Aseptic Germination Of Interspecific Hybridrization Seeds And Its Physiological And Biochemical Characteristics Of Cymbidium Hybrida×C. Sinese And C. Faberi

Orchid is one of the most popular flower in the world.In China there have been many Cymbidium species,such as the C.goeringii,C.faberi,C.ensifolium,C.sinense.Cymbidium is a genus belonging to Orchidaceae family.The traditional Chinese orchids have long been valued for their good genes.Many orchid species and hybrids have been successfully unsing to breed new varieties(C.hybrida).And we got a number of C.hybrida that have the characteristics of Chinese orchids.But now hybridization is still rely on the breeding of wild plants from the choice of traditional practices,the development of new varieties of orchids could not get the attention,There were even less cultivation of the economic value species.With the development of orchid industry,broad cross breeding will inevitably arise. Through inter-specific and specific hybridization we can enhance the quality of flower, extended flowering period,and so on.From these we may get a typical characteristic of the native species,and also has the advantage of Cymbidium species.Cross-breeding of orchids,the important position of National Cymbidium hybrization,Several investigators have reported successful cross breeding in and out of country,and few reports on the multi- species hybridization embryo culture and its hybrid technology.In the present paper we describe an aseptic germination protocol of seeds from interspecific hybridization between Cymbidium hybrida×C.sinense and C.faberi was established technique of proliferation and plant regeneration,and research on the development stage of physiological and biochemical and morphogenesis of hybrid cymbidium were also conducted,the results are as follows:1.An in vitro plant regeneration protocol of Cymbidium hybrida×C.sinense and C. faberi using hybrid seed as explants,The results were as follows:seed germination has undergo seed - protocorm- rhizome - seedling,germination rate is related to the maturity of the seed,we found that 240d after pollination of embryo has good germination capacity, The suitable medium for hybrid seed germination was half strength MS basal medium supplemented with 0.5 mg·L¹6-BA and 2.0 mg·L¹Î±-naphthalene acetic acid(NAA).And we also found that NAA 1.0 mg·L^-1combinate with 6-BA 1.0 mg·L^-1can be effective in promoting Rhizome division,70 d is the best time for rhizome subculture. 2.Taking PLBs and rhizomes germinated from 240 days after pollination seed as proliferation and differentiation testing material,the effects of differentiation condition on proliferation of PLBs and rhizomes were conducted.The results showed that:1/2MS was the suitable basal medium for PLBs proliferation and differentiation;The optimal medium for proliferation of PLBs was on basal medium supplemented with 0.5 mg·L¹ 6-BA and 2.0 mg·L¹ NAA,the development and plantlet regeneration of PLBs can be highly enhanced on medium containing 1.0 mg·L¹ 6-BA and 0.5～1.0 mg·L¹ NAA.the addition of 100ml·l^-1 coconut milk promoted PLBs proliferation rate,On the other hand,1/2MS was the suitable for rhizomes proliferation and differentiation;the basal meidum containing 0.5 mg·L^-1 6-BA and 1.0 mg·L^-1 NAA proliferation,and subsequently more than 80%of the apex of rhizomes developed into shoots within 70 days culture on the medium containing 1.0 mg·L^-1BA and 0.5 mg·L^-1 NAA.the addition of 100ml·l^-1 coconut milk promoted rhizomes proliferation rate andl.0g·L¹ activated charcoal improved on the rhizomes proliferation.3.Histological and morphogenesis characteristics studied PLBs and rhizomes formation and development,The results showed that,PLBs and rhizomes included epidermis,cortex and vascular cylinder.PLBs stage contain more starch than rhizomes. The proliferation of PLBs are developed from the apical meristem of the apical meristem which can grow into seedlings,rhizomes' epidemal and cortical cells could be divided and formed into lateral meristem under hormone treatment.Then apical and lateral meristem can develop into seedlings.PLBs and rhizomes were typical vegetative proliferation organ.4.Changes of soluble sugar,soluble protein,starch and the activity of three antioxidant enzymes(SOD,POD and CAT) correlated with the process of development of hybrid of Cymbidium.The results showed that:sugar and starch content is closely related in two ways they all showed decrease changes,The content of soluble sugar and starch during two ways were shown to decrease then rising,During the whole period,the accumulation in soluble protein and sugar,have a close relationship with the development of PLBs and rhizomes. It was shown that SOD,POD and CAT activities were closely related to the development of hybrid of Cymbidium.Two ways in the seedling stage of the physiological indicators are not differences.By SDS-PAGE analysis of protein components found that hybrid development and protein expression has a close relationship.