Study On Bamboo Vinegar Antifungal Activity And Applied Technology

In this paper, the bamboo vinegar and bamboo oil were tested for antifungal activities against seven species of plant pathogen, which included Botrytis cinerea isolated from tomato(A), strawberry(B) and hot pepper(C), Alternaria solani(Ellis et Martin) sorauer(D), Colletrichum gloeosporioides(E), Fusarium graminearum Schw(F) and Fusarium oxysporum Schlechtendahl(G), and the main activity compounds were isolated from bamboo oil guided by Botrytis cinerea(A), Fusarium graminearum Schw(F) and Fusarium oxysporum Schlechtendahl{G) in vitro, the practical application of the formulations of bamboo oil and its mixtures were studied. The main results were summarized as following:1 .The inhibition rate of bamboo oil against seven species of plant pathogen were better than bamboo vinegar in the same concentration. When the concentration were 1000mg/L in the culture medium, the inhibition rates of bamboo oil against all seven species of plant pathogen were 100%, while comes to bamboo vinegar, the inhibition rates were all under 30%. The value of EC₅₀ of bamboo vinegar and bamboo oil against (A), (B), (C), (D), (E), (F), (G) in vitro were between 2131mg/L to 4613mg/L and between 86.30mg/L to 270.4mg/L respectively; while the value of EC₉₀ of them were between 4941mg/L to 11962mg/L and between 292.4mg/L to 1741mg/L respectively. The results indicated that, all those EC₅₀ and EC₉₀ of bamboo oil were about 1/60 to 1/10 of that of the bamboo vinegar. The best antifungal activities were bamboo oil against (A), (B), (C) and (E).2.The value of EC₅₀ and EC₉₀ of mixtures of bamboo oil and prochoraz (the ratio were 10:1 and 20:1 ) against (E) in vitro were 0.2520mg/L, 0.8434mg/L and 0.6412mg/L, 2.1321mg/L respectively, the value of CTC of them were 107 and 80.0 respectively, these mixtures have summation action; but when the ratio increase to 50:1, 100:1 and 1000:1, the value of EC50 and EC90 of these mixtures against (E) in vitro were 1.635mg/L,7.897mg/L; 4.012mg/L,27.23mg/L and 65.01mg/L, 250mg/L respectively, and the value of CTC were 75.8,59.8 and 31.9 respectively, these mixtures have antagonistic action, these actions were getting more and more evidence in company with the increase of the ratio between bamboo oil and prochoraz. The value of EC₅₀ and EC₉₀ of mixtures of bamboo oil and bromothalonil (the ratio were 10:1, 20:1,30:1, 40:1 and 80:1) against (E) in vitro were 63.24mg/L, 236.0mg/L; 71.12mg/L, 172.2mg/L; 124.6mg/L, 249.8mg/L; 124.2mg/L, 308.2mg/L and 190.5mg/L, 417.8mg/L respectively, and the value of CTC were 79.7, 112, 80.5, 93.1 and 79.2 respectively, these mixtures all have summation action. The value of EC₅₀ and EC₉₀ of mixtures of bamboo oil and diniconazole (the ratio were 14:1 and 139:1) against (E) in vitro were 10.45mg/L, 36.10mg/L and 70.57mg/L, 161.1mg/L respectively, their CTC were 88.6 and 89.5 respectively, these mixtures have summation action.3. The value of EC₅₀ of mixtures of bamboo oil and pyrimethanil (the ratio were 10:1, 19:1, 39:1 and 59:1 ) against (A) in vitro were 16.35mg/L, 27.02mg/L, 23.60mg/L and 46.21mg/L respectively, and the value of CTC were 276, 340, 314 and 168 respectively, these mixtures have synergistic action. The value of EC₉₀ of them were 112.6mg/L, 128.4mg/L, 164.2mg/L and 237.5mg/L respectively, the ratio of 19:1 was the best at higher inhibition rate calculated by the ratio. According to these, it was selected as the final ratio of the mixture, it was used in formulation processing and the control efficiency in vivo.4.We have two difficulties in the formulation processing of bamboo oil and its mixtures. One was the screen of the solvent for them, in these papper, acetone, cyclohexanone, methyl-pyrrolidone were chosen as the solvent of bamboo oil and its mixtures, and the co-solvent were also screened, we chose the blended solvent S| as bamboo oil and its mixtures solvent after all the effect of these solvent to bamboo oil and its mixtures were tested. The other was the higher processing cost and use-cost of 15% bamboo oil·pyrimethanil EC, we can solve these problems by increasing the ratio of pyrimethanil in the mixtures and altering the formulations into water based ones.5.From the in vivo experiment of protective action and therapeutic action against gray mold of tomato and anthracnose of apple, we can say although bamboo oil has better toxicity in the in vitro experiment, while it has poor control efficiency of in vivo. There have significant difference between tomato inoculated with Botrytis cinerea immediately after treated with fungicides and treated after tomato inoculated with Botrytis cinerea two days latter. All of these indicated clearly that, the active compounds of bamboo oil have poor systematic characters, we can use it as a protective fungicide better than a therapeutic fungicide.6. In the field experiment of gray mold of hot pepper and tomato, 15% bamboo oil·pyrimethanil EC had very good control efficiency against the gray mold of these two plants, especially the protective efficiency of tomato was 83% when the using concentration was 4000mg/L, even better than 400mg/L 40% SCALA SC (the activity compound was pyrimethanil).In the field experiment of gray mold of hot pepper, the concentration of pyrimethanil in the treatment of B₁ was 150mg/L, only 37.5% of the treatment of D; in the field experiment of gray mold of tomato, the concentration of pyrimethanil in the treatment of B₁ was 200mg/L, only 50% of the treatment of D, but the control efficiency of these two B₁ treatments are better than their compare fungicides treatment of D, all of this showed that the mixture of bamboo oil and pyrimethanil had synergistic action both in vitro and in vivo, if we can solve the high processing cost and use-cost, the mixture of bamboo oil and pyrimethanil will have a promising application prospect.7. In the in vivo experiment of anthracnose of apple, we can saw significant control efficiency of the treatment D and the treatment E, their control efficiencies of apple inoculated with Colletrichum gloeosporioides immediately after soaked with fungicides were 69.7% and 79.9%, they all had significant differences from other treatments. The concentration of prochoraz in the treatment of D (the ratio between bamboo oil and prochoraz was 10:1) was 300mg/L, only half of the treatment of E, so we can tell that the mixtures of bamboo oil and prochoraz (the ratio was 10:1) had a really high synergistic action when it was put into practical use. The prospects of the bamboo oil and its mixtures were used as protective fungicides during the storage period of the apple and other fruits were quite optimism.8. Liquid-liquid extraction methods and liquid-solid column chromatography methods were used in this paper, we used bioassay-guided method to different fraction of bamboo oil during the isolating processing, the results shown that all the effluent of bamboo oil which were eluted by different polar elute solvents had certain activities against Botrytis cinerea in vitro, and that most of these fractions had the activities at the same level. The results showed that the active compounds of bamboo oil was not a single ones, the antifungal activities compounds of bamboo oil against different species of plant pathogen probably were the combined effects of several active compounds. After the second time of liquid-solid column chromatography processing, we got A3.2 fraction which antifungal activity against Botrytis cinerea was three times of crude bamboo oil and A3.3 fraction which antifungal activity against Fusariwn graminearum Schw and Fusariuni oxysporum Schlechtendahl were two times of crude bamboo oil, all these results showed that there had higher antifungal activity compounds in bamboo oil.