Expression And Functional Analysis Of LncRNA In Apple Under Drought Stress

The drought has become the major problems in agricultural production in China.The drought has caused sere plant,low-photosynthesis and the low yield of crops.Studies have shown that drought is the important factor which induces crop loss.Therefore,how to improve the drought resistance of plants has become one of the urgent problems that we need to solve.Gene research is the signficant way to figure out.LncRNA?long-noncoding RNA?is a kind of non-coding RNA in the organism,which plays an important role in plants and is related to the drought resistance of plants.Its length is longer than 200 nt.In the study of Arabidopsis thaliana,Populus trichocarpa and Musa nana Lour.,it is found that lncRNA is related to drought stress.This research attempts to elucidate the role of lncRNA in drought pathways and provide new ideas for drought resistance research.In this research,potential lncRNAs of the Malus hupenensis Rhed under different drought conditions were detected by high-throughput sequencing.The bioinformatics method was used to analyze the different expression of lncRNA in different tissues and different drought treatments.Besides,possible targets and biological functions are analysised by bioinformatics.Meanwhile,lncRNA with large difference was selected for real-time fluorescence quantitative PCR.The verified lncRNA was over-expressed and transgenic.On this basis,the drought resistance physiological indexes of transgenic Arabidopsis were measured to confirm the function of lncRNA.The main results are as follows:?1?Basised on sequenced results,17280 reliable lncRNAs were obtained.Analysising the expression characteristics of these lncRNA,we found that 61.9% of the lncRNA's length are more than or equal to 300 bp.And specific expression of lncRNA was in different tissues and under different drought conditions.Besides,we get the lncRNAs related to miRNAs by analysising sequences.?2?14 lncRNAs with high difference?P<0.01?and high expression?FPKM> 500?were screened by RT-PCR.The results were consistent with the sequenced results.The lncRNAs' target and target biological function were predicted.The results showed that the target could be related to Porphyrin and chlorophyll metabolism,Lipopolysaccharide biosynthesis,Metalloendopeptidase activity,proanthocyanidin biosynthetic process,Starch and sucrose metabolism and protein kinase activity.?3?9 of 14 lncRNAs were overexpressed.The lncRNA overexpression vector was successfully constructed by inserting lncRNAs into the plant expression vector pART27.TCONS₀0008531 and TCONS₀0052948 of transgenic Arabidopsis thaliana were obtained by infiltration of inflorescence by Agrobacterium tumefaciens.?4?The drought resistance of TCONS₀0008531 and TCONS₀0052948 transgenic Arabidopsis thaliana was studied by measuring the proline content,relative water content of plant tissue,malondialdehyde content and chlorophyll content.The result showed that after 10 days of drought treatment,the proline content,relative water content and chlorophyll content of transgenic plants were higher than those of wild type,while the content of malondialdehyde was lower than that of wild type.Our experiments indicated that lncRNA?TCONS₀0008531 and TCONS₀0052948?has a better effect on drought resistance of Arabidopsis thaliana.