Cellular and Metabolite Changes in the Secondary Phloem of Chinese Fir (Cuninghamia lanceolata (Lamb.) Hook.) during Dormancy Release

Abstract

Wood in the cold temperate zone is the product of the alternation of the growing season and the dormant period of trees, but our knowledge of the process of dormancy release in trees remains limited. Chinese fir (Cuninghamia lanceolata (Lamb.) Hook.) was used to investigate cellular and metabolite changes in the secondary phloem tissue during dormancy release. The sampling dates were 2 March, 28 March, and 13 April. The microsections of wood-forming tissue were prepared using the paraffin embedding technique to observe the formation of cambium cells; metabolites in secondary phloem cells were extracted using a methanol/chloroform organic solvent system. The results showed that the secondary phloem consists of phloem fibers, sieve cells and phloem parenchyma. The cells were regularly arranged in continuous tangential bands and were in the order of Phloem fiber-Sieve cell-Phloem parenchyma-Sieve cell-Phloem parenchyma-Sieve cell-Phloem parenchyma-Sieve cell-Sieve cell-Phloem parenchyma-. The Chinese fir cambium was in dormancy on 2 March and 28 March, while on 13 April, it was already in the active stage and two layers of xylem cells with several layers of phloem cells were newly formed. The width of the cambium zone increased from 18.7 ± 5.7 μm to 76.5 ± 3.0 μm and the average radial diameter of sieve cells expanded from 15.4 ± 7.5 μm to 21.5 ± 7.4 μm after dormancy release. The cambium zone width and the average radial diameter of sieve cells before and after dormancy release were significantly different (p < 0.01). The phloem parenchyma cells without resin were squeezed and deformed by the sieve cells, and the width of the phloem during the active period was 197.0 ± 8.5 μm, which was larger than that during the dormant period. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS)-based metabolomics was employed to analyze the secondary phloem of Chinese fir on 28 March and 13 April. Thirty-nine differential metabolites during dormancy release were detected. The results showed that the composition of Chinese fir metabolites was different before and after dormancy release. The relative increase in pyruvic acid and ascorbic acid contents proved that the rate of energy metabolism in Chinese fir increased substantially after dormancy release. Changes in cell development and the composition of metabolites revealed that the dormancy release of Chinese fir was at early April and the formation period of phloem tissue is earlier than xylem tissue.