| 摘要: |
| 【目的】比较不同抗褐化剂减轻福建柏组织培养过程中褐化现象的效果，以提高福建柏愈伤组织培养的增殖分化能力。【方法】以福建柏鳞叶和茎段为外植体，通过单因素试验，研究0.0～2.5 mg/L NAA和2,4 D及0.0～3.0 mg/L IBA对愈伤组织诱导率与生长状态的影响；以福建柏茎段为外植体，采用单因素和多因素组合试验，研究100～2 000 mg/L聚乙烯吡咯烷酮（PVPP）、100～1 000 mg/L活性炭（AC）和VC以及不同质量浓度VC与AC组合，对愈伤组织污染率、死亡或褐化率及诱导率的影响；以诱导的愈伤组织为材料，采用单因素试验，研究100～1 000 mg/L PVPP、100～300 mg/L VC、200～1 000 mg/L AC、30～100 mg/L VB对愈伤组织增殖生长量、增殖率、褐化率及PPO、POD活性的影响。【结果】① 不同质量浓度NAA、2,4-D和IBA对福建柏鳞叶愈伤组织的诱导率总体高于茎段，各生长素对福建柏鳞叶和茎段愈伤组织的诱导率均表现为NAA＞IBA＞2,4-D，所有处理中以1.0 mg/L NAA对福建柏鳞叶愈伤组织的诱导率最高，为83.33%；以0.5 mg/L NAA对茎段愈伤组织的诱导率最高，为61.11%；愈伤组织生长状态和抗褐化效果优劣依次为NAA、2,4-D、IBA。②仅100 mg/L VC+200 mg/L AC处理的愈伤组织污染率低于不添加抗褐化剂的对照；死亡或褐化率仅500 mg/L PVPP处理与对照相同，均为0.00%，其余处理均高于对照；诱导率则以不同质量浓度PVPP、1 000 mg/L VC、100 mg/L VC＋200 mg/L AC处理与对照相当或高于对照，其余处理均低于对照；愈伤组织生长状态总体以PVPP处理表现最优。③仅300 mg/L PVPP、200和400 mg/L AC处理的愈伤组织增殖率高于对照，增殖生长量仅100和200 mg/L PVPP、100 mg/L VC、200 mg/L AC处理高于对照；所有处理中以200 mg/L AC处理的愈伤组织增殖率和增殖生长量最高，分别为70.00%和10.07 g；各抗褐化剂对福建柏愈伤组织增殖效果的优劣依次为PVPP、AC、VC、VB。④不同抗褐化剂处理的愈伤组织褐变率均呈先增加后趋于稳定的变化趋势，但变化幅度存在显著差异；各抗褐化剂的抗褐化效果优劣依次为AC、PVPP、VC、VB。⑤不同质量浓度PVPP、VC、AC处理对福建柏愈伤组织PPO和POD活性的影响具有显著差异，但各处理PPO活性均与对照无显著差异，POD活性以500 mg/L AC处理显著高于其他处理。【结论】在福建柏愈伤组织诱导阶段，愈伤组织的褐化与生长素种类有关，生长素NAA、2,4-D适用于福建柏愈伤组织的诱导，IBA诱导的愈伤组织容易褐化，添加适量抗褐化剂能减少愈伤组织的褐化情况。在愈伤组织增殖培养阶段，适宜质量浓度的PVPP、AC作为抗褐化剂能有效减缓福建柏愈伤组织的褐化进程，保持愈伤组织的正常增殖生长，但高质量浓度的抗褐化剂会抑制愈伤组织的生长。 |
| 关键词: 福建柏 愈伤组织 组织培养 褐化 抗褐化剂 |
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| 基金项目:福建省林业种苗科技攻关七期项目（LZKG-202207） |
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| Healing tissue induction and anti-browning of Fokienia hodginsii |
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WANG Xiaomei,WU Xia,ZHANG Yuxin,et al
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| Abstract: |
| 【Objective】This study compared the effects of different anti-browning agents on reducing browning during tissue culture of Fokienia hodginsii to improve proliferation and differentiation ability of callus culture of F. hodginsii.【Method】The effects of 0.0-2.5 mg/L NAA and 2,4-D and 0.0-3.0 mg/L IBA on callus induction rate and growth were studied by single factor experiment using scale leaves and stem segments of F. hodginsii as explants.Using stem segments of F. hodginsii as explants,the effects of 100-2 000 mg/L polyvinylpyrrolidone (PVPP),100-1 000 mg/L activated carbon (AC) and ascorbic acid (VC) as well as different combinations of VC and AC on callus contamination rate,death or browning rate and induction rate were studied by single factor and multi-factor combination experiments.The effects of 100-1 000 mg/L PVPP,100-300 mg/L VC,200-1 000 mg/L AC and 30-100 mg/L riboflavin (VB) on proliferation,proliferation rate,browning rate and activities of polyphenol oxidase (PPO) and peroxidase (POD) of callus were also studied by single factor tests.【Result】① The induction rates of NAA,2,4-D and IBA at different concentrations on the callus of F. hodginsii scale leaves were higher than those of stems.The induction rates of each auxin on scale leaves and stems were in the order of NAA>IBA>2,4-D.The treatment of 1.0 mg/L NAA had the highest induction rate of 83.33% on scale leaf callus.The highest induction rate of stem callus was 61.11% in treatment of 0.5 mg/L NAA.NAA had the best growth and anti-browning effect,followed by 2,4-D and IBA sequentially.② Compared with the control without anti-browning agent,only the pollution rate of 100 mg/L VC＋200 mg/L AC treatment was lower.The death or browning rate of 500 mg/L PVPP treatment was the same as that of the control,which was 0.00%,while other treatments were higher.The induction rates of treatments with PVPP,1 000 mg/L VC and 100 mg/L VC＋200 mg/L AC were no lower than those of the control,while other treatments were lower.The growth of callus was the best in PVPP treatment.③ Only the callus proliferation rates of 300 mg/L PVPP,200 and 400 mg/L AC treatments were higher than those of the control,and the proliferation growth of 100 and 200 mg/L PVPP,100 mg/L VC and 200 mg/L AC treatments was higher than those of the control.The callus proliferation rate and proliferation growth of 200 mg/L AC treatment were the highest of 70.00% and 10.07 g,respectively.The effect of each anti=browning agent on the proliferation of callus of F. hodginsii was in the decreasing order of PVPP,AC,VC and VB.④ The browning rates of callus treated with different anti-browning agents increased first and then tended to be stable,but the change ranges were significantly different.AC had the best anti-browning effect,followed by PVPP,VC and VB in turn.⑤ The effects of PVPP,VC and AC at different concentrations on PPO and POD activities in callus of F. hodginsii were significantly different,while there was no significant difference in PPO activity.POD activity in 500 mg/L AC treatment was significantly higher than in other treatments.【Conclusion】In the induction stage of F. hodginsii callus,the browning of callus was related to auxin type.NAA and 2,4-D were suitable for the induction of F. hodginsii callus,and the callus induced by IBA was easy to browning.Adding an appropriate amount of anti-browning agent could reduce callus browning.In the stage of callus proliferation culture,PVPP and AC at appropriate concentrations as anti-browning agents could effectively slow down the browning process of F. hodginsii callus and maintain normal proliferation and growth of callus,but high concentrations would inhibit the growth of callus. |
| Key words: Fokienia hodginsii healing tissue tissue culture browning anti-browning agent |
