4个优良石榴品种叶片高频再生体系的建立

为提高石榴叶片再生率，增加再生芽数量及缩短再生周期，以‘泰山红’‘、泰山三白甜’‘、皮亚曼’‘、牡丹’4个石榴品种盆栽苗和无菌苗叶片为试材，研究培养基类型、生长调节剂配比、叶片来源和培养步骤对不定芽再生的影响。结果表明：MS是叶片再生最适培养基类型；无菌苗叶片比盆栽苗叶片更易再生不定芽；BA3.0mg/L+KT1.0mg/L+IBA0.3mg/L是叶片愈伤组织诱导、分化和不定芽形成过程中生长调节剂最佳配比；先在液体培养基上培养3~5天，再转至固体培养基比仅在固体培养基上效果更好，叶片再生率最高达98.47%，出芽数达7.51个。叶片不定芽的最佳单芽培养基和增殖培养基分别为B5+BA0.6mg/L+IBA0.5mg/L+椰汁100mL/L+PVP2.0g/L+GA31.0mg/L和B5+BA0.8mg/L+IBA0.5mg/L+椰汁100mL/L+PVP2.0g/L，增殖系数为5.56。适宜的生根培养基为B5+IBA1.0mg/L+椰汁100mL/L，生根率为89.63%。由此建立了石榴叶片高频再生体系。

Establishment of Adventitious Shoot Regeneration System from Leaves of 4 Pomegranate Cultivars in Vitro

Studies were carried out in order to improve regeneration rate of pomegranate leaves, increase the number of adventitious buds and reduce the regeneration cycle. The tested materials were the leaves of potted seedlings and aseptic seedlings from 4 pomegranate cultivars‘Taishanhong’ ,‘Taishansanbaitian’ ,‘Pyaman’ and‘Peony’ . The effects of medium type, mixture ratio of growth regulator, source of leaves, and training steps on adventitious bud regeneration were investigated. The results showed that MS medium was the most suitable for leaves regeneration, besides, leaves from aseptic seedlings produced shoot easier than that from potted seedlings. BA 3.0 mg/L+ KT 1.0 mg/L+ IBA 0.3 mg/L was the best mixture ratio of growth regulator in the process of leaves callus induction and differentiation and formation of adventitious buds. The leaves cultured in liquid medium for 3-5 days at first and then transferred to solid medium showed stronger regeneration ability than that cultured in the solid medium only. In this way, the ratio of leaf regeneration reached up to 98.47%, with an average of 7.51 adventitious shoots on each leaf. For leaves adventitious buds, the best single bud culture medium and proliferation medium were B5+ BA 0.6 mg/L+ IBA 0.5 mg/L + CW(coconut water)100 mL/L+ PVP 2.0 g/L+ GA3 1.0 mg/L and B5+ BA 0.8 mg/L+ IBA 0.5 mg/L + CW 100 mL/L+ PVP 2.0 g/L. The optimum rooting medium was B5+ IBA 1.0 mg/L+ CW 100 mL/L, of which the rooting rate was 89.63%. So the high frequency regeneration system from pomegranate leaves was established.