An integrated physiological indicator and transcriptomic analysis reveals the response of soybean buds to high-temperature stress.

Background: Under global warming, high temperature (HT) has become a major meteorological factor affecting soybean production. To explore the candidate genes and regulatory mechanism of the soybean bud response to HT stress, previously identified as HT-tolerant ('Handou14'; HD14) and HT-sensitive ('Jiadou36'; JD36) were treated for 5 days in an artificial climate incubator either with HT (43 °C (day)/ 33 °C (night), 16 h light/8 h darkness) or the non-stress growth condition (25 °C, 16 h light/8 h darkness) as the control at the bud stage were used as experimental materials in this study. After HT treatment, changes in physiological indicators including hypocotyl length, enzyme activity and hormone content were detected; at the same time, the cotyledons, hypocotyls, and main roots were collected for transcriptome sequencing analysis. Analyzing the mechanisms of HT stress response in the bud stage of HD14 and JD36 at physiological and transcription levels.

Results: Analysis of physiological indicator showed that the activities of superoxide dismutase (SOD) were significantly increased 47.4% and 41.2% in the cotyledon of HD14 and the main root of JD36, and the contents of peroxidase (POD) were significantly increased 61.5% and 125% in the hypocotyl of HD14 and JD36; the contents of malonaldehyde (MDA) were significantly increased 44.8% and 22.2% in the main root of HD14 and JD36 after HT treatment. The content of abscisic acid (ABA) were significantly increased 1.9 fold and 1.2 fold in the root of HD14 and JD36 in response to HT treatment, whereas the content of gibberellin (GA) were decreased 2.2 fold and 1.3 fold in the cotyledon and root, and increased 1.6 fold in the hypocotyl in HD14 (P < 0.05). Thus, higher SOD and POD activities, higher ABA content, and a smaller increase in MDA content may improve tolerance to HT stress. The HT-tolerant cultivar may have stronger GA signal transduction in the hypocotyl to combat the negative effects of HT. RNA-sequencing was performed to analyze the differential expression of genes in buds of the two cultivars under the HT treatment and control condition. In total, 3,633, 1,964, 9,934, and 3,036 differentially expressed genes (DEGs) were identified in the CH (control group of HD14) vs. TH (HT-treatment group of HD14), CJ (control group of JD36) vs. TJ (HT-treatment group of JD36), TJ vs. TH, and CJ vs. CH comparison groups, respectively. Bioinformatic analysis revealed that most DEGs were mainly involved in metabolic processes, catalytic activity, carbohydrate, energy transduction, and signaling pathways. The results of qRT-PCR validation (86.67%) and changes in physiological indicators were consistent with the RNA-sequencing data. Five DEGs were selected as candidate genes in the response to HT stress at the bud stage.

Conclusion: In summary, soybean cells are protected from oxidative damage by an increase in antioxidant enzyme activities and accumulation of hormone content under HT stress. Concomitantly, changes in the expression of crucial genes and signal transmission processes are induced, thus initiating adaptive and protective mechanisms. This study provides a theoretical basis for clarification of the physiological and molecular mechanisms in the response to HT stress of soybean bud.