Screening of Key Indices and the Gene Transcriptional Regulation Analysis Related to Salt Tolerance in Salix matsudana Seedlings

Abstract

Pot experiments were performed to comparatively study the differences in 16 salt tolerance indices between the seedlings of six Salix matsudana clones under the stress of various concentrations of NaCl (0, 0.1%, 0.3%, 0.5%, and 0.7%), including the salt injury index, shoot fresh weight, root fresh weight, leaf water content, relative conductivity, malondialdehyde content, and antioxidant enzyme activity. The salt-tolerant clones and key indices of salt tolerance were selected. Transcriptome sequencing analysis was performed on the selected salt-tolerant and salt-sensitive clones under salt stress, and the links between the physiological indices of salt tolerance and gene expression were analyzed. Results: (1) Superoxide dismutase (SOD), peroxidase (POD), chlorophyll, and net photosynthetic rate were closely related to the salt tolerance of Salix matsudana at the seedling stage. The regression equation was constructed as follows: salt tolerance index (y) = 0.224x10 + 0.216x11 + 0.127x12 + 0.191x7 − 0.187 (x10 = chlorophyll, x11 = SOD, x12 = POD, x7 = net photosynthetic rate). (2) The number of differentially expressed genes between the seedlings of salt-tolerant and salt-sensitive clones varied with the time of exposure (0 h, 4 h, 12 h, and 24 h) to 200 mmol·L−1 NaCl stress. The most differentially expressed genes in Sm172 were detected upon 24 h vs. 4 h of salt treatment, while the most in Sm6 were in the 24 h vs. 0 h comparison. Gene Ontology analysis and Kyoto Encyclopedia of Genes and Genomes analysis showed that several differentially expressed genes were involved in carotenoid biosynthesis and plant mitogen-activated protein kinase signaling pathways. The nine highly expressed transcription factor genes (Sm172-f2p30-2392, Sm172-f2p28-2386, Sm6-f8p60-2372, Sm6-f2p39-2263, Sm6-f16p60-2374, Sm6-f3p60-931, Sm6-f2p60-1067, Sm172-f3p54-1980, and Sm172-f3p54-1980) were closely correlated with the four key indices of salt tolerance. These genes could become genetic resources for salt tolerance breeding of Salix matsudana.