北美豆梨杂种幼胚挽救及再生体系的建立

Establishment of Immature Embryo Rescue and Regeneration System for Pyrus calleryana cv.‘Cleveland’

以北美豆梨(Pyrus calleryana cv.‘Cleveland’)杂种幼胚为材料,系统开展了杂种幼胚灭菌、愈伤组织诱导、不定芽增殖和生根培养基的筛选研究。结果表明,4℃低温贮藏21天的杂种幼胚萌发率最高,为67.23%;幼胚最佳灭菌处理为75%乙醇灭菌30秒,10%H2O2和0.1%HgCl2分别消毒10和14分钟;最适幼胚萌发培养基为1/2MS+4.0 mg·L^(–1)6-BA+0.5 mg·L^(–1)NAA,萌发率达89.67%;最适愈伤组织诱导培养基为1/2MS+1.0 mg·L^(–1)IBA+1.0 mg·L^(–1)6-BA,愈伤组织诱导率为93.33%;最佳分化培养基为1/2MS+0.2 mg·L^(–1)IBA+2.0 mg·L^(–1)6-BA,再生频率为87.44%;最佳继代增殖培养基为1/2MS+1.5 mg·L^(–1)6-BA+0.1 mg·L^(–1)IBA,增殖率为100%;最佳生根培养基为1/2MS+20 g·L^(–1)蔗糖+1.0 g·L^(–1)活性炭+1.5 mg·L^(–1)IBA+0.05 mg·L^(–1)NAA,生根率为82.63%。该研究为北美豆梨杂交种质资源高效繁殖提供了科学依据和指导。

In this study,the seed embryos of Pyrus calleryana cv.‘Cleveland’were used as materials to systematically study the sterilization of seed embryo explants,callus induction,adventitious bud proliferation and rooting medium.The results showed that the germination rate of young embryos stored at 4℃ for 21 d was the highest(67.23%)The suitable sterilization treatment of seed embryo explants was 75%alcohol(30 s)+10%H2O2(10 min)+0.1%HgCl2(14 min).The most suitable medium for seed embryo germination was 1/2MS+4.0 mg·L^(–1)6-BA+0.5 mg·L^(–1)NAA,and the germination rate was 89.67%.The optimum culture medium for callus induction was 1/2MS+1.0 mg·L^(–1)IBA+1.0 mg·L^(–1)6-BA,and the callus induction rate was 93.33%.The best differentiation medium was 1/2MS+0.2 mg·L^(–1)IBA+2.0 mg·L^(–1)6-BA,and the regeneration frequency was 87.44%.Best transgenerational proliferation medium was 1/2MS+1.5 mg·L^(–1)6-BA+0.1 mg·L^(–1)IBA,and the proliferation rate was 100%.The most suitable medium for root formation was 1/2MS+20 g·L^(–1)sucrose+1.0 g·L^(–1)active carbon+1.5 mg·L^(–1)IBA+0.05 mg·L^(–1)NAA,and the rooting rate was 82.63%.In summary,the seed embryo regeneration system of P.calleryang cv.‘Cleveland’was established,which provides scientific basis and guidance for the efficient reproduction of high-quality pear germplasm resources.