Study On Tissue Culture Of Lavandula Angustifolia Mill.

Lavandula angustifolia M.is a perennial subshrubs,it is a very precious,high ornamental value of natural spices,as an aromatic plant has important medicinal value.Lavandula angustifolia M.can be used for many purposes,its leaves can be used as a seasoning,external petals can be used as spices.The extracted essential oil is not only effective in anti-cancer,anti-tumor,reducing blood pressure,improving sleep and other aspects,but also use as raw material for making soap and cosmetics.At present,most of the propagation methods of Lavandula angustifolia M.are asexual propagation and seed propagation,with low reproduction rate.The use of plant tissue culture technology can quickly obtain a large number of sterile plantlets,cultivate excellent varieties and improve the amount of secondary metabolites.In order to establish a complete regeneration system of tissue culture of Lavandula angustifolia M.and provide technical support for mass production after Lavandula angustifolia M.and the extraction of essential oils provide raw materials。In this experiment,the stem segments of Lavandula angustifolia M.were selected as explants to establish the rapid propagation system of Lavandula angustifolia M.tissue culture.The research content and main results of this paper are as follows:(1)Headspace solid phase microextraction(HS-SPME)combined with GC-MS was used to detect the chemical composition and content of essential oil from native plants of Lavandula angustifolia M.HS-SPME extraction conditions for:60 min extraction time,extraction temperature 70℃,the fiber head for DVB/CAR/PDMS,parsing time 5 min.The main chemical constituents of 38 lavender essential oils were determined,among which camphor and borneol were the highest with an average content of 10.53%and 27.27%respectively,followed by linalyl acetate and linalyl alcohol with an average content of 5.95%and 8.28%respectively.(2)To establish the method of sterilization on the surface of Lavandula angustifolia M.explants and the induction of Lavandula angustifolia M.callusThe optimal disinfection time of explant is 75%alcohol sterilization for 25s and 0.1%HgCl2 for 8 min,and the explant survival rate can reach 78%.The medium for callus induction:MS+0.2mg/LTDZ+0.2mg/LNAA,the induction rate is 96%,and the callus is yellow-green and loose.(3)Establishment of adventitious bud system of Lavandula angustifolia M.callus differentiationThe medium for callus proliferation:MS+0.3mg/LTDZ+0.05-0.2mg/LNAA,and the callus have been transformed into green callus,and there is obvious dark green particles at the edge of the callus.The medium for callus differentiation of adventitious buds:MS+0.5mg/L6-BA+0.2mg/LNAA,the induction rate of adventitious buds is 90%,the length of adventitious buds is up to 0.6cm on average,and the number of adventitious buds is up to 8.0 on average.(4)To establish the reverse optimization system of hyperhydricity shoots and induced rootThe combination of 0.5mg/L 6-BA and 0.2mg/L NAA provided the growth power for the proliferation of adventitious buds.In order to overcome the difficulty in survival of hyperhydricity shoots,In this experiment,the effect of adjusting the composition of MS medium,the concentration of agar,the number of days of succession and adding PVP on the reversal of hyperhydricity shoots were studied,then this experiment mothod(RSM)was optimized by using response surface to find the optimal experimental conditions,hyperhydricity shoots a reversal of the four factor 3 levels orthogonal experiment design was carried out,the hyperhydricity reversal rate as index,finally it was concluded that the basic culture medium for 1/2 MS culture medium,ager concentration was 9.70g/L,transgenerational days for 21d,PVP concentration wasl.Og/L,adventitious bud hyperhydricity reversal rate was 66%.The rooting medium was 1/2MS+IBA(0.5mg/L),tissue culture shoots roots slowly,the root of slender,the seedling refining matrix was perlite:vermiculite:sand=1:1:2,and the plants in good growth state were moved to the greenhouse for culture。In this paper,through the disinfection of Lavandula angustifolia M.explants,the induction of callus,the proliferation and differentiation of callus,the reversal of hyperhydricity shoots,the rooting of shoots and acclimatization,the successful establishment of rapid propagation system of Lavandula angustifolia M.tissue culture,can provide a large number of plantlets into garden ornamental,provide large-scale reproduction for the production of essential oils,and provide materials and theoretical basis for the improvement of Lavandula angustifolia M varieties.