Study On Fungal Protein Elicitor PeaT1 Scale Fermentation Process And Product Quality Detecting Technology

PeaT1 was a protein elicitor extracted from Alternaria tenuissima,it could induce a variety of crops' systemic acquired resistance.Research showed that on rice,wheat,cabbage and celery,PeaT1 has obvious effect to promote growth,increase production,improve the quality,disease resistance and stress tolerance ability.In order to provide a stable and reliable preparation,we were research the factory fermentation conditions for further study,aim to lay a foundation for large-scale production.In this paper,we according to Alternaria tenuissima nutritional requirements,through orthogonal test for the culture medium is improved and optimized,rely on the 500 L,5T and 20 T fermentation step by step,we obtain the ideal protein preparation.By using HPLC,SDS-PAGE and Coomassie blue staining,the protein in the preparation was qualitatively and quantitatively tested.We used the preparation and its compound preparation to bioassay.The specific research results are as follows:1.We through the research of the mass production process of PeaT1,amplify the laboratory miniaturization fermentation of Alternaria tenuissima.And culture medium was determined by the orthogonal test: Corn starch 3.5%,soybean flour 1%,sugar 0.5%,peptone 0.5%,monosodium glutamate 0.5% ammonium sulfate 0.2%,magnesium sulfate 0.02% and ferrous sulfate 0.02%.2.Based on the 500 L,5T and 20 T fermentation,we mastered the fermentation time is 40 hours,26 hours and 20 hours.Through dynamic parameters in the fermentation process monitoring,we got the dynamic curve of pH,dissolved oxygen,temperature.3.By used HPLC and SDS-PAGE method for the preparation of spectra and electrophoresis analysis,identified PeaT1 ingredients in the products,and get the appraisal method of the active ingredients.By Coomassie blue staining for rapid detection of the effective composition content of product,it provides a quality control method for large-scale fermentation production.4.Stability of PeaT1 protein preparation prepared by industrial equipment was tested by laboratory bioassay.PeaT1 preparation was used for seed soaking treatment of wheat and corn seeds,and it was found that each batch was significantly better than the control group,and the difference between each batch was not significant,which proved that the preparation produced by industrialization was stable and reliable.5.Use of Jinggangmycin and PeaT1 compound preparation,it shows that PeaT1-Jinggangmycin by 3:4 mixed ratio had the most cost-efficient.Use Nicotiana benthamiana infect TMV-GFP by the way of friction,and observe virus infection situation under UV light.It significantly showed that the compound preparation's relative control effect reached 73.39% than spraying water.This compound preparation is applying for temporary license pesticides.