Establishment Of Tissue Culture And Rapid Propagation System Of Litsea Cubeba Stem Segment With Bud

Litsea cubeba is a traditional essential oil tree species in China.In recent years,it has also been used as an industrial raw material tree species.The essential oil extracted from its fruit contains extremely high citral content and is widely used in medicine,food,cosmetics and other fields.At present,Litsea cubeba is mostly reproduced by conventional methods such as seed seeding and cuttings.Its germination rate is low and the production cycle is long,which cannot meet my country’s excessive seedling germplasm demand.In this study,wild Litsea cubeba stems with buds were used as experimental materials,and orthogonal experimental design,analysis of variance and other methods were used to establish a complete tissue culture system for Litsea cubeba stems with buds.The best disinfection and sterilization method and the hormone combination of bud growth,clumping bud induction,clumping bud elongation induction and rooting.The main findings are as follows:1.In the process of disinfection and sterilization of axillary bud stems of Litsea cubeba spring and autumn shoots,the disinfection and sterilization methods of Litsea cubeba spring and autumn shoots are different.The axillary bud stems of the spring shoots of Litsea cubeba in early April were sterilized with 0.1%mercury liters for 4 min.The survival rate of the buds was 80.83%after 28 days of sterilization.In early September,the axillary bud stems of the autumn shoots of Litsea cubeba were best sterilized with 0.1%mercury liters for 8 min.The survival rate of the buds after 28 days was 79.17%.The sterilization of the spring and autumn shoot explants of Litsea cubeba varies The deepening of the degree of lignification can appropriately extend the sterilization time,and the combination of sodium hypochlorite and multiple disinfectants is not suitable for sterilization of Litsea cubeba2.Litsea cubeba spring and autumn slightly budded stems have different optimal bud growth induction media.Litsea cubeba spring and autumn budded stems are induced with MS as the basic medium,1.8mg·L-1 6-BA+0.5 mg·L-1 IBA The hormone combination of/L IBA has the best induction effect,and the bud germination rate is 88.75%;the stems with buds in the autumn of Litsea cubeba are using MS as the medium,and the optimal hormone combination is 2 mg·L-16-BA+0.5 mg·L-1 IB A,The bud germination rate is 85%.3.The induction of cluster buds of Litsea cubeba takes 1/2 MS as the basic medium,and the optimal combination of hormones is 0.5 mg·L-1 6-BA+0.5 mg·L-1 IBA,and the growth coefficient of cluster buds is 5.558.The growth of cluster buds is better,and the optimal combination of hormones is 0.5mg·L-1 6-BA+0.5mg·L-1 IBA.It is also relatively large.4.The growth and elongation induction of Litsea cubeba cluster buds is mainly affected by gibberellin GA3.Litsea cubeba cluster buds use MS as the basic medium,and the hormone combination 2 mg·L-1 6-BA+0.5 mg·L-1 IBA+0.5 mg·L-1 GA3 has the best induction effect for the growth and elongation of the cluster buds.The proportion of the bud height after induction is greater than 2.5 cm.It was 84.17%,and the stem growth was normal.5.Litsea cubeba is a kind of mixed rooting plant.The hormones IBA and IAA can induce the rooting of Litsea cubeba clump buds,but the effect of IBA is better than IAA.Low concentration of GA3(0.3 mg·L-1)can help Litsea cubeba clump buds.The stem segments break dormancy,thereby promoting rooting.When using 1/2 MS as the basic medium and 0.5 mg·L-1 IBA+1.0 g·L-1 AC to induce the rooting of the clump buds of Litsea cubeba,most of them are rooting from the bark,the rooting rate is 56.67%,and the average number of roots is 4.08.When WPM is used as the basic medium and the hormone combination 0.5 mg·L-1 IBA+0.3 mg·L-1 GA3+1.0 g·L-1 AC induces rooting,most of it is callus rooting,the rooting rate is 71.67%,and the average root number is 3.41.6.In the light culture room,the bottle cap of the tissue culture seedlings of Litsea cubeba was half-opened for 1 d,and then transferred to the artificial climate room for 1 d after fully opened.It grows in a matrix of acid yellow soil:nutrient soil:vermiculite(2:1:1)when transplanted to Hunan,with a maximum survival rate of 41.67%.