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黄毛草莓叶片愈伤组织诱导研究

来源:用户上传      作者:钱邵函 刘倩文 武春霞

  摘 要:为建立黄毛草莓叶片繁殖体系,筛选诱导愈伤组织的最适培养基,以黄毛草莓幼嫩叶片为外植体,研究了消毒时间、激素组合、光暗条件对黄毛草莓叶片愈伤组织诱导的影响。结果表明:黄毛草莓叶片最佳的外植体消毒处理为0.1%的升汞消毒6~8 min;黄毛草莓叶片愈伤组织褐化严重,经15%抗坏血酸(Vc)溶液浸泡20 min可显著降低褐化率,提高出愈率;愈伤组织诱导的最适培养基组合为MS+1.0 mg・L-1 6-BA+0.1 mg・L-1 NAA和MS+1.0 mg・L-1 TDZ+0.1 mg・L-1 IBA;暗培养10 d出愈率最高,可达到87.5%。因此,黄毛草莓叶片愈伤组织诱导最佳培养方法为:MS+1.0 mg・L-1 6-BA+0.1 mg・L-1 NAA+30 g・L-1 蔗糖+7 g・L-1 琼脂,培养基pH值为 5.6,15%Vc溶液浸泡20 min,出愈率可达到95.0%。
  关键词:黄毛草莓;叶片;愈伤组织;诱导
  中图分类号:S668.4 文献标识码:A DOI 编码:10.3969/j.issn.1006-6500.2022.06.008
  Study on Callus Induction of Fragaria nilgerrensis Leaves
  QIAN Shaohan, LIU Qianwen, WU Chunxia
  (College of Horticulture Landscape, Tianjin Agricultural University, Tianjin 300384, China)
  Abstract: In order to establish an efficient method for plant regeneration from leaf explant of Fragaria nilgerrensis plantlet the most suitable medium for inducing callus was screened. The leaves of Fragaria nilgerrensis plantlet were used for studying the effects of disinfection time, plant growth regulator concentration and ratio, different incubation time in dark, on callus induction in leaves were studied using the young and tender leaves of Fragaria nilgerrensis as explants. The results showed that the optimal explant disinfection treatment for 0.1% of the leaves of Fragaria nilgerrensis was 0.1% mercury disinfection for 6-8 minutes, and the callus of Fragaria nilgerrensis leaf was severely browned, and the browning rate and callus induction rate could be significantly reduced by soaking in 15% ascorbic acid (Vc) solution for 20 minutes. The optimal medium combination for callus induction was MS+1.0 mg・L-1 6-BA+0.1 mg・L-1 NAA and MS+1.0 mg・L-1 TDZ+0.1 mg・L-1 IBA for 10 d in dark, the callus induction frequency was up to 87.5%. The best cultivation method is MS+1.0 mg・L-1 6-BA+0.1 mg・L-1 NAA+30 g・L-1 sucrose+7 g・L-1 Agar-Agar, medium pH 5.6 and soaks in 15%Vc solution for 20 minutes, the callus induction frequency is up to 95%.
  Key words: Fragaria nilgerrensis; leaf; callus; induction
  S毛草莓(Fragaria nilgerrensis Schlecht. ex Gay)是蔷薇科(Rosaceae)草莓属(Fragaria L.)多年生二倍体草本植物,具有较强的抗病抗旱抗寒能力,在世界各地广泛栽培[1-2]。黄毛草莓中富含的氨基酸总量在野生草莓资源中相对较高,具有较高的营养价值,是培育具有高氨基酸含量果品的宝贵野生资源[3]。就目前草莓发展的趋势来看,野生种质资源在改善草莓品种特性上具有极大的应用潜力。
  匍匐茎分株法普遍用于草莓的无性繁殖,这种方法繁殖系数较低、速度较慢,并且特别容易感染病毒而使种苗退化[4],导致草莓的产量和品质下降,不利于优良品种的推广,不能满足大范围生产的需要。此外,在黄毛草莓引进后,其大规模种苗生产技术还不够完善,加上作物多倍体的特性,制约了传统的育种方法。草莓的组培技术则可以获得无毒苗以保持其品种的优良特性[5-6]。目前,关于国内草莓的组培研究大多集中于草莓的茎尖培养[7-9],对于叶片繁殖的研究较少,不少草莓生产基地对草莓组培和扩繁等进行了系统的研究。晁慧娟等[10]以‘甜查理’草莓叶片为试验材料,对其进行组培,得出一组适宜叶片诱导愈伤组织的激素组合水平,试验结果表明:叶片诱导愈伤培养基最佳组合为MS+1.0 mg・L-1 6-BA+0.2 mg・L-1 NAA出愈率92.5%。刘玲等[11]用TDZ与IBA激素的不同浓度组合诱导黄毛草莓离体叶片愈伤组织形成。试验结果表明,诱导叶片愈伤组织形成得最佳培养基为MS+1.0 mg・L-1 TDZ+0.1 mg・L-1 IBA出愈率95.4%。本试验设置了2种植物激素组合,6-BA和 NAA为一组、TDZ和IBA为一组,研究黄毛草莓叶片不同消毒处理、不同水平激素组合和添加15%抗坏血酸以及不同暗处理时间下愈伤组织诱导的状况,以期得到适合黄毛草莓叶片诱导出愈伤组织的植物激素组合水平,为黄毛草莓快速繁殖和育种奠定理论基础。

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