Studies Of Some Problems On Resource Utilization And Tissue Culture Of Polytrichum Commune Hedw.

Polytrichum commune Hedw.(Polytrichaceae,Musci),a wordwide moss species,was widely used in moss biological research for its large plant body and abundant biomass.The medicinal application,high effective NH₄⁺ assimilation and potential effects of P.commune on soil nitrogen circle have been attra(?)d more attention.The aims of the present researches are:1) to study the main chemical components of the plants of P.commune collected from the north snburb of Jinhua,Zhejiang province,China;2) to test the effects of wild plants of P. commune on the properties of its distributed soil;3) to find a suitable technology to culture P. commune in viro;4) to discover ideal conditions including culture type,pH,illumination time,temperature and so on for the spore germination and protonemal growth of P.commune in vitro;5) to investigate the effects of phytohormones on the protonemal growth and callus induction of P commune;6) to test the effects of nitrogen deposition on protonemal growth and gametophytes induction of P comune.According to the above researches,we expect to find the most optimal method to proliferate P commune for the exploration and utilization of medicine,and to providing experimental material for physiological,genetic,developing and evolutional researches of mosses.Our main contents are as follows.1.The extract of P.commune with ethers was analyzed by GC-MS technology and the results show that there are 61 types of compounds(with 80%matching degree) identified by searching the mass spectral library in computer.The relative content of each compound was confirmed according to the peak area of compound tested by area normalization method. 99.96%peak area was occupied by the composition content of identified compounds.The main compounds of ethers extract of P.commune include alkanes,alkenes,alcohols,ketones and ester compounds such as benzenes and acid.The identified compounds of comparatively high concentration were 9,12-Octadecadienoic acid[Z.Z](21.225%),Oleic-acid(20.988%), n-Hexadecanoic acid(10.118%),2,6.10,15,19,23-hexamethyl-[all-E]-2,6,10,14,18,22-Tetracosahexaene(8.275%),Arachidonic acid(3.6%),Naphthale -ne(2.544%) and so on. in the study two terpenoid([8beta,13beta]-kaur-16-ene and 8-propoxy-Cedrane) were discovered firstly in the extract of P.commune with ethers.There are large numbers of alkanes which the nun(?)er of carbon atom less than 23 in the extract of P.commune with ethers,such as Heptadecane, Octadecane and Nonadecane.2.The soil properties of bare ground and plots covered with P.commune were studied by mixed soil sampling method.The results showed that P.commune has significant effects on soil pH,but has little or no effects on electrical conductivity,water content and crganic matter content.The Urease and Calalase were not affected by P.commune except for Invertase, which could be inhibited by P.commune.There was no significant difference in soil respiratory intensity between bare ground and plots covered with P.cornmune after 48h:= 0.178),96h(P = 0.585),and 144h(P = 0.397) counted from soil sample placed in senled container.The growth of P.commune posed significant effects to the number of microorganisms.The number of the soil microorganism tested in plots covered with P. commune were all larger than that in bare ground except for actinomycetes,which was only 0.8 times in bare ground.The number of bacteria,fungi,azotabacter and nitrifying bacteria in plots covered with P.cornmune were 1.9,2.5,2.9 and 6.7 times of them in bare gronnd, respectively.3.The effects of culture medium types,content of sucrose,light intensities,temperatures and Pb²⁺ concentration on the spore goerminationo of P.commune were tested in virto and the results show:1) the spores of P.commmune were able to germinate on SM,MS,KM media,but not on NM and BM media,they were more sensitive to light in sucrose-only medium;2) their germination rates were lower in the media only with minerals or those only with sucrose at lower or higher concentration,but higher in the media with minerals and sucrose;3) the temperatures from 20℃to 30℃were appropriate for the spore germination of P.commune,4) Pb²⁺ had significant inhibitory effects on their spore germination;5) the concentrations of nutrients in media could affect the biopolar germination of the spores of P.commune.The responses of the spores of P.commune to the factors in vitro agree with their habitat characteristics in the field to an extent. 4.In vitro the influences of culture medium type,light intensity,temperature, photohormone,the aqueous extract from its own gametophyte,temperature and nitrogen and carbon concentrations on the protonemai deveiopment and growth of P.commune were determined.The results are:1) BM medium is optimal for its protonemal growth;2) the optimum light intensity and temperature were 200μEm^-2s^-1and 25℃,respectively;3) 2,4-D significantly facilitated the protonemal growth;but 6-BAP inhibited the protonemal growth and promoted the induction of shoots 5) the combination of the osmotic stress and some secondary compounds of aqueous extract from its own gametophytes posed some influences to its prootonemal growth;6) the protonemal growth was significantly affected by carbon and nitrogen concentrations.The media with a ratio of 10:0.2(sucrose:ammonium nitrate) was suitable for the growth of the protonemata,but the media with a ratio of below 10:0.2(sucrose: ammonium nitrate) or those without addition of nitrogen have inhibitory,effects on the growth of the protonemata.5.The effects of nitrogen source,nitrogen and carbon concentrations on the gametophyte induction from the explant of P.commune were investigated in vitro.We found that nitrogen source had no significant effects on the formation of the gametophytes from the explants,the gametophyte number ranging from 4.3±0.3 to 4.9±0.3 per explant.Carbon(sucrose) concentration(0 gl^-1,10 gl^-1,40 gl^-1),nitrogen(ammonium nitrate) concentration(0 gl^-1,0.2 gl^-1,0.4 gl^-1) and their combination had significant effects on the formation of gametophytes of P.commune,mean gametophyte number per explant ranging from 1.3±0.2 to 7.0±0.5. The induction of gametophytes which is improved by situablely high nitrogen concentration, was inhibited by the deficienty of nitrogen.The inhibition of high sucrose concentration on induction of gametophytes was more significant when in the media with low nitrogen concentration.