| Ramie(Boehmeria nivea (L.) Gaud) is a perennial bast fiber crop of the Urticaceae Boehmeria. Ramie fiber is known as the "king of natural fibers", one of the longest natural fiber in the world. Stem is composed of bast fiber cells, which is the major source of fiber. In this study, ramie stems on the top was used as the Tester, stem base was used as the Driver for subtractive hybridization to construct two subtractive Library P and Library U.The results were shown as:1. Library P preserved 1102 clones, in which 13 positive clones were obtained and sequenced.And BlastX and BlastN comparison analysis were conducted in Library P. Library U preserved 1152 clones, in which 185 positive clones were obtained and sequenced.And Blast analysis and bioinformatics analysis were conducted in Library U.2. There were 171 effective EST sequences of Library U, in which small fragments and vector sequences were removed. The 171 sequences were divided into 14 functional genes:there were 52 energy sequences, which was about 30%; there were 39 unclassified sequences, about 23%; there were both 14 protein synthesis and cell structure sequences, each about 8%; there were 13 unclear classified sequences, about 8%; there were 10 metabolism related sequences, which was about 6%; there were 8 protein degradation and storage related sequences, which was about 5%; there were 4 pieces of transcription, disease resistance and defense related sequences respctively,each about 2%; there were 3 pieces of cell growth and differentiation, intracellular transport, secondary metabolism related sequences respectively, each about 2%; there 2 pieces of transport, signal transduction related sequences respectively, each about 1%.3. In the Gene Ontology analysis,166 valid sequences were obtained from the Library U with 185 EST sequences.37 unisecquences were obtained after splicing, including 19 contigs and 18 singlets, which were analysised and classified from three aspects:Function at the molecular level 2:the binding related sequences, which was about 67%; the catalytic activity related sequences, which was about 45%; antioxidant related sequences,which was about 18%; molecular structure activity related sequences, which was about 8%; translational regulation activity related sequences, which was about 2%; transport activity associated sequences, which was about 2%.Function at the cellular component level 2:composition of cells sequences, which was about 43%; composition of the organelle related sequences, which was about 37%; composition of the macromolecular complex related sequences, which was about 32%; part of organelles related sequences, which was about 8%; composition of the extracellular related sequences, which was about 2%.Function at the biological process level 2:metabolic processes related sequences, which was about 62%; cellular processes related sequences, which was about 54%; stress related reaction sequences, which was about 21%; biological control processes related sequences, which was about 5%; development process related sequences, which was about 5%; the establishment of localization processes sequences, which was about 5%; localization processes related sequences, which was about 5%; the process of multi-celled organisms related sequences, which was about 2%; regulated control of biological processes related sequences, which was about 2%; negative control of biological processes related sequences, which was about 2%.4.9 fiber development related genes were selected by Blast, including:4 pieces of a-tubulin family gene; 1 piece ofβ-tubulin family gene; 1 piece of a calmodulin gene; 1 piece of chitinase gene/fiber glycosylhydrolase gene; 1 piece of pectin esterase gene; 1 piece of peroxidase gene.5.5 fiber development related genes were selected by GO, including:tubulinα-chain gene, actin gene, Xyloglucan endotransglucosylase/hydrolase protein 22 gene,Ⅴ-type proton ATPase 116 kDa subunit a isoform 4 and Programmed cell death protein 4. |
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