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Stigma and Stand Specificity in Fusarium Verticilloides Associated with Talbotiella Gentii

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Stigma and Stand Specificity in Fusarium Verticilloides Associated with Talbotiella Gentii ® The African Journal of Plant Science and Biotechnology ©2011 Global Science Books Stigma and Stand Specificity in Fusarium verticilloides Associated with Talbotiella gentii Daniel Dompreh1* • Michael D. Swaine2 • George Odamtten3 1 Department of Silviculture and Forestry Management, Faculty of Renewable Natural Resources, Kwame Nkrumah University of Science and Technology, Kumasi, Ghana 2 School of Biological Sciences, University of Aberdeen, Aberdeen, AB24 3UU, Scotland, UK 3 Department of Botany, University of Ghana, Legon, Accra, Ghana Corresponding author : * [email protected] ABSTRACT Pathogens, mainly fungus, were after fruit set, the most common cause of fruit abortion in Talbotiella. The measurement of stigmatic pollen load demonstrates that there is no shortage in the quantity of pollen flow within Talbotiella populations. However, germination of pollen grains on stigmatic surface of the species and growth of pollen tube down the style to fertilize ovules are inhibited by fungal hyphae. Air sampling at 3 populations of Talbotiella using 3 agar media was carried out to find out the prevalence of fungi. A total of 26 fungal species belonging to 17 genera was observed. The genera Aspergillus, Penicillium and Cladosporium recorded highest number of species. Penicillium aurantiogriseum and Fusarium verticilloides were the dominant species within Talbotiella populations. Fusarium verticilloides was found to mainly occupy the stigmatic surface of the Talbotiella species. _____________________________________________________________________________________________________________ Keywords: air-borne, fungi, pathogenic, population, spores INTRODUCTION Talbotiella gentii, a rare endemic species in Ghana, is under severe threat of extinction (Swaine and Hall 1981) therefore requires immediate conservation measures to prevent the few remaining populations from extinction. Though Talbo- tiella is a non-timber product in Ghana, it produces excel- lent charcoal of very high market demand therefore the spe- cies is preferentially exploited for charcoal. Periodic bush fires and farming activities have also contributed to the loss of Talbotiella populations. Little progress has been made in conservation of Talbotiella partly due to lack of information on its reproduction and genetic diversity. Information on the genetic diversity of the few isolated Talbotiella populations remaining will suggest which populations are more hetero- geneous that could be targeted for ex situ conservation. To date, no integrated plan for conservation of Talbotiella has been developed though the Forestry Commission has made some effort to reduce its exploitation for charcoal and fuel- Fig. 1 Records of Talbotiella populations in Ghana. Filled dots = extant wood. Records of Talbotiella are shown in Fig. 1. populations, empty dots = extinct populations. The number of flowers initiated by plants normally ex- ceeds the final number of mature fruits (Stephenson 1981; Bustan and Goldschmidt 1998) and fruit mortality due to produce various types of nutrient-rich secretions (nectar, internal (e.g. Nichols and Walmsley 1965; Udovic and Aker stigmatic exudates, and pollen exudates) that may serve as a 1981; Pías and Guitían 2006) and external (e.g. Louda substrate for colonization and infection by plant pathogens 1982; Arnold et al. 2003) factors can be a major drawback and saphrophytes. Plants produce flowers in exposed or in plant reproduction. Results from reproductive studies of elevated positions to facilitate pollen and /or seed dispersal. Talbotiella revealed that flowers and developing fruits are In out-crossing plants, increased height favours pollen dis- infested with spores and hyphae of fungi (Dompreh, unpub- persal by wind and insect pollinators (Carromero and Ham- lished). Pollen germination on the stigmatic surface may rick 2005; Rocha et al. 2005), therefore flowers are easily therefore be inhibited by fungal infection and could be the infested with fungal pathogens if present in the air. Flowers cause of pollination failure, low seed set or premature fruit serve as a pathway for infection of the developing seeds and abortion in T. gentii. Fungal spores which land on plants can fruits, with associated benefits of nutrient supply for the infest flowers and leaves causing plant diseases or limiting fungus. Nectar is rich in nutrients and would therefore light penetration and photosynthetic activities. Fungal infec- favour colonization and infection by plant pathogens and tion may also inhibit fruit and seed development resulting in saprophytes (Brysch-Herzberg 2004; Ngugi and Scherm premature fruit and ovule abortion. 2006). Thus, flower-infecting fungi can readily take advan- Flowers are considered excellent microbial habitats tage of the resources allocated to host reproduction. Flower- (Brysch-Herzberg 2004; Ngugi and Scherm 2006). They infecting fungi, by sporulating on the plant inflorescence, Received: 15 July, 2010. Accepted: 29 March, 2011. Original Research Paper The African Journal of Plant Science and Biotechnology 5 (1), 20-25 ©2011 Global Science Books are also in an ideal position to facilitate their dispersal (Clay Counted colonies were expressed as colony forming units per plate 1991; Antonovics 2005; Ngugi and Scherm 2006). The de- per hour (cfu/p/h). position of fungal spores on stigmatic surfaces of flowers may affect pollen performance by either physically redu- Identification of fungi cing the amount of stigmatic surface area available for pol- len germination since spores compete with pollen for ger- The species of fungi isolated were identified by their morpholo- mination space and /or by inhibiting pollen germination and gical, colour and other cultural characteristics using standard and tube growth down the style through chemical process. reference texts and identification manuals (Thom and Raper 1945; Airborne pathogenic and saprophytic fungi may be of Smith 1960; Von Arx 1970; Barnett and Hunter 1972; Raper et al. different types (species) and the level of pathogenic effect 1973; Neergaard 1983; Klich and Pitt 1988; Samson and Reenen- on host plants may differ from species to species. Some of Hockstra 1988; Samson et al. 2007). the most common airborne fungi belong to the genera Alter- naria, Aspergillus, Cladosporium, Penicillium, Candida, Isolation and identification of fungi on stigmatic Rhodotorula, Trichoderma, Cephalosporium and Chaeto- surface of flowers of Talbotiella mium. Fungal spores in air, whether pathogenic or not, compete with pollen grains for space and nutrients on stig- Two methods were employed using the modified method of Limo- matic surface, inhibiting pollen germination and may be nard (1966) and Tempe (1967). (i) A single flower with its stig- limiting seed set in Talbotiella. matic surface was placed on sterile Whatman No.1 filter paper in a In the present study, aeromycoflora within the popula- 9-cm diameter sterile Petri dish moistened with 5-10 ml of sterile tions of T. gentii were sampled, isolated, cultured on agar distilled water. Thirty replicates (10 from each population) were media and identified. Fungi on the stigmatic surface (patho- used. (ii) A single flower with its stigmatic surface was placed on genic or non pathogenic) surface of flowers of Talbotiella three solid media (PDA, DG-18, and OGYE), for each population were also isolated and identified. Comparison of fungal in- (total sample = 30). In both methods, samples were incubated at fection rates with pollen germination may offer an expla- 28-32°C in each instance (i and ii) until fungi grew (after 3 days). nation for differences in fruit abortion rates in Talbotiella. The number of colonies per flower was recorded. The fungi which appeared were identified using their cultural, colour and morpho- MATERIALS AND METHODS logical characteristics. In the case of flowers placed on the filter paper only, the species were observed and identified using a bino- Study site cular stereomicroscope (Leica Zoom, 2000 model, Germany). Both flowers and isolated stigmata were studied (data in tables Three populations (Yongwa, Sapawsu and Hotel) were selected for come only from the latter). the aeromycoflora studies in Talbotiella. Yongwa is about 17.5 and 16.2 km from Sapawsu and Hotel, respectively. Selection of these Data analysis sites was based on the level of pathogenic fungal infestation of the Talbotiella flowers recorded in a reproduction experiment by All statistical analyses were performed on MINITAB version 15. Dompreh (unpublished) and the concurrent rates of fruit abortion Analysis of variance (ANOVA) was performed to compare the dis- in T. gentii. The study was carried out during the peak of flowering tance from Talbotiella populations on fungal infestation and also period of Talbotiella. test for normality of the data before used for analysis. A t-test was used to discriminate between the means at P < 0.05. Sampling of aeromycoflora at the three populations RESULTS Five 9-cm diameter replicate Petri dishes of three media dichloran- The highest mean count of air-borne fungal colonies (389 ± glycerol agar (DG-18; Hocking and Pitt 1980), potato dextrose 2.1 cfu/p) from three populations was recorded for Yongwa agar (PDA; Merck 2002), oxytetracycline glucose yeast extract with 20 min exposure on PDA followed by Sapawsu. Con- agar (OGYE; Mossel et al. 1970) appropriate for the development sistent differences among sites, at all exposure times were of different fungal groups, and also to increase the chance of cap- recorded (Table
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