Indonesian J. Pharm. Vol. 24 No 1 : 22 – 29 ISSN-p : 0126-1037 Research Article IDENTIFICATION OF BAWANG SABRANG (Eleutherine americana Merr. ex K. Heyne) IN INDONESIA BASED ON CHROMOSOME CHARACTERS Budi Setiadi Daryono1*, Wenny Deisshinta Rahmadani1, and Sudarsono2 1Genetic Laboratory, ABSTRACT Biology Faculty, Bawang Sabrang (Eleutherine americana Merr. ex K. Universitas Gadjah Mada Heyne) is a plant belongs to Iris family (Iridaceae). Genetic study Jl. Teknika Selatan, Sekip of the Eleutherine species should be investigated to yield valuable Utara, Yogyakarta 55281, information for breeding program. The aim of this research was to Indonesia determine chromosome characters as a preliminary research on 2 Department of the genetic characterization of Bawang Sabrang. Squash method Pharmaceutical Biology, on the root tips was used for chromosome preparation of this Faculty of Pharmacy plant. The results showed that the time of cell division and Universitas Gadjah Mada prometaphase stages of Bawang Sabrang were occurred at about 55281,Indonesia 08.00-08.30 a.m. and 08.20 a.m., respectively. Chromosome number of Bawang Sabrang was 2n=12 and the karyotype Submitted: 17-07-2012 Revised: 29-09-2012 consisted of 8 (4 pairs) of metacentric chromosomes, 2 (1 pair) Accepted: 06-10-2012 submetacentric chromosomes and 2 (1 pair) subtelocentric chromosomes which have the longest of total length *Corresponding author chromosomes. Therefore, the karyotype formula of Bawang SAT Budi Setiadi Daryono Sabrang was 2n=12=8m+2sm+2st . Besides that, on the pair of subtelocentric chromosomes there was a satellite at each of the Email : chromosome. Analysis of chromosome characters exhibited that [email protected]. the long of total length chromosomes was about 1.687 ± 0.111 µm ac.id to 5.320 ± 0.716 µm. Based on the R value ( 3,65 ± 0,41), it revealed that there was variation of chromosome size on this Eleutherine species in Indonesia. Moreover, data of the chromosome characters is important to complete the database of Bawang Sabrang as a potential medicinal herb in Indonesia Keywords: Bawang Sabrang, Eleutherine americana, chromosome, karyotype INTRODUCTION Ph.D.). This plant has a quite a lot of local Iridaceae is one of family which most of names; for example in Kalimantan it is known the members are perennial herbs. One of its as bawang Sabrang or bawang Tiwai (Heyne, 1988). member, which is neither well known nor Morphology of this plant has bright red beneficial understand yet but has been underground storage organs of bulb like onion. examined is from the genus Eleutherine. This plant also has a pseudo-trunked, green According to Govaerts (2006), this genus sword-shaped leaves that overlap each other consists of 13 species. Most researchers transversely at the base which the apical and examine more Eleutherine species in the the base are pointed. Its inflorescence is southern America and southern Africa. This is definite composite; rhipidia shaped, white because they are scattered in these areas. color, and have 6 petaloid tepals which are However, Goldblatt (1991) reported that the arranged in a double circle. origin habitat of this plant is still unknown. Bawang Sabrang is a wild plant, but in One of its members growing in Indonesia, Java it is being kept as ornamental plants, especially in Borneo and Java is Eleutherine cultivated, and naturalized. Bawang Sabrang can americana Merr. ex K. Heyne. Currently, it is grow well in the altitude of 600 to 1500m said that Eleutherine bulbosa is the revised name above sea level, in the cool and cold areas as in of Eleutherine americana Merr. ex K. Heyne mountains. This plant is vegetative (personal discussion with Peter Goldblatt, reproductive using bulbs. Although it can grow 22 Volume 24 Issue 1 (2013) Budi Setiadi, et al. well in Indonesia, however cultivation of determine the active mitotic time. Stages Bawang Sabrang is still low. Cultivation is preparation chromosome is as follows: generally done in the household scale, Sample Preparation: Bawang Sabrang obtained meanwhile it has many benefits. Therefore, to from Pontianak, West Kalimantan. First, the improve the cultivation and also to preserve the bulb of bawang Sabrang soaked in a Petri dish fill germ plasma of the plant in both quantity and with aquadest to germinate the root. Roots will quality, conservation efforts are required. One grow ± 3 days after soaking and will be able to of the initial steps that can be done to realized use in sample preparation; Root cutting: Pieces it by knowing its genetic characterization, are taken from the root tip of the meristem because it can be further known its genetic region, which is ± 3 mm from the root at the identity. estimated mitotic time. At this research, Research on the molecular and cytology pretreatment were not been done because it of this plant has been done, but not yet in didn’t show much different; Fixation Root: tip Indonesia. One of the research on the cytology is inserted to the bottle flacon that has been Bawang Sabrang which was done by Goldblatt filled with glacial acetic acid 45% (45ml glacial and Snow (1991), states that the number of acetic acid added with 55 ml aquadest). Then chromosome Eleutherine americana (syn. the root tip of the snippet is stored at a Eleutherine bulbosa) is 2n = 12. Moreover, temperature of 4°C for 15 minutes. After that, research on the chemical constituents from the root-tips washed by aquadest; Hydrolysis by Eleutherine Americana in China has resulted 9 HCl : After fixation, the root-tips hydrolyzed compounds (Liu et al., 2009). It was reported by chloride acid (HCL) 1N (1 ml chloride acid that naphthoquinones, anthraquinones and plus 11 ml aquadest) and then than inserted in naphthalene was isolated from the Bulbs of the the incubator at a temperature of 55 ° C for 5 plant (Mahabusarakam, 2010). Whereas, an minutes. After that, the root-tips washed by active compound from the bulb of Eleutherine aquadest.; Tint : After fixation and hydrolysis, americana L. Merr. collected from East the root-tips cleaned and colored using aceto- Kalimantan, Indonesia, was tested for its orcein 1% (1gr orcein synthetic acid in 100 ml antidermatophyte and antimelanogenesis acetate 45%) and stored at room temperature activity (Kusuma et al., 2010). Therefore, the for 24 hours.; Squashing: Root- tips were object used of this research were to study the placed in a colored glass objects. Aceto-orcein mitotic time, the number of chromosome of the excess around the ends of the root-tips Bawang Sabrang, and the chromosome characters cleaned with clean tissue paper to avoid the that includes the shape and size of the preparation for the dirty. Then the root-tips chromosome. Data of the chromosome dropped by glycerol and covered with glass characters is expected to study a potential of cover: Labeling: To seal the glass cover, use Bawang Sabrang as a medicinal herb in paint nail clear glass cover on the sidewalk. The Indonesia. sample is stored in the box preparation and placed in the refrigerator at a temperature of 4º METHODOLOGY C until observed; Observation by Light Materials Microscope : Observation of mitotic phases Plant used in this research was Bawang and chromosomes number of bawang Sabrang Sabrang plants (Eleutherine americana Merr. ex K. are using a light microscope. After getting a Heyne) collected from Lambungmangkurat good preparation, picture were taken by digital University, South Kalimantan and herbal camera and microphotograph camera. nursery in Yogyakarta, Indonesia. Data Analyze Method Chromosome is calculated at the time of Preparation of chromosome was using observation and the results of the images have squash method on the plant root-tip (Jahier et formatted in digital. The number of al., 1996). It was conducted at 08.00-14.00 chromosome is calculated from the root of WIB, started with 30 minutes time interval to three different plants of Bawang Sabrang, each with two or three recycling cell that is on a Volume 24 Issue 1 (2013) 23 Identification of Bawang Sabrang prometaphase. Measurement the length of both chromosomes move toward the metaphase chromosomes short arm (p) and long arm (q) plate or division area. This phase takes place using a computer program, AutoCAD Map quickly so that it is become the most difficult 2000i. Measurement of Centromere Index was phase found during the observation. At this obtained to determine chromosomes form. The phase the form of the chromosomes are most making of Ideogram is done by Corel Draw X3 excellent, in the form of sister chromatid and based on the short arm length (p) and of long scattered in the cells, so this is most appropriate arm length (q) chromosome. To create a time to observe the chromosomes morphology, karyotype is done by cropping chromosome amount and size. images with Magnetic lasso tool in the The further phase is the metaphase. At application program, Adobe Photoshop CS2. this phase the chromosomes has been on metaphase plate in equator region, so that the RESULTS AND DISCUSSION chromosomes appears in a line and gather in Bawang Sabrang usually forms a large the middle of the cell. Position of the homolog clump. It is originally wild plants but can also chromosome in metaphase plate is random. At be kept as ornamental plants and medicinal the end of this phase, the spindle thread made plants in the house yard. Beside it can grow on prophase will be attached to the centromere. well in cool and cold region, Bawang Sabrang At the next stage, chromosomes also has capability to grow in hot areas such as centromere that has been attached by the in urban areas. spindle thread will split along the chromosome arms and vertically toward the threads spindle, Mitotic Time and Cell Cycle so that each similar chromatid will be On this research, chromosome separated.