A, Alpha-amylase Inhibition Of Protein Harvest Sprouting Elisa Detection Method To Establish Two Rapd Analysis Of Pathogenic Dsrna Commelina Grass Point Mold

Relationship betweenα-amylase inhibitor and pre-harvest sprouting and establishment of determination method of ELISAThe monoclonal and polyclonal antibodies ofα-amylase inhibitor were produced and the ELISA method which to determine the concentration of wheatα-amylase inhibitor was established; The resistances of pre-harvest sprouting were determined by petri dishes and mist chamber with intact spikes in different periods after flowering; The activities ofα-amylase were determined by gel diffusion assay method in different periods after flowering; The concentrations ofα-amylase inhibitor determined by the established sandwich ELISA and SDS-PAGE of different periods after anthesis were also determined. The results showed that there were significant difference among the resistances of pre-harvest sprouting, the activities ofα-amylase and theα-amylase inhibitor concentrations of these varieties. The resistances of red verities were higher than that of white. The method of mist chamber with intact spikes was effective at determination of pre-harvest sprouting. Theα-amylase activity was the highest in .25d after flowering, then decreased gradually. There were significant positive relationship between theα-amylase activity and the sprouting rate, whereas there were significant negative relationship between the falling number and the sprouting rate. Theα-amylase activity at the late stage after anthesis might be used as an index of pre-harvest sprouting resistance.α-amylase inhibitor could not inhibit the sampleα-amylase . There was not significant negative relationship between the concentrations ofα-amylase inhibitor and the sprouting rates, but theα-amylase inhibitor could inhibit the sprouting rates in some extent. Some varieties with low activity ofα-amylase and high concentration ofα-amylase inhibitor were selected to improved the resistance of pre-harvest sprouting . A model of pre-harvest sprouting was proposed: theα-amylase inhibitor influenced the pre-harvest sprouting by inhibiting the activities ofα-amylase, the remain activities of the verities were the main reason of pre-harvest sprouting. 1,观察了东北地区鸭跖草叶点霉的培养特性,根据培养特性可将东北地区鸭跖草叶点霉实步分为5个培养类型。2,鸭跖草叶点霉存在致病力分化现象,致病力强弱从南到北逐渐减弱。3,测定各菌株的dsRNA,在所测定的12个菌株中,有10个菌株含有dsRNA,2个菌株不含dsRNA。含有dsRNA的菌株中,dsRNA的分子量不同。4,菌株的致病力的强弱与dsRNA的菌株中,dsRNA的分子量不同。5,CTAB法是提取真菌DNA的一种有效的方法。5,根据各菌株的RAPD图谱,在类间距离15以下可将12个鸭跖草叶点霉分为5个类群。来自辽宁和吉林地区的菌株类间距离较小,其亲缘关系较近；来自黑龙江的菌株与辽宁和吉林的菌株亲缘关系较远些。The biological characters and pathogencity of 12 isolates of Phyllosticta commelimecola obtained from Northeast in china were observed. These strains could be divided into six cultural types preliminarily on the basis of colony morphology, color, growth rate and asexual sporulation. There were obvious difference in pathogencity of these strains, of which could be divided into high, medium and low types. Among 12 tested isolates, dsRNA was found in 10 isolated, however, 2 strains contained no dsRNA, there was no relationship between the pathogencity and existence of dsRNA. The method of CTAB is good at extraction of genomic DNA of fungi. The RAPD was used to analyze the polymorphism of 12 isolates of P. commelimecola, 18 random primers were used to amplify the DNA of 12 isolates, all the primers amplify DNA fragments in 12 isolates. The molecular weight of the fragments ranged from 3．0Kb to 0．1Kb. The polymorphism DNA were cluster analyzed by SPSS9．0, the results were that the isolates can be divided into 5 groups under the distance of 15．There were close relationship between the isolates of Jilin province and Liaolin province .3 Of the 4 isolates from Helongjiang province belong to group one, however, the other ,which has the longest distance with all other 11 isolates, belongs to another group of its own.