Functional Analysis Of Purple Acid Phosphatase GmPAP17 And Evaluation Of Germplasms With GmPAP14 In Soybean

Soybean is an important crop which needs more phosphorus,while the phosphorus in soils exists mainly in the form of organic and cannot be directly absorbed and utilized,which seriously affected soybean growth and development.Thus,mining and utilizing the high phosphorus efficient utilization related genes becomes an important strategy to solve this problem.In view of this,one purple acid phosphatase gene Gm PAP17 was cloned and analyzed based on the previous studies of soybean root RNA-seq data in this study,which could be used in plant phosphorus transgenic molecular breeding.Meanwhile,one transgenic soybean germplasm with another purple acid phosphatase Gm PAP14 obtained in our previous studies were also be evaluated under different phosphorus conditions,which could be used in soybean phosphorus efficient utilization molecular breeding in future.The main results are as follows:1.One purple acid phosphatase gene GmPAP17 was cloned in soybean.The total length of Gm PAP17 was 1832 bp,including 7 exons and 6 introns,with a 996 bp open reading frame which encoding 331 amino acid residuals.Real time quantitative PCR analysis showed that the expression levels of Gm PAP17 in different phosphorus efficiency soybean varieties were obvious different.Under the 28~42 d after phytate treatments,Gm PAP17 in high phosphorus efficient variety(Zhong-huang 15)was significantly higher than the phosphorus inefficient variety(Niu-mao-huang),which indicated that Gm PAP17 was closely related to the soybean phytate decomposition and utilization.2.The biological function of purple acid phosphatase GmPAP17 was studied.The enzyme activity analysis in vitro confirmed that Gm PAP17 had relatively higher acid phosphatase and phytase activities.More importantly,the over-expression transgenic Arabidopsis plants with Gm PAP17 showed significantly higher acid phosphatase and phytase activities than the wild-type control under the phytate conditions.Meanwhile,the phosphorus utilization relevant characteristics such as plant biomass and phosphorus concentration were also increased significantly than wild-type.In addition,the transgenic soybean hairy roots with Gm PAP17 also showed significantly higher acid phosphatase and phytase activities than wild-type control.All these results demonstrated that Gm PAP17 could increase phytase activity around the plant rhizosphere,thus could improve the phytate utilization efficiency in soils.3.The yield related characteristics of transgenic soybean germplasm with another purple acid phosphatase Gm PAP14 was evaluated.The PCR and q PCR identification results showed that Gm PAP14 could stably inherited and expressed in transgenic soybean germplasm,and the pod numbers per plant,seed numbers per plant,hundred-seeds weight and plant biomass of transgenic soybean germplasm increased significantly than the wild-type control,and it could be used in soybean phosphorus efficient utilization molecular breeding in future.Based on results above,the conclusion are as follows: One purple acid phosphatase Gm PAP17 with 996 bp open reading frame was cloned,and the encoding protein Gm PAP17 showed higher acid phosphatase and phytase activities.Under the phytate conditions,the expression levels of Gm PAP17 in high efficient phosphorus soybean variety were significantly higher than in the inefficient variety.The transgenic Arabidopsis plants and soybean hairy roots with Gm PAP17 showed significantly higher acid phosphatase and phytase activities than wild-type,and the phosphorus utilization relevant traits such as plant biomass,phosphorus concentration were also increased significantly,which indicated Gm PAP17 could increase phytase activity and phytate utilization efficiency around the plant rhizosphere.Meanwhile,another purple acid phosphatase Gm PAP14 could inherited and express stably in transgenic soybeans,and the pod numbers,seed numbers per plant,hundred-seeds weight and plant biomass of transgenic soybean increased significantly than the wild-type control,and it could be used in soybean phosphorus efficient utilization molecular breeding in future.