柳葉石櫟胚培養及體胚形成

Embryo Culture and Somatic Embryogenesis from Embryo Culture of Pasania dodonaeifolia Hayata

本試驗從浸水營及歸田地區採取柳葉石櫟母株種子，在濕冷層積後去殼，進行組織培養。累積三年結果顯示柳葉石櫟胚培養可以誘發非胚性及胚性組織並有體胚發生，而體胚發生可有直接體胚及間接體胚二種型式。由胚起始培養至明顯體胚發生平均約需50至60日。以歸田1號株為例，發芽率為19.8%，培養後約30日長根，50日萌芽，70天正常枝葉才明顯長成。去殼種子介於4 mm至12mm均可能誘導形成胚性及非胚性癒合組織，小於及等於3 mm則易褐化死亡。堅果採集後，溼冷貯存，可以維持其培養活性約2個月，第3個月後培養污染率高，且組織再生率低。以MS為基本培養基加BA 0.02，0.1，0.2，0.5，1，2 mg／L配合NAA 0，0.01，0.1，1 mg／L，計24處理以複因子設計分析時，發根與胚性癒合組織之發生率在NAA濃度間，可達10%顯著差異，以低濃度較佳；BA處理間差異不顯著。以單因子分析時，發根率、胚性癒合組織及非胚性癒合組織發生率在植物生長調節劑處理間差異顯著。胚性癒合組織與非胚性癒合組織發生比例亦受種子大小影響，種子越大比例明顯愈高。培養之胚體需大（等）於10 mm才有發根潛能，大於12 mm才有正常發芽的機會。母株來源似與胚性癒合組織及體胚誘導率有關，初步以浸水營5號株，誘導率最高可達27.2 %，而浸水營3號株最低，未能形成體胚。 After cold stratification and husking, the seeds of Pasania dodonaeifolia Hayata, which were collected from Chin Shui Ying and the Kuei Tien area, were cultured on MS medium supplemented with growth regulators for three years. The Pasania dodonaeifolia Hayata seeds produced both non-embryogenic callus and embryogenic callus . Embryogenic callus tissue came out for two types: direct embryo and indirect embryo. On the average, it took about 50 to 60 days from the beginning of embryo culture to obvious somatic embryogenesis. Take the ‘Kuei Tien No. 1’ seed for example, the germination rate was 19.8%. Root emerged after the seed was cultured for 30 days, shoots came out after 50 days, and normal leaves and twigs grew out after 70 days. Husked seeds whose diameter was from 4 mm to 12 mm could induce both embryogenic and non-embryogenic callus tissues; the seed whose diameter was smaller than 3 mm would die after being culture. Seeds were viable only for 3 months after cold stratification. When BA and NAA were used in a 4×6 factorial design experiment, the concentration of NAA significantly influenced rooting and embryogenic callus formation, and the most efficacious formulas were 1 mg/L NAA with 0.1 to 1 mg/L BA. Seed diameters and collection areas also significantly influenced the performance of tissue culture.