The Expression of Millettia pinnata Chalcone Isomerase in Saccharomyces cerevisiae Salt-Sensitive Mutants Enhances Salt-Tolerance

Abstract

The present study demonstrates a new Millettia pinnata chalcone isomerase (MpCHI) whose transcription level in leaf was confirmed to be enhanced after being treated by seawater or NaCl (500 mM) via transcriptome sequencing and Real-Time Quantitative Reverse Transcription PCR (QRT-PCR) analyses. Its full length cDNA (666 bp) was obtained by 3′-end and 5′-end Rapid Amplification of cDNA Ends (RACE). The analysis via NCBI BLAST indicates that both aminoacid sequence and nucleotide sequence of the MpCHI clone share high homology with other leguminous CHIs (73%–86%). Evolutionarily, the phylogenic analysis further revealed that the MpCHI is a close relative of leguminous CHIs. The MpCHI protein consists of 221 aminoacid (23.64 KDa), whose peptide length, amino acid residues of substrate-binding site and reactive site are very similar to other leguminous CHIs reported previously. Two pYES2-MpCHI transformed salt-sensitive Saccharomyces cerevisiae mutants (Δnha1 and Δnhx1) showed improved salt-tolerance significantly compared to pYES2-vector transformed yeast mutants, suggesting the MpCHI or the flavonoid biosynthesis pathway could regulate the resistance to salt stress in M. pinnata.

The Chalcone Isomerase Family in Cotton: Whole-Genome Bioinformatic and Expression Analyses of the Gossypium barbadense L. Response to Fusarium Wilt Infection

“…More recent investigations suggested an involvement of CHI genes in the abiotic stress responses and biotic stress responses. The expression of Millettia pinnata CHI gene (MpCHI) in saccharomyces cerevisiae salt-sensitive mutants enhances salt-tolerance, so MpCHI regulates the resistance of Saccharomyces cerevisiae to salt treatment in Millettia pinnata [12]. The change of Deschampsia antarctica CHI gene (DaCHI) expression was regulated by salinity stress, so the DaCHI gene may have a unique function of regulating salt stress in the phenylalanine metabolism pathway in Deschampsia antarctica [13].…”

Mechanisms of Soybean Roots' Tolerances to Salinity Revealed by Proteomic and Phosphoproteomic Comparisons Between Two Cultivars

“…Hence, significant amounts of research have been stimulated to elucidate the biosynthetic networks of flavonoids (90,91). However, there are very few reports on the contribution of chalcone metabolism to plant salt tolerance (92,93). Recently, a cytochrome P450 monooxygenase mutant was shown to be involved in a series of abiotic stresses including ABA and salt in Arabidopsis (94).…”

Pathway enzyme engineering for flavonoid production in recombinant microbes

“…A 926 bp-CHI from Ginkgo biloba L. was successfully expressed in E. coli and its activity was confirmed by the formation of flavonoids (Cheng et al., 2011). The expression of CHI from Millettia pinnata was conducted in S. cerevisiae , which enhanced yeast’s tolerance to salt treatment (Wang et al., 2013). Additionally, overexpression of CHIs has been used in hairy roots to confirm their functions and their enhancing effects on flavonoid levels (Li et al., 2006).…”