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欧洲李叶片再生体系的建立

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【目的】为建立稳定、高效的李再生体系。【方法】以欧洲李叶片为外植体,探究不定芽诱导、增殖、生长和生根的最适条件。【结果】不定芽诱导的最适培养基为WPM+噻苯隆(TDZ)2.0 mg · L-1+2,4-二氯苯氧乙酸(2,4-D) 0.2 mg · L-1 ,茎中部成熟叶是最佳外植体,暗培养14 d时再生率最高为33.4%;最适增殖培养基为MS+6-苄基腺嘌呤(6-BA)2.0 mg · L-1+3-吲哚丁酸(IBA)0.1 mg · L-1 ,增殖倍数为7.03;最佳生长培养基为:MS+6-BA 0.2 mg · L-1+IBA 0.1 mg · L-1;以MS+IBA 0.5 mg · L-1进行生根培养,可获得74%的生根率;驯化移栽后幼苗成活率达83.3%。【结论】以欧洲李女神叶片为外植体建立了完整的再生体系:再生率最高为33.4%,增殖倍数为7.03,生根率为74%,移栽成活率为83.3%,为进一步利用欧洲李叶片开展遗传转化奠定了基础。
Establishment of leaf regeneration system for European plum
【Objective】To construct a stable and efficient regeneration system for plum, the leaves from European plum (Prunus domestica) were used as explants to study the optimum conditions for adventi-tious shoot regeneration, proliferation, growth and rooting. Using the leaves as explants has the advan-tages of convenient collection, abundant materials and low cost.【Methods】Leaves from branches of four P. domestica cultivars Startovaya, Victoria, France and Bluebyrd were surface-sterilized in 0.5%so-dium hypochlorite (NaClO) solution for 10 min and rinsed thrice in sterile distilled water, and then sur-face-sterilized in 0.1% mercuric chloride (HgCl2) solution for 6 min and again rinsed 5-6 times with sterile distilled water. The main vein of leaves was slightly cut and plated to Murashige and Skoog (MS) media supplemented with different concentrations of 6-Benzylaminopurine (6-BA) and Indole-3-butyric acid (IBA) for regeneration. Leaves from in vitro cultured seedlings after subculture for 30 days were cut in the same way and placed on the woody plant (WPM) media supplemented with different concentrations of Thidiazuron (TDZ) and 2, 4-Dichlorophenoxyacetic acid (2, 4-D) for regeneration. The regenerated shoots from 40-day-old cultures were placed on MS media supplemented with different concentrations of 6-BA and IBA for proliferation. The small individual shoots from proliferation culture were transferred to MS media supplemented with different concentrations of 6-BA and IBA for stem elongation. Then the elongated adventitious shoots with 3-4 leaves were transferred for rooting. All of each experiment repeated for three times.【Results】For four P. domestica cultivars, the adventitious shoots were only regenerated from Victoria with a regeneration rate of 5.4%, and then the following re-searches were carried out for P. domestica cultivar Victoria. Firstly, WPM medium was used as basal culture medium and the effect of plant growth regulators, different explant and dark incubation time onadventitious shoot regeneration was investigated. The concentrations of TDZ and 2, 4-D were 2.0 mg · L-1 and 0.2 mg · L-1, which were best for adventitious shoot regeneration and the regeneration rate reached 18%. The intact leaves rather than petioles, middle part of leaves and leaves tip from the middle part of tissue culture seedling were the best explants for adventitious shoot regeneration. The regeneration rate was further raised up to 25.6%. The five different dark incubation times were set and the intact leaves incubated in dark for 14 days had the highest adventitious shoot regeneration rates (33.4%). Secondly, MS medium was used as basal culture medium and the effect of plant growth regulators on adventitious shoot proliferation was investigated. When the concentration of 6-BA was 3 mg · L-1, it had a higher pro-liferation rate than the other two concentrations (2 mg · L-1 and 1 mg · L-1) and the state of regenerative shoot was yellowing, curly and vitrified. When the concentration of 6-BA was 2 mg · L-1, a small quanti-ty of regenerative shoot was vitrification. When the concentration of 6-BA was 1 mg · L-1, the state of re-generative shoot was yellowing. Thirdly, MS medium was used as basal culture medium and the effect of different concentrations of 6-BA was investigated on the growth of regenerative shoot. When the con-centrations of 6-BA and IBA were 0.2 mg · L-1 and 0.1 mg · L-1, the stem of regenerative shoot was robust and grew faster than others. Finally, the effect of different concentrations of IBA was analyzed on root-ing rate, root length and rooting coefficient. There were no roots from regenerative shoot without IBA addition. When the concentration of IBA was 0.5 mg · L-1, the highest rooting rate achieved 74%with an average root length of 14.7 cm and highest rooting coefficient of 4.69.【Conclusion】A stable regenera-tion system for European plum using leaves was established. The concrete steps include: the mature leaves in mid part were inoculated on WPM media supplemented with 2.0 mg · L-1 TDZ and 0.2 mg · L-1 2, 4-D with dark-incubated for 14 days, and the highest frequencies of adventitious shoot regeneration was 33.4%. The optimal adventitious shoot proliferation medium was MS media supplemented with 2.0 mg · L-1 6-BA and 0.1 mg · L-1 IBA, and the proliferation rate was 7.03. The optimal growth medium was MS media supplemented with 0.2 mg · L-1 6-BA and 0.1 mg · L-1 IBA. The highest rooting rate of 74%was observed on MS media supplemented with 0.5 mg · L-1 IBA. Then the seedling survival rate was 83.3%after seedlings were transplanted into the pots.

European plumLeaf regenerationAdventitious shoot

张郎郎、张洁、吕虹霖、谭彬、王伟、程钧、冯建灿

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河南农业大学园艺学院,郑州 450002

欧洲李 叶片再生 不定芽

国家自然科学基金青年基金国家重点研发计划项目国家自然科学基金联合项目

320020142019YFD1000104U1804114

2022

果树学报
中国农业科学院郑州果树研究所

果树学报

CSTPCDCSCD北大核心
影响因子:1.486
ISSN:1009-9980
年,卷(期):2022.39(10)
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