| Mentha crispata is a perennial herbaceous plant of the genus Lentiaceae.The plant contains volatile aromatic substances such as esters,alcohols,aldehydes and ketones.The essential oil of Spearmint can be widely used in medicine and food production.The daily chemical industry has a wide range of application values and has a good market prospect.In this paper,the regenerative system of Fragaria sylvestris L.was established by three rapid propagation methods:cutting propagation,axillary bud propagation and callus differentiation.The chemical constituents of volatile oil from the cultured seedlings of Brassica chinensis and callus were studied by GC-MS.analysis.The purpose of this paper is to provide theoretical basis and technical support for the cultivation,development and utilization of Fragaria frondosa.The main research results obtained in this paper are as follows:1.Stem segment propagation:cutting propagation was carried out on the stem segments of Fragaria sylvestris L.,and the results showed that the rooting rate of stem segment cutting was up to 100%after treatment with 50 mg/L NAA for 30 min.2.Axillary bud propagation:The stalks of the stalks of the leaves of the scented leaves are sterilized in different ways.The experimental results show that the best explant sterilization method is:first soak for 5min with 0.5%washing powder solution,then rinse under water for 1～2h,then sterilize with 70%ethanol for 30s,then sterilize with 0.1L for 8min.This sterilization method has the best effect,the pollution rate is 25%,and the germination rate is 71.67%.The proliferation medium of the stems of the leaves of Cymbidium sinensis was MS+agar 7.5g/L+sucrose 30g/L+6-BA0.5mg/L+NAA0.01 mg/L.The growth coefficient of shoots reached 3.1,and the buds sprouted early,the number of effective shoots was large and the growth was robust,and the height of the shoots was 3.5 cm.3.Establishment of callus induction and regeneration system:The best explants induced by callus of Fragaria serrata were tip stem segments.The optimal callus induction medium was MS+TDZ2mg/L agar 6.5g/L+sucrose 30g/L.+25%coconut milk,the callus induction rate reached 99.05%,the optimal callus differentiation proliferation medium was MS+6-BA0.5mg/L+NAA0.02mg/L+agar 6.5g/L+sucrose 30g/L,the more The injury differentiation rate was 58%.The optimal callus culture conditions were as follows:after inoculation,the cells were darkly cultured for 7 days and then irradiated.The culture temperature was(25±1)℃,the light intensity was 2000-2500 lx,the illumination time was 16 h/d,and the darkness was 8 h/d.The relative humidity is 70%to 80%4.Rooting culture and seedling transplanting:The wrinkled leaves are more susceptible to rooting,and the low concentration of hormones can make the wrinkled leaves leave the roots of orchids.1/2MS+agar 7.0g/L+sucrose 20g/L+IBA0.5mg/L is the best medium for rooting of Fragaria frondosa.The rooting rate can reach 100%.The most suitable transplanting substrate for seedling transplanting is fine sand:perlite:humus soil(2:1:1).The survival rate can reach 99%.5.Volatile essential oil extraction and GC-MS analysis:Menthrica crisp cultivated seedlings(A)in greenhouse and tissue cultured seedlings(B)produced from callus culture were used as experimental materials to extract essential oils from plants by steam distillation.(volatile oil)components were analyzed by GC-MS gas chromatography-mass spectrometry.Among the volatile oils extracted from greenhouse seedlings(A),25 chemical components were identified,while the volatile oil extracted from callus seedlings(B)was identified.a chemical composition.There are 19 chemical constituents in the two types of Fragaria.The most important chemical components are fragrant dihydrocarvone,carvone acetate,medlar oil,β-phellandene,a-pinene,β-myrcene,(3-caryophyllene,phytol,d-Limonene and the like. |
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