以黑莓(Rubus spp.)品种‘Arapaho’无菌苗叶片为外植体,通过正交和单因素实验分别研究了基本培养基类型、6-BA 和IBA 质量浓度以及暗培养时间、外植体的叶位和接种方式对不定芽诱导的影响,并研究了IBA 质量浓度对不定芽生根的影响; 在此基础上,初步建立了黑莓品种‘Arapaho’离体叶片的再生体系。正交实验结果表明:基本培养基类型对叶片不定芽诱导率及平均不定芽数的影响最大,而IBA 质量浓度对叶片不定芽诱导率及6-BA质量浓度对平均不定芽数的影响较小;适宜‘Arapaho’叶片不定芽诱导的最佳培养基为含有2. 0 mg·L^-1 6-BA 和1. 0 mg·L^-1 IBA 的MS 培养基。单因素实验结果表明:暗培养时间、外植体的叶位及接种方式对不定芽诱导率有显著影响;最适宜的暗培养时间为21 d;植株中、上部叶片的再生能力较强,其中第3 和第4 位叶的不定芽诱导效果最佳;叶面朝上接种更有利于不定芽的诱导。在含0. 2 mg·L^-1 IBA 的MS 培养基中,不定芽生根率达100. 0%,且根数多、长势良好。黑莓品种‘Arapaho’离体叶片的再生体系为:以无菌苗的第3 和第4 位叶为外植体,经过适当修剪后叶面朝上接种于含有2. 0 mg·L^-16-BA 和1. 0 mg·L^-1 IBA 的MS 培养基上,暗培养21 d 后置于光照条件下培养30 d;将不定芽转接到含有0. 5 mg·L^-16-BA 和0. 3 mg·L^-1 NAA 的MS 培养基上进行继代培养;当不定芽高约2 cm 时转接到含有0. 2 mg·L^-1 IBA 的MS 培养基上进行生根培养,最终获得完整植株。

Using in vitro leaves from aseptic seedling of blackberry (Rubus spp.) cultivar ‘Arapaho’as explants, effects of basic medium type and concentrations of 6-BA and IBA, dark culture time, leaf position and inoculation mode of explant on adventitious bud induction and effect of IBA concentration on adventitious bud rooting were studied by orthogonal and single factor experiments. And on this basis, regeneration system of in vitro leaves of cultivar ‘Arapaho’was preliminarily established. The orthogonal experiment result shows that influence of basic medium type on induction rate and average number of adventitious bud is the most, while influence of IBA concentration on induction rate and that of 6-BA concentration on average number of adventitious bud are both less. And the optimal medium being suitable for adventitious bud induction of cultivar ‘Arapaho’is MS medium containing 2. 0 mg·L^-1 6-BA and 1. 0 mg·L^-1 IBA. The results of single factor experiments show that effects of dark culture time, leaf position and inoculation mode of explant on induction rate of adventitious bud are significant. The optimal time of dark culture is 21 d. The regeneration capacity of leaves at middle and upper parts of a plantlet is stronger, in which, induction effect of adventitious bud from leaves at the 3rd and 4th positions is the best. Inoculating with foliar facing upwards on medium is more beneficial to induce adventitious bud. Rooting rate of adventitious bud is 100. 0% in MS medium containing 0. 2 mg·L^-1 IBA with more roots and well growth vigor. The regeneration system of in vitro leaves of cultivar ‘Arapaho’as follows: using the 3rd and 4th position leaves as explants, after proper pruning, inoculating with foliar facing upwards on MS medium containing 2. 0 mg·L^-1 6-BA and 1. 0 mg·L^-1 IBA, dark culturing for 21 d then light culturing for 30 d, transferring the adventitious bud to MS medium containing 0. 5 mg·L^-1 6- BA and 0. 3 mg·L^-1 NAA for subculture, transferring to MS medium containing 0. 2 mg·L^-1 IBA for rooting when height of adventitious bud reaches about 2 cm, finally, obtaining complete plantlets.