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Phytophthora root and stem rot caused by the oomycete pathogen  Phytophthora sojae  is a destructive disease of soybean worldwide. Plant dirigent proteins (DIR) are proposed to have roles in biosynthesis of either lignan or lignin-like molecules, and are important for defense responses, secondary metabolism, and pathogen resistance. In the present work, a novel  DIR  gene expressed sequence tag is identified as up-regulated in the highly resistant soybean cultivar ‘Suinong 10’ inoculated with  P. sojae . The full length cDNA is isolated using rapid amplification of cDNA ends, and designated  GmDIR22  (GenBank accession no. HQ_993047). The full length  GmDIR22  is 789 bp and contains a 567 bp open reading frame encoding a polypeptide of 188 amino acids. The sequence analysis indicated that GmDIR22 contains a conserved dirigent domain at amino acid residues 43–187. The quantitative real-time reverse transcription PCR demonstrated that soybean  GmDIR22  mRNA is expressed most highly in stems, followed by roots and leaves. The treatments with stresses demonstrated that  GmDIR22  is significantly induced by  P. sojae  and gibberellic acid (GA 3 ), and also responds to salicylic acid, methyl jasmonic acid, and abscisic acid. The GmDIR22 is targeted to the cytomembrane when transiently expressed in Arabidopsis protoplasts. Moreover, The GmDIR22 recombinant protein purified from  Escherichia coli  could effectively direct  E -coniferyl alcohol coupling into lignan (+)-pinoresinol. Accordingly, the overexpression of  GmDIR22  in transgenic soybean increased total lignan accumulation. Moreover, the lignan extracts from  GmDIR22  transgenic plants effectively inhibits  P. sojae  hyphal growth. Furthermore, the transgenic overexpression of  GmDIR22  in the susceptible soybean cultivar ‘Dongnong 50’ enhances its resistance to  P. sojae . Collectively, these data suggested that the primary role of GmDIR22 is probably involved in the regulation of lignan biosynthesis, and which contributes to resistance to  P. sojae .

A Novel Soybean Dirigent Gene GmDIR22 Contributes to Promotion of Lignan Biosynthesis and Enhances Resistance to Phytophthora sojae