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

Abstract

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 (GA3), 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.

Molecular Characterization, Evolution, and Expression Profiling of the Dirigent (DIR) Family Genes in Chinese White Pear (Pyrus bretschneideri)

“…Among them, GmDIR22 has been shown to be involved in the biosynthesis of pinoresinol (the dimer of coniferyl alcohol) and the overexpression of GhDIR1 can increase lignin content in cotton. These two genes are also related to plant resistance ( Shi et al, 2012 ; Li et al, 2017b ). Therefore, PbDIR1, 2 and 3 (which has a close relationship with them) may have similar biological functions to them, which is related to the stress response of pear and directly or indirectly participates in the metabolic process of lignin and pinoresinol.…”

“…Lignan can directly transport to the cell wall and converted into lignins ( Yin et al, 2013 ). DIRs improve plant stress resistance by regulating the metabolism of lignin and lignan ( Thamil Arasan et al, 2013 ; Ma and Liu, 2015 ; Li et al, 2017b ; Paniagua et al, 2017 ). Therefore, DIRs plays an important role in the process of plant stress resistance.…”

Integrated transcriptome, small RNA and degradome sequencing approaches provide insights into Ascochyta blight resistance in chickpea

“…These genes were also found to positively regulate defense responses against AB stress in previous studies (Coram and Pang, ; Mantri et al ., ). Dirigent proteins are known to be involved in biosynthesis of lignan and are important for secondary metabolism and pathogen resistance (Li et al ., 2017b). The induced expression of plant dirigent proteins in resistant genotypes as compared to susceptible genotypes might be an indication of their involvement in AB resistance.…”

Plant evolution and environmental adaptation unveiled by long-read whole-genome sequencing ofSpirodela

“…4A), compared to the array of two copies at most in maize (33). Dirigent proteins are also reported to be involved in defense response against fungi and insects (34). They were also induced by abiotic stress, such as drought, low temperature, and abscisic acid (35).…”