Functional Analysis Of The Novel Mycorrhiza-induced Phosphate Transporter Genes From Fragaria Vesca

Arbuscular mycorrhizal(AM)fungi form mutualistic symbiosis with plant.The fungi could promote the development and resistance of the plants.It allows plants to forage a larger soil area mainly for soil-immobile phosphorus but also for other nutrients,significantly promote the phosphorus content of the plants.Once the arbuscular mycorrhizal symbiosis were formed,relevant genes associated with nutrition element absorption were activated,and their coding proteins located in PA membrane and transferred the nutrients to host plant.The strawberry can form endomycorrhiza.Then the mycorrhiza phosphate transporter genes(MPT)was activated.It is greatly improved the phosphorus content of plants.The woodland strawberry(Fragaria vesca)has a small genome(240 Mb).The genome of the woodland strawberry {Fragaria vesca)has been sequenced with 34,809 genes predicted and approximately 25,050 genes annotated in its version 1.0.Under phosphorus starvation,it could inhibit the growing and development of woodland strawberry and had a bad effect on biomass and quality.It is about to find new MPT genes and functional analysis in Fragaria vesca.The main findings are as follows:(1)Using RNA-seq to find the differential expression of transporter genes.The total reads were 56,159,070 and average Cycle Q20 up to 100%.63%-79%of reads from each sample were mapped to strawberry reference genome.A total of 3112 genes were identified by IDEG6(with false discovery rate(FDR)<0.01,fold change log2(FC)>1).The DEG proteins were compared and analyzed,three PTs(FvPT4,FvPT5 and FvPT8)expressed specifically to mycorrhiza root proved by RNA-seq analysis.And using Phylogenetic relationship and qRT-PCR we found that FvPT4,FvPT5 and FvPT8 expressed specifically to mycorrhiza roots.The primary structure and tertiary structure and transmembrane domain were consisted with Phtl family.(3)The functional analysis of the FvPT4,FvPT5 and FvPT8 genes.In these three RNAi lines,the fungus was able to penetrate the roots,but all appeared a new phenotype compared to the controls.A lot vesicles were visible existed in the three PTs RNAi transgenic roots.According to the method in troouvelot et al,we calculated the mycorrhizal rate in the stained roots of RNAi lines and the controls.The results revealed that knockdown of FvPT4,FvPT5 and FvPT8 by RNAi reduces the formation of mycorrhizal,they do play an important role in the mycorrhizal formation.