Castanea henryi is a monoecious plant with a low female-to-male ratio, which limits its yield. The phytohormone cytokinin (CK) plays a crucial role in flower development, especially gynoecium development. Here, the feminizing effect of CK on the development of C. henryi was confirmed by the exogenous spraying of N-(2-chloro-4-pyridyl)-N’-phenylurea (CPPU). Spraying CPPU at 125 mg·L⁻¹ thrice changed the male catkin into a pure female catkin, whereas at 5 mg·L⁻¹ and 25 mg·L⁻¹, only a part of the male catkin was transformed into a female catkin. A comparative transcriptome analysis of male catkins subjected to CPPU was performed to study the mechanism of the role of CKs in sex differentiation. Using Pearson’s correlation analysis between hormone content and hormone synthesis gene expression, four key genes, LOG1, LOG3, LOG7 and KO, were identified in the CK and GA synthesis pathways. Moreover, a hub gene in the crosstalk between JA and the other hormone signaling pathways, MYC2, was identified, and 15 flowering-related genes were significantly differentially expressed after CPPU treatment. These results suggest that CK interacts with other phytohormones to determine the sex of C. henryi, and CK may directly target floral organ recognition genes to control flower sex.

Camellia oleifera Abel. is an important woody oil plant, and its pollination success is essential for oil production. We conducted this study to select the best pollinator candidates for C. oleifera using principal component analysis and multi-attribute decision-making. Field observations of the flower-visiting characteristics of candidate pollinators were conducted at three sites. The insect species that visited flowers did not considerably differ between regions or time periods. However, the proportion of each species recorded did vary. We recorded eleven main candidates from two orders and six families at the three sites. The pollen amount carried by Apis mellifera was significantly higher than that of other insects. However, the visit frequency and body length of Apis mellifera were smaller than those of Vespa velutina. Statistical analysis showed that A. mellifera is the best candidate pollinator; Eristaliscerealis is a good candidate pollinator; Phytomia zonata, A. cerana, and V. velutina were ordinary candidate pollinators; and four fly species, Episyrphus balteatus, and Eristalinus arvorum were classified as inefficient candidate pollinators. Our study shows that flies and hoverflies play an important role in the pollination system. Given the global decline in bee populations, the role of flies should also be considered in C. oleifera seed production.

Background: In seed plants, the ovule is the precursor to the seed. The process of ovule development and differentiation is regulated by multiple factors, including starch metabolism and endogenous hormones. Castanea henryi produces nuts with high nutritional value. However, the high proportion of empty buds restricts the commercial use of the tree. Previous studies have shown that the empty bud phenotype is closely related to ovule abortion. If none of the ovules in the ovary expand rapidly and develop in 7–8 weeks after pollination, an empty bud will form. Therefore, we studied the development and molecular mechanisms underlying single seed formation in C. henryi. Results: We found that 49 days after pollination (DAP) is a critical period for the formation of fertile and abortive ovules. The morphology and starch distribution of the fertile and abortive ovules differed significantly at 49 DAP. The fertile ovules were smooth and round in appearance, with a large amount of starch. In contrast, abortive ovules were smaller with only a small amount of starch. The embryo sac of the abortive ovule proceeded to develop abnormally, and the entire ovule lacked starch. We identified 37 candidate genes involved in metabolism with potential roles in the regulation of starch levels. Three ADP-glucose pyrophosphorylase (AGPase) genes, one granule-bound starch synthase (GBSS) gene, and two beta-amylase genes could affect starch accumulation. The levels of auxin, cytokinins, gibberellins, and jasmonic acid in fertile ovules were higher than those in abortive ovules. In addition, the levels of endogenous abscisic acid and salicylic acid in abortive ovules were higher than those in fertile ovules of the same age, consistent with the expression patterns of genes related to the synthesis of abscisic and salicylic acid and signal transduction. We identified and mapped the differentially expressed genes associated with hormone synthesis and signal transduction. Conclusions: These results improve our general understanding of the molecular mechanisms underlying single seed development in C. henryi and the phenomenon of empty buds, providing directions for future research.

The normal development of anthers and the formation of functional pollen are the prerequisites for successful pollination and fertilization. In this study, we observed dynamic changes in inflorescence and anther development in the chinquapin ( Castanea henryi ) using stereomicroscopy, light microscopy, and transmission electron microscopy. We found that cytokinesis during meiosis in microsporocytes was of the simultaneous type, and that the tetrads were mainly tetrahedral. Mature pollen grains contained two cells with three germ pores. The anther wall was of the basic type and composed of epidermis, endothecium, middle layers, and tapetum. Mature anthers had no middle layer and tapetum. The tapetum was of the glandular type. At the early microspore stage, a large number of starch granules appeared in the endothecium, which was deformed at the late microspore stage. Lipid droplets appeared in tapetum during the early microspore stage, and a few lipid droplets were still found during tapetum degeneration. The mature pollen accumulated a large amount of starch and lipids. These findings demonstrated that the anther wall provides nutrients and protection for pollen development. There is relatively stable correspondence between the external morphological characteristics of male flowers and internal structure of anther development.