Comparative analysis of element and hormone content in zygotic embryos of Pinus elliottii and P. elliottii × P. caribaea

Abstract

Somatic embryogenesis is a crucial method for achieving clonal forestry in conifers. Understanding the development of zygotic embryos is essential not only for enhancing the efficiency and quality of somatic embryogenesis, but also for advancing forestry breeding programs. This study investigated dynamic changes of element and hormone contents during ZE development of Pinus elliottii and its hybrid P. elliottii × P. caribaea. Significant differences in embryo development speed among different clones were observed. Elemental analysis was conducted using inductively coupled plasma mass spectrometry (ICP-MS) and identified 68 elements, including major, minor, and beneficial elements. In both species, the contents of potassium (K), calcium (Ca), iron (Fe), boron (B) and five beneficial elements decreased during early ZE development, while phosphorus (P) and copper (Cu) increased. Significantly higher levels of K, Ca and Fe at the initial stage, and sulfur (S) and nickel (Ni) decreased at later stages were detected in P. elliottii than in the hybrid. For the other elements, except for very few significant differences at certain stages, most differences between the two species did not reach a significant level. The contents of endogenous hormones were determined and different accumulation patterns were detected in most hormones between the two species, except abscisic acid (ABA) which simultaneously decreased with developments by stage 8. Significant differences were found in indole-3-acetic acid (IAA) contents at most stages between species, while higher levels of total cytokinin (CK) at each stage were detected in the hybrid in comparison with those in P. elliottii. As a result, lower IAA to CK ratios in the hybrid than in P. elliottii. Methyl jasmonate (JA-me) and gibberellin A3 (GA3) contents showed a similar pattern and exhibited an M-shaped fluctuation in the hybrid. Furthermore, JA-me, GA3, gibberellin A4 (GA4) and brassinolide (BR) showed significantly higher levels in the hybrid than in P. elliottii. K-means clustering and correlation analyses were used to explore relationships between elements and hormones during embryo development, revealing complex interplay in both species. These data indicate different requirement in element and hormone contents for embryogenesis and suggest species-specific media composition for each step in somatic embryogenesis. The findings provide insights into their developmental processes and informing future research and applications in somatic embryogenesis and forestry breeding.