油用牡丹良种‘凤丹NL10’组织培养与快繁初步研究

Primary Study on Tissue Culture and Propagation of A Variety ’Fengdan NL10’ Rich in Edible Oil

牡丹(Paeonia suffruticosa Andr.)属于毛莨科(Ranunculaceae)、芍药属(Paeoniaceae)多年生落叶灌木,为中国特有的木本花卉。除了药用和观赏价值,近年来人们又发现牡丹具有油用作用,并有凤丹牡丹(P.ostii)和紫斑牡丹(P. rockii)两大类栽培品种群。目前油用牡丹的繁殖主要以种子繁殖为主,后代个体之间差异较大,性状不稳定,不能保持母本的优良性状。植物组织培养和快繁技术具有能保证原有品种的优良性状、繁殖系数高以达到产业化等特点。本研究利用‘凤丹NL10’为试验材料,选取凤丹的鳞芽和种胚等材料进行试验,主要研究不同消毒剂对外植体消毒、不同基本培养基及植物生长调节剂对鳞芽和愈伤组织的诱导、种胚生根等的影响。研究结果如下:(1)以鳞芽为外植体,先用75%的无水乙醇消毒30 s,再用2%的NaClO消毒20 min,杀菌效果最好。以种胚为外植体,采用75%的无水乙醇消毒30 s、5%的NaClO消毒20 min,杀菌效果最好。(2)鳞芽初代培养的培养基为WPM+6-BA 0.5 mg/L+GA30.4 mg/L。(3)鳞芽增殖培养的培养基为WPM+6-BA 0.5 mg/L+GA30.4 mg/L+PIC 0.6 mg/L+PVP 4 mg/L。(4)种胚在MS培养基中萌发率最高,6-BA 1.0 mg/L有利于种胚成苗,TDZ 0.5 mg/L有利于种胚膨大。适宜种胚子叶诱导愈伤组织的培养基为MS+6-BA 1.0 mg/L+2,4-D 2.0 mg/L。(5)种胚生根的培养基为1/2MS+IBA 1.0 mg/L+GA30.2 mg/L。

Paeonia suffruticosa Andr., belonging to Ranunculaceae and Paeonia, is a perennial deciduous shrub and unique woody flower in China. In addition to its medicinal and ornamental value, recently it has been discovered that this peony is highly rich in edible oil. And there are two major varieties of cultivar groups: P. ostii and P.rockii. At present, the reproduction of oil peony is mainly based on seed reproduction. The genetic variation among the offspring are large, and the characteristics are not stable, leading to the loss of the good characteristics of the female parent. Plant tissue culture technology has the characteristics of ensuring the excellent traits of the original varieties and high reproduction coefficient to achieve industrialization. Previously, we found ’Fengdan NL10’ is highly rich in edible oil among peony germplasm we collected. To extend practical usage of this variety, in this study, ’Fengdan NL10’ was used as the test material. The scaly buds and seed embryos were selected for the test.This study is mainly divided into scaly bud culture and embryo culture. This paper mainly studies the effects of different disinfectants on various explants. It also studies the influence of different basic medium and plant growth regulators on the induction of scale buds and callus, the rooting of embryo. The main results are as follows:(1) The best disinfection effect was obtained by using scaly buds as explant, 75% absolute ethanol for 30 s and 2% sodium hypochlorite for 20 min. With embryo as explant, 75% absolute ethanol for 30 s and 5% sodium hypochlorite for20 min, the effect of disinfection is the best.(2) A suitable medium for primary culture of scaly buds was WPM+6-BA 0.5 mg/L+GA30.4 mg/L.(3) A suitable medium for the proliferation of scale buds was WPM+6-BA 0.5 mg/L+GA30.4 mg/L+PIC 0.6 mg/L+PVP 4 mg/L.(4) The germination rate of seed embryos was the highest in MS medium.1.0 mg/L 6-BA was propitious to seeding formation. 0.5 mg/L TDZ was propitious to embryo expansion. Suitable medium for seed cotyledon-induced callus was MS+6-BA 1.0 mg/L+2,4-D 2.0 mg/L.(5) Suitable medium for seed embryo rooting was 1/2 MS+IBA 1.0 mg/L+GA30.2 mg/L.