Trollius Haploid Induction Culture

Trollius chinensis Bunge is a kind of traditional Chinese medicine with high ornamental value inbashang area of zhangjiakou.As an ornamental material of urban greening,ornamental material of bashang scenic spot and medicinal plant resources,Trollius chinensis Bunge has been paid much attention to.However,there are fewer varieties of Trollius chinensis Bunge,single color,and poor resistance.In addition to natural distribution,the application area is small.Therefore,how to improve the stress resistance,yield and quality,and breeding new varieties are the main objectives of Trollius chinensis Bunge breeding.Anther culture technique is needed to obtain haploid plants in order to breed homozygous varieties of Trollius chinensis Bunge.Anther culture belongs to organ culture,in which buds developed to a specific stage are plucked,anthers are removed and inoculated into callus inducing medium through aseptic operation to change their original development path and induce embryoids or callus,so as to develop into complete plants.It is the main method of haploid culture because it is easier to operate than other methods.In this study,the anthers of Trollius chinensis Bunge were used as test materials,and the observation of bud development,anther callus induction,callus differentiation into seedlings,and ploidy identification of regenerated plants were systematically studied.In order to further obtain homozygous plants and provide new materials for breeding new varieties of Trollius chinensis Bunge,it provides a theoretical basis for the establishment of a perfect system for the cultivation of Trollius chinensis Bunge haploid.The results show:1.When the Trollius chinensis Bunge bracts are gradually loosened,the anthers are differentiated,and the anthers and flower buds are yellow,most of the microspores are in the single-nuclear edge period,and the transverse diameter is 11.35¹3.35mm;2.It was found that different culture conditions had different effects on callus induction of Anther of Trollius chinensis Bunge.Plant growth regulators play a key role in anther culture,MS+2.0mg·L^-16-BA+1.0 mg·L^-12,4-D is more suitable for callus induction of Trollius chinensis Bunge;pre-treatment of Trollius chinensis Bunge at 4℃for 2 days is the best;0.3 mol·L^-1 mannitol solution immersion for 3 days is the best;0.1mol·L^-1sucrose solution immersion for 60 minutes has the highest induction rate of Trollius chinensis Bunge;basic culture;In the medium screening test,all the five media could induce callus,but MS medium had the best induction effect;there was no significant difference in the induction effect of maltose and sucrose in the carbon source screening,but the induction rate of maltose was slightly higher than sucrose,which was more suitable for callus induction of Trollius chinensis Bunge;the inhibitory effect of vitamin C at the concentration of 50 mg·L^-1 in the medium additives on anther browning of Trollius chinensis Bunge was the most obvious.3.The callus was transferred into adventitious bud differentiation medium supplemented with0.5mg·L^-1NAA and 1.0 mg·L^-1KT,and the differentiation effect of adventitious bud of Trollius chinensis Bunge was the best.4.The adventitious buds were transferred to 1/2MS rooting medium supplemented with 0.8 mg·L^-1IBA.The adventitious buds were successfully rooted,and the rooting rate was 72.5%.5.75 regenerated plants were obtained by anther culture of Trollius chinensis Bunge.The proportion of aneuploidy plants was 4.16%,haploid plants were 4,the proportion was 8.33%,and diploid plants were42,the proportion was 87.5%.