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Ramie (Boehmeria nivea L. Gaud.) Nettle (Urticaceae) the Boehmeria (Boehmeria) perennial herb, is our specialty, known as \Ramie is one of the important textile materials, with good performance fiber textile and wearing apparel performance, is recognized as the \Ramie leaf, stem, root can carry out a variety of comprehensive utilization. China has Ramie resource distribution, introduction and cultivation, system selection and crossbreeding heterosis male sterility, mutation breeding, asexual micropropagation, tissue culture and other aspects to do a more in-depth study. Ramie breeding practice difficult to solve two problems remain: First, the traditional breeding approaches the breeding cycle is too long, the lower quality varieties mixed raw jute. Biological engineering, compared with the transformation of biological varieties has opened up new avenues, some high-yield, resilience, or other good traits gene into the original does not have these traits in vivo, can be reached in a short time the purpose of improved varieties. The purpose of this project is to be major factors affecting the ramie protoplast culture through research to establish a set of protoplast culture until efficient plant regeneration system for the future import of the genetic manipulation of the target gene and cell fusion, and ultimately create excellent varieties to lay good foundation. Its main contents and results are as follows:. Different sources protoplast free material differences ramie seeds need to add concentrations up to about 3.0mgL 2,4-D or KT can induce callus, but these more The poor state of the injured tissue, could not be further used as enzymatic free protoplast material. Different hormone combinations ramie cotyledons, the hypocotyl callus induction rate between the difference reached a very significant level, KT and 2,4-D 0.5mg / L combination for ramie cotyledons, hypocotyls start dedifferentiation good results. 2,4-D0 .5 mg / L and KT 1.0mg / L combination, can induce callus proliferation stable and light yellow callus particles loose under the microscope the cells are small and round, strong cytoplasmic , easy formation of embryogenic cell clusters, and may further be used to free protoplasts. 2 the genotypes of ramie callus induction Chinese limonene 4, Xiang limonene, No. 2, No. 1, Qian limonene, Ruichang Egeria green, Liuyang pumilum green, Enshi Tsing Ma, 1690 hypocotyls could induce produce callus, but the callus rate differences highly significant, and callus state is different. China limonene the 4th most suitable for the formation of callus induced 2,4-D0 .5 mg / L, KT1.0mg / L, GA 3 0.2mg / L of MSB medium hypocotyl and callus in good condition. 3. Suspension cell lines following the culture in order to maintain the activity of the shaking culture suspension cell lines 7d is necessary to conduct a subculture. After 3 to 4 times subculture, in the material of the first 4 to 6d each cultured subculture cycle most suitable for isolation of protoplasts. 4 protoplast isolation the ramie cotyledons and suspension cell lines, respectively, with cellulose the enzyme 3.0 percent macerozyme 0.5% pectinase and cellulase 5.0% 1.0% processing 9h the protoplast yield can reach 10 6 / ml, the vitality of 88.6% -90.6%. 5 protoplast culture
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