Establishment Of Liquid Cuture Method For Anthurium Somatic Embryo And Some Physilogical Characteristics During Somatic Embryogenesis

Anthurium(Anthurium andraeanum Lind.) is a popular ornamental plant of Araceae. At present,it's seedling production is mainly through organogenesis pathway which has many disadvantages such as long time callus induction,degeneration and mutation, contrastly,somatic embryogenesis can overcome these disadvantages.If large scale somatic embryo liquid culture system was established,it will be a valid method for automatic anthurium seedling production.Based on the present research,this study investigated the main facts of anthurium embryonic callus liquid culture and the related physiological characteristics were also studied during embryo development,maturation,and germination.The results are as follows:1.Using embryonic callus of cv 'Amigo' as material,the effects of different basal culture medium,2,4-D and sucrose,flask volume,organic additive,inoculation weight on the proliferation of embryonic callus in liquid culture were investigated,the result shows that the best conditions for embryonic callus proliferation are 1/2MS + 2,4-D 1.0mg.L^-1 + KT 0～0.5 mg.L^-1 + sucrose 3%,pH 5.5,vibrate speed 100 rpm.Dark culture for 30d, proliferation coefficient can up to 4～5;different basal medium and flask have no notable difference on proliferation coefficient;the weight variation curve of different inoculate weight treatments is about "S" during the 60d continuous liquid culture.The embryonic callus bengin to develop along with the gradul reduction of sucrose,plant growth regulator and nutrition concentration in the liquid culture medium.For 1g embryonic callus,the best inoculation liquid medium volume is 75ml and subculture cycle is 25d.2.Using embryonic callus of cv 'Amigo' as material,the effect of different concentration of NH₄NO₃,nutrition and sucrose,organic additive on somatic embryo development and the effect of different culture method and plant growth regulator concentration on anthurium somatic embryo germination are investigated,result indicates that low concentration of NH₄NO₃ in the medium is benefit for somatic emryo development,concentration of nutrition and sucrose is main factor for somatic embryo development,low concentration of them in medium can make somatic embryo development fast and well.The optimal medium for the development of somatic embryo in this experiment is 1/8MS + sucrose 1%, pH 5.5.The developed somatic embryo number is about 75 per 0.5g culture callus after 30d culture.1000mg/L Myoinositol or 500mg/L Casein Hydrolysate can improve the quality of the somatic embryo.Best germination medium is 1/SMS + sucrose 1%.Put the Anthurium somatic embryos on a filter paper with 4 layers of absorbent paper which saturated with liquid medium under it,afer 30d culture,the germination rate is about 50%.The addition of plant growth,regulators(GA₃,KT,BA ) inhibit germination.3.Using cv 'Amigo' as material,some physiological and biochemical characteristics of different developmental stages of somatic embryogenesis were studied.The activity of POD and CAT maintains at a high level at the stage of embryonic callus,and SOD activity presents a high level at the later stage of somatic embryogenesis.The content of soluble protein is the highest at the stage of callus stage,and the content of starch is down first and up later but the content of soluble sugar up first and down later contrarily during the somatic embryogenesis.SDS-PAGE analysis showed that there are some specific proteins and high expression proteins at the stage of embryonic callus.It concluded that the embryonic callus formation stage is important for the regulation of somatic embryogenesis in Anthurium andraeanum Lind.