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Control of Lasiodiplodia Theobromae (PAT) on Rhizophora Racemosa

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Control of Lasiodiplodia Theobromae (PAT) on Rhizophora Racemosa AMERICAN JOURNAL BIOTECHNOLOGY AND MOLECULAR SCIENCES ISSN Print: 2159-3698, ISSN Online: 2159-3701, doi:10.5251/ajbms.2013.3.1.1.7 © 2013, ScienceHuβ, http://www.scihub.org/AJBMS Control of Lasiodiplodia theobromae (PAT) on Rhizophora racemosa using plants extracts *Ukoima, H.N ; Ikata,M and Pepple,G.A *Department of Forestry and Environment, Faculty of Agriculture , Rivers State University of Science and Technology,p.m.b.5080, Port Harcourt, Nigeria E-mail :[email protected] ABSTRACT Laboratory experiments were conducted to ascertain fungal isolates of Rhizophora racemosa .Studies on controlling L. theobromae using plant extracts was also carried out. Fungal pathogens were isolated by cutting the infected portions of the leaves and aseptically placing in glass Petri- dish for 7 days. Pathogenicity test was done in-situ with spore suspension inoculated on Rhizophora racemosa seedling leaves and observed for 15 days in the green house. Different concentrations ranging from (20%, 40%, 60%, 80% and 100%) of extract derived from the bark of Rhizophora racemosa, leaves of Aloe vera and Jatropha curcas were tested on the isolated fungi. The results showed that Lasidiplodia theobromae, Penicillium citrinum, Aspergillus niger and Paecilomyces lilacinus were isolated from the leaves of Rhizophora racemosa. Pathogenicity test revealed that Lasidiplodia theobromae was pathogenic on Rhizophora racemosa. Aloe vera and the bark of Rhizophora racemosa were inhibitory at 100% on Lasidiplodia theobromae (0.1645g and 0.2946g mycelial dry weight respectively). Jatropha curcas had les inhibitory effect at 80% concentration on the tested fungus (0.9118g mycelial dry weight). Biochemical test showed that Quinone, terpenoid and saponin from Aloe vera, bark of Rhizophora racemosa and Jatropha curcas respectively were suspected to account for the inhibition of the tested fungus (Lasidiplodia theobromae). INTRODUCTION have been under utilized in the Niger Delta of Nigeria. They protect the shoreline and are particularly Red Mangrove thrives under a range of inter-tidal important in coastal areas that are subjected to major wetland conditions including high salinity levels from tropical storm tides during hurricanes and typhoons seawater to fresh water (Connell and Lowman 1989). (Basire PLAT, 2005). They tolerate a range of flooding, soil types and other physical site factors. Typically, they are common in Also, the mangrove serves as marine food chain. the middle to low inter-tidal zone above mean sea They play a vital role in protecting and supporting level, extending often along the seaward margin of marine food chains. The species create crucial mangrove stands. habitat needed for marine and estuarine invertebrate nesting grounds and food sources for wading birds They are medium to tall trees; they may reach 20- (Lucien-Brun, 1997). It is also a resource base for 50m in height. Although, commonly much shorter local inhabitants. It is a place they live and derive around 5-8m. Height growth is rapid after their means of livelihood (Ukoima et al, 2000). establishment on the substrate while food reserves are taken up from the hypocotyls of established Rhizophora spp have also been found as good seedlings. They tend to be of shorter statue and material for tannin and dyes. Extractives derived from more spreading in shape on the seaward edge of mangrove barks and hypocotyls for local industries stands or in sites of higher salinity. Taller, single include tannin and dyes. Tannin is a phenotic stemmed trees are found just behind the seaward substance used for the processing of leather, and to edge of stand (Connell and Lowman, 1989). produce dyes ranging from red, brown to black. Its importance is usually seen in the production of In Niger Delta, three red mangrove species are viscose rayon for all normal textile needs, tyre cords, recognized; Rhizophora mangle, Rhizophora industrial pulp (Bawayan, 1971). harrisonii and Rhizophora racemosa. Rhizophora harrisonii is a hybrid of the other two species (Wilcox- Some mangrove species have medical and Enwaraye, 1985). Mangroves are very important but pharmaceutical value. Red mangrove bark is used to Am. J. Biotechnol. Mol. Sci., 2013, 3(1): 1-7 treat angina, boils and fungal infections. The leaves various methods used to control plant diseases, use and bark have been used as an antiseptic and to of chemical fungicides is very common and very treat diarrhea, dysentery, fever, malaria and leprosy. effective. However in view of the complexities arising For example, it is used to cure throat cancer with from the use of chemical pesticides, such as harmful gargles of mangrove bark (Bandaranayale, 1999). effect on environment and non-target organisms including man, domestic animals, beneficial insects, It is a source for timber production. The wood of red wild/life, the use of biocides has provided a very mangrove is widely used for structural components of promising alternative and less hazardous method for traditional homes. For example, poles, beams, plant diseased control. flooring, well-cladding, rafters and others such as fencing, cabinet works, tool handles and boat anchor This research is therefore aim at isolating and etc. (Kohlmeyer, 1971). identifying fungi associated with leaf spot and mangrove seedlings and to confirm their Red mangroves serve as habitat for a wide range of pathogenicity. To evaluate the effects of some plant terrestrial and arboreal wild life. It provides shelter extracts such as Jatropha curcas, Aloe vera, bark of and food for a number of associated Fauna, including Rhizophora racemosa on Lasiodiplodia theobromae birds, small animals, shell fish, and other marine life. under in-vitro conditions. They stabilize soils with their network of sturdy overlapping prop roots which deepen water MATERIALS AND METHODS movement and promote sedimentation in areas that Location/Climate of Study Area: Laboratory might otherwise be eroded (Mehlig, 2006). experiments were carried out at the Rivers State In the Niger Delta mangrove ecosystem of Nigeria, University of Science and technology, Faculty of fungal pathogens of mangrove plants have been Agriculture located at latitude 4.43oS and 4.54oN and reported by researchers, (Ukoima, 1996). These longitude 6.56oW and 7o.03E Fodiboye, (1990). The fungi include Lasiodiplodia theobromae, field experiment was conducted at Ogonokom Paecilomyces lilacinus, Colletotrichum, Gloesporides, mangrove swamp in Abua/odual Local Government Fusarium moniliforme, Aspergillus flavus, Aspergillus Area of Rivers State, Latitude 04.150S, and 7.20oN niger, Alternaria solani, Penicillium citrinum, and longitude 5.5oW and 90o301E in the rainforest Botryodiplodia theobromae, Phoma sp. Rhizopus zone of Nigeria. The study area lies in the zone of stolonifer e.t.c. These fungi were isolated from humid tropical climate which has two seasons. The Rhizophora racemosa, Rhizophora mangle, wet season extends from March to October and dry Rhizophora harrisonii and Avicennia africana season extends from November to February. The (Ukoima 1996; 2000; 2006; 2007). A similar work has mean annual rainfall varies between 400mm - also been done in southern Florida (Newell, 1976). 1700mm. annual temperature range from 21oC and Botryodiplodia theobromae is known to cause seed 29oC (Fodiboye, 1990). rot, wilting and necrosis on mangrove plant in India Isolation and Identification of Fungi: Diseased (Mishra, 2002). Pestalotia palmarium have been leaves were collected from Ogonokom area of identified as causal agent of leaf spot at Batel-nut in Abua/Odual Local Government Area from parent Bangladesh (Islam, 2001). plants of Rhizophora racemosa and surface sterilized Efficacy of some plant extracts and plant products with methylated spirit and rinsed in distilled water. against powdery mildew of pea has been The infected portions of the leaves were cut into experimentally demonstrated. Ajoen, a constituent of pieces (1cm2), then placed on sterile filter paper in garlic at 100ppm was found to be inhibitory to some glass Petri-dish and incubated at room temperature fungi namely Aspergillus and saprophytic (27 2oC) for 7 days. The hyphal tips were Talaromyces flavus. (Singh et al 2000). Rhizome transferred into Potato dextrose agar (PDA) plate to powder of ginger gives a good control as sulfex and obtain pure cultures. The organisms were identified Bavistin. (Singh,2004). Different plant extracts like on the basis of morphological characteristics such as neem (Azadirachate indica), Tulshi (Ocimum colony form and color, type of mycelium, fruiting sanctum), Garlic (Alium sativium) Aloe vera, Bitter bodies and spores (Singh, 2000 and Toussoun and leaf (Veronica amygdalina), Scent leaf Cocimum Nelson, 1988). gratussima etc. have been recommended for the control of Pestalotia spp. and other fungi (Pandey et al, 1983; Ukoima and Okah 2006). Among the 2 Am. J. Biotechnol. Mol. Sci., 2013, 3(1): 1-7 Pathogenicity Test: A total of forty red mangrove (R. The plant materials were washed several times with racemosa) plants were used for this experiment. distilled water and dried at room temperature. 100g of Plants to be inoculated were chosen randomly from plant part were separately crushed to a powder using 35-65cm height saplings of red mangrove, collected sterilized mortar and pestle. These crushed from Abua/Odual Local Government Area. materials were extracted sequentially into 300ml Inoculations were done in green house seedlings and distilled water. Phyto-chemical screening test was the leaflets
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