Regeneration In Dendrocalamus Asper And Genetic Transformation Of Bamboo

Bamboo genetic and breeding research is relatively weak. In this study, using mature zygoticembryos as explants, we firstly developed an efficient and consistent regeneration system for plantletsof the bamboo species Dendrocalamus asper. On the basis of this and establishing efficientregeneration system of D.hamiltonii in our group, we carry out the transgenic research. The bamboogenetic transformation system was preliminary builted, which lay a good foundation for geneticimprovement of bamboo. The main results showed as follows:1. D.asper regeneration system establishment(1) The optimal callus induction medium: Murashige and Skoog (MS) supplemented with3mg/L2,4-dichlorophenoxyacetic acid (2,4-D),500mg/L proline (Pro),500mg/L glutamine (Gln),500mg/L casein hydrolysate (CH),30g/L sucrose and agar8g/L (typeA). with the frequency ofcallus and granular and compact callus to more than85%and about30%respectively.(2) The optimal maintenance culture medium: MS medium supplemented with0.5mg/L2,4-D,500mg/L Pro,500mg/L Gln,500mg/L CH,30g/L sucrose and agar8g/L (typeA).(3) Optimal callus differentiation and subsequent shoot growth were obtained in MS mediumsupplemented with2mg·L-1benzyladenine (BA),1mg·L-1KT, and0.5mg·L-1naphthaleneacetic acid(NAA),30g/L sucrose and8g/L Type A agar, with the highest differentiation rate of about45%. Inthe experiment, when callus subculture times was2times (6weeks), callus differentiation rate washighest. when callus subculture times was4or5times, the callus differentiation rate was significantlyreduced, and it was easy to differentiate the albino.(4) The optimal rooting medium was1/2MS medium supplemented with3mg/L IBA,30g/Lsucrose and0.3%gelrite. Rooting rate reached90%, the average number of root per plant was6-7, upto a maximum of approximately15, the induced average length of about3.39cm.(5) Rooted plantlets were transferred to artificial mixture (peat: vermiculite: perlite=1:1:1) with95%shoot survival.2. Bamboo transgenic researchUsing callus of D. hamiltonii as receptors, pCAMBIA1301plasmids related DNA fragments weretransformed into callus of D. hamiltonii by Agrobacterium-mediated transformation, ten greenplantlets of D.hamiltonii were obtained with the conversion rate of35.7%. By PCR and electrophoresis products were sequenced, the results demonstrate that HPT gene had beentransferred to the bamboo genome.In transgene reseachs of D. asper, calli were cultured in screening medium supplemented withHyggromycin, after six weeks they were transferred to differentiation medium, two albinos wereobtained successfully and green spots appeared in a callus, which are yet to be detected.