Green tea (Camellia sinensis, Cs) abundantly produces a diverse array of phenylpropanoid compounds benefiting human health. To date, the regulation of the phenylpropanoid biosynthesis in tea remains to be investigated. Here, we report a cDNA isolated from leaf tissues, which encodes a R2R3-MYB transcription factor. Amino acid sequence alignment and phylogenetic analysis indicate that it is a member of the MYB4-subgroup and named as CsMYB4a. Transcriptional and metabolic analyses show that the expression profile of CsMYB4a is negatively correlated to the accumulation of six flavan-3-ols and other phenolic acids. GFP fusion analysis shows CsMYB4a’s localization in the nucleus. Promoters of five tea phenylpropanoid pathway genes are isolated and characterized to contain four types of AC-elements, which are targets of MYB4 members. Interaction of CsMYB4a and five promoters shows that CsMYB4a decreases all five promoters’ activity. To further characterize its function, CsMYB4a is overexpressed in tobacco plants. The resulting transgenic plants show dwarf, shrinking and yellowish leaf, and early senescence phenotypes. A further genome-wide transcriptomic analysis reveals that the expression levels of 20 tobacco genes involved in the shikimate and the phenylpropanoid pathways are significantly downregulated in transgenic tobacco plants. UPLC-MS and HPLC based metabolic profiling reveals significant reduction of total lignin content, rutin, chlorogenic acid, and phenylalanine in CsMYB4a transgenic tobacco plants. Promoter sequence analysis of the 20 tobacco genes characterizes four types of AC-elements. Further CsMYB4a-AC element and CsMYB4a-promoter interaction analyses indicate that the negative regulation of CsMYB4a on the shikimate and phenylpropanoid pathways in tobacco is via reducing promoter activity. Taken together, all data indicate that CsMYB4a negatively regulates the phenylpropanoid and shikimate pathways.Highlight: A tea (Camellia sinensis) MYB4a is characterized to encode a R2R3-MYB transcription factor. It is shown to repressively control the phenylpropanoid and shikimate pathway.
Main conclusion
LARs promoted the biosynthesis of catechin monomers and inhibited their polymerization. The accumulation of catechin monomers and polymers was increased by up-regulating the expression of
NtLAR
and
NtANR
s in CsMYB5b transgenic tobacco.
Tea is rich in polyphenolic compounds, and catechins are the major polyphenols in tea. The biosynthesis of polyphenols is closely related to the expression of the leucoanthocyanidin reductase (LAR) and anthocyanidin reductase (ANR) genes. In this paper, an evolutionary analysis and functional characterization of three CsLARs were performed. The phylogenetic tree showed that plant LARs could be grouped into three, including gymnosperms, monocotyledons and dicotyledons (clusters I and II). The eighth amino acid residue in a conserved LAR-specific motif is changeable due to a transversion (G → T) and transition (G → C) that occur in the corresponding codon. Therefore, plant LARs can be classified as G-type, A-type and S-type LARs due to this variable amino acid residue. Although (2R, 3S)-trans-flavan-3-ols were the products of recombinant CsLARs proteins expressed in Escherichia coli, both (2R, 3S)-trans and (2R, 3R)-cis-flavan-3-ols were detected in tobacco overexpressing CsLARs. However, a butanol/HCl hydrolysis assay indicated that overexpression of the CsLARs caused a decrease in polymerized catechins. A hybridization experiment with CsLARc + AtPAP1 also showed that no polymers other than epicatechin, catechin and glycoside were detected, although the accumulation of anthocyanins was markedly decreased. CsMYB5b promoted the biosynthesis of both flavan-3-ols and proanthocyanidins (PAs). Therefore, LARs promoted the biosynthesis of catechin monomers and inhibited their polymerization. The accumulation of catechin monomers and polymers was increased by up-regulating the expression of the NtLAR and NtANRs in CsMYB5b transgenic tobacco.Electronic supplementary materialThe online version of this article (doi:10.1007/s00425-017-2771-z) contains supplementary material, which is available to authorized users.
In this article, we study the joint pricing and inventory control problem for perishables when a retailer does not sell new and old inventory at the same time. At the beginning of a period, the retailer makes replenishment and pricing decisions, and at the end of a period, the retailer decides whether to dispose of ending inventory or carry it forward to the next period. The objective of the retailer is to maximize the long‐run average profit. Assuming zero lead time, we propose an efficient solution approach to the problem, which is also generalized to solve three extensions to the basic model. A feature of the present study is that we consider explicitly the influence of perishability on the demand. Among the insights gathered from the numerical analysis, we find that dynamic pricing aids extending shelf life and when disposal incurs a lower cost, or even a positive salvage value, the retailer is induced to dispose earlier since the benefit of selling new inventory offsets the loss due to disposal. We also observe that the faster the perceived rate of deterioration, the lower the threshold of the ending inventory for disposal. Perhaps a bit counter‐intuitive, maximizing profits does not mean eliminating disposals or expirations.
Abstract:In this work, we study manpower allocation with time windows and job-teaming constraints. A set of jobs at dispersed locations requires teams of different types of workers where each job must be carried out in a preestablished time window and requires a specific length of time for completion. A job is satisfied if the required composite team can be brought together at the job's location for the required duration within the job's time window. The objective is to minimize a weighted sum of the total number of workers and the total traveling time. We show that construction heuristics used with simulated annealing is a good approach to solving this NP-hard problem. In experiments, this approach is compared with solutions found using CPLEX and with lower bounds obtained from a network flow model.
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