The Research Of Anti-mildew Properties And The Diversity Of Anti-fungal Protein Of Different Harness Wheat

Wheat is the most widely planted area in the crop of the world. In the northern areas of China, wheat is one of the main food crops. Its storage resistance is very strong. In the process of wheat storage, the biological conditions and non-biological conditions have influenced in wheat yield and quality, such as the skin tissue of wheat and the ability of its hygroscopic. When the relative humidity was in a high level or wheat was in a higher water content, wheat is easier to produce the mildew than the other crops such as rice, corn. These mold damage the internal structure of wheat,reduced the production of wheat and even bring the toxin, this all harm to the health.Because wheat don’t have the immune system, wheat will caused the following lesions in physiologically and modal when they were infect by the variety germs.Therefore, it is very important to knowing the antibacterial property of wheat and studying the antifungal proteins’ difference of wheat breeding. In this paper, the ordinary planting wheat is the main study object, and did the following research work :(1) simulated storage varieties of wheat in different harness and analyzing the mildew differences and the antifungal differences of the wheat under the same storage condition.(2) Using the two-dimensional electrophoresis technique and mass spectrum technique to analysis the antifungal protein differences of wheat in different harness.(3) Researching the gene cloning, sequencing analysis and the difference of antimicrobial properties on the different harness of the wheat. We receive the following study result:(1) The mildew differences and the antifungal differences of wheat on the same storage conditionIn order to study the difference of mould growth in red wheat in different hardness, two soft red wheat and two hard red wheat were stored under the same moisture content and temperature conditions after their moisture contents modified.The results showed that the increase of mould counts in hard red wheat cultivars Yangmai 16 and Zhenmai 10 appeared significantly higher than in soft red wheat cultivars Yangmai 15 and Yangfumai 4(p < 0.01) after stored under the conditions of 13.5% moisture contents at 28 °C. The two soft red wheat cultivars could significantly resist the growth of Aspergillus glaucus stored at 13.5% moisture content. After stored under the conditions of 15.0% moisture contents at 28 °C, the two soft red wheat,Yangmai 15 and Yangfumai 4, could also resist the mould growth to a certain extent,and the mould counts in the two soft red wheat were significantly lower than in hard red wheat cultivars Yangmai 16 and Zhenmai 10. Moisture absorption tests were performed under 80% relative humidity condition and the mould counts of Aspergillus glaucus in two hard red wheat were above six times more than in two soft red wheat. Moreover, the mould growth on the wheat testa was observed using microscope and it was found that the mould were mainly grown on the germ part of hard red wheat kernel. Therefore, the difference of the mould growth both in soft and hard red wheat exists during storage, and the soft red wheat could resist the mould growth under the same storage condition.(2) Studing on the antifungal discrepancy of different harness of wheat germ By separating each organization of wheat germ(yangmai No.15 and yangmai No.16)and extracting protein. Using the two gel electrophoresis and mass-spectrometric technique to analysis the difference between the protein of the two kinds of wheat.The results show that there have 56 specific proteins in soft wheat germ and 18 in hard wheat germ’s. Recycling these specific proteins and hydrolyzing them by trypsase. Analysing the peptide which has been cleavaged of all the specific proyeins by mass-spectrum method. It turn out that soft wheat germ have more antifungal proteins than hard wheat germ’s. Beside, researchingthe antifungal proteins on different harness wheat germ have obtained that there have 5 antifungal proteins been reported already.(3)the analysis of different hardness of wheat antifungal protein activityUsing the DNS method to test the lucanase activity, peroxidase activity and alpha amylase inhibitors activity in different hardness of wheat. The results show that the higher enzyme activity in wheat, the stronger its antibacterial ability.(4) the cloning and expression of wheatWe have cloned 6 gene of wch by using the PCR method. Put these gene into expression vector, and switch into Ecol R I(BL21), using the 0.5mmol/L IPTG inducing 3 h, and test the expression result by SDS-PAGE. This result indicate that the gene of wch was potential target gene in plant which prevent fungal disease.This study has analysed the mildew differences and the antifungal differences of the wheat, and understanded the character of storage wheat, in order to provide basis for the antimidew character of antifungal protein of wheat.