84K杨叶片再生系统的建立

选用84K杨的叶片作为外植体,MS作为基本培养基,采用不同浓度组合的BA和NAA对84K杨再生系统进行了研究.结果表明,在MS培养基中单独添加2.0mg/L BA时,不定芽分化率为90%,单独添加0.3mg/L NAA时,不定根产生率最高(90%),但无不定芽生成.采用1.5mg/LBA 0.3mg/LNAA组合搭配时,不定芽分化率最高(95%).在此基础上,采用M1(MS 1.5mg/L BA 0.3mg/L NAA)培养基,研究了不同着生位置的叶片、同一着生位置叶片的不同切段以及叶片在培养基上的放置方式对84K杨叶片再生的影响,结果表明带有叶柄的下切段比其余叶片切段不定芽再生率高,幼嫩叶片比老化叶片再生率高,叶片近轴端向下接触培养基比远轴端向下接触培养基再生率高.叶片再生系统的建立为开展84K杨的工厂化育苗和遗传转化奠定了基础.

Establishment of High Frequency Regeneration System of Poplar 84K

Using the leaf of poplar 84k as the explant, MS medium as the basal medium, several important factors affecting the efficiency of regeneration of poplar 84k, such as concentration of plant growth regulators,leaf explant orientation, leaf growth sequence and segment,were studied. The results indicated that the differentiation rate of random shoots was 90% on basal MS medium only supplemented with 2.0mg/L BA and reached the highest level(95%)when 1.5mg/L BA and 0.3 mg/L NAA were added to MS medium. 90% of differentiation rate of random shoots were obtained when 0.3mg/L NAA was only added to MS medium, it was also found that more random shoots were regenerated from lower segment of leaf(with petiole)than the other segments, the number of random shoots decreased from top to base of leaf growth sequence and the percentage of random shoots induction with adaxial side downward was higher than that with adaxial side upward. The results achieved laid a sound foundation for the regeneration of poplar 84k on an industrial basis and Agrobacterium tumefaciens-mediated genetic transformation.