‘凤丹白’成熟胚愈伤诱导、增殖及组织学观察

Callus induction,proliferation and histological observation of Paeonia ostii cv.‘Phoenix White’mature embryo

【目的】探讨植物生长调节剂对‘凤丹白’成熟胚愈伤组织诱导与增殖的影响,鉴定具分化潜能的愈伤组织,为‘凤丹白’再生技术体系的完善与遗传改良提供参考。【方法】以‘凤丹白’成熟胚为外植体,进行愈伤组织诱导和增殖培养,并对不同类型的愈伤组织进行外观形态与细胞组织学观察。【结果】在改良MS+0.5 mg/L水解酪蛋白+6 mg/L PIC+0.5 mg/L TDZ+30 g/L蔗糖的培养基上,‘凤丹白’成熟胚愈伤组织诱导率最高,达到70.61%;在改良MS+0.5 mg/L水解酪蛋白+4 mg/L 2,4-D+30 g/L蔗糖的培养基上,愈伤组织诱导率最低,为21.88%。诱导出的初代愈伤组织经增殖培养后,出现6种状态的愈伤组织,即松散透明、致密白色、致密绿色、致密片状、絮状披针形和透明褐色。石蜡组织切片观察结果表明,松散透明、致密白色和致密绿色愈伤组织为胚性愈伤组织,致密片状、絮状披针形和透明褐色愈伤组织为非胚性愈伤组织。在改良MS培养基+0.5 mg/L水解酪蛋白+6.0 mg/L PIC+0.5 mg/L NAA+30 g/L蔗糖的培养基上,愈伤组织扩增率最高,为97.78%,扩增系数为19.97;在改良MS培养基+0.5 mg/L水解酪蛋白+0.4 mg/L PIC+30 g/L蔗糖的培养基上,愈伤组织扩增率最低,为13.46%,扩增系数为2.97。【结论】诱导14 d后,成熟胚开始形成愈伤组织,PIC和TDZ为‘凤丹白’成熟胚愈伤组织诱导和扩增的适宜激素,经继代培养后松散透明、致密白色和致密绿色的愈伤组织具有分化潜能。

【Objective】To establish Paeonia ostii cv.‘Phoenix White’regeneration system which laid the foundation for genetic improvement,the effects of plant growth regulators on the callus induction and proliferation of P.ostii cv.‘Phoenix White’mature embryo were investigated.In addition,callus with different differentiation potential was identified.【Method】Callus induction and proliferation were performed using the mature embryo of P.ostii cv.‘Phoenix White’as an explant.Morphological and histological identification were conducted on different callus types.【Results】The optimum medium for callus induction was MS+hydrolyzed casein 0.5mg/L+PIC 6 mg/L+TDZ 0.5 mg/L+sucrose 30 g/L,and the callus induction rate reached to 70.61%.The worst callus induction medium was MS+hydrolyzed casein 0.5 mg/L+2,4-D 4 mg/L+sucrose 30 g/L,and the callus induction rate was 21.88%.There were 6 different types of callus after the primary callus proliferation culture,including loose and transparent callus,dense white callus,dense green callus,dense flaky callus,flocculent lancet callus and transparent brown callus.Tissue section observation showed that loose and transparent callus,dense white callus and dense green callus were embryogenic callus,and dense flaky callus,flocculent lancet callus and transparent brown callus were non-embryonic callus.The highest callus proliferation rate was 97.78%when cultured on MS+hydrolyzed casein 0.5mg/L+PIC 6.0mg/L+NAA 0.5mg/L+sucrose 30 g/L,the proliferation index was 19.97.The lowest callus proliferation rate was 13.46%when cultured on MS+hydrolyzed casein 0.5mg/L+PIC 0.4 mg/L+sucrose 30 g/L,the proliferation index was 2.97.【Conclusion】The callus began to form callus after 14 days culture.PIC and TDZ are suitable hormones for the callus induction and proliferation from mature embryos of‘Phoenix White’.After callus subculture,loose and transparent callus,dense white callus and dense green callus are embryogenic callus with differentiation potential.