A Phytophthora sojae gene of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) induced in host infection and its anti-oxidative function in yeast

Abstract

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a multifunctional protein well defined in eukaryotes, especially in mammalian and Saccharomyces cerevisiae. Using the method of suppression subtractive hybridization (SSH), we identified a Phytophthora sojae cDNA coding GAPDH, which was up-regulated during the early stage of soybean infection. The termed PsGapdh gene possessed three copies in the P. sojae genome. Its amino acid sequence harbored overall conserved domain of GADPH, homologous closest to GapC1 of Achlya bisexualis (oomycete) and adjoined to GapC2s of Odontella sinensis and Phaeodactylum tricornutum (diatom), on the C-branch of subfamily Ⅱ GapC in phylogeny tree of GAPDH. The transcriptional level of PsGapdh was up-regulated throughout early infection. Heterogenous expression of PsGapdh in the yeast tdh1-deleted mutant could rescue growth arrest under continuous exposure to H 2 O 2 . These results indicated active roles of PsGapdh in pathogen-host interaction and anti-oxidation.

Plant-mediated gene silencing restricts growth of the potato late blight pathogen Phytophthora infestans

“…Besides the green fluorescence protein ( GFP ) marker gene, sequences of the following genes were used: G-protein β-subunit ( PiGPB1 ) ( Latijnhouwers and Govers, 2003 ), cellulose synthase A2 ( PiCESA2 ) ( Grenville-Briggs et al , 2008 ), pectinesterase ( PiPEC ) ( Ospina-Giraldo et al , 2010a ), and the constitutive glyceraldehyde 3-phosphate dehydrogenase ( PiGAPDH ) gene. The choice of target genes relied on published data on PiCESA1 and PiGAPDH ( Zeng et al , 2006 ; Grenville-Briggs et al , 2008 ) together with our analysis of PiGPB1 and PiPEC in infected potato plants (see Supplementary Fig. S1 at JXB online).…”

Expression of γ-tocopherol methyltransferase gene from Brassica napus increased α-tocopherol content in soybean seed

“…The targeted fragment of BnTMT (424 bp) was amplified in 50 mm 3 reaction mixture containing 5 mm 3 template, 1.25 U ExTaq (TaKaRa), 1× PCR buffer and 0.2 mM of dNTP each by using TMT-5 (5′-AATGAAAGCGACTCTCGCACC-3′) and TMT-6 (5′-CCTTGAGCTTCCTCCGATCC-3′) as primers. A 225 bp fragment of soybean actin was also amplified, as the quantitative control of RT-PCR reaction, under the same conditions with actin-R (5′-TTAGAAGCA CTTGCGGTGCACG-3′) and actin-F (5′-GTACTGCAA CATCGTGCTGTCG-3′) (Zeng et al 2006) as primers.…”