ABSTRACT
Triterpenoids are crucial active ingredients of Ganoderma lucidum (G. lucidum) with various health benefits. Yet the low biosynthesis greatly restricts their industrial applications, novel metabolic engineering strategies are needed to further enhance Ganoderma triterpenoids production. Transcription factors play vital roles in the metabolic regulation of terpenoids, which are still scarce to study in G. lucidum. Herein, a transcription factor GlbHLH5 (GenBank No. MZ436906.1) potential for metabolic regulation of Ganoderma triterpenes was identified for the first time. MeJA could increase Ganoderma triterpenoids biosynthesis, and GlbHLH5 significantly responded to MeJA induction, suggesting GlbHLH5 is a new target for Ganoderma triterpenoids overproduction. The regulatory effect of the newly identified target was further validated by homologous gene overexpression and silence in G. lucidum. It's demonstrated that overexpression of GlbHLH5 significantly increased triterpenoids accumulation and the key enzyme genes transcription in the biosynthetic pathway, while silencing it displayed the opposite effect, indicating GlbHLH5 could positively regulate the triterpenoids biosynthesis by activating the synergistic expression of key enzyme genes in the biosynthetic pathway. Consequently, GlbHLH5 was identified as a positive regulator and novel metabolic target for Ganoderma triterpenoids biosynthesis, it sheds new lights on the regulatory effect regulation and synthetic biology of Ganoderma triterpenoids.
ABSTRACT
Dirigent-jacalin (D-J) genes belong to the plant chimeric lectin family, and play vital roles in plant growth and resistance to abiotic and biotic stresses. To explore the functions of the D-J family in the growth and development of Moso bamboo (Phyllostachys edulis), their physicochemical properties, phylogenetic relationships, gene and protein structures, and expression patterns were analyzed in detail. Four putative PeD-J genes were identified in the Moso bamboo genome, and microsynteny and phylogenetic analyses indicated that they represent a new branch in the evolution of plant lectins. PeD-J proteins were found to be composed of a dirigent domain and a jacalin-related lectin domain, each of which contained two different motifs. Multiple sequence alignment and homologous modeling analysis indicated that the three-dimensional structure of the PeD-J proteins was significantly different compared to other plant lectins, primarily due to the tandem dirigent and jacalin domains. We surveyed the upstream putative promoter regions of the PeD-Js and found that they mainly contained cis-acting elements related to hormone and abiotic stress response. An analysis of the expression patterns of root, leaf, rhizome and panicle revealed that four PeD-J genes were highly expressed in the panicle, indicating that they may be required during the formation and development of several different tissue types in Moso bamboo. Moreover, PeD-J genes were shown to be involved in the rapid growth and development of bamboo shoots. Quantitative Real-time PCR (qRT PCR) assays further verified that D-J family genes were responsive to hormones and stresses. The results of this study will help to elucidate the biological functions of PeD-Js during bamboo growth, development and stress response.
