33 Morphological and Anatomical Changes by Colchicine in Seedling of Impatiens balsamina L. Ni Made Sastriyani Wiendra and Made Pharmawati* Biology Study Program, Faculty of Mathematics and Natural Science, Udayana University Kampus Bukit Jimbaran, Badung, Bali * Corresponding author: made_pharmawati@unud.ac.id Abstract. Impatiens balsamina L. is a plant that is widely cultivated in Bali. The flower of this plant is used as a component of offering in Hindu religious ceremonies. The flower petals are thin and wilt easily, therefore genetic modification is needed to get better quality. This research aimed to study the effect of 0.01% colchicine with soaking periods of 4 hours, 8 hours, 12 hours and 24 hours to seedlings of I. balsamina. Seeds of I. balsamina were germinated and sprouted seeds were treated with 0.01% colchicine for 4 hours, 8 hours, 12 hours and 24 hours. The sprouted seeds were then transfer to polybag with soil media. The experiment was designed with randomized blok design with five replicates. Observations were done on plant height, length and width of leaf, stem diameter, number of stomata as well as observation on plant stem cortex cells. Results showed that colchicine reduced plant height at three weeks after planting, while plant stem diameter increased. Control plants had the smallest leaf length and width. The number of stomata decreased at plants derived from colchicine treated sprouted seeds and the lowest stomatal number observed was at soaking duration of 24 hours. Keyword: colchicine, growth, Impatiens balsamina, stomata I. INTRODUCTION 1 Impatiens balsamina is a plant whose flowers are used as ceremonial material by the Hindu community in Bali. The colour of the flowers varies greatly but is very thin and wilt quickly. Therefore, genetic improvement of I. balsamina is necessary to improve flower quality. Genetic improvement can be done conventionally or non-conventionally. Conventional way is done by artificial crossing or artificial pollination. The results of crossing are expected to bring superior traits. Non-conventional genetic improvement is done by mutant selection, somatic hybridization and gene transfer [1]. One way to improve genetic of I. balsamina is by inducing polyploid. Polyploid plants are plants that have three or more sets of chromosomes in their cells [2]. The morphological characteristic of polyploid plants is bigger plants including bigger plant parts such as roots, stems, leaves, flowers and fruit [3] Polyploid plants can be induced by various chemical compounds, one of which is colchicine. Colchicine is a toxic alkaloid extracted from the Colchinum autumnale plant. Colchicine is a mitotic inhibitor. It acts by preventing tubulin polymerization, so the separation of chromosome sets is disrupted when mitosis and cells have multiplied chromosomes [4,5]. Induction of polyploid plants with colchicine has been widely carried out. In 'Sumenep' onion, the soaking period and colchicine concentration affected the effectiveness of polyploid induction. Colchicine caused shorter onion plants, reduced the number of leaves but the leaves became thicker, reduced the amount of stomata but there was an enlargement of stomata [6]. Polyploid of Phalaenopsis amabilis seedlings can be produced by dripping colchicine 1000, 3000, 4000 and 5000 mg L -1 (0.1%, 0.3%, 0.4% and 0.5%). It was found that the most effective colchicine concentration was 5000 mg L -1 (0.5%). Polyploid P. amabilis seedlings had larger stomata size than diploid seeds, and lower stomata density [7]. This research aimed to evaluate various soaking periods of colchicine 0.01% to induced morphological changes in seedlings of I. balsamina. II. RESEARCH METHODS Plant Materials Seeds of I. balsamina were harvested from I. balsamina plantation di Sading Village, Badung Regency, Bali, Indonesia. The viability of the seeds was tested by soaking seeds in water and floating seeds were discarded while sinking seeds were treated. Advances in Tropical Biodiversity and Environmental Sciences 3(2): 33-36, September 2019 ISSN: 2622-0628 DOI: 10.24843/atbes.v03.i02.p04 Available online at: https://ojs.unud.ac.id/index.php/ATBES/article/view/53950