In Vitro Culture And Establishment Of The Regeneration System In Melastoma Affine

Melastoma affine is a forever green shrub,which belongs to Melastoma Gene,Melastoma Family.It grows in Fujian,Guangdong,Yunnan,Guizhou,Taiwan etal.It has good adaptation,high appreciation value.The fruit can be used as food and the whole plant can be used as medicinal herbs.It is a wild plant which has various value.This experiment chose Melastoma affine's seeds which grew in natural state as basic material,researching about every steps of seeds axenic bourgeon,proliferation,root induction,transplantion etc.At the same time,the experiment took the leaves of Melastoma affine as explant for the induction of calli and calli proliferation followed. The main results were described as follows:The axenic system of Melastoma affine was establishment.Compared the different disinfection methods used on the Melastoma affine's seeds.The results showed that the method which 70%alcohol disinfected 30 seconds first,axenic water washed 2 times followed,then deeped in 0.1%HgCl₂,and axenic water washed 5 times last was the best.It can control the pollution rate below 15%.Next,the seeds were inoculated to the medium of MS and B₅.Recorded 30d later,it kept vigorous growth of seeds.The proliferation ofMelastoma affine axenic seedlings.Considered the influence of different plant growth hormones on the proliferation,which orthogonal design,screened out the medium was MS supplemented with 6-BA1.0mg/L,sugar 20g/L,agar 6.0g/L and the proliferation rate could reach 3.476 by 30d later.In the second repetitious cultivation,the medium was MS supplemented KT0.5mg/L,BA1.0mg/L,agar 6.0g/L and sugar 20g/L,and the proliferation rate could reach 4.118 by 30d later.In the third and more times proliferation,the medium was MS+ agar 6.0g/L +sugar 20mg/L +KT0.2mg/L+IBA3.0mg/L.Rootage of Melastoma affine.Thinked about the influence of different kind and concontraction hormones on the axenic seedlings root rate,screened the medium was MS supplemented with NAA0.2mg/L,IBA0.2mg/L,agar 6.0g/L and sugar 20mg/L.The records showed that this medium was superiority than others on rooting rate,rooting length and average rooting numbers.The transplantion of Melastoma affine.Analysis the results of the survival rate of the different Transplanted stroma of M.affine,it showed that the sand was best and the survival rate can reach 85%.The explant disinfection and calli induction.Collect the leaves of Melastoma affine which grew in wild state,70%alcohol disinfected 20 seconds,axenic water washed 2 times,deeped in 0.1%HgCl₂ for 8 minutes,axenic water washed 5 times.The pollution rate could be controlled below in 20%.Explant calli induction medium was MS supplemented with BA 1.0mg/L,NAA0.1mg/L,sugar 20g/L and agar 6.0g/L,induction rate was 46.7%.The influence to calli of different light intensity.When the light was 0lx,the calli was easy to be browning.When the light was 2400lx,the calli grew well.