Cloning And Functional Analysis Of PeNAC10 Gene In Phyllostachys Edulis

Moso bamboo（Phyllostachys edulis）is one of the most important economic bamboo species in China.Drought,salinity and other stresses not only affected the yield of bamboo,but also restricted its distribution.NAC（NAM,ATAF1/2 and CUC）gene family is a unique and one of the largest transcription factor families in plants,which plays an important role in the plant growth,development and responding to stress.In this study,the homologous gene PeNAC10 of OsNAC10 in rice was selected from the moso bamboo NAC gene family for cloning,and its function was further explored through transgenic means.The main research results are as follows:1.According to the moso bamboo genome database,152 NAC genes were identified.An evolutionary tree was constructed among moso bamboo,Oryza sativa and Arabidopsis thaliana NAC aimno acid sequences,and 25 subfamilies were obtained.Then,we isolated 6 members from the moso bamboo NAP subfamily,named PeNAP-1、PeNAP-2、PeNAP-3、PeNAC10、PeNAC047-1 and PeNAC047-2.All of them were identified as typical NAC transcription factors containing NAM domain.Subcellular localization experiments confirmed that PeNAP-1,PeNAP-2,PeNAP-3,PeNAC10,PeNAC047-1 and PeNAC047-2 proteins were localized in the nucleus.Transcription activation experiments showed that they all had transcriptional activation activities except PeNAP-2 and PeNAC10.2.The expression levels of PeNAP-1、PeNAP-2、PeNAP-3、PeNAC10、PeNAC047-1 and PeNAC047-2 were analyzed by qRT-PCR under different stress conditions and hormone treatments（high salt,drought,high temperature,ABA,MeJA and GA₃）and different tissues.The results found that these 6 genes had certain conserved functions.They all had the highest expression levels in leaves,and responded to a variety of stress and hormones to different degrees.3.Yeast two-hybrid experiment showed that PeNAC10 protein interacted with PeNAP-1,PeNAP-2,PeNAP-3,PeNAC10,PeNAC047-1 and PeNAC047-2 proteins.To investigate the function of PeNAC10gene,it was transformed into rice.And then we treated T2 seedlings under the stress.After,We found that the survival rate of GM rice was higher than WT after 9 days treatment and 10days rehydration with high salt（150 mmol/L）and drought（20%PEG6000）.The antioxidant enzymes activity（SOD,CAT,POD,APX and GR）and osmotic substance（proline and soluble sugar）content of transgenic rice were higher than WT,and the contents of MDA,H₂O₂ and O₂^-were lower than WT.The above results showed that PeNAC10 gene,a member of Moso bamboo NAC gene family,had regulatory function in abiotic stress,and the overexpression of it could enhance the salt and drought tolerance in rice.In this paper,the research provided a theoretical basis for clarifying the stress resistance mechanism of Moso bamboo.