宜昌百合胚性愈伤组织诱导及植株再生体系的研究

doi:10.7525/j.issn.1673-5102.2019.03.003

摘要/Abstract

摘要： 采用L₉（3⁴）正交试验等方法，以野生宜昌百合（Lilium leucanthum）鳞片为外植体，分别在光照和黑暗培养条件下，研究了TDZ、NAA、2，4-D不同浓度配比对宜昌百合的胚性愈伤组织诱导效果的影响，并采用石蜡切片技术对愈伤组织进行了的组织学观察。在此基础上，探讨了NAA对体胚苗壮苗生根的影响。结果表明：在光照条件下诱导胚性愈伤组织的最佳培养基为MS+TDZ 0.5 mg·L^-1+NAA 1.0 mg·L^-1+2，4-D 0.05 mg·L^-1，诱导率为62.690%；黑暗条件下诱导胚性愈伤组织的最佳培养基为MS+TDZ 0.5 mg·L^-1+NAA 1.5 mg·L^-1+2，4-D 0.1 mg·L^-1，诱导率为59.423%。胚性愈伤组织黄绿色或深绿色，颗粒状明显；非胚性愈伤组织松散易碎、呈水渍状，经石蜡切片观察可观察到胚性愈伤组织细胞核大、胞质浓，且细胞内含丰富淀粉粒，细胞直径为50~80 μm。非胚性愈伤组织细胞较大，未见明显细胞核，直径约为100~180 μm。体胚苗壮苗生根的最适宜培养基为1/2MS+NAA 0.2 mg·L^-1+AC 1 g·L^-1，培养60 d后，幼苗移至草炭：蛭石：珍珠岩=1：1：1基质中，成活率为90%。本试验成功建立了宜昌百合鳞片胚性愈伤组织的再生体系，为宜昌百合利用胚状体进行无性育种以及种质资源的创新奠定了基础。

关键词: 宜昌百合, 胚性愈伤组织, 再生体系

Abstract: With Lilium leucanthum Bakeri bulbs as explants, L₉(3⁴) orthogonal test was used to study the effects of different hormones and their concentrations on its callus induction, bulblet differentiation and rooting culture. Histological observation of embryogenic callus was carried out by paraffin sectioning technique. The optimal medium combinations for inducing callus of L.leucanthum under light and dark conditions were MS+TDZ 0.5 mg·L^-1+NAA 1.0 mg·L^-1+2, 4-D 0.05 mg·L^-1 and MS+TDZ 0.5 mg·L^-1+NAA 1.5 mg·L^-1+2, 4-D 0.1 mg·L^-1, with the induction rates of 62.690 and 59.423, respectively. The medium suitable for bulblet rooting was 1/2MS+NAA 0.2 mg·L^-1+1 g AC. After 60-day culture, the seedlings were transferred to peat:vermiculite:perlite=1:1:1 in a medium and cultured in a light incubator for 35 days with the survival rate of 90%. Our experiment successfully established the regeneration system of squamous callus in L.leucanthum, which laid a foundation for the research of tissue culture transgenic seedling and the innovation of germplasm resources.

Key words: Lilium leucanthum, embryonic callus, regeneration system

中图分类号:

Q949.71

张璐, 潘远智, 刘柿良, 陈源, 申理嘉. 宜昌百合胚性愈伤组织诱导及植株再生体系的研究[J]. 植物研究, 2019, 39(3): 338-346.

ZHANG Lu, PAN Yuan-Zhi, LIU Shi-Liang, CHEN Yuan, SHEN Li-Jia. Embryonic Callus Induction and Plant Regeneration of Lilium leucanthum[J]. Bulletin of Botanical Research, 2019, 39(3): 338-346.

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