Recalcitrant chestnut seeds are rich in γ-aminobutyric acid (GABA), which negatively regulates adventitious root development by altering carbon/nitrogen metabolism. However, little is known regarding the role of this metabolite in chestnut seeds. In this study, we investigated the effects of GABA changes on the germination of chestnut seeds treated with exogenous GABA and vigabatrin (VGB, which inhibits GABA degradation). Both treatments significantly inhibited seed germination and primary root growth and resulted in the considerable accumulation of H₂O₂, but the endogenous GABA content decreased before germination at 48 h. Soluble sugar levels increased before germination, but subsequently decreased, whereas starch contents were relatively unchanged. Changes to organic acids were observed at 120 h after sowing, including a decrease and increase in citrate and malate levels, respectively. Similarly, soluble protein contents increased at 120 h, but the abundance of most free amino acids decreased at 48 h. Moreover, the total amino acid levels increased only in response to VGB at 0 h. Accordingly, GABA and VGB altered the balance of carbon and nitrogen metabolism, thereby inhibiting chestnut seed germination. These results suggested that changes to GABA levels in chestnut seeds might prevent seed germination. The study data may also help clarify the dormancy and storage of chestnut seeds, as well as other recalcitrant seeds.

Quercus mongolica Fisch. ex Ledeb. and Q. variabilis Blume are two main oak species in China, producing large amounts of acorns every year. However, the trait variations in acorns, as a promising energy crop material, are not fully understood, hence we compared the traits of acorns from the different populations with the altered geographic distribution in this study. Thirteen acorn traits, including phenotype, proximate compositions and functional compounds, were analyzed in both Quercus L. species collected from 44 populations across China. The results showed that, except large differences found among accessions in acorn sizes, the starch varied from 140.96–297.09 mg/g in Q. mongolica and 130.99–306.28 mg/g in Q. variabilis, indicating its substantial differences among populations. The total polyphenols, total flavonoids and soluble tannins varied from 41.76–158.92, 23.43–91.94, and 15.11–17.81 mg/g, respectively, in Q. mongolica, 89.36–188.37, 50.59–116.07, 15.24–17.33 mg/g, respectively, in Q. variabilis, demonstrating their large variations in the levels of polyphenols among populations. Moreover, the acorns of Q. mongolica in North China and Q. variabilis in Southwest China had higher levels of starch and polyphenols. As the geographical location approached in the distribution of two Quercus species, the difference in acorn sizes gradually increased, while that in polyphenols were opposite. Principal component analysis and cluster analysis further revealed that the acorn sizes became larger and polyphenols became less with the increasing latitudes in both species. In North China, the acorns of Q. mongolica had small sizes and high polyphenols, which was contrary to those in Q. variabilis. These findings indicated that acorn traits were closely associated with the geographical distribution. Thus, our results will provide references for the selection breeding of acorn with the high starch, high or low polyphenols in the different regions.