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多胺对荔枝胚性愈伤组织增殖与体胚分化的影响 |
王 果1,刘耀婷1,2,李焕苓1,王树军1,李芳1,王家保1*
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1. 中国热带农业科学院环境与植物保护研究所·农业农村部儋州农业环境科学观测实验站,海南 儋州 571737;2. 海南大学热带作物学院,海口 570228
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摘要: |
为探讨外源多胺(PAs)对荔枝胚性愈伤组织(EC)增殖及体胚分化的影响机制,该研究以“妃子笑”荔枝EC为材料,采用单因素法在增殖培养基中添加腐胺(Put)、亚精胺(Spd)及精胺(Spm),分析了不同PAs处理后EC形态、结构、内源PAs含量及相关酶指标的变化。结果表明:(1)外源Put、Spd和Spm处理均显著提高了EC增殖率,降低了体胚诱导及萌发数量。经外源PAs处理增殖的EC胚性细胞大小较一致,染色深且均匀,多细胞原胚减少,可见已经分化完全的早期子叶胚。(2)外源PAs处理均显著提高了EC中内源PAs含量,其中Put处理EC中各类内源PAs及总PAs含量最高;当在含外源PAs培养基上增殖的EC转入不含外源PAs的培养基上增殖时(恢复培养),EC中的Put含量仍然显著高于对照,内源Spd和Spm则显著降低。(3)外源Put处理显著提高了EC中的鸟氨酸脱羧酶(ODC)、精氨酸脱羧酶(ADC)和二胺氧化酶(DAO)活性,而外源Spd、Spm处理显著降低了EC中的ODC及ADC活性,外源Spd显著提高了多胺氧化酶(PAO)活性;恢复培养后,EC中ADC和DAO活性比恢复培养前显著降低,ODC、PAO无显著性差异。综上认为,外源PAs可以通过调节PAs代谢相关酶活性影响内源PAs含量,进而影响荔枝EC增殖和体胚诱导。该研究结果为进一步研究PAs调节荔枝体胚发生机制,提高荔枝离体再生效率提供了基础。 |
关键词: 荔枝,离体再生,组织切片,多胺,酶活性 |
DOI:10.11931/guihaia.gxzw202308066 |
分类号: |
基金项目:海南省自然科学基金(320QN312);国家现代农业产业技术体系(CARS-32);海南省自然科学基金面上基金(323MS100)。 |
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Influences of polyamines on callus proliferation and somatic embryogenesis in Litchi chinensis |
WANG Guo 1, LIU Yaoting 1,2, LI Huanling 1, Wang Shujun 1, Li Fang 1, WANG Jiabao 1*
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Abstract: |
To explore the effect of exogenous polyamines (PAs) on embryogenic callus (EC) proliferation and somatic embryogenesis of litchi, the morphology, structure, endogenous PA content and related enzyme activities were systematically investigated using the ‘Feizixiao’ ECs as materials subcultured on the medium supplemented with various PAs. The results were as follows: (1) The exogenous putrescine (Put), spermidine (Spd), and spermine (Spm) treatment significantly increased the EC proliferation rate and reduced the amount of induced somatic embryos and regenerated plantlets. The proliferated embryogenic cells after exogenous PA treatments were more consistent in size and stained deeply and evenly. Furthermore, multicellular proembryos in EC were reduced, and fully differentiated early cotyledon embryos could be seen. (2) All the exogenous PA treatments significantly increased the endogenous PA content in EC. Among them, Put treatment had the highest content of each endogenous PA component and total PA. When the EC proliferated on the medium containing exogenous PAs was transferred to the medium without exogenous PAs (M3) for proliferating, the Put content in the EC was still significantly higher than the control, however, the endogenous Spd and Spm were significantly decreased. (3) Exogenous Put treatment significantly increased the activities of ornithine decarboxylase (ODC), arginine decarboxylase (ADC), and diamine oxidase (DAO) in EC, while exogenous Spd and Spm treatments significantly reduced the activities of ADC and DAO in EC, and exogenous Spd significantly increased PAO activity. When transferred to the M3 medium, the ADC and DAO activities of newly proliferated EC were significantly lower than those of EC cultured with exogenous PAs, but there was no significant difference in ODC and PAO activities. In summary, the exogenous PAs can affect endogenous PA content by regulating the activity of enzymes related to polyamine metabolism, thereby affecting EC proliferation and somatic embryo induction in litchi. These results would provide a basis for further study on the mechanism of PAs regulating litchi regeneration in vitro. |
Key words: litchi (Litchi chinensis), plant regeneration, histological section, polyamines, metabolic enzymes |