[Objective] The differential expression analysis was performed for FaGF14- B and FaGF14-C genes in tall fescue so as to provide certain basis for follow-up functional analysis of genes. [Method] The sequence fragments...[Objective] The differential expression analysis was performed for FaGF14- B and FaGF14-C genes in tall fescue so as to provide certain basis for follow-up functional analysis of genes. [Method] The sequence fragments of FaGF14-B and FaGF14-C obtained from reverse transcription were used as templates, and the full- length cDNA sequences of FaGF14-B and FaGF14-C were amplified using the 5＇ RACE use 3＇RACE techniques. They were named as FaGF14-B and FaGF14-C, and used for nucleic acid sequence analysis, encoded protein analysis, protein con- served domain analysis, phylogenetic analysis and differential expression analysis. [Result] The FaGF14-B gene has a full length of 1 548 bp. It has a complete open reading frame （ORF, 449-1 228 bp）, and encodes a protein composed of 261 amino acids. The FaGF14-C gene has a full length of 1 250 bp. It also has a complete open reading frame （ORF, 66-848 bp）, and encodes a protein composed of 261 amino acids. The GF14-B and GF14-C proteins all have a typical domain 14-3-3, and their secondary structures all contain 9 conserved co-helical structures and non-conserved N- and C- terminals. The phylogenetic analysis showed that the FaGF14-B and FaGF14-C from tall rescue have high similarities with GF14 protein from gramineous plants, and they are divided into the same clade with closer ge- netic relationship. The real-time fluorescence quantitative PCR analysis showed that the expression of FaGF14-B and FaGF14-C genes is all sensitive to nitrogen stress. [Conclusion] This study will lay a theoretical basis for further screening of low nitrogen-tolerant genes and breeding of low nitrogen-tolerant grass germplasms.展开更多
基金Supported by National Natural Science Foundation of China(31360576)~~
文摘[Objective] The differential expression analysis was performed for FaGF14- B and FaGF14-C genes in tall fescue so as to provide certain basis for follow-up functional analysis of genes. [Method] The sequence fragments of FaGF14-B and FaGF14-C obtained from reverse transcription were used as templates, and the full- length cDNA sequences of FaGF14-B and FaGF14-C were amplified using the 5＇ RACE use 3＇RACE techniques. They were named as FaGF14-B and FaGF14-C, and used for nucleic acid sequence analysis, encoded protein analysis, protein con- served domain analysis, phylogenetic analysis and differential expression analysis. [Result] The FaGF14-B gene has a full length of 1 548 bp. It has a complete open reading frame （ORF, 449-1 228 bp）, and encodes a protein composed of 261 amino acids. The FaGF14-C gene has a full length of 1 250 bp. It also has a complete open reading frame （ORF, 66-848 bp）, and encodes a protein composed of 261 amino acids. The GF14-B and GF14-C proteins all have a typical domain 14-3-3, and their secondary structures all contain 9 conserved co-helical structures and non-conserved N- and C- terminals. The phylogenetic analysis showed that the FaGF14-B and FaGF14-C from tall rescue have high similarities with GF14 protein from gramineous plants, and they are divided into the same clade with closer ge- netic relationship. The real-time fluorescence quantitative PCR analysis showed that the expression of FaGF14-B and FaGF14-C genes is all sensitive to nitrogen stress. [Conclusion] This study will lay a theoretical basis for further screening of low nitrogen-tolerant genes and breeding of low nitrogen-tolerant grass germplasms.
