金樱子(Rosa laevigata Michx.)叶片直接再生不定芽体系的建立

以金樱子组培苗的划伤叶片为外植体,研究了不同植物生长调节剂配比组合、暗培养时间、添加物L-脯氨酸、叶片不同部位对不定芽直接再生的影响。结果表明:当TDZ浓度为1.5 mg·L^-1、NAA浓度为0.000 5 mg·L^-1时,不定芽直接发生率最高,为9.5%;不同暗培养时间对不定芽直接诱导具有一定影响,暗培养时间为10 d时不定芽的直接发生率最高,为10.5%;添加了不同浓度的L-脯氨酸后不定芽的诱导率不增反降,所以以不添加L-脯氨酸为宜;叶片不同部位的再生率比较中,仅带叶叶柄可直接诱导出不定芽。综上所述,叶片直接再生不定芽的诱导方法是,以金樱子带叶叶柄为外植体,在直接诱导培养基上1/2MS+TDA 1.5 mg·L^-1+NAA 0.000 5 mg·L^-1+CH 100 mg·L^-1+AgNO3 10 mg·L^-1+蔗糖30 g·L^-1+琼脂7.5 g·L^-1,暗培养10 d后,转至正常光周期下培养3周左右,不定芽诱导率最高,为10.5%。所得不定芽在增殖培养基MS+NAA 0.1 mg·L^-1+6-BA 1.0 mg·L^-1生长3周后,增殖系数可达到3.5左右;将丛生芽切割成单芽后转入不含任何激素的MS培养基壮苗3周,幼苗可长高至3~5 cm;再转入MS+NAA 0.1 mg·L^-1培养基中生根,1个月后生根率达95%。

Direct Regeneration of Adventitious Buds From Leaves of Rosa laevigata Michx.

The scratched leaves of tissue culture seedlings of Rosa laevigata Michx.were used as explants,and the effects of different combinations of plant growth regulators,dark culture time,additives L-proline and different parts of leaves on the direct regeneration of adventitious buds were studied.The results showed that TDZ concentration was 1.5mg·L-1 and NAA concentration was 0.000 5 mg·L^-1,the direct incidence of adventitious buds was the highest,which was 9.5%.Different dark culture time had certain influence on the direct induction of adventitious buds.The direct incidence of adventitious buds was the highest when dark culture time was 10 days,which was 10.5%.The induction rate of adventitious buds did not increase or decrease after adding different concentrations of L-proline.In comparison of regeneration rate of different parts of leaves,adventitious buds could be directly induced only with petioles.In conclusion,the method of inducing adventitious buds from leaves to regenerate directly was as follows:1/2MS+TDA 1.5 mg·L^-1+NAA 0.000 5mg·L^-1+CH 100mg·L^-1+AgNO3 10mg·L^-1+sucrose 30g·L^-1 +agar 7.5g·L^-1 on the direct induction medium with petioles.After 10 days of dark culture,the induction rate of adventitious buds was the highest(10.5%).The adventitious buds in the proliferation medium MS+NAA0.1mg·L^-1+6-BA 1.0mg·L^-1 grew after 3 weeks,the proliferation coefficient could reach about 3.5.Then the cluster buds were cut into single buds and transferred to MS medium to sound seedlings for 3weeks.Subsequently,the resulted 3-5cm seedlings were then transfer to MS+NAA 0.1mg·L-1 medium for rooting,andthe post-rooting ratereached 95% one month later.