Cloning And Function Analysis Of Leucine-rich Repeat Extension Protein And Its Antisense Transcripts

Heavy metal Cu(Cu)is a transition metal element with redox activity.It is a necessary trace element to participate in plant life process as a cofactor of catalytic redox reaction.However,it is difficult to play a role when the copper ion overdose.LRX protein is a kind of cell wall proteins,extend LRX extensin is composed of five structural domain,probably including N the signal peptide,the structure of the variable domain,9 conservative LRR structure domains that are rich in leucine repeated sequences,rich in cysteine structure domain,the C-EXT hydroxyproline glycoprotein structure domain including rich in typical Ser-Hyp4 motif.Lnc40 is a long chain non-coding RNA with a length of more than 200 nt and no protein coding potential.The analysis of the results of the sequencing of the transcripts of Chinese elsholtzia transcripts shows that Lnc40 may respond to Cu.This experiment is to extensin LRX and natural antisense transcript as the research object,the comparative study on the organization of LRX and lnc40 positioning,stretch LRX family protein subcellular localization,the interaction between LRX and lnc40,lnc40 at the transcription level of response mechanism and lnc40 transgenic plants divalent metal ions in the phenotypic analysis of the main conclusions in this study are as follows:(1)Organizational positioning analysis results show that LRX3 in petals,sepals,young Indus leaves,stem leaf,pod at both ends,the stigma of expression,the seeds,pollen grains were not expressed,cauline leaf petioles,stems;LRX4 is expressed in stem leaves,petals,sepals,rosette leaves,and fruit pods;it is not expressed in seeds or stems;LRX5 is expressed in the petals,sepals,leaves,ends of the pod,and in the main stem,not expressed in seeds and pollen grains.Lnc40 on both ends of petals,sepals,leaf,fruit pods,grow plants of 3 days of leaf and root of vascular bundle in the expression of the pollen grain,seed and Indus leaves leaflets,growth is not expressed in the 3 days of seedling root tip.(2)LRX extended protein subcellular localization observed that LRX was located on the cell wall.In the absence of signal peptide structure domain,the subcellular localization changes,and the cell wall is transferred to the cytoplasm.When the EXT domain is missing,the location is even transferred to the nucleus.(3)The LRX natural antisense transcript lnc40 expression and RNAi transgenic strains grow under normal conditions,including wild type and transgenic lines were no obvious differences,but add 40 ?M Cu on the solid medium,the roots of the transgenic plants are better than wild type long.The results of ion content determination showed that the accumulation of wild type Cu in the aboveground was almost constant,and the Cu accumulation of transgenic plants increased,and the contents of the wild type and transgenic plant Cu in the underground part decreased.(4)Fluorescence quantitative PCR results showed that the expression levels of lnc40 and LRX5 of the wild-type arabidopsis thaliana were improved after the treatment of 50 ?M Cu,so lnc40 and LRX5 could be resistant to Cu.LRX5 gene and lnc40 in WT-BCS occurs,the expression of the upper mass were higher than the underground part,and aboveground is about 2 times the amount of underground part expression,lnc40 and LRX5 genes in WT-Cu in the upper part and underground part of the expression quantity is consistent,so can be concluded that between LRX5 and lnc40 is synergy.