香蕉胚性細胞懸浮培養的維持、增生、體胚發生與小植株建立

‘北蕉’、‘Robusta’、‘三尺蕉’、‘Cavendish BF’(於1993年命名為台蕉二號)、‘粉蕉’等由雄花序衍生之胚性細胞之懸浮培養細胞團，以60目以下之細胞團稀釋30倍，取1 c.c散播於含4.5 mL SH修正培養基(SH鹽類加上0.2 mg•L-1 NAA、0.2 mg•L-1 2ip、0.1 mg•L-1 kinetin、0.05 mg•L-1 zeatin)及棉花濾紙墊之9 cm培養皿中培養，其體胚正常度均依接種細胞團之主要表現型態而定。未成結構體的單或雙聚細胞團於撒種後，只能長成癒合組織，聚合或游離之多胞結構體，於撒種後可以形成叢聚或單獨體胚，數量多但不易轉白成熟；而游離或聚集的團胞、球狀體、橢圓體、平頭體(初期)均可於撒播後形成正常之體胚，而且植株轉換率高；平頭體(中晚期)於撒播後會形成具有明顯生長點突起的體胚，植株轉換易生成矮胖植株；平頭伸長體、初期暗化伸長體、暗化伸長體於撒播後依次分別形成具有漏斗狀的畸形體胚、具有各種小胚聚生的根狀體、梨形畸形胚；梨形暗化體Ⅰ、Ⅱ、Ⅲ於撒播後形成褐色細胞團或癒合組織化的細胞團；重組體則於撒播後形成部分小胚叢生的癒合組織。另懸浮細胞長期繼代培養基用硝酸態氮與銨態氮比為1:1或1:2時，可以使懸浮細胞長期維持在非極化生長之胚性細胞群或小細胞群的狀況，而2:1時則可誘使細胞群極化生長朝向具球狀體細胞結構的分化，而有利平板培養產生正常体胚及小植株的建立。

Embryogenic cell suspension cultures derived from male inflorescences of cultivars including 'Bei Jiao', 'Robusta', 'Sanchi Jiao', 'Cavendish BF' (renamed as Taijiao No. 2 in 1993), 'Fen Jiao' and others were used. Cell clusters smaller than 60 mesh were diluted 30-fold, and 1 mL of the diluted suspension was spread onto 9 cm Petri dishes containing 4.5 mL of modified SH medium (SH salts supplemented with 0.2 mg·L⁻¹ NAA, 0.2 mg·L⁻¹ 2ip, 0.1 mg·L⁻¹ kinetin, 0.05 mg·L⁻¹ zeatin) and a cotton filter paper pad for culture. The normality of somatic embryos was determined by the main phenotypic traits of the inoculated cell clusters. Single or double cell clusters without distinct structures could only develop into callus after inoculation. Aggregated or free multicellular structures could form clustered or individual somatic embryos, which were abundant but difficult to turn white and mature. Free or aggregated cell clusters, globular bodies, ellipsoidal bodies, and flat-top bodies (early stage) could all form normal somatic embryos after inoculation, with a high plant conversion rate. Flat-top bodies (middle and late stages) developed into somatic embryos with distinct shoot apical meristem protrusions after inoculation, and plant conversion tended to produce stunted, stout plantlets. Flat-top elongated bodies, early-stage darkening elongated bodies, and darkening elongated bodies sequentially formed funnel-shaped abnormal somatic embryos, rhizomes with various clustered small embryos, and pear-shaped abnormal embryos respectively after inoculation. Pear-shaped darkening bodies I, II, and III formed brown cell clusters or callus-like cell clusters after inoculation. Recombinant bodies developed into callus tissues with partial clustered small embryos after inoculation. Additionally, when the long-term subculture medium for suspension cells had a nitrate nitrogen to ammonium nitrogen ratio of 1:1 or 1:2, the suspension cells could be maintained for long periods as non-polarized embryogenic cell clusters or small cell clusters; a ratio of 2:1 induced polarized growth of cell clusters towards differentiation into globular cell structures, which facilitated the generation of normal somatic embryos and the establishment of plantlets via plate culture.