甜樱桃砧木离体叶片愈伤组织诱导及不定芽再生

Callus induction and adventitious bud regeneration from leaves of rootstock of sweet cherry in vitro

叶片再生效率的高低直接影响目的基因转化的成功率,为建立稳定、高效的樱桃不定芽离体再生体系,以30～40d苗龄的甜樱桃砧木ZY-1的组培继代苗为试材,取上部幼嫩叶片或不含腋芽的茎段,分别从培养基生长调节剂配比、接种材料类型、叶片接种部位、接种方式以及培养条件等方面进行了不定芽再生技术研究。结果表明,培养基中添加7.0mg/L的6-BA与0.5～1.0mg/L的IBA配比时不定芽再生率和出芽数均较高,TDZ和NAA不适于诱导ZY-1叶片再生不定芽;接种继代苗茎段比叶片再生率高;接种叶片以选择嫩叶横切2刀、远轴面向下接触培养基的方式为好;叶片不同部位处理以带叶柄的基部叶块最易再生;叶片接种后在25℃室温条件下,先暗培养3周再照光,利于不定芽的再生。

The rate of adventitious bud regeneration from explants is highly correlated to the efficiency of gene transformation. In order to set up the stable and high efficient system of adventitious bud regeneration in cherry, the factors affecting adventitious bud regeneration from leaves and stems in vitro were studied in the ZY-1 （Prunus cerasus L.）, which is used as a rootstock of sweet cherry, including the types and concentration of plant growth regulators applied in the culture media, the types of explants, the laying modes of leaves, and the incubating condition. The young leaves sampled from the top of shoots and stems without buds attached were used, and both of them were collected from the plantlets sub-cultured in vitro for 30- 40 d. The results showed that the rate of adventitious bud regeneration and the number of adventitious buds per leaf were higher on the media that supplied 7.0 mg/L 6-BA and 0.5-1.0 mg/L IBA. The positive results was not obtained by using TDZ and NAA in term of the adventitious bud regeneration in this experiment. The stems as explants generated more adventitious buds than the leaves. The young leaves, which were cut two times and placed on the medium with the lower surface touching the medium, showed higher regeneration rate. The adventitious buds were easier to generate from the leaf disks that were cut from the base part of the leaves and had the petioles attached. The conducting of dark incubation for three weeks prior to light culture in culture rooms at 25℃ tends to benefit the adventitious bud regeneration from the leaf disks.