Effects of Cold Acclimation on Morpho-Anatomical Traits of Heteroblastic Foliage in Pinus massoniana (Lamb.) Seedlings

Cold acclimation before winter has been shown to enhance the cold tolerance of evergreen conifers, including Pinus massoniana Lamb., a characteristic heteroblastic foliage tree in the conifer. In the initial growing season of P. massoniana, both primary needle seedlings (PNSs) and secondary needle seedlings (SNSs) are generated. While previous research has highlighted differences in the morphological structure and photosynthetic physiological functions of primary and secondary needles, their response to cold acclimation remains poorly understood. This study aimed to investigate the changes in morpho-anatomical structure, starch grain accumulation, and lignin deposition in the roots, stems, and leaves of PNSs and SNSs during cold acclimation using solid potassium iodide and hydrochloric acid phloroglucinol double-staining techniques. The results revealed that, during cold acclimation, the leaves and stems of PNSs exhibited sensitivity to low-temperature stress, resulting in noticeable shrinkage and fracture of mesophyll and cortical parenchyma cells. Furthermore, the early stages of cold acclimation promoted the accumulation of starch grains and lignin in the seedling tissues. In contrast to PNSs, the leaves and stems of SNSs exhibited a shorter cold acclimation period, attributed to the hydrolysis of starch grains in the epidermal cell walls and the transformation of xylem lignin, which supports cell structure stability and enhances cold resistance. In conclusion, these findings suggest that SNSs displayed a superior cold resistance potential compared to PNSs following cold acclimation, providing a significant theoretical basis for the further screening of cold-tolerant germplasm resources of P. massoniana and the analysis of cold resistance traits in heteroblastic foliage.