Genome-Wide Characterization and Functional Validation of the ACS Gene Family in the Chestnut Reveals Its Regulatory Role in Ovule Development.

Ovule abortion significantly contributes to a reduction in chestnut yield. Therefore, an examination of the mechanisms underlying ovule abortion is crucial for increasing chestnut yield. In our previous study, we conducted a comprehensive multiomic analysis of fertile and abortive ovules and found that ACS genes in chestnuts (CmACS) play a crucial role in ovule development. Therefore, to further study the function of ACS genes, a total of seven CmACS members were identified, their gene structures, conserved structural domains, evolutionary trees, chromosomal localization, and promoter cis-acting elements were analyzed, and their subcellular localization was predicted and verified. The spatiotemporal specificity of the expression of the seven CmACS genes was confirmed via qRT-PCR analysis. Notably, CmACS7 was exclusively expressed in the floral organs, and its expression peaked during fertilization and decreased after fertilization. The ACC levels remained consistently greater in fertile ovules than in abortive ovules. The ACSase activity of CmACS7 was identified using the genetic transformation of chestnut healing tissue. Micro Solanum lycopersicum plants overexpressing CmACS7 had a significantly greater rate of seed failure than did wild-type plants. Our results suggest that ovule fertilization activates CmACS7 and increases ACC levels, whereas an overexpression of CmACS7 leads to an increase in ACC content in the ovule prior to fertilization, which can lead to abortion. In conclusion, the present study demonstrated that chestnut ovule abortion is caused by poor fertilization and not by nutritional competition. Optimization of the pollination and fertilization of female flowers is essential for increasing chestnut yield and reducing ovule abortion.

Transcriptomic and Metabolic Profiling Reveal the Mechanism of Ovule Development in Castanea mollissima.

Ovule abortion, which is the main cause of empty burs in the Chinese chestnut, affects the formation of embryos and further reduces yield; therefore, it is important to study the mechanism of ovule abortion. In this study, we analyzed the transcriptomic and metabolomic data of ovules at critical developmental stages to explore the key regulatory networks affecting ovule development. The metabolites were enriched mainly in pathways involved in phytohormone signaling, energy metabolism, and amino acid synthesis in the endoplasmic reticulum. Analysis of the differentially expressed genes (DEGs) revealed that the HSP genes were significantly down-regulated during fertilization, indicating that this process is extremely sensitive to temperature. The hormone and sucrose contents of ovules before and after fertilization and of fertile and abortive ovules at different developmental stages showed significant differences, and it is hypothesized that that abnormal temperature may disrupt hormone synthesis, affecting the synthesis and catabolism of sucrose and ultimately resulting in the abortive development of Chinese chestnut ovules. At the pollination and fertilization stage of chestnuts, spraying with ethylene, ACC, and AIB significantly increased the number of developing fruit in each prickly pod compared to CK (water) treatment. These results indicated that both ethylene and ACC increased the rate of ovule development. This study provides an important theoretical molecular basis for the subsequent regulation of ovule development and nut yield in the Chinese chestnut.