Establishment Of Regeneration System And Preliminary Studies On Agrobacterium Mediated Genetic Transformation In Platanus Spp.

The Platanus spp. is a notable street tree and an important landscape tree used for improving the urban environment, it has the big crown and large shade, with the perfect effect for shady, dust proof, noise proof and purifying the air; its wood can also be used as a building and furniture. However, when it transfigure the city, its flosses from fruits and pollens spread everywhere, which brought great harm to people’s life. The plane trees produce floating things within flocculent or capillaceous which can cause people to diseases such as asthma, skin allergies. The researchers conducted a great deal of research both in order to solve this problem.The present research tried to induce callus by using platanus seedling as explants; to induce adventitious shoot by using leaf and stalk of asepsis seedlings as explants, established regeneration system of Platanus spp. and studied in detail for conditions that can affect platanus callus and adventitious shoot induction,such as explants selection, leaf age, hormone levels and so on, which laid a foundation for plane trees transgenic system.The platanus seedling is the best explant for callus induction. Immersing seedling in0.1%HgCl2for8min was the best method for disinfecting explants; chossing1/2MS culture medium with0.1mg/L IBA and0.1mg/L NAA for platanus seedling callus induction, obtained high callus induction rate and short period.Stalk of asepsis seedlings is the best explant for adventitious bud induction. Induced effect is best when we select leaf age of30-50days, it need7days dark culture first after putting explants on the culture medium, because this can help to adventitious shoot induction, choosing MS culture medium with0.1mg/L IB A,1.0mg/L6-BA and1.5mg/L KT for adventitious bud induction, adventitious bud induction rate is highest, clump shoot growing best.According to transformation factors of Platanus spp.,we performed the preliminary research on transformation conditions. The whole Platanus spp. seedling was used to be intruded for36hours by (OD600=1.0) agrobacterium, and then joined AS to the liquid culture medium when seedlings co-cultured, GUS transient expression rate reached18%that was the highest. The paltanus callus was used to be intruded for40min by (OD600=0-8) agrobacterium, and then joined AS to the liquid culture medium when calluses got co-culture, GUS transient expression rate reached62%that was the highest. This experiment preliminarily determined the plane tree genetic transformation conditions, it laid the foundation for later obtain transgenic Platanus spp..