Evaluation the effectiveness of sheath blight disease controlling on rice caused by Rhizoctonia solani in green house condition by applying some biofungicides45239

Evaluation the effectiveness of sheath blight disease controlling on rice caused by Rhizoctonia solani in green house condition by applying some biofungicides Ha Thi Thanh Tuyen (1), Luu Ba Hoa(1)((*) (1) Kien Giang University, Kien Giang, Vietnam *Correspondence: lbhoa@vnkgu.edu.vn Abstract: This research was conducted at green house of Plant Protection Department, College of Agriculture and Applied Biology, Can Tho University The aim of this study find out the timing application of biofungicides (Validan 5SL and Trico-DHCT) for best effectiveness in controlling sheath blight disease (Rhizoctonia solani) on rice The result found that all of the bio-fungicide treatments were capable of controlling sheath blight disease on rice in green house condition with different levels Among these bio-agents, Trico-DHCT treatment sprayed before pathogen inoculation was to take a higher disease controlling effectiveness than the other treatments Keywords: Sheath blight disease; controlling disease; biofungicide Introduction Agriculture plays a very important role in the Vietnamese economy in which food production is the most important sector to contribute to Vietnamese GDP Vietnam is the second largest rice exporter in the world and also plays an important role in world food security It is located in a tropical climate to have a lot of conditions for paddy cultivation and also become the favorable conditions for the development of many pests In which, sheath blight disease (SBD) is one of main factors to damage the rice areas across Vietnam This disease can cause high yield losses a massive 25-50% (Ou, 1985; Agarwal et al., 1989; Gnanamanickam, 2009) In recent years, the trend of rice production following the monoculture production, increasing the number of crops and using high yielding varieties are causes for more and more diseases Becides, sheath blight disease is one of the most important one to destroy the states’ rice area which affected yield crop, environment and human health when applying the pesticides indiscriminately In countryside, farmers always apply pesticides overdose and continuously to make a high pressure on the pathogen resistance in the crop In order to control sheath blight disease, many methods had researched, until 2010, American institute of Agriculture had discovered the ability of gen which was resistant sheath blight disease However, that is application in the research and non-application in the wide area for planting rice Currently, there are many biofungicides to control sheath blight disease on rice which some of them were made of biological active ingredients (a.i) consist of Validan 5SL (Validamycin A) and Trico-DHCT which is two important a.i in plant disease controlling This topic focus on the effect and timing application of these biofungicides in control sheath blight disease on rice So, the study “Evaluation the effectiveness of sheath blight disease controlling on rice caused by Rhizoctonia solani in green house condition by applying some biofungicides” is researched Methodology 2.1 Objectives Determining the timing application of biofungicides for the effectiveness on controlling SBD in greenhouse condition 2.2 Design experiment Experiments were followed by the completely randomized design with two factors including 12 treatments and replications Factor A consists of three biofungicides such as Validan 5SL (Validamycin A),Trico-DHCT and Evitin 50SC (Hexaconazole 50g/l) And, factor B includes three periodic treatments (spraying biofungicides at days before pathogenetic inoculation, spraying biofungicides at days after pathogenetic inoculation and combine spraying before and after days inoculation with the pathogen) Inoculated method: Using Jasmine variety of paddy was sowed in each pot under greenhouse condition and inoculated at 40 days after sowing The fungi of Rhizoctonia solani cultured in a PDA medium, multiplied the microbial masses on rice husk medium, then adding 5g medium contain Rhizoctonia solani on each paddy bust and were transferred to a greenhouse at ambient environmental condition under high humidity Application of biofungicides and chemical fungicide: follow the recommendation of the produce Data recording: - Percentage of infection (%) = (the number of infected tillers/the total of observing tillers) x 100% - Relative lesion height (RLH) = (lesion length/plant length) x 100% - Disease index (DI) 1 n1    n3    n5     n7     n9   DI (%)   100% tn  Where: n(1), n(3), n(5), n(7), and n(9) = numbers of plants showing a reaction in a scale (1), (3), (5), (7) and (9) respectively (IRRI, 1996); tn = total number of plants scored 2.3 Data analysis The SPSS software was applied to calculate by using Duncan test Results 3.1 Percentage of infection 3.1.1 Percentage of infection at days after inoculation (dai) The ability of SBD prevention and control on Jasmine 85 variety by using biofungicides (Validan 5SL and Trico-DHCT) and chemical fungicide (Evitin 50SC) in greenhouse condition was evaluated by infected percentage, disease index and the yield components at harvest Table 3.1 Percentage of infected tiller (%) caused by SBD at dai Timing application (B) Percentage of infected tiller (%) Average factor (A) Spraying before Spraying before and Spraying after inoculation after inoculation inoculation Validan 5SL 45.72 abc 31.78 cd 56.58 ab 44.69 BC Trico-DHCT 54.68 ab 34.30 bcd 64.44 a 51.14 B Evitin 50SC 43.49 abc 13.10 de 44.98 abc 33.86 C Control 60.26 a 68.18 a 67.10 a 65.18 A Average factor 51.04 A 36.84 B 58.28 A F(A)** F(B)** F(AB) ns Factor (A) (B) Level of significance CV(%) 43.51 Note: Means in the same column followed by the same letter are not significantly different by Duncan test, ns: non-significant difference; **: significant difference at the 0.01 level Data was converted by arcsine x before analysis The result showed that, all three treatments gave to reduce SBD through the percentage of infected tiller and significant difference compared with the control at the 0.01 level Analyzed spraying with combined before and after pathogen inoculation (averaging 34.86% percentage of infected tiller) gave lower the percentage of infected tiller significantly than the other treatments The interaction between fungicides and timing application, there were two treatments ò biofungicides (Validan 5SL and Trico-DHCT) with spraying combined before and after pathogen inoculation gave lower the percentage of infected tiller significantly, however non-significantly with Evitin 50SC 3.1.2 Percentage of infection at days after inoculation (dai) All treatments gave the percentage of infected tiller arranged from 37.92% - 77.30% The treatment applied different fungicides were lower than and significantly different by the percentage of infected tiller compared with the control (Table 3.2) Following at dai, the method of using different fungicides by spraying with combined before and after pathogen inoculation were lower than the applied spraying before pathogen inoculation or spraying after pathogen inoculation by percentage of infected tiller Table 3.2 Percentage of infected tiller (%) caused by SBD at dai Timing application (B) Percentage of infected tiller (%) Spraying Spraying before Spraying after Average before and after inoculation factor (A) inoculation inoculation Validan 5SL 54.31 ab 32.43 bc 54.77 ab 47.17 B Trico-DHCT 59.30 a 40.79 ab 65.30 a 55.13 B Evitin 50SC 49.14 ab 19.33 cd 45.28 ab 37.92 C 77.30 A Factor (A) Control 73.04 a 79.90 a 78.97 a Average factor (B) 58.95 A 43.11 B 61.08 A Level of significance F(A)** F(B)** F(AB) ** CV (%) 46.91 Note: Means in the same column followed by the same letter are not significantly different by Duncan test, ns: non-significant difference; **: significant difference at the 0.01 level Data was converted by arcsin x before analysis Considered the interaction between the different fungicides and the timing application In which, only the treatment was applied with Validan 5SL by spraying with combination before and after (32.43% percentage infection) was better than the control (percentage infection of 61.97%) through percentage infection and was equivalent to the treatment with Evitin 50SC (percentage infection of 19.33%) 3.1.3 Percentage of infection at days after inoculation (dai) The result showed in Table 3.3, the treatment with Trico-DHCT and Validan 5SL reduced SBD by decreased percentage of infected tiller of 64.99% and 60.52%, respectively However, this treatment was higher than the treatment with Evitin 50SC (4923%), but lower than the control (98.09%) About timing application, the treatment of spraying with combined before and after (average of 53.96% percentage of infection) continued to show higher percentage of infected tiller and significant difference compared to spraying before pathogen inoculation or spraying after pathogen inoculation a massive 74.53% and 76.13% respectively Analyzed the interaction between some fungicides and timing application, the best treatment consists of Trico-DHCT and Validan 5SL were lower percentage of infection than the other timing application by spraying of combination before and after pathogen inoculation the same at dai Table 3.3 Percentage of infected tiller (%) caused by SBD at dai Timing Percentage of infected tiller (%) Average application (B) Spraying Factor Spraying before Spraying (A) before and after after Factor (A) Validan 5SL Trico-DHCT inoculation inoculation inoculation 71.90 ab 72.72 ab 38.54 c 49.56 b 71.11 ab 72.70 ab 60.52 B 64.99 B Evitin 50SC Control 57.89 cd 95,60 a 27.77 d 100.00 a 62.03 bc 98.68 a 49.23 C 98.09 A Average Factor (B) 74.53 A 53.96 B 76.13 A Level of significance F(A)** F(B)** F(AB) ** CV (%) 16.41 Note: Means in the same column followed by the same letter are not significantly different by Duncan test, ns: non-significant difference; **: significant difference at the 0.01 level Data was converted by arcsine x before analysis 3.1.4 Percentage of infection at 14 days after inoculation (dai) The figure of two treatments such as Trico-DHCT and Validan 5SL (percentage inoculation of 71.90% and 66.04%, respectively) were lower than the control (98.09%), and significant difference at the 0.01 level Besides, the treatment with Validan 5SL nonsignificant difference by percentage of infected tiller (Table 3.4) Table 3.4 Percentage of infected tiller (%) caused by sheath blight disease at 14 dai Timing application (B) Percentage of infected tiller (%) Average Factor (A) Spraying before Spraying before and Spraying after inoculation after inoculation inoculation Validan 5SL 78,16 b 47,02 c 72,91 bc 66,04 BC Trico-DHCT 80,63 b 58,43 b 76,65 b 71,90 B Evitin 50SC 67,57 c 37,90 d 63,80 d 56,42 CD Control 95,60 a 100,00 a 98,68 a 98,09 A Average Factor 80,49 A 60,84 B 78,01 A F(A)** F(B)** F(AB) ** Factor (A) (B) Level of significance CV (%) 12,20 Note: Means in the same column followed by the same letter are not significantly different by Duncan test, ns: non-significant difference; **: significant difference at the 0.01 level Data was converted by arcsine x before analysis 3.2 Disease index (DI) 3.2.1 Disease index at days after inoculation (dai) The result in Table 3.5 showed that, using two biofungicide and chemical fungicide gave reduce disease index of sheath blight disease on rice through the evaluation of disease index between the treatments Table 3.5 Disease index (%) of sheath blight disease at dai Timing application (B) Disease index Average Spraying before Spraying before and Spraying after inoculation after inoculation inoculation Factor (A) Factor (A) Validan 5SL 4.00 cd 4.50 de 4.61 cd 4.17 C Trico-DHCT 5.83 bc 5.00 bcd 6.25 abc 5.69 B Evitin 50SC 3.39 de 3.37e 3.23 de 3.33C Control 8.37 a 8.32 a 8.30 a 8.33 A Average Factor 5.40 5.30 6.22 (B) Level of F(A)*, F(B)ns, F(AxB)* significance CV (%) 14.22 Note: Means in the same column followed by the same letter are not significantly different by Duncan test, ns: non-significant difference; **: significant difference at the 0.01 level Data was converted by arcsin x before analysis At this time, average disease index of all treatments such as spraying before, spraying after, spraying before and after pathogen inoculation (disease index of 5.40%, 6.22% and 5.30%, respectively) had not significant difference, there was no the effect of treatment clearly by timing application In general, the majority of treatments showed that reducing ability was expressed by disease index and was different from the control In the timing application, the disease index of the Trico-DHCT (5.83%) was equivalent to that of the Validan 5SL (4.00%) but higher and different from that of the Evitin 50SC (3.39%) At the next spraying after pathogen inoculation, Validan 5SL had disease index of 4.61, equivalent to Evitin 50SC (3.23%) In the spraying before and after pathogen inoculation, the disease index of low Validan 5SL treatment (4.50%) and no significant difference compared to Evidin 50SC (3.37%) and Trico- ĐHCT; However, Trico-DHCT treatment gave higher disease index and significantly different with the Evitin 50SC treatment 3.2.2 Disease index at days after inoculation (dai) The treatments were lower the average of disease index than the control In particular, the treatment with Validan 5SL (DI: 12.41%) non-significant difference the treatment with Evitin 50SC (DI: 12.08%) and lower than significantly the treatment with Trico-DHCT (DI: 26.25%) (Table 3.6) At this time, there was a difference between the average of the timing application, the average of disease index at the spraying before pathogen inoculation (16.11%) and spraying before and after pathogen inoculation (15.68%) showed that effective treatment of disease was not significant different compared with the others, that is was higher and different from the treatment of spraying after pathogen inoculation (20.15%) Table 3.6 Disease index (%) of sheath blight disease at days after inoculation Timing application (B) Disease index (%) Spraying Spraying before Spraying Average before and after after Factor inoculation inoculation inoculation (A) Validan 5SL 11.42 bc 11.25 bc 14.58 b 12.41 C Trico-DHCT 14.58 b 14.17 b 26.25 a 18.33 B Evitin 50SC 12.28 bc 11.08 c 12.88 bc 12.08 C 26.25 A Factor (A) Control 26.14 a 26.25 a 26.36 a Average Factor (B) 16.11 B 15.68 B 20.15 A Level of significance F(A)*, F(B)*, F(AxB)* CV(%) 7.59 Note: Means in the same column followed by the same letter are not significantly different by Duncan test, ns: non-significant difference; **: significant difference at the 0.01 level Data was converted by arcsin x before analysis Similar to the previous time, the treatments showed that the effect of different fungicides was at different levels and higher than the control Specifically, the spraying before pathogen inoculation, all three treatments showed that the effect on controlling sheath blight disease was high similar (DI: 11.25%, 14.58% and 12.28%, respectively) and significant different compared to the control In spraying after inoculation, only Validan 5SL treatment with disease index was 14.58%, the non-significant difference compared to the Evitin 50SC treatment (DI: 12.88%) In the spraying combination before and after pathogen inoculation, the Validan 5SL treatment (DI: 11.42%) showed that the high effect as well as Tricho-ĐHCT (DI: 14.17%) and Evitin 50SC treatment (DI: 11.08%) 3.2.3 Disease index at9 days after inoculation (dai) At this time, the Validan 5SL treatment and Trico-DHCT treatment gave the effect on controlling the fungi by the average of disease index was 22.50% 25.56%, respectively, and was lower and significant difference compared to the control (37.92%), but this treatment was higher than the Evitin 50SC treatment (19.45%) (Table 3.7) About the average of the timing application, all the treatments showed the efficiency compared with the control The spraying of combination before and after pathogen inoculation and spraying before pathogen inoculation were equally effective in managing the disease through the disease index was 25.32% and 24.08%, respectively and was higher than the spraying after pathogen inoculation (29.67%) The analyzed treatments in the interaction between the applied fungicides and timing application Three treatments of applied fungicides gave high the average disease index was 22,80 %, 21.77% and 18.75%, respectively, it was lower and significantly different the control at 37.95% In the spraying after pathogen inoculation, the Validan 5SL treatment was lower the average disease index at 24.17% and was not significant difference the Evitin 50SC treatment (23.20%) In the spraying of combination before with after pathogen inoculation, the Validan 5SL treatment and Tricco-ĐHCT treatment continue to maintain the ability to control the disease only after the Evitin 50SC treatment, with the average of disease index 21.50% and 21.57%, respectively, was lower than and significant different the control (37.81%) Table 3.7 Disease index (%) of the affected sheath blight disease at dai Timing application (B) Disease index (%) Average Spraying Spraying before Spraying Factor before and after after (A) inoculation inoculation inoculation Validan 5SL 22.80b 21.50 b 23.20 b 22.50 B Trico-DHCT 21.77b 21.57 b 33.33a 25.56 B Evitin 50SC 18.75 bc 15.42 c 24.17b 19.45 C Control 37.95 a 37.81 a 37.99 a 37.92 A Average Factor (B) 25.32 B 24.08 B 29.67 A Factor (A) Level of significance F(A)*, F(B)*, F(AxB)* CV(%) 8.73 Note: Means in the same column followed by the same letter are not significantly different by Duncan test, ns: non-significant difference; **: significant difference at the 0.01 level Data was converted by arcsin x before analysis 3.2.4 Disease index at 14 days after inoculation (dai) All treatment by applied biofungicide and chemical fungicide was the affected inhibition of fungi through the average of disease index which was lower than the control (51.25%) (Table 3.8) Table 3.8 Disease index (%) of the affected sheath blight disease at 14 dai Timing application (B) Factor (A) Disease index (%) Spraying Spraying before Spraying before and after after inoculation inoculation inoculation Validan 5SL 34.07 b 33.27 b Trico-DHCT 37.50 b 35.83 b Evitin 50SC 30.83 b 25.75 c Control Average Factor 51.00 a (B) Average 50.25 a 38.26 AB 36.50B Level of significance F(A)*, F(B) *, F(AxB)* CV(%) 6.81 35.17 b Factor (A) 34.17 CD 47.08 a 40.14 B 34.75 30.44D b 52.50 a 51.25 A 41.88 A Note: Means in the same column followed by the same letter are not significantly different by Duncan test, ns: non-significant difference; **: significant difference at the 0.01 level Data was converted by arcsine x before analysis At the time of treatment, the average of timing application showed the affected inhibition of fungal pathogens Among them, the spraying of combination before with after pathogen inoculation has the highest effectiveness (36.50%) However, non-significant difference compared with the spraying before pathogen inoculation (38.26%), but was differ from the spraying after pathogen inoculation (41.88%) Similar the previous time, analyzed the interaction between applied fungicide and timing application showed that three treatment gave reduce sheath blight disease with the average of disease index (arranged from 30.83 - 34.17%) Besides, this treatment was lower than and different significantly compared to the control (51.00%) In the spraying after pathogen inoculation, the Validan 5SL treatment of disease index (34,75%) was not significant difference compared to the Evitin 50SC treatment (34.75%) In the spraying of combination before with after pathogen inoculation, the Evitin 50SC treatment gave the best effect to control sheath blight disease, the second treatment of Validan 5SL and Trico-DHCT were maintain the effect well and was lower than the control (50.25%) Conclusions and discussion Validan 5SL and Trico-DHCT had affected on the management of sheath blight disease significantly under greenhouse condition with the different applications Among them, spraying of Validan 5SL at before pathogen inoculation or before and after pathogen inoculation and spraying of Trico-DHCT at before and after pathogen inoculation gave higher effectiveness than the others treatment In addition, the effect of Validan 5SL had no significant difference Evitin 50SC at the time observation In short, the evaluation by percentage of infection and disease index at the observable time illustrated the biofungicide gave the affected inhibition against Rhizoctonia solani at different level At and 14 dai, the Validan 5SL treatment was reduce the affected tiller caused by sheath blight disease and was not significant different the Evitin 50SC treatment In addition, at 5, and 14 dai, the validan 5SL treatment also showed the effect of control was high compared with Evitin 50SC by evaluation of disease index That is two fungicide contain antifungal agents Rhizoctonia solani, which is commonly used in production today Vailidan 5SL fungicide was contain Validamycin A, which is produced from the Streptomyces sp that is a antibiotic for control Rhizoctonia solani on rice and some other crops (Vu Trieu Man, 2007a, b, Nguyen Manh Chinh et al., 2016) In addition, the Evitin 50SC fungicide contains active Hexaconazole, a fungicidal antibiotic that prevents and treats many diseases, including sheath blight disease in rice (Nguyen Manh Chinh et al., 2016) So in this study used two fungicide consist of Validan 5SL fungicide and Evitin 50SC fungicide gave the effect on treat sheath blight disease caused by Rhizoctonia solani Moreover, using Trico-DHCT was also reduce the tiller affected by sheath blight disease compared to the control and non-significant difference Vailidan 5SL fungicide Although, this treatment gave lower the effect than Evitin 50SC fungicide The same Prasad and Kumar (2011) reported spraying with Trichoderma spp to controlling sheath blight disease on rice The result, the percentage tiller was affected by Rhizoctonia solani which was lower than the control untreated and was higher than the control treated by chemical fungicide In addition, the investigation of the timing application has been shown to be effective against pathogen suppression, but there was an interaction with each fungicide at different levels Specifically, spraying before pathogenic inoculation gave more the effect in disease management than spraying after pathogen inoculation to dai, the spraying of combination before with after pathogen inoculation the disease to achieve in control sheath blight disease on rice until 14 dai Among them: Treated with Validan 5SL or Trico - DHCT before pathogen inoculation, showed that these treatments decreased the percentage of infection of the affected tiller at 5, and dai but was not significantly different the disease index with the Evitin 50SC fungicide at 7, and 14 dai Fungal Trichoderma spp growing up so fast in the soil, they grow faster than other fungi (Saksena, 1960) So when treated before pathogen inoculation, Trichoderma ssp have much time as well as they are easy to grow and increase the titer rapidly enough to against Rhizoctonia solani which was inoculated later Klein and Eveleigh (1998) reported that Trichoderma spp directly attacked by curling up the harmful fungi and release chitinase enzymes that is destroy the cell wall of harmful fungi into small molecules that are easily absorbed, and helps plants resist disease Validan 5SL fungicide and Tricho- ĐHCT fungicide of spraying combination before with after pathogen inoculation showed the ability to limit the percentage of affected tiller This result is the same with the study by Nguyen Van Hung (2010) that biological control caused by R solani by using Trichoderma viride The experiment was designed include applied the treated T viride before and inoculated R solani the same time for the percentage of infected plants which was lower than the use of T viride after R solani inoculated On the other hand, evaluation of the disease index, two bio-fungicide consists of Validan 5SL and Tricco-ĐHCT with spraying combination before with after pathogen inoculation showed that the ability of control is a maintain until at 14 dai; Validan 5SL has reduced the disease index and was not significant difference compared with Evitin 50SC at and dai This reason may be explained by the fact that both Validan 5SL and Trico-DHCT gave the ability of prevention well when treated before inoculation as well as analyzed discussed above so that applied fungicide at days after pathogen inoculation that was contributed to the increased ability to manage of sheath blight disease A B C D E F Figure The level of sheath blight disease on rice at 14 days after inoculation A-Control; B-Spraying Trico-DHCT before pathogen inoculation; C-Spraying Validan 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applying some biofungicides” is researched Methodology 2.1 Objectives Determining the timing application of biofungicides... blight disease on rice through the evaluation of disease index between the treatments Table 3.5 Disease index (%) of sheath blight disease at dai Timing application (B) Disease index Average Spraying... after pathogen inoculation to dai, the spraying of combination before with after pathogen inoculation the disease to achieve in control sheath blight disease on rice until 14 dai Among them: Treated