Studies on cell suspension culture and somatic embryogenesisi of Musa formosana
Date Issued
2009
Date
2009
Author(s)
Liu, Shi-Hua
Abstract
Summaryhe young male inflorescence of Taiwan native diploid banana (Musa formosana) was used as explant, and cultured in modified MS medium supplemented with complex auxin including IAA 1 mg/L, NAA 1 mg/L, and 2,4-D 1 mg/L and Picloram 3 mg/L suitable for callus formation. The top male hand portions ranged from the 13th to 17th sections were capable to give rise 64-82% callus formation on the medium as above. The zygote embryos were aptly to generate embryogenic callus on MS medium added with NAA 1 mg/L, IAA 1 mg/L combined Picloram 1 mg/L or Dicamba 1 mg/L in 2-3 months after inoculation. In further, the callus proliferation could be subcultured on the above medium but supplemented Picloram or Dicamba as the sole auxin. The male inflorescence-derived callus was subcultured in liquid TB5 medium ( Ma, 1988), and obtained homogenous cell population about 2 month after suspension culture. he extracellular pH level in TB5 medium of the cell suspension culture was varied in the range pH 4.45±0.37, and also concordantly associated with growth phase change. The addition of MES 10 g/L could upgrade the extracellular pH in the stable level 4.51±0.17, and also induce the preembryogenic cells destine to polar growth depicted by the asymmetric division. Controlling extracellular pH at 5.7 and culture in SH3、SH3 with MES led up to the polar growth. The cells culture in SH3 with MES were polarized earlier, and remain extracellular pH over 5.46. After polarization treatment for 7 days, the embryogenic cells could formed proembryo and globoids. Used PIPES with SH3 medium could also be induced to polarization and formed globoids with complete protoderm Acidic treatment (pH 4.0-4.5) promoted globoids to disunite small mass and release single cells. After acidic treatment for 7 and 14 days, the bicellular trended to do symmetric division. Controlling extracellular pH at 5.7 and culture in SH3、SH3 with MES led up to the polar growth. The cells culture in SH3 with MES were polarized earlier, and kept extracellular pH over 5.0. After polarization treatment for 7 days, the embryogenic cells could formed proembryo and globoids. Used PIPES with SH3 medium could also be induced to polarization and formed globoids with complete protoderm Before plating, the suspension prembryogenic cells were pretreated on TB5, SH3 medium with or without was consistent 10 g/L MES. The SH3 pretreated cells regenerated higher quantity of somatic embryos on plating medium than those on TB5, TB5+MES and SH3+MES pretreated. The size of suspension cell cluster was separated with <60 or 30-60 meshes and were diluted 1/30 c.c PCV proceeding to plat on 8ml SH3 medium with filter paper and cotton as the culture bridge, could development normal somatic embryos than agar, gelrite and 4 ml SH3 addition. Somatic embryo inoculated on 1/2 MS medium supplement with 10 mg/L BA induced 69% of shooting. The rooting rate did not increase, when medium supplement NAA. However, the shooting rate increase of plantlets on medium complex added NAA with BA. The increase of shooting rate and leave length in medium GA, but the leaves had exceptional discoloration and elongation.
Subjects
Musa formosana
banana
tissue
somatic embryos
Type
thesis
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