Abstract

Lotus (Nelumbo nucifera Gaertn) is a wetland vegetable famous for its nutritional and medicinal value. Phenolic compounds are secondary metabolites that play important roles in the browning of fresh-cut fruits and vegetables, and chemical constituents are extracted from lotus for medicine due to their high antioxidant activity. Studies have explored in depth the changes in phenolic compounds during browning, while little is known about their synthesis during the formation of lotus rhizome. In this study, transcriptomic analyses of six samples were performed during lotus rhizome formation using a high-throughput tag sequencing technique. About 23 million high-quality reads were generated, and 92.14% of the data was mapped to the reference genome. The samples were divided into two stages, and we identified 23,475 genes in total, 689 of which were involved in the biosynthesis of secondary metabolites. A complex genetic crosstalk-regulated network involved in the biosynthesis of phenolic compounds was found during the development of lotus rhizome, and 25 genes in the phenylpropanoid biosynthesis pathway, 18 genes in the pentose phosphate pathway, and 30 genes in the flavonoid biosynthesis pathway were highly expressed. The expression patterns of key enzymes assigned to the synthesis of phenolic compounds were analyzed. Moreover, several differentially expressed genes required for phenolic compound biosynthesis detected by comparative transcriptomic analysis were verified through qRT-PCR. This work lays a foundation for future studies on the molecular mechanisms of phenolic compound biosynthesis during rhizome formation.

Highlights

  • Lotus (Nelumbo nucifera Gaertn), a popular aquatic vegetable in Asian countries [1], is famous for its nutritional and medicinal value [2,3]

  • Sacred lotus has been cultivated in China for over 2000 years and has gradually developed into two ecotypes due to its different biological properties [5]; the temperate type produces an enlarged rhizome and the tropical type has a long period of florescence

  • Our study describes a complex genetic crosstalk-regulated network involved in the biosynthesis of phenolic compounds during lotus rhizome development and identifies the transcription levels of key enzymes in phenol biosynthesis

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Summary

Introduction

Lotus (Nelumbo nucifera Gaertn), a popular aquatic vegetable in Asian countries [1], is famous for its nutritional and medicinal value [2,3]. Nonenzymatic browning results from a chemical process called the Maillard reaction Another aspect is that the phenolic substances provide the lotus rhizome with anti-oxidation, anti-mutagen, anti-aging, and anti-inflammatory properties and pathogen resistance. Erythrose-4-phosphate reacts with phosphoenolpyruvate (PEP) by glycolysis for further reaction through the shikimate pathway, which converts sugar phosphates to aromatic amino acids (shikimate–chorismate–cinnamate–phenolics) This pathway is involved in the synthesis of other secondary metabolites (e.g., lignins, salicylates, flavonoid phytoalexins and pigments) that play a critical role in plant growth, development, defense and metabolism. The tuber part of the lotus rhizome was used as the object to explore the accumulation of phenolic substances at different growth and development stages (S1, S2, S3, S4, S5, S6). The specific genes involved in phenolic anabolic pathways during the growth and development of the lotus rhizome were analyzed. Gene expression levels in S1 and S2 are intensive, while in other samples, the expression levels are higher and unconsolidated

Genes Expression Pattern during Rhizome Formation
Key Enzymes Related to Phenols Synthesis during Rhizome Development
Plant Materials and Determination of Total Phenolic Content
RNA Isolation and Sequencing
Analysis and Mapping of RNA-Seq Reads
Differential Expression Analysis
Gene Ontology and KEGG Ortholog Enrichment Analysis
Findings
Quantitative Real-Time PCR Validation of RNA-Seq Data
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