‘雪花梨’扩繁和叶片再生体系的建立  

Research on the establishment of rapid propagation and leaf regeneration system of pear ‘Xuehuali’

作　　者：闫允青 姜桦韬 谷超[1] 吴俊[1] 

机构地区：[1]南京农业大学园艺学院/梨工程技术体系研发中心,江苏南京210095

出　　处：《南京农业大学学报》2017年第1期68-75,共8页Journal of Nanjing Agricultural University

基　　金：国家科技支撑计划项目(2013BAD02B01-2);国家梨产业技术体系(CARS-29);江苏省"六大人才"高峰项目(2014-NY-025)

摘　　要：[目的]本文旨在获得高效的‘雪花梨’扩繁体系和叶片再生体系。[方法]以‘雪花梨’茎段为材料,通过调控培养基中6-苄氨基嘌呤(6-BA)和吲哚丁酸(IBA)的配比,统计其芽增殖率和植株高度,筛选出适宜‘雪花梨’扩繁的培养基;以‘雪花梨’叶片为材料,通过比较基础培养基种类、叶片接种方式、激素种类和浓度配比及暗培养时间对‘雪花梨’叶片再生率、愈伤发生率、平均每叶再生芽数和褐化率的影响,以获得高效的叶片再生体系。[结果]适宜‘雪花梨’扩繁的培养基为MS+1.0 mg·L^(-1)6-BA+0.3 mg·L^(-1)IBA,芽增殖率为4.1,植株高度为2.66 cm。适宜‘雪花梨’叶片再生的培养基为NN69+1.5 mg·L^(-1)TDZ(噻重氮苯基脲)+0.2 mg·L^(-1)IBA,再生率达到70.83%,平均每叶再生芽数为2.06,愈伤发生率达到100%,褐化率为0;远轴面朝下接种有利于提高叶片再生率;能使芽再生率最高的暗培养时间为21 d;缩短暗培养时间和提高细胞分裂素与生长素含量能有效防止褐化;适宜再生芽初代培养的培养基为MS+0.2 mg·L^(-1)IBA+1.0 mg·L^(-1)6-BA。[结论]本研究获得了‘雪花梨’高效扩繁和再生体系,为‘雪花梨’快繁和遗传转化研究提供了有效的理论和实践依据。[ Objectives]The purpose of this research is to obtain an efficient rapid propagation and leaf regeneration system of pear. [ Methods ] Taking stems of pear variety ＇ Xuehuali＇ as materials, the optimal medium for propagation was evaluated by bud proliferation rate and plant height through adjusting the 6-benzyladenine （6-BA）and indole-3-butytric acid （IBA）ratio. The leaves of ＇ Xuehuali ＇ were used as explants, a high-efficiency system for leaf regeneration was constructed by comparing the effects of basal medium types, leaf inoculation ways, hormone types and concentration, and dark incubation time on leaf regeneration rate, callus incidence, average regeneration-bud number per leaf and browning rate. [ Results ] The results showed that the optimal medium for＇ Xuehuali＇ propagation was MS medium containing 1.0 mg·L-1 6-BA and 0.3 mg·L-1 IBA,the bud proliferation rate was 4.1 and the plant height was 2.66 cm. The optimal medium for leaf regeneration of＇ Xuehuali＇ was NN69 medium containing 1.5 mg· L-1 TDZ （thidiazuron） and 0.2 mg· L-1 IBA. The leaf regeneration rate was achieved at 70.83%, the average regeneration-bud number per leaf was 2.06,coupled with 100% of callus incidence and zero of browning rate. The leaf regeneration rate was improved when the abaxial side of leaf touched the medium, and the highest regeneration rate was observed under the treatment of 21 days dark incubation. The tissue browning could be effectively prevented through shortening the time of dark incubation and increasing the content of cytokinin and auxin. The optimal medium for the primary culture of regeneration-bud was MS medium containing 0.2 mg· L-1 IBA and 1.0 mg·L-1 6-BA. [ Conclusions] This research achieved a high-efficiency propagation system and leaf regeneration system,which laid an effectively theoretical and practical basis for rapid propagation and genetic transformation of ＇ Xuehuali＇ and provided a valuable reference to the solution of the problem of browning in leaf regeneration

关 键 词：'雪花梨’ 扩繁体系 再生体系 叶片 增殖率 再生率 

分 类 号：S661.2[农业科学—果树学]