Study On Tissue Culture And Acclimation Of Endangered Mongolian Medicine Plant Gymnadenia Conopsea

Due to its remarkable curative effect and diverse uses,the demand of Gymnadenia conopsea is increasing day by day.,However,due to long-term blind excavation and its self-reproductive difficulties,the wild Gymnadenia conopsea resources are nearly exhausted.In this paper,the methods of plant resources were used to investigate the resources of Gymnadenia conopsea in Inner Mongolia to master the distribution and the reserves of Gymnadenia conopsea resources,and domesticate and cultivate them,observe their morphological characteristics,record the phenological period and make a comparative study with wild Gymnadenia conopsea.the experiment tissue culture provides a reference for the establishment of tissue culture and rapid propagation system of Gymnadenia conopsea.The specific results are as follows :By applying the method of plant resource science from 2021 to 2022,a total of 7distribution areas of Gymnadenia conopsea resources in Inner Mongolia were investigated..and the content of soil nutrient elements was detected in some areas.They are Inner Mongolia Gogestai Hanwula National Nature Reserve(GG),Inner Mongolia Saihanwula National Nature Reserve(SH),Bayanchagan town of Hexhigten League(BY),Denglonghezi Grassland of Ongnuud county(DL),Inner Mongolia Teginhan Mountain National Nature Reserve(TJ),Baogd Mountain Forest Farm of East Wuzhumqin county of Xilingol League(BG)and Amgolang Town of Xinbargo Left League of Hulun Buir City(AM).The average number of Gymnadenia conopsea in GG and BG was the largest in 7 regions,and the average number of Gymnadenia conopsea in DL and AM was the least.The dominant companion species in GG area was Koenigia divaricata,the dominant companion species in SH area was Hypochaeris ciliata,the dominant companion species in BY area was Veronicastrum tubiflorum,the dominant companion species in DL area was Ligularia mongolica,and the dominant companion species in TJ area was Ranunculus japonicus.Scutellaria scordifolia was the dominant companion species in BG,and Echinochloa crus-galli was the dominant companion species in AM.The average abundance of BG is the most,GG,SH,BY and TJ are more,DL and AM are the least.The average coverage of GG and BG areas is the largest,SH,BY and TJ areas are larger,DL and AM areas are smaller.In this study,a comprehensive investigation of the distribution of Gymnadenia conopsea in Inner Mongolia was carried out,and the distribution and resources of Gymnadenia conopsea in Inner Mongolia were preliminarily understood,as well as the differences in the growth environment between different regions,including the detection of organic matter in soil.The morphological characteristics of Gymnadenia conopsea in different regions are different due to the different organic matter content contained in the soil.On the basis of the previous field investigation,the morphological characteristics and phenological period of Gymnadenia conopsea(GG area)and transplanted Gymnadenia conopsea were compared,and the seedling stage,elongation stage,leaf expansion stage,flowering stage,fruiting stage,dead stage,germination stage and overwintering stage of the two places were observed and recorded.The results showed that the tuber,plant height,leaf length,inflorescence and floret number of transplanted Gymnadenia conopsea were smaller than those of wild,and the whole growth cycle(seedling stage-dead stage)was about 30 days shorter than that of wild Gymnadenia conopsea.From the results,the growth of transplanted wild was significantly lower than that of wild Gymnadenia conopsea due to the different climate environment and soil.This study provides a strong scientific basis for the study of alternative habitats and artificial cultivation of Gymnadenia conopsea in the future.First,using MS and 1/2MS medium as the basic medium,NAA,2,4-D,6-BA,ZT,KT,adenine were added separately or in combination to prepare the medium,then immature seeds were selected for culture.As a result,no germination was found.Secondly,by means of plant tissue culture,MS medium was used as the base medium The MS medium was used as the basic medium to add different concentrations of plant growth regulators such as 2,4-D,6-BA and NAA to try to obtain the induction conditions of Gymnadenia conopsea organogenesis.In the screening experiment of explants,all the stem segments and leaves were browned after 2-3 days of inoculation,and the browning rate was 100 %.After 60 days of subculture on MS + NAA0.25 mg / L + Vc2 g / m L medium,adventitious buds were grown from the roots.then the adventitious buds were transferred to rooting medium,and NAA 0.1,0.5,1.0 mg/L was added to MS and 1/2MS respectively.The optimal effect of 1/2MS + 0.5 mg/L NAA was found..The experimental results showed that it was feasible to establish the rapid propagation system of Gymnadenia conopsea by tissue culture,and the results of this study will provide a strong reference for the establishing the rapid propagation system of Gymnadenia conopsea.