303 American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) ISSN (Print) 2313-4410, ISSN (Online) 2313-4402 © Global Society of Scientific Research and Researchers http://asrjetsjournal.org/ Integrated Management of Anthracnose (Colletotrichum capsici (Syd.)): Implications to Disease Reactions, Quality and Growth Parameters of Three Genotypes of Chili Serawit Handisoa*, Tesfaye Alemub a,bAddis Ababa University, College of Natural Sciences, Department of Microbial, Cellular and Molecular Biology, Po Box: 1176, Addis Ababa, Ethiopia aEmail: serawithandiso@gmail.com, bEmail: tesfayealemu932@gmail.com Abstract Anthracnose (Colletotrichum spp ) is one of the most important disease that decimate chili production in Ethiopia. The efficacy of three Trichoderma isolates viz., AAU-37, AAU-Th and AAU-69 with aqueous leaf extracts Onion, Garlic, Neem and Cassia spp were harmoniously applied on Oda Haro, Mareko Fana and Melka zala pepper varieties, with the aim to manage chili anthracnose in rainy season of 2013. The treatments were arranged in RCBD replicated thrice. Data on disease reaction, quality and growth characteristics of chili had been collected. Analysis of data was carried out using ANOVA. The lowest plant infection (12.8%), leaf infection per plant (15.2%), percent diseased leaf area(15.2%)and infected fruits per plot (17.4%) was observed on combined application of isolates Trichoderma spp, plant extracts and Ridomil in Maraqo fana variety. Regarding the growth parameters, viz. the highest Mean Percent establishment (81.67), mean days to 50% flowering (65.33), mean days to 50% maturity (82) days to first harvest (106.3) in was observed in T16, T16, T4, and T8, respectively. From the quality parameters, the highest mean number of branches per stem (9), mean canopy diameter (24.8), mean number of flowers per plant (9.6) and mean plant height (61.4) in T10, T15, T6 and T7, respectively. Both negative and positive control showed higher incidence and severity as compared to single and combined application of isolates Trichoderma spp, plant extracts and Ridomil. Therefore, integrated use of Trichoderma spp and plant extracts can be recommended. Conventional fungicides will be replaced by antagonists and botanicals. This will improving crop quality and growth maximize profitability of chili and ultimate sustenance. KeyTerms: Anthracnose; infection; Quality; Growth Parameters; Integrated Disease Management; Plant Extracts. ------------------------------------------------------------------------ * Corresponding author. http://asrjetsjournal.org/ American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) (2017) Volume 32, No 1, pp 303-315 304 1. Introduction Chili (Capsicum frutscenes L.) is a highly profitable cash crop popular among farmers and their markets [1], but its production poses significant risks. Peppers continue to grow under substantial pressure from pests and diseases. It is widely consumed in home, fresh seasoning, and as a cooking ingredient. Farmers mainly produce their crop for commercial marketing. Ethiopia’s share in the world, however, is insignificant (5%) compared to India (36%) and China (11%) with a production of 1.25, 0.39 and 0.17 million tones [3]. The decline of hot pepper production (0.4 tones fruit yield/ha) is attributed to the prevalence of fungus among others [1, 4]. The reason for decline of hot pepper production is attributed to poor varieties, poor cultural practices, the prevalence of fungal (blights) and bacterial as well as viral diseases [4]. It is a serious threat to crop productivity during rainy weather [1]. This can be doubled or tripled through in the absence of appropriate disease management coupled with good agricultural practices. According to [6], the present situation indicates that in the study area there is no improved hot pepper varieties but there is one local variety named “Mita Mito” by local growers, the green pod yield (3 ton per hectare) of this local variety is very low compared to national average yield. As a result, information for the improvement of the crop for high fruit yield and quality in the existing agro-ecology is insufficient [5]. There has also been no research on evaluation of hot pepper which enables the growers to select the best performing varieties in the study area [6]. Conversely, the dose and frequency of fungicides being applied to control the disease is costing too much for the small scale farmers often without significant benefit. The effectiveness of fungicides against chili anthracnose was lower or comparable to antagonists and botanicals [7-10]. Even though botanicals are safe, cheap and obtainable [11], their application is far below it was supposed to be. However, limited efforts have been made to screen plants that are suspected to possess antimicrobial properties for effect against C. capsici. Higher plants may contain secondary compounds that could effectively control plant diseases [12]. Yet, options have been identified to address these challenges. Conventional synthetic fungicides need to be replaced by bio-fungicides as the former lost their effectiveness due to pesticide treadmill [13, 14]. Environment friendly control tactics gained impetus due to growing socio-economic concerns. In pursuit of finding replacement of toxic pesticides scientists working in these lines started trying botanical extracts and such other substitutes. In the past decades, therefore, quite a few scientific published information became available on this subject. This research was, therefore, initiated with objective of managing chili anthracnose through integrated use of antagonists and plant extracts under field conditions. 2. Materials and Methods 2.1 Experimental site The experiment was conducted at the FTC site in Alaba which is one of the most important pepper growing locations that have altitude of 1680 above sea level, is characterized by dry sub humid climate during June, 2013. Alaba has monthly mean minimum and maximum air temperature of 15°C and 29.5°C, respectively, and rain fall of 900-1300mm/year. This location is a hot spot area for anthracnose (Colletotrichum spp) and wilts [2, 15, 16]. American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) (2017) Volume 32, No 1, pp 303-315 305 2.2 Treatments A total of 16 treatments, viz., Untreated (T0), combination of all (Extracts + Trichoderma spp.+ Ridom)(T1) Neem (T2), Cassia spp (T3), Onion (T4), Garlic (T5), Neem + Onion+ Cassia spp (T6), Neem + Onion + Garlic (T7), Ridomil (T8), AAU-Th (T9), AAU-37 (T10), AAU-69(11), AAU-Th + Garlic (T12), AAU-69 + AAU-Th + Onion(T13), AAU-37 + AAU-Th +Neem (T14), AAU-69 + AAU-37 +Neem(T15), and AAU-69 + AAU-37 + AAU-Th (16) had been used in this experiment. 2.3 The Fungal Pathogen Sweet pepper fruits with anthracnose lesions were collected from farmers’ fields in main chili growing areas in September 2012. Sections of 3-5 mm were cut from the margin of the infected lesions and sterilized for one minute in 1.0% sodium hypochlorite solution and rinsed in three changes of sterile distilled water (SDW). The sterile pieces were blotted dry using sterile filter papers and placed on Potato Dextrose Agar (PDA) in 9cm Petri dishes. The dishes were incubated at ambient conditions of light and temperature (30 ± 2°C) for 7 days after which cultures with salmon-pink sporulation typical of Colletotrichum spp were sub-cultured to obtain pure cultures [12]. Culture identification was confirmed by microscopic examination and comparison with reference cultures [14]. From these, virulent isolate of C. capsici, A38, was obtained from the Laboratory of the Department of Microbial, and Cellular and Molecular Biology, College of Natural and Computational Sciences, Addis Ababa University, Addis Ababa, Ethiopia. The fungal pathogen was maintained on potato dextrose agar (PDA) slants at 4O C [14]. 2.4 Preparation of Trichoderma spp isolates for the Experiment Three isolates of Trichoderma spp with high biocontrol efficacies were cultured in Potato dextrose broth (PDB) and cells were collected by centrifugation at 3,000 rpm for 20 min. Trichoderma cells were washed twice with sterile distilled water and re-suspended. Then 20 µl of cell suspension of each strain of Trichoderma spp at concentration of 5x106 cells/ml was added to the wound of 30 treated chili fruits. After air drying, 20 µl of C. capsici, 5x106 cells/ml was added to the wound. Disease severity, as indicated by increased wound diameter, was counted after 5 days of inoculation. The ability to reduce disease incidence of each Trichoderma spp strain was observed and compared [17]. The most effective Trichoderma spp strain was selected for field studies. 2.5 Layout and Design of the Experiment The experiment was laid in Randomized Complete Block Design (RCBD) and divided into three blocks and each block was divided into 17 plots. The plot size was 5m x 1.2m. Treatment was assigned to each block at random. The space between the blocks and between the plots was 1.00 m and 0.50 m, respectively. The first spray was given on the 21st DAT as leaf spots could be located the day before. All the treatments (T0-T17) were administered as foliar spray. Plant extracts were formulated at 1:10 suspensions, T6 and T7 as 1:10 suspension cocktail. Ridomil (T8) was administered @ 0.2% suspension. Usually in the evening calm weather light spraying was done so that neighboring plots cannot share a wrong treatment. As the symptom bearing leaves/infected plants serve as a source of inoculum, field combination of all (Extracts + Trichoderma spp + Ridomil) was used American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) (2017) Volume 32, No 1, pp 303-315 306 as a treatment (T2). In this case all the fallen or hanging diseased leaves were collected removed and destroyed. 2.6 Data collection Number of infected plants per plot, Number of infected leaves per plant, number of spots per leaf, Percent diseased leaf area (DLA %), were data collected on disease reactions. Data collected on Growth Parameters included Plant height, days to 50% flowering, number of flowers per plant, Days to first harvest, Canopy diameter (cm), Number of branches per stem and Dry weight content per plant. Data collected on Quality factors were fruit pericarp thickness (mm), Fruit dry weight content (g), fruit length (cm), and fruit diameter (cm). 2.7 Analysis of data The data were statistically analyzed with the help of Analysis of variance (ANOVA). To compare the means, Fisher’s Least significance Difference (FLSD) was used to compare the effect of the treatment at p<0.05 [18]. 3. Results The obtained data on different parameters are presented in the Tables (1-3). The effect differences of the treatments significantly varied from one another and gave a clear picture about the effects on disease reaction, quality and growth factors . 3.1 Effect of IDM on Disease reaction 3.1.1 Plant Infection The results of this study revealed that the treatments (T1-T16) reduced the incidence of anthracnose of chili disease significantly compared to the control treatment through the observation period, i.e., Bako local, Oda Haro and Maraqo Fana. In all the observed varieties the control (T0) plots had the highest percent plant infection. The highest percentage of plants showing anthracnose of chili symptoms on Maraqo Fana was 85.33 in T0 (Control) and it was the lowest (12.80) in combinations of Trichoderma spp isolates, plant extracts and Ridomil (T1). Though numerically different, percent plant infection in T16 (AAU-69+AAU-37+AAU-Th) was in a statistically similar level of significance with T1 (combination of Trichoderma spp isolates, plant extracts and Ridomil). On Oda Haro variety, the highest percent plant infection was found in To (control) which was 89.6 and the lowest percent plant infection was found in T1 (Trichoderma spp isolates, plant extracts and Ridomil) which was 15.33. On Bako local variety, the highest (85.2) and the lowest (19.6) percent plant infection was observed in T0(control) and T1(Trichoderma isolates, plant extracts and Ridomil). The AAU-Th and AAU-37 Trichoderma spp isolates (T9 and T10) in the same level of significance with 22.6 and 23.6, respectively. Overall, percent plant infection was found to be higher in aqueous plant extracts than isolates of Trichoderma spp. The infection in Trichoderma spp treated plots, on the other hand, was significantly lower than Ridomil (T8) treatment. American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) (2017) Volume 32, No 1, pp 303-315 307 Table 1: Percent plant and Leaf infection per plant due to at Anthracnose o f chili at different days after transplanting as in influenced by some management practices Treat- Ments Percent Plant Infection* Leaf Infection Per Plant % Diseased Leaf Area (%DLA) % Fruit Infection Plant MF OH BL MF OH BL MF OH BL MF OH BL T0 85.33k 89.6 i 85.2k 87.4g 89.9f 85.2g 87.4i 89.9 f 87.4g 89.9f 85.33f 81.8g T1 12.8a 15.33a 19.6a 15.2a 17.4a 19.9a 15.2a 17.4a 19.9a 17.4a 19.9a 15.33a T2 71.66j 73.7 i 78.66j 22.8d 25.33d 29.6e 32.8e 35.33d 39.6f 36.33d 49.6e 23.7d T3 27.62f 44.54g 54.2i 23.66d 23.7c 28.66de 33.66e 33.7c 38.66e 35.7cd 48.66de 24.54de T4 44.44i 49h 53.25i 20.62c 24.54cd 26.2cd 30.62d 34.54cd 36.2d 35.54c 46.2d 29f T5 40.66h 46.22gh 51.23i 24.44df 29e 29.25e 34.44e 39e 39.25ef 39.11e 49.25e 26.22e T6 33.14g 35.88f 41.56gh 20.66c 26.22d 28.23d 30.66d 36.22d 38.23e 37.22d 48.23d 25.88e T7 27.66f 32.31ef 42.2h 22.14d 25.88d 26.56d 32.14de 35.88d 36.56d 37.88de 46.56d 22.31cd T8 22.43d 30.6e 37.2g 24.66f 22.31bc 22.2a 34.66e 32.31c 32.2b 36.31d 42.2c 20.6b T9 23de 19.19b 22.6a 21.43cd 20.6b 27.2d 31.43d 30.6b 37.2de 36.6d 47.2d 19.19b T10 18.3c 19.9b 23.6ab 14 a 19.19b 22.6ab 24b 29.19b 32.6bc 29.99b 42.6c 19.9b T11 22.6d 26.53d 29.9cd 17.3b 19.9b 23.6bc 27.3c 29.9b 33.6c 29.9b 43.6c 26.53e T12 27.6f 29.9de 31.8d 23.6d 26.53d 29.9ef 33.6e 36.53de 39.9f 37.53d 49.9e 20.9b T13 17.7c 20.3c 25.6bc 15.6a 20.9b 31.8f 25.6b 30.9bc 31.8b 36.9d 41.8c 20.3b T14 21.6d 30e 35.5fg 15.7a 20.3b 25.6c 25.7bc 30.3b 35.6cd 36.3d 45.6cd 20b T15 24e 28.2d 32.2df 18.6bc 20b 25.5c 28.6cd 30b 35.5c 38e 45.5c 21.2bc T16 15ab 20.15bc 25.6bc 17ab 20.2b 23.2b 23b 30.2b 31.2b 33.2bc 38.2b 26.33e LSD5% 2.25 4.2 4.4 3.1 1.53 2.94 3.59 3.3 2.2 2.6 3.4 2.3 CV 15.8 16.1 10.6 18.2 13.3 15.5 14.3 18.1 15.4 18.7 18.5 21.7 ** Significant at 5% level; Means followed by the same letter (s) in a column did not differ at 5% level by LSD, MF=Maraqo Fana, OH=Oda Haro, BL= Bako local Statistically identical results were recorded in Maraqo Fana plots in T10 (AAU-37) and T13 (AAU-69+Th + Onion) with values of 17.7 and 18.3, respectively. The next highest percent plant infection in Oda Haro was observed inT9 and T10, 19.19 and 19.9, respectively. Moderate infection was observed in T13(with value of 20.3) and T16 (with 20.15). In Bako local variety, the second lowest (29.9, 25.6 and 25.6 ) was observed in T11 (AAU-69), T13(AAU-69+AAU-Th+Onion) and T16 (AAU-69+AAU-Th+Onion).But treatment T8 (Ridomil) was chemical which was not environment friendly which also significantly differed from To(Control), had a value of 22.43, 30.6 and 37.2 percent plant infection, in Maraqo fana, Oda Haro and Bako local varieties. This higher infection may be due to the fact that Ridomil is a very toxic and injurious to human health and not an environment-friendly product it would have destroyed the beneficial microorganisms too. 3.1.2 Leaf Infection American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) (2017) Volume 32, No 1, pp 303-315 308 Effect of different treatments o n percent leaf infection are had revealed that there is statistical difference among the treatments at p<0.05. On Maraqo Fana variety, the highest percentage of leaf infection 87.4 % was observed in the treatment T0 (control). The result clearly indicated that the treatments had significant effect on percent leaf infection. The lowest (14) percent leaf infection was observed in treatment T10(AAU-37 ), T2 (combination of Trichoderma isolates, plant extracts and Ridomil), T13(AAU-69+AAU-Th+Onion ), T14(AAU-37+AAU-Th+Neem) and T16 (AAU-69 + AAU-37+ AAU-Th) with values of 15.2, 15.6, 15.7 and 17 percent leaf infection, respectively (Table 1). On Oda Haro variety, the highest percentage of leaf infection, 8 9 . 9 , was observed in the treatment To (control) and this was significantly higher than under any other treatments. The result clearly indicated that the treatments had significant effect on percent leaf infection. The lowest (17.4) percent leaf infection was observed in treatment T2 (combination of Trichoderma isolates, plant extracts and Ridomil). The second lowest(19.19 and 19.9) percent leaf infection was observed in T10 (AAU-37) and T11(AAU-69) which were statistically similar to T 1 3 ( AAU-69+AAU-Th+Onion) , T15(AAU-69+AAU-37+Neem), T16 (AAU- 69+AAU-37 + AAU-Th), T14 (AAU-69+AAU-Th+Neem), T9 (AAU-Th ) and T8 (Redomil) having of 20.9%, 20, 20.2, 20.3, 20.6 and 22.31% respectively (Table 1 ). The result clearly indicated that, on Bako local variety, the treatments had significant effect on percent leaf infection. T he highest leaf infection, 85.2% , was observed in treatment To (control). The lowest (19.9 %) percent leaf infection was recorded in treatment T2 (combination of isolates Trichoderma spp, plant extracts and Ridomil). Percent leaf infection was statistically similar in T8 (Ridomil) and T10 ( AAU-37) having of 22.2% and 22.6%, respectively. Regarding the second lowest infection, T16 (AAU-69+AAU-37 + AAU- Th), and T11 (AAU-69) were statistically similar having 2 3 . 2 % and 23.6%, respectively. The rest of t h e treatments had shown statistically significant effect (Table 1). 3.1.3 Percent of Diseased Leaf Area (%DLA) The result clearly indicated that the treatments had significant effect on percent diseased leaf area. On Maraqo Fana variety, the highest percent diseased leaf area was observed in To (control) having value 87.4 % and the lowest diseased leaf area was in T2 (combination of Trichoderma isolates, plant extracts and Ridomil). Many treatments had fallen in the second statistical significance category. Treatment T16 (AAU-69+AAU-37 + AAU-Th ), T10 (AAU-37), T13 (AAU-69+AAU-Th+Onion) and T14 (AAU-69+AAU-Th+Neem) showed statistically similar significant effect having values 23.0 %, 24.0 % , 25.6% and 25.7 %, respectively. The rest of the treatments also showed significant effect (Table 1). Single (T9-T11) and combined (T9-T16) treatments of Trichoderma spp, that includes AAU-Th, AAU-37and AAU-69; and plant extracts which include extracts of Neem, Onion and Garlic as mixture spray was found more efficient than Ridomil. The relative efficiencies of treatments suffered only a slight change on other varieties probably due to environmental factors and the growth stage physiology of the plants. The disease development in the infected leaves at this period was rather low Maraqo fana compared to the Oda Haro variety. On Oda Haro variety, the highest percent diseased leaf area was observed in To (control) having value 89.9 % American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) (2017) Volume 32, No 1, pp 303-315 309 and lowest diseased leaf area was in T1 (combination of isolates of Trichoderma spp, plant extracts and Ridomil) having value 17.4 %. On the other hand, Treatment T9 (Th), T10 (37), T11 (69), T13 (69+Th+O), T14 (37+Th+N ), T15(69+37+N )and T16 (69+37+Th ) showed statistically similar effect having values 30.6 %, 29.19 %, 29.19%, 30.9%, 30.3%, 30% and 30.2 % respectively. The rest of the treatment also showed significant effect (Table 1). On Bako local variety, the highest percent diseased leaf area was observed in T0 (control) having value 87.4 % and the diseased leaf area was in T1(combination of Trichoderma isolates, plant extracts and Ridomil) having value 19.9 %. The second least infection was observed in Treatment 16 (AAU-69+AAU- 37 + AAU-Th), T13 (AAU-69+AAU-Th+Onion ),T10 (AAU-37) and T8 (Ridomil) with statistically similar effect having values 31.2 %, 31.8%, 32.6% and 32.2 %, respectively. The rest of the treatment also showed significant effect (Table 1). 3.1.4 Infected Fruits per plot Effect of different treatments on percent fruit infection had shown that in Maraqo fana variety, all the treatments showed statistically significant effect on reducing percent fruit infection compared to control. The highest percent of fruit infection was observed in control plot To (89.9%) and the lowest percent of fruit infection was recorded in T1(17.4%).The treatments 16 (AAU-69 + AAU-37 + AAU-Th), T11(AAU-69) and T10 (AAU-37) had shown statistically similar significant effect with values of 32.2%, 29.9% and 29.99%, respectively (Table 1). On Oda Haro variety, the treatments also showed statistically significant effect on percent fruit infection. The highest percent of fruit infection was observed in control plot To (85.33%) and the lowest percent of fruit infection was recorded in T1 (19.9%).The treatments 16 (AAU-69 + AAU-37 + AAU-Th) had shown the second lowest infection with value of 38.2%. On Bako local variety, the highest percent of fruit infection was observed in control plot To (81.8 %) and the lowest percent of fruit infection was recorded in T1(15.33%). The treatments T8 (Ridomil), T9 (AAU-Th), T10 (AAU-37), T12 (AAU-Th + Garlic), T13 (AAU-69 + AAU- Th + Onion), T14 (AAU-37 + AAU-Th + Neem), T15(AAU-69+ AAU-37+Neem) and T16 (AAU69+ AAU-37 + AAU-Th) showed statistically similar effect having values 20.6 %, 19.19 %, 19.9%, 20.9%, 20.3%, 20% and 21.2 %, respectively (Table 1). Control (T0) where no treatment was given, the % fruit infection on Bako local became less as compared to the situation observed on Maraqo fana and Oda Haro varieties. But the plants which received treatment, whichever it may be, the spread of the disease on the fruit was considered low. The highest fruit infection was observed in T0 (Control) and significantly small difference in other treatments. From the tested Trichoderma spp isolates applied singly or in combination, the lowest infection was observed on AAU- 69+AAU-37+AAU-Th. However, the efficacy of antagonists, extracts and fungicides, i. e, AAU-Th + Neem + Ridomil, appeared to be compatible and effective. Among the botanical extracts the strongest anti-Colletrotrichum capsici (Syd) reaction has been shown in terms of percent fruit infection by Garlic leaf extract (39.11, 49.25 and 26.25, on Maraqo Fana, Oda Haro and Bako local varieties, respectively), which was lower than Cassia spp extract (35.7, 48.66 and 24.54, on Maraqo Fana, Oda Haro and Bako local varieties, respectively). Ridomil's effect in reducing % fruit infection was not as anticipated. It was lower than the antagonists and leaf extracts. American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) (2017) Volume 32, No 1, pp 303-315 310 Throughout the experiment in all the parameters taken into account Integrated use of Antagonists, plant extracts and 0.2% application of Ridomil as foliar spray performed excellent. This was the expected result too. Ridomil treatment was a positive control treatment as no treatment was considered as a control treatment. The aim was to compare the effects of the antagonist (Trichoderma spp isolates) and organic (botanicals) treatments with both. The results and their analyses revealed that the test treatments, antagonists and botanical extracts, are capable of reducing anthracnose of chili in the cultivars Maraqo fana, Oda haro and Bako local quite significantly even when applied in combination and singly. However, combination of antagonists and botanical extracts were more strong and effective even to a level that with minimum risk such treatment can replace a highly effective chemical fungicidal treatment. 3.2 Effect of IDM on Growth Performance of Hot pepper There were significant variations among the tested genotypes in terms of the percent establishment, days to 50% maturity and days to first harvest (Table 2). The result showed a range of 48.67 (T0) to 82.67(T11), 48.00 (T0) to 83.33(T11) and 49.67(T0) to 89.67(T4) for percent establishment; 43.0(T0) to 67.33(T13), 41.00 (T0 and T4) to 53.33(T10) and 36.33(T0) to 53.67(T5) for days to 50% flowering; 62.0(T0) to 82.0(T4), 65.33 (T0) to 79.67(T13) and 40.67(T0) and 81.0 (T14) for days to 50% of maturity; 78.67(T0) to 106.0(T8), 52.67 (T0) to 108.33(T13) and 51.67(T0) to 107.33(T13) for days to first harvest for Maraqo fana, Oda haro and Bako local varieties, respectively (Table 2). Quality parameter trial, the result revealed that significant variations existed among the tested genotypes in terms of number of branches per stem, canopy diameter (cm), number of flowers per plant and plant height (Table 3). The result showed a range of 1.0 (T0) to 9.0(T10), 1.0 (T0) to 8.0(T4) and 2.0 (T0 and T13) to 8.0 (T11) for branches per stem; 5.80(T0) to 24.8(T15), 4.8 (T0) to 12.8 (T11) and 5.6 (T0) to 12.6 (T2) for canopy diameter (cm); 5.26 (T12) to 9.4 (T2), 4.7 ( T1) to 9.6(T3) and 5.8(T0) to 10.8(T4) for number of flowers per plant, 13.9 (T0) to 61.4 (T7), 15.5 (T0) to 78.8 (T15) and 12.8 (T0) to 68 (T8) for plant height, on Maraqo Fana, Oda Haro, and Bako local varieties, respectively. 3.3 Effect of IDM on Quality Performance of Hot pepper On another quality parameter trial, the result revealed that significant variations existed among the tested genotypes in terms of fruit diameter(in cm), Fruit pericarp thickness (mm), fruit length (cm) and fruit dry weight content (g) (Table 3). The result showed a range of 0.5 (T0) to 3.3(T1), 0.6 (T0) to 1.9(T10) and 0.8(T0) to 3.6 (T14) fruit diameter(in cm); 0.35 (T0) to 1.38 (T5), 0.31 (T0) to 1.6 (T7) and 0.32 (T0) to 1.55(T6) for fruit pericarp thickness (mm); 5.25 (T0) to 9.7(T4), 4.69 ( T1) to 9.59 (T3) and 6.3(T10) to 10.79(T4) for fruit length (cm); 4.9(T9) to 9.2 American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) (2017) Volume 32, No 1, pp 303-315 311 (T12), 6.74 (T13) to 11.0 (T1) and 6.5 (T13) to 10.7 (T3) for fruit dry weight content (g) on Maraqo Fana, Oda Haro, and Bako local varieties, respectively (Table 4). Table 2: Effect of IDM on growth parameters of Hot pepper varieties in 2013 cropping season Var. *Establishment % Days to 50% Flowering Days to 50% Maturity Days 1st Harvest Treat MF OH BL MF OH BL MF OH BL MF OH BL T 0 48.67a 48.00a 49.67a 43.00a 41.00a 36.33a 62.00a 65.33a 40.67a 78.67a 52.67a 51.67a T 1 69.33b 72.00b 73.00b c 65.33f 48.00d 42.67b 70.00b 73.33c d 78.00e 100.00 e 101.33b 102.00c T 2 71.00bc 81.33d 74.00c 60.67e 42.00ab 43.33b 72.33b c 72.33b 76.67d 98.00c d 104.33c 100.33b T 3 75.33d 77.33c 74.00c 54.00b c 42.67b 50.67f 78.67g 72.00b 70.00b 97.67c 102.33b 99.67b T 4 74.67d 77.00c 89.67e 63.00ef 41.00a 46.33cd 82.00h 75.33d 80.67g 98.67d 102.67bc 98.67b T 5 74.00cd 82.33d 71.67b 51.67b 46.33c 53.67g 79.67g 73.00b c 71.00bc 98.67d 106.00d 102.33c T 6 81.67f 73.67b 73.00b c 56.33c d 46.33c 50.00df 73.67c d 77.00f 75.67d 97.67c 104.33c 102.67c T 7 81.00f 81.33d 70.67b 57.33d 47.00d 44.00b 81.67h 78.33g 78.00e 100.0e 104.67cd 102.00c T 8 81.33f 81.33d 73.00b c 59.67d 56.00g 44.67b 75.00d e 78.33g 80.33g 106.3f 104.33c 102.67c T 9 77.00de 82.33d 78.00d 58.00d 48.33d 50.00df 75.67e 79.00g 76.00d 97.00c 102.33b 100.33b T 10 81.00f 81.67d 76.00d 56.00c 53.33f 45.00bc 76.67f 76.67ef 76.33d 95.33b 104.33c 106.00c T 11 82.67f 83.33f 73.00b c 59.67d e 51.67ef 45.67c 72.33b c 75.67e 80.33g 98.67d 104.00c 104.33c T 12 81.33f 81.67d 74.00c 64.00f 52.00f 45.00bc 76.00e 79.33g 79.67fg 101.0e 105.33d 106.33c T 13 80.00e 80.67d 71.33b 67.33g 46.33cd 47.67d 73.00c 79.67g h 79.00ef 100.0e 108.33e 107.33cd T14 80.67ef 82.67e 76.00d 62.33e 45.33bc 43.33b 75.67e 75.67e 81.00gh 102.7e 106.00de 105.67c T 15 72.00c 82.00d 70.67b 53.00b 52.33f 45.67c 79.00g 72.33b 73.33c 100.6e 104.33c 105.33c T 16 81.67f 80.00d 75.33c d 65.33f 48.33de 51.33fg 76.00ef 72.67b 79.00ef 102.7e 101.67b 101.67bc LSD(5%) 3.11 2.51 2.78 3.32 2.60 2.42 1.43 1.14 1.32 1.32 2.41 5.62 CV% 11.1 28.4 16.7 18.7 13.1 22.5 18.9 16.8 17.4 11.00 22.4 23.2 *Means followed by the same letter in the same column are not significant difference at P<0.05; MF= Maraqo Fana, OH= Oda Haro, BL= Bako local American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) (2017) Volume 32, No 1, pp 303-315 312 Table 3: Effect of IDM Quality Parameters of three chili accessions at Alaba 2013 Treat- ments Number of Branches Per Stem* Canopy Diameter (cm) Number of Flowers Per Plant Plant Height (cm) MF OH BL MF OH BL MF OH BL MF OH BL T0 1a 1a 2a 5.8a 4.8a 5.6a 6.7b 4.7a 5.8a 13.9a 15.5a 12.8a T1 4. 3b 7de 6. 3c 12b 11.6c 12.6e 9.4e 4.7a 7.3ab 47.6d 61i 67.3g T2 7d 2a 2.1a 13.8b 10.44b 11.3c 6.8b 8.2d 9.7cd 26.5b 47.2d 49.5d T3 5.3c 2a 2.2a 12.8b 10.6b 11.1b 8.5d 9.6d 6.6a 37.5c 49.1e 40.7c T4 2.5b 8ef 2.2a 24.6f 10.9b 10.7b 9.6e 8.2d 10.8d 37.1c 30.5b 58.9f T5 3.6b 5bc 6bc 19.9d 11.4bc 10.89b 6.87b 7.1c 7.5b 36.7c 52.7f 57.99e T6 4.7bc 4b 2.3a 16.8c 12.5d 10.84b 6.9bc 6.5bc 8.7bc 60.6h 59.4h 67.34g T7 9e 7de 2.1a 19.6d 12.4d 12.1d 5.6a 7.9d 9.5c 61.4h 38c 60f T8 2.6b 2a 3a 21.3de 12.6d 11.9d 6.7b 6.1b 9.1c 58.8g 49.3e 68g T9 6.7cd 6cd 2.3a 18.7cd 11.8c 11.6c 7.6cd 7.6cd 7.8b 54.9e 69.4k 49.2d T10 9e 9f 7.4c 19.6d 12.3d 11.7cd 6.3ab 8.2d 6.3a 55.8f 38.6c 49.9d T11 3.33b 6cd 8cd 11.8b 12.8d 11.6c 6.7b 6.4b 7.8b 56.9f 49.5e 48d T12 6c 5bc 4.2b 13.8b 11.9cd 11.8d 5.26a 7.3c 6.6a 59.2gh 57.2fg 39.1b T13 8de 7de 2a 22.7e 11.3b 10.9b 5.44a 5.6a 7.5b 58.7g 67.4j 56.5e T14 6c 6cd 3.6ab 16.8c 11.9c 12.2de 8.7de 5.8ab 8.2b 58.8g 47.6d 58ef T15 5c 6cd 5.6b 24.8f 11.68c 10.96b 6.2a 7.8de 7.6b 56.4f 78.8m 58.76f T16 4b 4b 6bc 23.3ef 11.7c 11.2bc 5.5a 8.5d 9.3c 57.1fg 72 l 59.7f LSD(5%) 2.2 1.4 2.1 2.33 0.94 0.59 1.3 1.2 1.6 2.4 1.3 2.1 CV% 18.1 19.6 16.2 10.3 14.5 17.3 10.1 12.4 13.7 18.5 16.7 17.8 *Means followed by the same letter in the same column are not significant difference at P<0.05, MF=Maraqo Fana, OH=Oda Haro, BL= Bako local Table 4: Effect of IDM Quality Parameters of three chili accessions at Alaba 2013 Treat. *Fruit Diameter (cm) Fruit Pericarp Thickness (mm) Fruit Length (cm) Fruit dry weight Content (g) MF OH BL MF OH BL MF OH BL MF OH BL T0 0.5a 0.6a 0.8a 0.35a 0.31a 0.32a 6.7b 6.4b 7.8c 6.9b 9.5cd 8ab T1 3. 3d 1.7b 2. 3cd 1.11b 0.95a 1.11b 9.4d 4.7a 7.3b 7.6cd 11d 7.3a T2 1.2b 1.2a 2.1c 1.15b 1a 1.15b 6.8b 8.2d 9.7ef 6.5b 7.24a 9.5b T3 1.4bc 1.0a 2.2c 1.13b 1.33b 1.13b 8.5c 9.6e 6.6b 7.5c 9.1bc 10.7c T4 1.2b 1.8b 2.2c 1.21b 1.1b 1.21b 9.6d 8.2d 10.8f 7.1bc 10.5d 8.9b T5 1.3b 1.5ab 1.6b 1.38bc 1.28b 1.38bc 6.87b 7.1c 7.5b 6.7b 8.27b 7.99a T6 1.4bc 1.4a 2.3cd 1.55c 1.45b 1.55c 6.9bc 6.5bc 8.7d 6.6b 9.4c 7.34a T7 1.9c 1.7b 2.1c 1b 1.6b 1b 5.6a 7.9d 9.5e 6.4b 8b 10bc T8 1.1b 1.2a 2.3cd 1.32b 1.52b 1.32b 6.7b 6.1b 9.1de 5.8ab 9.3c 8ab T9 1.0b 1.6b 2.3cd 1.22b 1.32b 1.22b 7.6c 7.6c 7.8c 4.9a 9.4c 9.2b T10 1.9c 1.9b 2.4d 1.37b 1.17b 1.37b 6.3ab 8.2d 6.3a 5.8ab 8.6b 9.9b T11 1.0b 1.6b 2.8e 1.21b 1.11b 1.21b 6.7b 6.4b 7.8c 6.9b 9.5cd 8ab T12 1.3b 1.5ab 2.2c 0.88ab 0.98a 0.88ab 5.26a 7.3c 6.6b 9.2d 7.25a 9.1b T13 1.8c 1.7b 2.0c 0.91b 0.87a 0.91b 5.44a 5.6a 7.5b 8.7d 6.74a 6.5a T14 1.6c 1.6b 3.6f 1.13b 1.23b 1.13b 8.7cd 5.8ab 8.2cd 8.8d 10.6d 8ab T15 1.6c 1.6b 1.6b 0.89b 0.9a 0.89b 6.2a 7.8cd 7.6bc 6.4b 7.88ab 8.76b T16 1.4bc 1.4a 2.2c 0.94b 0.99a 0.94b 5.5a 8.5de 9.3e 7.1bc 7.2a 9.7b LSD(5%) 0.4 0.98 0.31 0.56 0.68 0.57 1.11 1.12 1.1 1.4 1.23 2.21 CV% 10.1 11.6 15.2 12.3 16.5 19.3 20.1 18.4 14.5 13.5 14.7 19.8 *Means followed by the same letter in the same column are not significant difference at P<0.05, MF=Maraqo Fana, OH=Oda Haro, BL= Bako local American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) (2017) Volume 32, No 1, pp 303-315 313 4. Discussions This study revealed that antagonists and plant extract had very effective strong fungicidal effect. In some cases, the reported effect is more than that of chemical pesticides. Out of the many such reported promising plants, on the simple basis of availability Garlic, Neem, Onion and Cassia have been selected to assess their combat ability against Collectrotrichum capsici, that is the ability against anthracnose of chili. Different integrated practices like A total of 16 treatments, viz., Untreated (T0), combination of all (Extracts + Tricho.+ Ridomil) (T1) Neem (T2), Cassia spp (T3), Onion (T4), Garlic (T5), Neem + Onion+ Cassia spp (T6), Neem + Onion + Garlic (T7), Ridomil (T8), AAU-Th(T9), AAU-37( T10), AAU-69(11), AAU-Th + Garlic (T12), AAU-69 + AAU-Th + Onion (T13), AAU-37 + AAU-Th +Neem (T14), AAU-69 + AAU-37 + Neem (T15), and AAU-69 + AAU-37 + AAU-Th(16) had been used in this experiment. Data on disease reaction, quality and growth parameters had been collected. Plant infection, leaf infection, leaf area diseased, and fruit infection, were the parameters for disease reaction. The highest average fruit yields however were observed under the treatment T6 and T7 and the lowest under the treatment To. As a result low yield was found in control plots and high in the plots where combined treatments were applied. The combined treatment had a highly significant effect on the fruit yield. The results obtained by Serawit and Tesfaye [11,19], reference [20] with the use of garlic and Kabir [21] with Neem (Azadirachta indica) leaf and Neem seed extracts nicely corroborate with the present findings. The results obtained through this experiment indicated that a judicially designed combined organic treatment even may be profitable than a chemical fungicide treatment of all the fruit collection from the respective plots. The chili test plants which have received Ridomil @ 0.2% had the lowest disease intensity as well as severity parameters. Plot treated with antagonists, plant extracts and fungicides ( T1) has shown the highest disease control potential but significantly superior to the control(To). The plant extract treatments have shown significantly better control than the T8, though they varied widely amongst themselves. The combined treatments T6 (Neem + Onion + Cassia ) and T7 (Neem + Onion + Garlic) have shown very strong response though lesser than the T8 (Ridomil) in controlling anthracnose of chili. This is in conformity with [22,13]. The highest average fruit diameter however were observed under the treatment T1 and T10 and the lowest under the treatment To. As a result, low diameter was found in control plots and high in the plots where combined treatments were applied. The combined treatment had a highly significant effect on the fruit diameter. The results obtained by [21] with the use of garlic and Serawit and Tesfaye [11] with Neem (Azadirachta indica) leaf and Neem seed extracts nicely corroborate with the present findings. 5. Conclusions and Recommendation The results obtained through this experiment indicated that a judicially designed combined organic treatment even may be profitable than a chemical fungicide treatment. Thus it can be concluded that the incidence and severity of Anthracnose of chili disease can significantly be reduced by the combined use of (Trichoderma spp + Plant Extracts) 1: 10 dilution suspension foliar spray in order to have a higher profitable yield and American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) (2017) Volume 32, No 1, pp 303-315 314 eventual higher economic return with minimum health risk as well as environmental pollution. However, meticulous with higher potency or efficacy must go on as the pathogenecity of the causal agent is quite dynamic. Therefore, the farmers may be advised to take an integrated approach which should include antagonists and plant extracts to raise a profitable production without polluting the environment and adding toxins in the food chain. Acknowledgement The authors thank Addis Ababa University and Wolaita Sodo University for their support. References [1]. Amusa, N.A, Kehinde, I.A. and Adegbite, A.A., “Pepper fruit anthracnose in the humid forest region of south-western Nigeria”. Nutrition and Food Science 34(3): 130 -134. 2004. [2]. Tameru, Alemu., Hamacher, J. and Dehne, H.W. “The increase in importance of Ethiopian Pepper mottle virus (EPMV) in the rift valley part of Ethiopia” Time to create Awareness among researchers an extension workers. Pepper presented at Deutsches Tropentage, October 18-21, 2003. Gottingen, Germany. [3]. Faisal H. and Muhammad A. “Pests and Diseases of Chili Crop in Pakistan: A Review”. Int. J. Biol. Biotech., 8 (2): 325-332, 2011. [4]. Fekadu, M. and Dandena, G. “Status of Vegetable crops in Ethiopia”. Ugandan Journal of Agriculture, 2006, 12(2): 26-30. 2006. [5]. Haileslassie Gebremeskel, Haile Abebe, Wakuma Biratu, Kedir Jelato. “Performance evaluation of hot pepper (Capsicum annum L.) varieties for productivity under irrigation at Raya Valley, Northern, Ethiopia”. Basic Research Journal of Agricultural Science and Review Vol. 4(7) pp. 17. 2015. [6]. Melaku, F.T., Alemayehu, T. G., and Lidet, B. T. “Adaptation Trail of Different Improved Hot Pepper (Capsicum species) Varieties under Gedeo Zone, Dilla, Ethiopia”. International Journal of Life Sciences. Vol. 4. No. 4. Pp. 216-220. 2015. [7]. Deeksha, L., Tripathi, H. S. and Joshi, D. “Effect of Indofil M-45 in disease severity of anthracnose in Urdbean”. J. Mycology and P.Path.32(1): 86-87. 2002. [8]. Ekbote, S.D. Bio-efficacy of Copper Hydroxide(coxid) against anthracnose of chili. Kamataka J.Agril.Sci.Agril.Res.Sta.,Haveri, India.15(4):729-730. 2002. [9]. Khoda, S. K., Hosna, K. and Khan, M. A. “Application of foliar fungicides to control Alternaria blight of cauliflower seed crop”. Bangladesh J. Plant Path. 19(1/2):33-37. 2003. [10]. Rahman, M.K.; Islam, M.R. and Hossain, I. “Effect of 'Bion, Amistar and Vitavex on anthracnose of chili”. J.Food Agriculture and Environment. (2):210-217. 2004. [11]. Serawit Handiso and Tesfaye Alemu. “Evaluation of Extracts of some noxious plants against coffee berry disease (Colletotrichum kahawae L.)”. International Journal of Basic and Applied Research. Volume16. Jordan. 2014. [12]. Nduagu, C, Ekefan E. J. and Nwankiti, A.O. “Effect of some crude plant extracts on growth of Colletotrichum capsici (Synd) & Bisby, causal agent of pepper anthracnose”. Journal of Applied Biosciences 6(2): 184 –190. 2008. American Scientific Research Journal for Engineering, Technology, and Sciences (ASRJETS) (2017) Volume 32, No 1, pp 303-315 315 [13]. Rashid, M.M., A.B.M. Ruhul Amin and F. Rahman, “Eco-Friendly management of Chilli Anthracnose (Colletotrichum capsici)”. Int. J. Plant Pathol., 6 (1): 1-11, 2015 [14]. Fekadu Alemu and Tesfaye Alemu. “Pseudomonas fluorescens isolates as an inducer of physiological activities of faba bean (Vicia faba)”. African Journal of Agricultural Research,8(38): 4864-487. 2013. [15]. Belete, N., Alemayehu, C., Girma, T.,G., G.E., Teferi. “Evaluation of farmers’ “Markofana–types” pepper genotypes for powdery mildew (Leveillula taurica) resistance in Southern Ethiopia.” International Journal of Basic and Applied Sciences Vol. 1 No. 2. 2012. [16]. Simon, A., Tameru, A., Ferdu. A. and Temesgen, A. “Population dynamics of aphids and incidence of Ethiopian pepper mottle virus (EPMV) in the rift valley part of Ethiopia”. crop protection. 28: 443-448. 2009. [17]. He, D., Zheng, X.D., Yin, Y.M., Sun, P., and Zhang, H.Y. “Yeast application for controlling apple postharvest diseases associated with Penicillium expansum.” Botanical Bulletin Academia Sinica 44: 211–216. 2003. [18]. Obi, I.U. “Statistical Methods of Detecting Differences between Treatment Means and Research Methodology Issues in Laboratory and Field Experiments”. 2nded. AP Express Publishers, Nsukka. 117pp. 2002. [19]. Ngullie, M., L. Daiho and D. Upadhyay, “Biological management of fruit rot in the world's hottest chilli (Capsicum chinense Jacq.)”. J. Plant Prot. Res., 50: 269-273. 2010. [20]. Nashwa, S.M.A. and K.A.M. Abo-Elyousr, “Evaluation of various plant extracts against the early blight disease of tomato plants under greenhouse and field conditions”. J. Plant Prot. Sci.,48: 74-79. 2012. [21]. Kabir, M.H., M.M. Rashid, M.R. Bhuiyan, M.S. Mian, M. Ashra fuzzaman, M.Y. Rafii and M.A. Latif, “Integrated management of alternaria blight of broccoli”. J. Pure Applied Microbiol., 8: 149-158. 2014. [22]. Ademe, A., A. Ayalew and K. Woldetsadik, “Evaluation of antifungal activity of plant extracts against papaya anthracnose (Colletotrichum gloeosporioides)”. Plant Pathol. Microb., Vol. 4. 2013. Integrated Management of Anthracnose (Colletotrichum capsici (Syd.)): Implications to Disease Reactions, Quality and Growth Parameters of Three Genotypes of Chili Serawit Handisoa*, Tesfaye Alemub aEmail: serawithandiso@gmail.com, bEmail: tesfayealemu932@gmail.com Abstract 1. Introduction 2. Materials and Methods 2.1 Experimental site 2.5 Layout and Design of the Experiment 4. Discussions