Laris even under normal growth conditions without salt stress.MethodsPlant samples and culture conditionsSalix linearistipularis seeds

Laris even under normal growth conditions without salt stress.MethodsPlant samples and culture conditionsSalix linearistipularis seeds were harvested from saline soil of Anda experimental site (Anda, Heilongjiang, China) in May. The seeds were cultured in 0.8 agar and 2.5 sucrose 1/2MS solid medium at 22 ? 1 with a photoperiod of 12 h light and 12 h dark until five true leaf growth. The salt treatment conditions of S. linearistipularis seedlings (SLH-treated group) were 0, 3, 6, 12, 16 and 24 h in 100 mM NaCl added at 1/2MS medium, and compared with no NaCl-treated control (SLH-control group).Conductivity measurementsConclusions 1. The genomic sequence and functions of S. linearistipularis were noted by using transcriptome data in this study. This paper provided for the first time the genetic information of S. linearistipularis with gene expressions and their functions. The analysis results showed that 85 of the genes overlapped with the Comospore genome, and the 85 of genes that overlapped with other species including Ricinus communis, Vitis vinifera, Prunus RR6 chemical information persica, P. trichocarpa, P. deltoids, and others. This suggests that the genome of S. linearistipularis contains a different gene and the DEGs contributed to the adaptation to the salt stress. Of the 53,362 all-unigenes, the 3134 DEGs ( logRatio 0.584, PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/28298493 FDR 0.05), included 1397 upAfter S. linearistipularis seedlings were treated to 3, 6, 12, 16 and 24 h with an additional 50, 100, 200 mM NaCl in 1/2MS solid medium, REC was measured and compared with the SLH-control group. The conductivities were measured according to Yao et al. [35].RNA extraction and quality assessmentThe CTAB method was used to extract total RNAs of S. linearistipularis. Every sample was selected separately for each treatment consistent with the size of six S. linearistipularis seedlings. Using isopropanol precipitation, the DNA was digested with DNaseI to obtain total RNAs of S. linearistipularis. The RNA samples were used to build a database that met with the quality requirements using NanoDrop (Thermo Scientific, USA). Additionally, using the Agilent 2100 (Agilent, USA) or Caliper Lab Chip GX (Caliper Lifescience Inc., USA) detection, the database of the total RNA samples was refined to meet theNan et al. J of Biol Res-Thessaloniki (2016) 23:Page 12 ofrequirements. The 28S/18S concentration ratio and RIN value of SLH-treated group were 1474 ng -1, 1.3 and 7.7 ng -1, respectively, while those of the SLH-control group were 1870, 1.6 and 8.6 ng -1, respectively.Library construction and sequencing of cDNAAn alternated CTAB method was used for the total RNA extraction of the S. linearistipularis. After the DNaseI treatment, magnetic beads with Oligo (dT) were used to isolate the mRNA (for eukaryotes) or by removing rRNAs from the total RNA (for prokaryotes). By mixing the fragmentation buffer in the Thermomixer?(Eppendorf AG, Germany), the mRNA was segmented into short fragments. Then cDNA was synthesized using the mRNA fragments as templates. Double stranded cDNA was then synthesized. Short fragments were purified and resolved with EB buffer for end reparation and single nucleotide A addition. Subsequently, the short fragments were connected with adapters. Suitable fragments were then selected for the PCR amplification as templates. During the QC steps, the Agilent 2100 Bioanaylzer (Agilent, USA) and the ABI StepOnePlus Real-Time PCR System (ABI, USA) were used in the quantification of the sampl.