Copyright © 2018 Authors. This 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. International Journal of Engineering & Technology, 7 (4.14) (2018) 107-111 International Journal of Engineering & Technology Website: www.sciencepubco.com/index.php/IJET Research paper Callus Formation on Endosperm from Immature and Mature Fruits of Barringtonia Racemosa D S M Soder 1 , D N A A Khalid 2 , A Saleh 3 , F Pardi 4 , N J Sidik 5 * Faculty of Applied Sciences, Universiti Teknologi MARA, 40450 Shah Alam, Selangor, Malaysia * Corresponding author E-mail: norri536@salam.uitm.edu.my Abstract Barringtonia racemosa is mangroves type of plant which had been extensively utilized in conventional practices for relieving ailments of pain and inflammation. Many studies have been done on ethnobotanical profiles, pharmacological activities and chemical compounds in Barringtonia racemosa. However, there is a limited study on callogenesis of this plant particularly from different maturity stage of fruits. The present study is to identify the callogenesis of Barringtonia racemosa from endosperm explants of immature and mature fruits in MS medium supplemented with different concentrations of hormones 2,4-Dichlorophenoxyacetic acid (2,4-D) (0, 0.5, 1.0, 1.5 and 2.0 mg/L) and Kinetin (KIN) (0, 0.5, 1.0, 1.5 and 2.0 mg/L). The optimum hormone combination was found in callus grown on endosperm of immature fruits in MS medium supplemented with 1.5 mg/L 2,4-D and 1.0 mg/L KIN. It was also found that the callus in this treatment grew profusely with highest fresh weight (0.513 ± 0.022 g), 100% callus induction and friable callus texture. The callus fresh weight on endosperm explants was higher in immature fruits compared to mature fruits for all the hormone combinations. Therefore, callogenesis were found more efficient from endosperm explant of immature fruits in Barringtonia racemosa species. Keywords: Barringtonia racemosa, callogenesis, endosperm, immature fruits, mature fruits 1. Introduction Medicinal plants have been used extensively by natives or medical practitioner to treat diseases. Barringtonia racemosa Linn. (Family Lecythidaceae) is one of the medicinal plants possess various of bioactivity that engaged to habitat in a wet and watery area such as by the side of fresh swamps, river banks, lakes, shore of backwaters and banks of paddy fields. Barringtonia racemosa fruits have been used to treat a cough, asthma as well as diarrhea in Sri Lanka, and treat malaria in Africa [1]. Meanwhile, in Malaysia, the shoots of Barringtonia racemosa is widely consumed perceiving to gain health benefits. There is also a practice use a paste of the pounded leaves to reduce the itching skin [2]. Many studies have been done on different parts of Barringtonia racemosa towards pharmacological activities such as antibacterial, anti-tumor, anti-nociceptive, antioxidant, anti- inflammatory, an alpha-glucosidase inhibitor, anti-fungal, anti- tuberculosis, anti-arthritic and anti-diarrhoeal [3]. The previous study also supported that Barringtonia racemosa is a plant- derived anti-gouty arthritis remedy [4]. On the other hand, the ethnomedicinal uses of the Barringtonia racemosa fruits are mitigating the pain and inflammation [5] as well as desirable anti-tumour effect in seed extracts [6]. Locally in Malaysia, the fruits of Barringtonia racemosa with bitter taste are eaten fresh as a salad or cooked with turmeric coconut stew. The immature fruits are easier to be eaten fresh due to soft flesh meanwhile mature fruits have hard fibrous flesh unpalatable to be eaten. The use of in vitro culture technique is important in the conservation of plant germplasm and in securing valuable products of medicinal and commercial importance. One of the in vitro cultures that suitable for mass production is callus culture. In plant tissue culture, the supplemented nutritional medium with single or multiple hormones is important in directing the callus growth [7]. Cytokinins and auxins have extensively been used to induce callus from plant explants in in vitro medium [8]. The suitable hormone concentrations used of cytokinin and auxin as well as source of explant is important in the study of callogenesis or callus formation. Nevertheless, there are yet no reports on the comparison of callus formation from endosperm explants of immature and mature fruits in Barringtonia racemosa species. Hence, determining the optimum callus culture is essential in a mass production of cells that can be used in the further study of somatic embryogenesis and suspension culture for long-term plant conservation and supply. 2. Methodology 2.1. Plant materials The endosperm explants source was chosen from 2 type fruits maturity. Immature and matured fruits of Barringtonia racemosa were collected at Universiti Kebangsaan Malaysia (UKM), Bangi, Selangor. The voucher specimen (SK 3191/17) of the sample was deposited in the Herbarium, Department of Biology, Universiti Putra Malaysia (UPM), Serdang, Selangor. The immature fruit selection was chosen from big size ranged 6-8 cm with soft texture of flesh and pale yellow seeds. The matured fruit selection was categorized by big size ranged 6-8 cm with hard, fibrous texture of flesh and pale yellow seeds. The matured fruits were carefully plucked from the tree before fruits reach over mature (dry fibrous flesh).