| At present,the plant configuration in some northern areas is single,which can not meet the public’s demand for beauty,so it is necessary to introduce new garden ornamental plants.Verbena hybrida and Verbena bonariensis of Verbenaceae and Verbena genus are popular because of their high ornamental value and long flowering period.Because the seeds of Verbena hybrida and Verbena bonariensis are small,the seed skin of Verbena hybrida is thick and hard,which is difficult to germinate;The seeds of Verbena bonariensis have a long dormancy period and limited timely breeding,which leads to the difficulty of seedling raising and lack of market supply.In order to improve the utilization rate of their seeds,it is of great significance to establish a tissue culture rapid propagation system with seeds as explants.To provide technical support for genetic engineering breeding of two plants,and establish their regeneration system.In this thesis,the seeds,leaves and stem segments of Verbena hybrida and Verbena bonariensis were used as explants.Using tissue culture methods,different disinfection methods,different growth hormone and concentration at different growth stages,the best treatment was selected,and two tissue culture rapid propagation systems of ornamental plants were established.The test results were statistically and analyzed by data analysis method.The results are as follows:（1）Before the experiment,the seeds of Verbena hybrida and Verbena bonariensis were soaked in distilled water for 12 hours,and the germination effect was better.The results showed that both seeds needed to cut the length of 0.01 cm at both ends to expose the embryo.If the seed coat was not destroyed,the seeds could not emerge.（2）The best disinfection method was 75%alcohol for 30 s and 2%sodium hypochlorite for 17 min,and the survival rate was 61.11%;The optimum medium for seed germination was MS+6-BA 1.5 mg/L+NAA 0.05 mg/L+GA30.5 mg/L,and the induction rate was 46.7%;The optimal proliferation medium was MS+6-BA 2.0mg/l+NAA 0.05 mg/l+GA30.5 mg/l,and the proliferation coefficient was 3.30;The best rooting medium is 1/2 MS+NAA 0.05 mg/L,which has good rooting effect,strong plants and 100%rooting rate;The seedlings of Verbena hybrida were transplanted into the mixed matrix of nutrient soil:Perlite:vermiculite=4∶2∶1,and the survival rate was 82.35%.（3）When the seeds and leaves of aseptic seedlings of Verbena hybrida were used as explants to induce callus,the optimum induction medium and subculture medium of the two materials were MS+6-BA 0.5 mg/L+2,4-D 0.5 mg/L.The leaves of the two materials grew the fastest callus and the highest induction rate,and the structure was dense,The induction rate was 96.75%.The optimum differentiation medium was MS+6-BA 4 mg/L+IBA 0.10 mg/L,The differentiation rate was 27.4%.（4）The best disinfection method was 75%alcohol for 30 s and 2%sodium hypochlorite for 15 min,and the survival rate was 36.70%;The optimum medium for seed germination was MS+6-BA 1.0 mg/L+NAA 0.05 mg/L,and the induction rate was 66.67%;The optimal proliferation medium was MS+6-BA 0.30 mg/L+NAA 0.01mg/L,and the proliferation rate was 3.56;The best rooting medium was MS+6-BA0.10 mg/L+NAA 0.10 mg/L,and the rooting rate was 100%;The seedlings of Verbena bonariensis cultivated by rooting were transplanted into the mixed matrix of nutrient soil:Perlite:vermiculite=4∶2∶1,and the survival rate was 90.18%.（5）When callus was induced from seeds,leaves and stem segments of Verbena bonariensis,the optimum induction medium of the three explants was MS+6-BA 0.5mg/L+2,4-D 0.5 mg/L,and the optimum culture environment was dark culture.The leaves produced the fastest callus and the highest induction rate from the three explants,The induction rate was 100%,but compared with the callus cultured from seeds and stems,the callus from leaves was loose and the callus induced from seeds and stems was dense.The optimum subculture medium was MS+6-BA 0.5 mg/L+NAA 0.05 mg/L;The optimum differentiation medium was MS+6-BA 0.5 mg/L+NAA 0.1 mg/L,The differentiation rate was 37.76%. |
