Search

Lycoris longituba is an important ornamental and medicinal plant and contains various Amaryllidaceae alkaloids such as galanthamine (GAL), lycorine (LYC), and lycoramine (LYCM), which have been reported to exhibit medicinal values. However, the effects of light quality on L. longituba are unknown. The present study aimed to explore the effects of different light emitting diode (LED) light quality on growth, physiological characteristics, and alkaloid accumulations in the seedlings. Lycoris longituba seedlings were cultured under different ratios of blue and red LEDs. The results showed that compared to CK, RB (1:2) treatment improved growth, biomass accumulation, and the synthesis of photosynthetic pigments and reduced membrane lipid peroxidation. Blue light increased GAL, LYC, and LYCM contents which were 2.45-, 1.74-, and 1.92-fold higher than CK, respectively. Most of Amaryllidaceae alkaloids biosynthetic pathway genes including PAL, C4H, NBS, TYDC, OMT, and CYP96T1 were upregulated under B treatment. In general, RB (1:2) was the optimal light quality condition for the growth of L. longituba seedlings. B treatment could promote the accumulation of alkaloids and related gene expressions. This study has established the theoretical foundation and technical support for the use of LED light quality control technology in the production of high-quality L. longituba seedlings.

Lycoris is a genus of plants with flowering bulbs that have high ornamental, medicinal, and ecological value. The slow growth of Lycoris bulbs under natural conditions is a constraint to the Lycoris industry. Thus, accelerating the development of Lycoris bulbs is extremely important. Starch provides the energy and material basis for bulbs to enlarge. Previous studies have shown reduced starch metabolism capacity in Lycoris bulbs when dormant, but changes within bulbs during the vegetative growth stage remain poorly understood. In our present study, four groups of developing Lycoris radiata bulbs during the vegetative growth phase and with the same genetic background were created and used to investigate changes in carbohydrate content, starch synthesis, metabolic enzyme activity, and the gene expression patterns of several genes encoding these enzymes. Increases in enzyme activity during starch synthesis, especially for AGPase, were observed and correlated with the expression levels of the AGPase genes. Starch levels initially increased and then declined. This may be due to the increasing activity of amylase, which resulted in a rapid increase in sugar concentration during bulb development. We also detected changes in starch synthesis and metabolism from different layers of the bulbs during development, with increases in starch synthesis and starch degradation activity being more significant in the middle scales. The sugar within the middle scales may be transported to the inner scales, which increases the inner scale sugar content, promotes the expression levels of CycD genes, and accelerates cell division during bulb development.

Lycoris species have great ornamental and medicinal values; however, their low regeneration efficiency significantly restricts their commercial production. Exogenous hormone application is an effective way to promote bulblet development, but their effect on Lycoris radiata has not been verified to date. In the present study, we examined the effect of different exogenous hormones on bulblet development in L. radiata, and found that gibberellic acid (GA) significantly inhibited, whereas paclobutrazol (PBZ), abscisic acid (ABA), and ethrel promoted bulblet development, especially PBZ, a GA biosynthesis inhibitor. Furthermore, GA reduced endogenous cytokinin (CK) content, as well as the activities of carbohydrate metabolism enzymes, including sucrose synthase (SUS) and glucose-1-phosphate adenylyltransferase (AGPase), by downregulating the expression levels of LrSUS1, LrSUS2, and genes encoding AGPase large and small subunits. This resulted in the decrease in carbohydrate accumulation in the bulblets, thus hindering their development. PBZ had the opposite effect to GA on carbohydrate metabolism; it decreased endogenous GA15 and GA24, thereby promoting bulblet development. ABA promoted endogenous auxin content and the activities of starch synthesis enzymes, especially soluble starch synthase (SSS) and granule-bound SS (GBSS), through the up-regulation of the expression levels of LrSS1, LrSS2, and LrGBSS1 genes, which could also result in the accumulation of carbohydrates in the bulblets and promote their development. In addition, ethrel application partly promoted bulblet development by promoting endogenous CK content. Although the accumulation of carbohydrates and the activity of starch enzymes were increased by ethrel treatment, we hypothesized that the effect of ethrel on regulating carbohydrate metabolism may be indirect. Our results could provide a basis for improving the propagation efficiency of L. radiata for production, as well as propose some directions for future research.

Lycoris longituba is a traditional medicinal plant containing the bioactive compound galanthamine (Gal), a type of amaryllidaceous alkaloid and can be used to treat Alzheimer’s disease. However, research on its genome or transcriptome and associated genes in the biosynthetic pathway is incomplete. In this study, we estimated the nuclear genome size of this species to be 29.47 Gb by flow cytometry. Then, RNA sequencing of the leaves, roots, and bulbs of L. longituba was carried out. After de novo assembly, 4,74,589 all-transcripts and 3,33,440 all-unigenes were finally generated. In addition, the differentially expressed genes (DEGs) were identified, and genes involved in the galanthamine metabolic pathway encoding tyrosine decarboxylase (TYDC), phenylalanine ammonia-lyase (PAL), cinnamate 4-hydroxylase (C4H), p-coumarate 3-hydroxylase (C3H), norbelladine synthase (NBS), norbelladine 4'-O-methyltransferase (OMT), noroxomaritidine synthase (CYP96T1), and N-methyltransferase (NMT) were detected and validated by real-time quantitative PCR analysis. One candidate gene, Lycoris longituba O-Methyltransferase (LlOMT), was identified in the proposed galanthamine biosynthetic pathway. Sequence analysis showed that LlOMT is a class I OMT. LlOMT is localized in the cytoplasm, and biochemical analysis indicated that the recombinant LlOMT catalyzes norbelladine to generate 4′-O-methylnorbelladine. The protoplast transformation result showed that the overexpression of LlOMT could increase the Gal content. Our results indicate that LlOMT may play a role in galanthamine biosynthesis in L. longituba. This work provides a useful resource for the metabolic engineering of amaryllidaceous alkaloids.

Background Lycoris species have great ornamental and medicinal values; however, their low regeneration efficiency seriously restricts their commercial production. Understanding the mechanism of bulblet propagation in this genus, which has remained underexplored to date, could provide a theoretical basis for improving the reproductive efficiency. Therefore, we studied the bulblet initiation and developmental processes in Lycoris radiata. Results We found that bulblets are formed on the junctions of the innermost layers of scales and the basal plate, and initially present as an axillary bud and gradually develop into a bulblet. We also determined the changes in carbohydrate and endogenous hormone contents during bulblet initiation and development, as well as the expression patterns of genes involved in carbohydrate metabolism and hormone biosynthesis and signaling through transcriptome analysis. Soluble sugars derived from starch degradation in the outer scales are transported to and promote bulblet initiation and development through starch synthesis in the inner scales. This process is mediated by several genes involved in carbohydrate metabolism, especially genes encoding ADP glucose pyrophosphorylase, a crucial starch synthesis enzyme. As for hormones, endogenous IAA, GA, and ABA content showed an increase and decrease during bulblet initiation and development, respectively, which were consistent with the expression patterns of genes involved in IAA, GA, and ABA synthesis and signal transduction. In addition, a decrease in ZR content may be down- and up-regulated by CK biosynthesis and degradation related genes, respectively, with increasing auxin content. Furthermore, expression levels of genes related to BR, JA, and SA biosynthesis were increased, while that of ethylene biosynthesis genes was decreased, which was also consistent with the expression patterns of their signal transduction genes. Conclusions The present study provides insights into the effect of carbohydrate metabolism and endogenous hormone regulation on control of L. radiata bulblet initiation and development. Based on the results, we propose several suggestions to improve L. radiata propagation efficiency in production, which will provide directions for future research.