| In order to study the differentially expressed genes of Maize under the condition of full film coverage.In this experiment,separated differentially expressed gene fragments in Maize under full film coverage by cDNA-AFLP technology,and the differently expressed fragments of the related genes were screened,isolated,sequenced and functionally analyzed under the condition of full film coverage and not covered with plastic film.The results were as follow:1.Establishing the method for extraction of total RNA from roots and leaves of maize seedlings,and the total RNA was reverse transcribed into double stranded cDNA.Enzyme digestion of double stranded cDNA,by using of PstI and MseI two kind of enzymes,design of joints and the corresponding nine pairs of selective amplification primers for the specific fragment amplification and sequencing.2.The use of cDNA-AFLP double enzyme digestion technique,there were bands of 1481,the differentially expressed fragments of 336,accounting for about 22.7%of the total bands.Then the sixty fragments of them were sequenced to obtain fifty-eight sequencing results.3.Homology analysis and functional prediction of 58 sequencing results by NCBI system,the results were as follow:thirteen have a high homology with plants and most of the protein function;six have a high homology with animals;twenty have a high homology with microorganisms;In addition,nineteen differentially expressed fragments were unable to find homologous sequences.4.Thirteen have a high homology with plants,there are three have a high homology functional genes in wheat,sorghum and citrus,the other ten were highly similar to some of the cloned genes in maize.The acquisition of these genes laid the foundation for further study on the differentially expressed genes of Maize under the condition of film mulching.5.The results of sequencing showed that twenty genes with a high homology to microorganisms were obtained,and the role of these genes in maize growth needs further study. |
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