Transcriptomics-based Analysis of the Response of Sugar Content in Litchi Pulp to Foliar Calcium Fertilizer Treatment

Abstract

Ten 16-year-old trees were used as test materials to investigate the effect of foliar calcium fertilizer on the sugar content of ‘Feizixiao’ litchi (Litchi chinensis Sonn.) pulp. The experiment began 35 days after anthesis (DAA) in 2020 and 2021, and the treatment was a foliar spray application of 0.3% CaCl2 aqueous solution, whereas the control was a foliar spray application of water. The sugar content, sucrose-metabolizing enzymes, and ATP-dependent phosphofructokinase (PFK) activities in pulp were measured in 2020 and 2021. Transcriptome sequencing (RNA-seq) was performed on RNA samples from treatment and control fruit pulps at 35, 63, and 69 DAA (full mature stage) in 2020, and 10 genes were chosen for confirmation by real-time polymerase chain reaction (PCR) in 2020 and 2021. At full maturity, the soluble sugar content in the calcium-treated group was extremely significantly or significantly higher than that in the control group. After 63 DAA, the net sucrose-metabolizing enzyme activity in the calcium-treated group was significantly higher than that in the control group. Furthermore, at full maturity, the calcium-treated group had significantly higher sucrose synthase cleavage activity and significantly lower PFK activity than the control group. Fifty-four highly expressed genes in the glycolytic pathway (EMP) were screened from transcriptome data, including hexokinase, PFK, and pyruvate kinase genes; 87% of these genes were downregulated in the treatment group compared with the control group at 69 DAA in 2020. The linear regression between RNA-seq and real-time PCR results was significant in 2020 (r = 0.9292) and 2021 (r = 0.8889). When the fruit is fully ripe, calcium treatment increases net sucrose-metabolizing enzyme activity by increasing sucrose synthase cleavage activity, promoting the accumulation of reducing sugars, and it downregulates phosphofructokinase gene expression in EMP, promoting sugar accumulation.