Pak. J. Bot., 51(1): 201-204, 2019. DOI: 10.30848/PJB2019-1(19) IN VITRO ANTIFUNGAL, ANTIBACTERIAL, PHYTOTOXIC, BRINE SHRIMP, INSECTICIDAL ACTIVITIES AND COMPOSITION OF ESSENTIAL OIL OF TAGETES MINUTA FROM DIR-KOHISTAN, PAKISTAN ABDUL KHALIQ JAN 1* , ALI HAZRAT 2 , SHUJAAT AHMAD 1 , TOUR JAN 2 AND GUL JAN 3 1 Chemistry Department, Shaheed BB University, Sheringal, Pakistan 2 Department of Botany, University of Malakand, Chakdara Dir Lower, Pakistan 3 Department of Botany, Abdul Wali Khan University Mardan, Pakistan * Corresponding author’s email: abdukhaliq@gmail.com Abstract The essential oil of the flowering shoot of Tagetes minuta L., collected from Dir Kohistan of Pakistan, was investigated for biological activities and chemical composition. The GC and GC/MS analysis data showed that among twenty two compounds, verbenone (25%), Unknown(11.6%),4-(5-methyl-2-furanyl)-2-butanone (8.8%) and D-limonene (8.4%) were the major constituents. The biological activity results showed that out of six bacteria stains, the oil possessed inhibitory activities against S. flexenari and S. aureus with a zone of inhibition 22mm and 20mm respectively. Among five fungal strains, only Aspergillus flavus and Candida albicans were susceptible to oil, with 60% and 30% inhibition respectively. In insecticidal tests, the oil was noted for its effectiveness toward Sitophilus oryzae, with 50% mortality. Although the oil showed no cytotoxicity against Artemia Salina, while moderate phytotoxic activity was observed for Lemna minor. Key words: Essential oil; Tagetes minuta L., verbenone; D-limonene, Antibacterial, Antifungal cytotoxicity, Phytotoxicity, Insecticidal activities. Introduction The term of medicinal plants incorporates different kinds of plants utilized as a part of Herbalism and a portion of these plants have a therapeutic exercise. These restorative plants, consider as a rich asset of fixings which can be utilized as a part of medication advancement and blend. Other than that these plants assume a basic part in the advancement of human societies around the entire world. In addition These plants incorporate ginger, green tea, walnuts and some other Medicinal plants used in different cultures, traditions and folklore medicine for the treatment of various ailments (Khalil et al., 2014. The synthetic antibiotics that have been using in the recently, are accociated with several side effects, high cost and increasing resistance, therefor further investigation on the clinical findings and traditional medicines is needed to obtain new sources of therapeutic agents. Recently most of the scientists are focusing of the natural source of drugs for curing various ailments and have keen interest in the clinical trails of such medicines to investigate its potency and side effects. One of such traditional remidies is the essential oils (EOs) that has recently attracted the focus of practioners and researchers (Aneta et al., 2018; Cowan 1999; Jan et al., 2018; Jan et al., 2017; Kalemba et al., 2003; Noor et al., 2018). Recent literature reveals about 3000 essential oils (Thosar et al., 2013), and it was found that EOs have various biological activities (Baratta et al., 1999; Hammer et al., 1999; Guleria et al., 2013).Our country has a greater potential of indigenous herbal resources and most of the medicinal plants of the country are yet to be explored (Shinwari, 1996; Shinwari, 2010) for its essential oil and bioactivities. The present research is based on the biological activities and components of the essential of Tagetes minuta L. it is locally known as Zangali Dambar Guley an annual plant of the Asteraceae family, distributed in northern areas of Pakistan in a wide range of climatic conditions (Sadia et al., 2013). Marigold (Tagetes minuta L.) oil has a good market in perfumery and flavor industry in India, cultivated in main growing areas like Himachal Pradesh, Jammu and Kashmir and Uttar Pradesh, producing the net profit estimated as 30-40 thousand rupees per hectare (Kumari et al., 2000). It has been found that limonene, which is the main component of essential oil is responsible for the mosquito repellent effects (Gillij et al., 2008). It has been reported that the essential oil of T. minuta L. contains z-β-ocimene and Dihydroxyacetone that is the active compounds for the control of nematode i.e. Meloidogyne incognita (Adekunle et al., 2007). Tagetes has been used as a source of carotenoids, also used as a colorant in poultry feed, citrus juices, dairy products, confectionery, pasta, margarine, salad dressing, ice cream, vegetable oil and baked materials. T. minuta essential oil has been found to have antimicrobial, cytotoxic (Mohsen et al., 2014), antioxidant, anti-inflammatory (Karimian et al., 2014 ) and diuretic activities (Ambasta SP 1986). The literature survey reveals the differences in the composition and qualities of the essential oil of the T. minuta L. depending on the various locations, growth stages and environmental conditions (Gili et al., 2000; Omidbaigi et al., 2008; Meshkatalsadat et al., 2010; Craveiro et al., 1988; Rao et al., 2006; Moghaddam et al., 2007; Chamorro et al., 2008). Due to variation in the composition from locality to locality and expected variation in medicinal potential, the composition and biological activities of the essential oil of Tagetes minuta L., was selected for investigation. Materials and Methods Plant material: Flowering shoot biomass of Tagetes minuta L. (Asteraceae) was collected from the vicinity of Shaheed Benazir Bhutto University Sheringal, Pakistan (26th November 2013). The Plant materials were