以蝴蝶蘭白葉片誘導擬原球體與植株再生

Plant Regeneration from Protocorm-like Bodies Induced in Etiolated Leaves of Phalaenopsis aphrodite Rchb. f.

本試驗以蝴蝶蘭Phal. aphrodite Rchb. f.之實生苗，經黑暗處理所獲得之白化葉片來誘導擬原球體及植株再生，以建立有效率之無性繁殖體系。葉片分化程度、蔗糖濃度與生長調節劑為影響擬原球體發生之主要因子，白化葉片具有幼嫩細胞，比綠色葉片逆分化能力強且容易產生擬原球體。誘導擬原球體最佳的蔗糖濃度為0.3%，高濃度之蔗糖不利於擬原球體發生與增殖，且易導致培植體褐化。誘導擬原球體發生與增殖最佳的生長調節劑濃度為0.5mg/l BA與0.5mg/l TDZ組合，BA與較高濃度2,4-D組合會抑制擬原球體發生與增殖。將擬原球體繼代於不含生長調節劑之培養基中可以持續產生擬原球體。由葉片誘導之擬原球體移植至再生培養基，照光培養3週，可獲得再生植株，移植至溫室生長良好。 Etiolated leaves of in vitro seedlings from Phalaenopsis. aphrodite Rchb. f. grown in the dark were usedas explants to induce PLBs and subsequent plant regeneration for efficient vegetative propagation. Thedevelopmental stage of leaves, sucrose concentration and plant growth regulator are main factors effectingPLBs formation. The etiolated leaves are much easier to dedifferentiate into PLBs than green leaves. Theoptimal concentration of sugar for PLB formation is 0.3%. High concentration of sucrose was inhibitory tothe formation and proliferation of PLBs since explants turned browning easily. Explants on mediumsupplemented with 0.5mg/l BA and 0.5mg/l TDZ produced a large number and proliferation of PLBs. BAcombined with higher level 2,4-D inhibited PLBs formation and proliferation. PLBs could multiply furtheron growth regulator free medium. Development of normal plantlets was obtained after transferring PLBs ontoregeneration medium for three weeks under light and subsequent transfer to greenhouse.