title: [Protoplasts isolation, purification and plant regeneration of Pinellia cordata].
abstract: The main factors which affected the isolation, purification and cultivation of Pinellia cordata protoplasts from leaves were studied. The results indicated that the optimum enzyme solution for P. cordata leaves was 13% CPW + 1.0% Cellulose +0.1% Pectolase, at pH 6.0, temperature (25-28 degrees C ) for 4 h. The sucrose density gradient centrifugation was adopted to purificate the protoplasts collected, when 25% sucrose was used as mediator, centrifugating at 500 rpm for 10 min. When the protoplasts were shallow liquid and liquid-solid double layer cultured on the medium of MS + 0.5 mg x L(-1) 6-BA + 0.25 mg x L(-1) NAA + 13% mannitol at the density of 2.5 x 104 protoplasts/mL, or fed and nursed cultured at the density of 100-500 protoplasts/mL, cell division could be observed for 3 days; granular calli appeared for 30 days. Calli was proliferated on the medium of MS + 0.5 mg x L(-1) 6-BA + 0.25 mg x L(-1) NAA solidified by 0.55% agar, and differentiated and regenerated after 5-6 months. Plant generation of P. cordata is successfully established.
authors.forename: Xian
authors.initials: X
authors.lastname: Yang
authors.forename: Dan-Dan
authors.initials: DD
authors.lastname: Ma
authors.forename: Fu-Sheng
authors.initials: FS
authors.lastname: Jiang
authors.forename: Ni-Pi
authors.initials: NP
authors.lastname: Chen
authors.forename: Bin
authors.initials: B
authors.lastname: Ding
authors.forename: Li-Xia
authors.initials: LX
authors.lastname: Jin
authors.forename: Chao-Dong
authors.initials: CD
authors.lastname: Qian
authors.forename: Zhi-Shan
authors.initials: ZS
authors.lastname: Ding
mesh_terms: D002469:Cell Separation / Q000379:methods*; D003470:Culture Media; D030021:Pinellia / Q000502:physiology*; D011523:Protoplasts / Q000502:physiology*; D012038:Regeneration
chemical_list: D003470:Culture Media