Establishment Of Rapid Propagation And Genetic Transformation System Of Actinidia Arguta

Kiwiberry(Actinidia arguta)is a large deciduous liana plant of Actinidiaceae,which is a new kind of small berry in recent years.It is not only rich in nutrition,but also has high ornamental value,medicinal value and ecological value.At present,the wild resources of kiwiberry are gradually reduced,and the development of planting industry is slow.With the increase of its consumption,the insufficient production is becoming a serious problem.With the increase of planting area,shortage of cultivated varieties,poor environmental adaptability,and short shelf life are the main constraints on the development of kiwiberry planting industry.In this paper,a variety of rapid propagation methods of kiwiberry were explored,including the methods of obtaining a large number of seedlings in a short period of time from seedling,layering,multiple bud clumps,etc.,and a fast propagation system suitable for kiwiberry was established;the tissue culture regeneration efficiency of a variety of explant materials was compared through experiments,and an efficient tissue culture regeneration system was established;the transformation and the genetic transformation system of kiwiberry was established.The main conclusions are as follows:1.Kiwiberry can be proliferated by three methods.Seedling:the germination rate of seeds was 82%when the seeds were bathed in 45? water for 30 min and soaked in 0.07%gibberellin for 24 h;multiple bud clumps:the young stem segment of kiwiberry was used as explants,the stem proliferation medium was MS basic medium+6-BA(3 mg/L)+NAA(0.1 mg/L),and the proliferation coefficient was 5.45;the rooting rate of shoots was higher when the shoots were soaked in 0.02%Rooting Powder.2.The young stems segment of kiwiberry were used as explants,and were soaked in 75%alcohol for 30 s and 0.2%mercuric chloride for 4 min.the survival rate was 80%.Leaves,petioles,stem segments and cotyledons were used as explants.Taking MS(4.43 g/L)+sucrose(30 g/L)+agar(7 g/L)as the basic medium,the callus induction medium was MS basic medium+ZT(1 mg/L)+NAA(0.1 mg/L),the induction rate of stem segments and cotyledons was 100%,that in leaves was 83.33%,and that in petioles was 63.33%;the medium for callus differentiation was MS basic medium+6-BA(3 mg/L)+NAA(0.1 mg/L),the differentiation rate of stem segments and cotyledons was 96.7%,that in leaves was 10%,and that in petioles was 6.67%;the medium for rooting induction was 1/4 MS(2.21 g/L)+sucrose(10 g/L)+agar(7 g/L),the rooting rate was 96.7%.3.The expression vector pCAMBIA3301-121 was integrated into the genome.The stem segments were immersed in Agrobacterium EHA105(OD600=0.6,containing AS 20 mg/L)for 20 min,then cultured for 4 days(20±2?,dark),washed with MS liquid medium(containing Carb,500 mg/L),inoculated to medium containing Carb(400 mg/L)for callus induction for about 20 days,and then put it into the selection medium for screening,and then change the proliferation medium every 20 days and gradually reduce the concentration of carb until the complete plant is obtained.