The CRISPR/Cas9 Application Study In Rice Xylan Side Chains

Relative to the non-graminaceous monocots and dicots,the most prominent feature of the important crop of grass and economy is their cell walls contain abundant heterogeneous polysaccharides.Differences in Gramineae is the key content and structure of hemicelluloses.Hemicellulose consists mainly of xylan,Arabinoxylan,galactomannan chitosan,xyloglucan.There are different types of hemicelluloses in plant cell walls,from different cell wall of plant species and different plant tissues have different composition.Xylan is constituted of β(1,4)xylan pyranose chain.L-Arab sugar is D-xylose epimer,the extensively distributed in plant kingdom.It exists in pectin,hemicellulose and hydroxyproline rich glycoprotein,Arabia galactose.Xylan as the important gramineous hemicellulose contains β(1,4)xylose skeleton and α(1,2)or α(1,3)link Arabinoxylan contains sugar.Xylan also contains the main non-cellulose composition in secondary wall of angiosperms.Therefore,second only to cellulose xylan in quantity.Xylan is very valuable for human and animal,contains the main components of the corn dietary.CRISPR/Cas9 system is found in a system of archaea and bacteria.It can be modified to cut target gene after transformation,so as to achieve the effect of genetic mutations.In this paper,by constructing the CRISPR/Cas9 carrier to Os UXE and Os XAT gene knockout,observation and analysis of the differences between wild type and mutant,preliminary understanding xylan structure change in the side chain.(1)Use fluorescence quantitative PCR analysis Os UXE and Os XAT expression in different tissue volume,results show that the high expression in the stem.And then according to different period FPKM value analysis expression in different tissue volume,found in bud expression quantity and tissue is high,there is no other period and the expression of the tender stem volume is much higher than other parts.(2)Construe CRISPR/Cas9 vector to knockout gene of rice,get different mutant strain.Recognize the different strains of mutant,found no big difference from the performance;Formation of xylan structure may have no influence on plants structure.(3)Using bioinformatics analysis different the knockout mutant site information,analysis of the differences between them from the base level,found after different strains in different bases broken there is no obvious difference.(4))through the comparison between different mutant and wild type structure of stem.By using toluidine blue staining,found that different in different circumstances knockout mutant strain,compared with wild type,no obvious structural differences.