Effects Of Appling Trichoderma Viride With High-Carbon-based Fertilizer On Growth And Quality Of Flue-cured Tobacco

The effects of Trichoderma viride high-carbon-based fertilizer on growth,yield and quality of flue-cured tobacco were investigated in order to provide theoretical reference for agricultural application of Trichoderma viride and biochar.The effects of agricultural application of Trichoderma viride and high-carbon-based fertilizer was studied in field experiment.The combined application of Trichoderma viride and high-carbon-based fertilizer field experiment with 'Zhongyan 100' as the sample variety using a randomized block experiment design was conducted in Xuchang,Henan province in 2015,which included 4 fertilizer treatments viz.,without Trichoderma viride and high-carbon-based fertilizer(CK),Trichoderma viride(T1),high-carbon-based fertilizer(T2),combined application of Trichoderma viride and high-carbon-based fertilizer(T3).In order to increase the spore yield of Trichoderma viride YCG-1,the response surface methodology(RSM)was used to optimize the solid state fermentation conditions.The fermentation conditions optimization was conducted in Xuchang,Henan province in 2016.The main results were as follows.1.Effects of appling Trichoderma viride with high-carbon-based fertilizer on growth of flue-cured tobaccoa.Growth and biomassThe results showed that combined application of Trichoderma viride and high-carbon-based fertilizer can improve plant height,stalk width,leaf area per plant and biomass of flue-cured tobacco at early stage.During 30 to 60 days after transplant,plant height of T3 and T2 significantly increased by 24.79%?31.41%and 10.06%?13.43%respectively compared with control.Stalk width of T3 and T2 significantly increased by 20.35%?28.75%and 8.96%?17.81%respectively compared with control in the same period.Leaf area per plant of T3 significantly increased by 43.54%,41.98%and 41.98%respectively compared with control at 30,75 and 90 days after transplant,which indicated the promoting effect of combined application on leaf area per plant occurred at early and late growth stage of flue-cured tobacco.Root shoot ratio of T3 significantly increased by 12.50%and 23.81%respectively compared with control at 60 and 75 days after transplant.Combined application of Trichoderma viride and high-carbon-based fertilizer also increased root stalk leaf biomass and total biomass of flue-cured tobacco,while no significant different between Trichoderma viride treatment and control.b.Photosynthetic charactersTotal chlorophyll content in leaves of T3 significantly increased by 20.92%,58.03%and 37.98%respectively compared with control at 60,75 and 90 days after transplant,which indicated the promoting effect of combined application on photosynthetic characters occurred at middle and late growth stage of flue-cured tobacco.Total chlorophyll content of T2 increased by 17.56%?37.68%compared with control,while no significant different between T1 and control.Combined application and application of high-carbon-based fertilizer can improve carotenoid content in tobacco leaves,while no significant different between T1 and control.During 45 to 90 days after transplant,combined application of Trichoderma viride and high-carbon-based fertilizer increased stomatal conductance and decreased intercellular CO2 concentration of tobacco leaveas so that the net photosynthetic rate of tobacco leaves increased.The net photosynthetic rate of T3 and T2 significantly increased by 16.32%?32.22%and 9.44%?36.02%respectively compared with control.Combined application of Trichoderma viride and high-carbon-based fertilizer increased Fv/Fm,ETR,qp and ?PS?of tobacco leaves at middle and late growth stage of flue-cured tobacco,which indicated photosynthetic activity of tobacco leaves could be effectively improved by combined application of Trichoderma viride and high-carbon-based fertilizer.c.Morbidity and disease index relative to tobacco diseases and control effectThe results indicated that combined application of Trichoderma viride and high-carbon-based fertilizer significantly reduced the morbidity and disease index relative to tobacco mosaic virus disease and tobacco brown spot disease compared with control treatment.Disease index and morbidity relative to tobacco mosaic virus disease of T3 significantly decreased by 74.89%and 74.70%respectively compared with control,while disease index and morbidity relative to tobacco brown spot disease significantly decreased by 70.66%and 31.74%respectively.Disease index and morbidity relative to tobacco mosaic virus disease of T2 decreased by 35.99%and 30.85%respectively compared with control,while disease index and morbidity relative to tobacco brown spot disease decreased by 34.17%and 9.58%respectively.No significant different between T1 and control on disease index and morbidity relative to two tobacco diseases.The control effect on tobacco mosaic virus disease and tobacco brown spot disease of T3 increased by 120.03%and 107.66%respectively compared with T2.d.Mineral elements uptakeNitrogen,phosphorus,potassium,magnesium,ferrum and manganese uptake in leaves and plant of T3 significantly increased at middle and late growth stage of flue-cured tobacco compared with control,which indicated combined application of Trichoderma viride and high-carbon-based fertilizer improved absorption of mineral elements of flue-cured tobacco.2.Effects of appling Trichoderma viride with high-carbon-based fertilizer on quality of middle leavesa.Chemical components contentThe total sugar content in middle tobacco leaves of T2 and T3 significantly increased 15.45%and 29.14%respectively compared with control,while reducing sugar content increased by 19.98%and 39.01%respectively,which indicated combined application of Trichoderma viride and high-carbon-based fertilizer improved carbon metabolism of flue-cured tobacco.Total nitrogen and nicotine content in middle tobacco leaves of T3 significantly increased by 51.66%and 29.31%respectively compared with control.Potassium content in middle tobacco leaves of T2 and T3 significantly increased by 6.96%and 32.17%respectively compared with control.There was no significant difference in chlorine content among treatments.Ratio of reducing sugar to total sugar,ratio of total nitrogen to nicotine and ratio of potassium to chlorine of T3 significantly increased by 8.97%,16.92%and 36.89%respectively compared with control.b.Petroleum extracts contentPetroleum extracts content in middle leaves of T3 significantly increased by 27.51%compared with control,while petroleum extracts content in middle leaves of T2 increased by 11.11%.There was no significant difference in petroleum extracts content between T1 and control,as well as T2 and control.c.Polyphenol contentChlorogenic acid content in middle leaves of T3 significantly increased by 43.84%?44.15%and 23.91%respectively compared with control,T1 and T2,while no significant difference among control,T1 and T2.Scopoletin content in middle leaves of T3 and T2 significantly increased by 41.38%and 20.69%respectively compared with control,while no significant difference between T1 and T2,as well as T1 and control.Rutin content in middle leaves of T3 significantly increased by 79.17%?81.45%and 25.69%respectively compared with control,T1 and T2.Total polyphenol content in middle leaves of T3 significantly increased by 53.76%,54.39%and 32.58%respectively compared with control,T1 and T2.d.Organic acid contentMalic acid and total organic acid content in middle leaves of T3 significantly increased by 31.71%and 18.24%respectively compared with control.Malic acid content in middle leaves of T2 significantly increased by 10.92%compared with control.There was no significant difference in total organic acid content in middle leaves among control,T1 and T2,as well as in malonic acid,stearic acid and linoleic acid content among all treatments.Citric acid and oxalic acid content in middle leaves of control and T1 were higher than T2 and T3.e.Aroma component contentCarotenoid catabolites,products of browning reaction,neophytadiene,total content of aroma component and total content of aroma component except neophytadiene in middle leaves of T3 significantly increased by 25.17%,21.70%,37.78%,36.77%and 32.06%respectively compared with control.Carotenoid catabolite,aromatic amino acid catabolites,Solanone,products of browning reaction,neophytadiene,total content of aroma component and total content of aroma component except neophytadiene in middle leaves of T2 significantly increased compared with control.3.Fermentation conditions optimization of Trichoderma viride YCG-1 strainTrichoderma viride YCG-1 as original strain,optimization of solid sate fermentation conditions was performed to improve the spore yield in the flask.Through the single factor experiment,Plackett-Burman experiment,the steepest ascent experiment and the response surface analysis,loading sample volume,temperature and pH were identified as the most important factors to influence the spore yield.The optimal conditions are as follow:the optimal initial water content of medium was identified as 75%,the optimal ratio of major medium to minor medium was identified as 10:0.2,the optimal loading sample volume was identified as 10.2 g per 250 mL flask,the optimal temperature was identified as 25.7?,the optimal initial pH of medium was identified as 7.07 and the optimal inoculum amount was identified as 1×106 CFU/g major medium.The highest spore yield has been achieved 8.28 × 109 CFU/g initial dry substrate(IDS)under the optimized solid state fermentation conditions,which is 1.72 times higher than the initial medium.In summary,the combined application of Trichoderma viride with high-carbon-based fertilizer(high-carbon-based fertilizer 1500 kg·hm-2,Trichoderma viride 3 kg·hm-2)could improve the biomass,photosynthesis and mineral elements uptake of flue-cured tobacco.Field experiment showed that the control effect on tobacco mosaic virus disease and tobacco brown spot disease of combined application was effective.The optimal soli state fermentation conditions were identified through the response surface methodology(RSM).