JIA-2018-09

1936 HAN Jiao et al. Journal of Integrative Agriculture 2018, 17(9): 1932–1945 wild rice and the transgenic rice. Differences in genetic screening conditions were indicated by the difference in multiples ≥2 and q-value (or FDR)≤0.01. GOseq was used for GO enrichment analysis of differentially expressed genes, and GO analysis with p-value≤0.05 were assigned as differentially expressed genes (Young et al . 2010). Pathway enrichment analysis of DEGs was performed by the KEGG (the major public pathway-related database). 3. Results 3.1. The McPht gene is a member of Pht3 subfamily and located to the mitochondria McPht cDNA encodes a protein of 357 amino acids with the molecular weight of 37.77 kDa and isoelectric point of 9.30 (accession no: AGC00815.1). The McPht phosphate transporter is composed of six conserved domains, in which the 1st, 2nd, 5th, and 6th transmembrane domains are separately located outside the membrane (Fig. 1-A). PHYRE2 analyses showed that the McPht phosphate transporter has three tandem domains with different hydrophilicity regions and six alpha helices (Fig. 1-B). Multiple alignment showed that the McPht phosphate transporter exhibits more than 80% identity with the Pht3 members from 15 dicotyledonous plants (Fig. 1-C), and is a typical homolog of mitochondrial phosphate transporters (Fig. 2), indicating that the McPht gene is a member of the Pht3 subfamily. To localize the McPht protein, the 16318hGFP vector and the McPht::GFP fusion construct under the control of the 35S promoter were separately transferred into Arabidopsis protoplasts by PEG-mediated method. As shown in Fig. 3, the transformant carrying the McPht::GFP construct seems A C B TMHMM posterior probabilities for WEBSEQUENCE 50 100 150 200 250 300 350 Transembrane Inside Outside 1.2 1.0 0.8 0.6 0.4 0.2 0 Probability Mesembryanthemum crystallinum (gb|JQ343214.1) Betula pendula (emb|Y08499.1) Manihot esculenta (gb|KM396289.1) Manihot esculenta (gb|KM396291.1) Manihot esculenta (gb|KM396288.1) Manihot esculenta (gb|KM396290.1) Populus trichocarpa (gb|FJ465175.1) Populus trichocarpa (ref|XM_002300107.2) Ciecer arietinum (emb|AJ275306.1) Medicago truncatula (ref|XM_003624430.2) Glycine max (ref|NM_001250375.1) Glycine max (ref|NM_001248723.1) Lotus japonicus (dbj|AB077283.1) Prunus persica (gb|KP122948.1) Solanum lycopersicum (gb|JQ582707.1) Solanum lycopersicum (ref|NM_001279338.2) Fig. 1 Structural profiles of McPht protein and phylogenetic analysis of McPht gene. A, transmembrane profiles of McPht phosphate transport protein. B, structural characteristics of the McPht protein. C, phylogenetic tree analysis.