Abstract
Flavonoids are widely distributed secondary metabolic products with many biological functions in plants. Further elucidation of the accumulation and localization patterns of its biosynthesis enzymes will broaden our understanding of flavonoids biosynthesis and regulation. Chalcone isomerase (CHI, EC 5.5.1.6) is an early-step enzyme in the flavonoids biosynthesis pathway. In this study, using an antibody specifically developed against grapevine CHI enzyme, we found that the accumulation of CHI protein exhibited temporal and spatial specificity. In grape berries, CHI was investigated mainly in the outer hypodermis cells of exocarp tissues, in the vascular bundles of mesocarp; and in the integument and the cells around the raphe of seeds. At the subcellular level, CHI was visualized in the cytoplasm, nucleus, and plastids (chloroplasts) of the exocarp cells, while only in the cytoplasm of mesocarp vascular bundle cells. In grapevine vegetable organs, the leaf mesophyll and phloem of leaf veins, the pith ray and primary phloem of stems, the primary phloem and endoderm of roots, and the young leaves, leaf primordium, and the growth point of leaf buds were CHI signal-positive. In these tissue cells, CHI was primarily observed in the cytoplasm, cell wall, and nucleus. The distinct localization patterns of CHI suggested the complexity of flavonoids biosynthesis in grapevine.
Key Message
The distribution of CHI protein exhibited temporal and spatial specificity. Different subcellular localization patterns were observed in grape berries and grapevine vegetable tissues.
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Abbreviations
- PAL:
-
Phenylalanine ammonia-lyase
- C4H:
-
Cinnamate 4-hydroxylase
- CHI:
-
Chalcone isomerase
- CHS:
-
Chalcone synthase
- ANS:
-
Anthocyanidin synthase
- IgG:
-
Immunoglobulin fractions
- BSA:
-
Bovine serum albumin
- PBS:
-
Phosphate-buffered saline
- BS:
-
Blocking solution
- RSR:
-
Regular salt rinse solution
- DFAB:
-
Days after full bloom
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Acknowledgements
This study was funded by the special fund of China Agriculture Research System (CARS-29), the National Natural Science Foundation of China (No. 31601712), Beijing Municipal Natural Science Foundation (No. 6182007), and the Natural Science Foundation of Beijing Academy of Agriculture and Forestry Science (No. QNJJ201604).
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Weidong Huang and Haiying Xu designed the experiments and revised the manuscript; Huiling Wang, Wei Wang, Jicheng Zhan performed the experiments; Huiling Wang and Wei Wang processed statistical analysis; Huiling Wang prepared the manuscript.
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Wang, H., Wang, W., Zhan, J. et al. Tissue-specific accumulation and subcellular localization of chalcone isomerase (CHI) in grapevine. Plant Cell Tiss Organ Cult 137, 125–137 (2019). https://doi.org/10.1007/s11240-019-01557-y
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DOI: https://doi.org/10.1007/s11240-019-01557-y