Research Article
Determination of Suitable Microspore Stage and Callus
Induction from Anthers of Kenaf (Hibiscus cannabinus L.)
Ahmed Mahmood Ibrahim, Fatimah Binti Kayat, Zeti Ermiena Surya Mat Hussin,
Dwi Susanto, and Mohammed Ariffulah
Faculty of Base Industry (FIAT), Universiti Malaysia Kelantan, Jeli campus, Locked Bag 100, 17600 Jeli, Kelantan, Malaysia
Correspondence should be addressed to Ahmed Mahmood Ibrahim; a ibrahim1965@yahoo.com
Received 1 December 2013; Accepted 23 January 2014; Published 13 March 2014
Academic Editors: J. E. Barboza-Corona and N. K. Tripathi
Copyright © 2014 Ahmed Mahmood Ibrahim et al. Tis is an open access article distributed under the Creative Commons
Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is
properly cited.
Kenaf (Hibiscus cannabinus L.) is one of the important species of Hibiscus cultivated for fber. Availability of homozygous parent
lines is prerequisite to the use of the heterosis efect reproducible in hybrid breeding. Te production of haploid plants by anther
culture followed by chromosome doubling can be achieved in short period compared with inbred lines by conventional method
that requires self pollination of parent material. In this research, the efects of the microspore developmental stage, time of fower
collection, various pretreatments, diferent combinations of hormones, and culture condition on anther culture of KB6 variety of
Kenaf were studied. Young fower buds with immature anthers at the appropriate stage of microspore development were sterilized
and the anthers were carefully dissected from the fower buds and subjected to various pretreatments and diferent combinations of
hormones like NAA, 2,4-D, Kinetin, BAP, and TDZ to induce callus. Te best microspore development stage of the fower buds was
about 6–8 mm long collected 1-2 weeks afer fower initiation. At that stage, the microspores were at the uninucleate stage which was
suitable for culture. Te best callus induction frequency was 90% in the optimized semisolid MS medium fortifed with 3.0 mg/L
BAP + 3.0 mg/L NAA.
1. Introduction
Kenaf (Hibiscus cannabinus L.) belongs to the Malvaceae
family and section Furcaria. It is closely related to cotton,
okra, hollyhock, and roselle. Nowadays, it has been cultivated
in 20 countries worldwide and its total production (kenaf
and allied crops) is 352,000 tons (2010/2011). Kenaf is an
annual fber crop cultivated for numerous uses (paper pulp,
fabrics, textile, building materials, biocomposites, bedding
material, oil absorbents, etc.) [1]. In Malaysia, this plant is
considered new and cultivated to replace tobacco plantation
that has been no longer supported by the government [2].
Kenaf grows quickly and will achieve 5 to 6 m in height and
2.5 to 3.5cm in diameter within 5 to 6 months. Kenaf has a
unique combination of long bast and short core fbers which
makes it suitable for a range of paper and cardboard products.
Fify-fve percent of dried kenaf stalks are used to make paper.
Waste products from the process can be made into fertilizer
and feed binder. When kenaf is grown in home gardens for
fber, the more tender upper leaves and shoots are sometimes
eaten either raw or cooked [3]. Alternative uses, the top leafy
portion of the kenaf plant is not useful for pulping. Terefore,
this part of the plant would be useful as forage if harvest
equipment could be practically adapted to a dual collection
operation. One of the most popular methods for production
of haploids is through anther and microspore culture on the
artifcial culture medium. Haploids are plants (sporophytes)
that contain a gametic chromosome number (n) as in the
egg and pollen cell. Each pollen mother cell (PMC) in the
anther produces 4 microspore or pollen grains. Tus, anther
with microspore or isolated pollen grains can be cultured on
artifcial medium to raise haploid plant. Haploid production
through anther culture has been referred to as androgenesis
[4].
Te frst time haploid plants were discovered in Datura
stramonium by A.D Bergner in 1921. Afer the initial reports
of successful production of haploids from anther culture
in Datura [5], haploids have been obtained in more than
Hindawi Publishing Corporation
e Scientific World Journal
Volume 2014, Article ID 284342, 5 pages
http://dx.doi.org/10.1155/2014/284342