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蝴蝶兰的微繁殖

Micropropagation of Phalaenopsis Blume.

作者信息

Sinha Pinaki, Alam M Firoz, Hakim M Lokman

机构信息

Department of Botany, Rajshahi University, Rajshahi, Bangladesh.

出版信息

Methods Mol Biol. 2010;589:77-85. doi: 10.1007/978-1-60327-114-1_8.

Abstract

For high frequency regeneration of Phalaenopsis, young leaf segments are cultured on gelrite-gelled (1/2) MS medium supplemented with 2% sucrose, 2.0 mg/L BA, 0.5 mg/L NAA, 10% coconut water (CW), 2 g/L peptone and 1 g/L activated charcoal. Cultures are incubated at 24 +/- 2 degrees C under fluorescence light 50 micromol/m(2)/s for 16 h photoperiod per day. The PLBs (protocorm like bodies) are induced within 12 weeks of culture and are subcultured to proliferate on the fresh nutrient culture medium for 8 weeks. The PLBs clumps are dissected and cultured on (1/2) MS medium containing 2% sucrose, 10% coconut water (CW), 2 g/L peptone, 150 mg/L L-glutamine and 1 g/L activated charcoal. The PLB sections elongate to form shoots and new PLBs are induced from the base within 8 weeks of culture. For plantlets formation the shoots are cultured on (1/2) MS medium supplemented with 2% sucrose, 10% CW, 2 g/L peptone, 1 g/L activated charcoal and 50 g/L banana pulp. The regenerated plants are acclimatized and cultivated in the nursery, where they bloom within 2 years.

摘要

为实现蝴蝶兰的高频再生,将幼嫩叶片切段接种于添加了2%蔗糖、2.0毫克/升苄氨基腺嘌呤(BA)、0.5毫克/升萘乙酸(NAA)、10%椰汁(CW)、2克/升蛋白胨和1克/升活性炭的凝胶状(1/2)MS培养基上。培养物在24±2摄氏度、光照强度为50微摩尔/(平方米·秒)、每天光照16小时的荧光灯下培养。原球茎状小体(PLBs)在培养12周内诱导形成,并转接至新鲜的营养培养基上增殖8周。将PLBs团块切割后接种于含有2%蔗糖、10%椰汁(CW)、2克/升蛋白胨、150毫克/升L-谷氨酰胺和1克/升活性炭的(1/2)MS培养基上。PLB切段在培养8周内伸长形成芽,并且从基部诱导产生新的PLBs。为了形成完整植株,将芽接种于添加了2%蔗糖、10%椰汁(CW)、2克/升蛋白胨、1克/升活性炭和50克/升香蕉果肉的(1/2)MS培养基上。再生植株经炼苗后在苗圃中栽培,2年内即可开花。

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