Dataset Information

Comparative transcriptome analysis of roots, stems, and leaves of Pueraria lobata (Willd.) Ohwi: identification of genes involved in isoflavonoid biosynthesis.

ABSTRACT:

Background

Pueraria lobata (Willd.) Ohwi is a valuable herb used in traditional Chinese medicine. Isoflavonoids are the major bioactive compounds in P. lobata, namely puerarin, daidzin, glycitin, genistin, daidzein, and glycitein, which have pharmacological properties of anti-cardiovascular, anti-hypertension, anti-inflammatory, and anti-arrhythmic.

Methods

To characterize the corresponding genes of the compounds in the isoflavonoid pathway, RNA sequencing (RNA-Seq) analyses of roots, stems, and leaves of P. lobata were carried out on the BGISEQ-500 sequencing platform.

Results

We identified 140,905 unigenes in total, of which 109,687 were annotated in public databases, after assembling the transcripts from all three tissues. Multiple genes encoding key enzymes, such as IF7GT and transcription factors, associated with isoflavonoid biosynthesis were identified and then further analyzed. Quantitative real-time PCR (qRT-PCR) results of some genes encoding key enzymes were consistent with our RNA-Seq analysis. Differentially expressed genes (DEGs) were determined by analyzing the expression profiles of roots compared with other tissues (leaves and stems). This analysis revealed numerous DEGs that were either uniquely expressed or up-regulated in the roots. Finally, quantitative analyses of isoflavonoid metabolites occurring in the three P. lobata tissue types were done via high-performance liquid-chromatography and tandem mass spectrometry methodology (HPLC-MS/MS). Our comprehensive transcriptome investigation substantially expands the genomic resources of P. lobata and provides valuable knowledge on both gene expression regulation and promising candidate genes that are involved in plant isoflavonoid pathways.

SUBMITTER: Wang C

PROVIDER: S-EPMC7906042 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Pueraria lobata is an important medicinal and edible homologous plant that is widely cultivated in Asian countries. However, its production and quality are seriously threatened by its susceptibility to pseudo-rust disease. The underlying molecular mechanisms are poorly known, particularly from a transcriptional perspective. Pseudo-rust disease is a major disease in pueraria, primarily caused by Synchytrium puerariae Miy (SpM). In this study, transcriptomic profiles were analyzed and compared between two pueraria varieties: the disease-resistant variety (GUIGE18) and the susceptible variety (GUIGE8). The results suggest that the number of DEGs in GUIGE18 is always more than in GUIGE8 at each of the three time points after SpM infection, indicating that their responses to SpM infection may be different, and that the active response of GUIGE18 to SpM infection may occur earlier than that of GUIGE8. A total of 7044 differentially expressed genes (DEGs) were identified, and 406 co-expressed DEGs were screened out. Transcription factor analysis among the DEGs revealed that the bHLH, WRKY, ERF, and MYB families may play an important role in the interaction between pueraria and pathogens. A GO and KEGG enrichment analysis of these DEGs showed that they were mainly involved in the following pathways: metabolic, defense response, plant hormone signal transduction, MAPK signaling pathway-plant, plant pathogen interaction, flavonoid biosynthesis, phenylpropanoid biosynthesis, and secondary metabolite biosynthesis. The CPK, CESA, PME, and CYP gene families may play important roles in the early stages after SpM infection. The DEGs that encode antioxidase (CAT, XDH, and SOD) were much more up-regulated. Defense enzyme activity, endogenous hormones, and flavonoid content changed significantly in the two varieties at the three infection stages. Finally, we speculated on the regulatory pathways of pueraria pseudo-rust and found that an oxidation-reduction process, flavonoid biosynthesis, and ABA signaling genes may be associated with the response to SpM infection in pueraria. These results expand the understanding of pueraria resistance and physiological regulations by multiple pathways.