1 GA₃. All plantlets produced roots in hormone free medium. There was no significant difference among varieties in terms of callus induction (number of callus) and plant regeneration (number of plantlets). This protocol is useful to be used for the development of gene transformation protocol of kenaf through callus.

Keywords: Callus; in vitro; kenaf (Hibiscus cannabinus L.); leaf explants; plant regeneration

ABSTRAK

Kenaf (Hibiscus cannabinus L.) merupakan sejenis tumbuhan berdaya saing yang mempunyai pelbagai guna. Kegunaannya berjulat daripada pemakanan haiwan kepada pembangunan pelbagai produk biokomposit seperti kertas dan pulpa serta plastik diperkuat gentian. Bahan tanaman yang diperbaiki secara genetik adalah diperlukan untuk memenuhi keperluan tertentu industri. Pembangunan kaedah regenerasi tumbuhan melalui kalus amat penting bagi manipulasi genetik secara in vitro untuk tumbuhan kenaf. Buat masa kini, pembangunan kaedah transformasi genetik pada kenaf hanya dapat dijayakan melalui kaedah transformasi in planta. Regenerasi melalui pembentukan pucuk in vitro telah berjaya dihasilkan dengan mengunakan eksplan daun daripada varieti V36 dan G4 yang dikulturkan dalam tiga kombinasi N6 Benzil adenin (BA) dan asid Indol-3-butirik (IBA). Penghasilan kalus segar dalam peratusan yang tinggi terbentuk dalam media MS yang mengandungi campuran 1.5 mgL^-1 BA and 0.5 mgL^-1 IBA. Manakala 68.7% pembentukan anak pokok telah berjaya dihasilkan dalam media MS yang dicampurkan dengan 0.3 mgL^-1 GA₃. Plantlet membentuk akar dalam media tanpa hormon. Pembentukan kalus (bilangan kalus) dan regenerasi tumbuhan (bilangan anak pokok) tidak menunjukkan perbezaan yang bererti antara varieti. Protokol ini boleh digunakan untuk membangunkan transformasi kenaf melalui pembentukan kalus.

Kata kunci: Eksplan daun; in vitro; kalus; kenaf (Hibiscus cannabinus L.); penjanaan semula tumbuhan

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