Nucleoside 5′-Phosphoramidates Control the Phenylpropanoid Pathway in Vitis vinifera Suspension-Cultured Cells

Abstract

It is known that cells contain various uncommon nucleotides such as dinucleoside polyphosphates (NpnN’s) and adenosine 5′-phosphoramidate (NH2-pA) belonging to nucleoside 5′-phosphoramidates (NH2-pNs). Their cellular levels are enzymatically controlled. Some of them are accumulated in cells under stress, and therefore, they could act as signal molecules. Our previous research carried out in Arabidopsis thaliana and grape (Vitis vinifera) showed that NpnN’s induced the expression of genes in the phenylpropanoid pathway and favored the accumulation of their products, which protect plants against stress. Moreover, we found that NH2-pA could play a signaling role in Arabidopsis seedlings. Data presented in this paper show that exogenously applied purine (NH2-pA, NH2-pG) and pyrimidine (NH2-pU, NH2-pC) nucleoside 5′-phosphoramidates can modify the expression of genes that control the biosynthesis of both stilbenes and lignin in Vitis vinifera cv. Monastrell suspension-cultured cells. We investigated the expression of genes encoding for phenylalanine ammonia-lyase (PAL1), cinnamate-4-hydroxylase (C4H1), 4-coumarate:coenzyme A ligase (4CL1), chalcone synthase (CHS1), stilbene synthase (STS1), cinnamoyl-coenzyme A:NADP oxidoreductase (CCR2), and cinnamyl alcohol dehydrogenase (CAD1). Each of the tested NH2-pNs also induced the expression of the trans-resveratrol cell membrane transporter VvABCG44 gene and caused the accumulation of trans-resveratrol and trans-piceid in grape cells as well as in the culture medium. NH2-pC, however, evoked the most effective induction of phenylpropanoid pathway genes such as PAL1, C4H1, 4CL1, and STS1. Moreover, this nucleotide also induced at short times the accumulation of N-benzoylputrescine (BenPut), one of the phenylamides that are derivatives of phenylpropanoid and polyamines. The investigated nucleotides did not change either the lignin content or the cell dry weight, nor did they affect the cell viability throughout the experiment. The results suggest that nucleoside 5′-phosphoramidates could be considered as new signaling molecules.

Efficient genome editing in grapevine using CRISPR/LbCas12a system

“…2 F, Supplementary Table S 5 ). The results obtained here provide a promising way to produce valuable compounds by modifying metabolic pathways using CRISPR/LbCas12a in suspension-cultured grape cells, which have been extensively used to synthesize a variety of secondary metabolites (Pietrowska-Borek et al 2021 ; Badim et al 2022 ). Given that both sugars and flavonoids contribute greatly to grape berry quality, the results of this study suggest that CRISPR/LbCas12a could be an important tool for trait improvement in grapevine.…”

Advances in the Synthesis and Analysis of Biologically Active Phosphometabolites

“…The structures of phosphoramidates, which are characterized by a single covalent bond between nitrogen and phosphorus with an oxidation state of +5 and coordination number 4, can be classified according to the type of amine bound to phosphorus (see Figure 9 ) into three different classes of N -phosphorylated compounds [ 21 ] where phosphorus is covalently bonded to the nitrogen (a) of a mono-substituted NH 2 -group, such as phosphagens or N -phosphorylated amino acids [ 40 ], (b) of a di-substituted NH 2 -group, and (c) of a free NH 2 -group, such as nucleoside 5′-phosphoramidates [ 41 ].…”