| 摘要: |
| 【目的】探究福建柏不同外植体再生过程中最适激素配比，为建立福建柏组织培养体系提供技术支持。【方法】以福建柏鳞叶和茎段作为外植体，在饱和洗衣粉液浸泡20 min和体积分数75%酒精浸泡30 s条件下，研究体积分数0.1% HgCl2浸泡不同时间(5,8,10,12,15 min)后的消毒效果；以福建柏鳞叶和茎段为材料，通过单因素试验,研究生长素（NAA和2,4-D）对不同外植体愈伤组织诱导的影响，再通过正交试验研究NAA、2,4-D、6-BA不同配比对外植体愈伤组织诱导的影响；以福建柏愈伤组织为材料，采用正交试验研究NAA、2,4-D、6-BA、KT对愈伤组织继代增殖的影响；以福建柏带芽茎段为材料，研究细胞分裂素6-BA对芽诱导和萌发的影响；以福建柏分化苗为材料，采用正交试验研究6-BA、2,4-D、KT对分化苗继代生长的影响；以福建柏无根苗为材料，研究无根苗扩增繁殖的效果。【结果】（1）福建柏鳞叶最适消毒配方为体积分数75%酒精30 s+体积分数0.1% HgCl2 10 min，污染率为16.67%；茎段为体积分数75%酒精30 s+体积分数0.1% HgCl2 15 min，污染率为23.33%。（2）鳞叶愈伤组织诱导最适配方为MS培养基+NAA 2.0 mg/L+2,4-D 1.0 mg/L+6-BA 1.0 mg/L，诱导率为81.67%；诱导出的愈伤组织白色近透明，长势良好。而茎段为MS培养基+NAA 0.5 mg/L+2,4-D 0.5 mg/L，诱导率为78.33%，诱导出的愈伤组织淡绿色，长势良好。（3）愈伤组织最适增殖配方为MS基本培养基+2,4-D 1.0 mg/L+6-BA 2.0 mg/L+KT 0.5 mg/L，增殖质量为1.984 g，褐化率23.33%。（4）芽诱导萌发最适配方为MS基本培养基+6-BA 2.0 mg/L，平均诱导芽数2.05个，平均萌动数1.70个，分化率100%。（5）分化苗生长和扩繁的最适配方为1/2MS基本培养基+2,4-D 0.5 mg/L+KT 1.0 mg/L，平均生长长度为2.241 cm，无褐化，扩繁成活率为95.50%，扩繁系数为3.18。【结论】福建柏以带芽茎段作为芽增殖再生途径外植体，鳞叶作为愈伤组织再生途径的外植体进行组织培养效果最佳，长势好且稳定。 |
| 关键词: 福建柏 愈伤组织 离体再生 诱导与增殖 |
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| 基金项目:福建省科技重大专项(2018NZ0001-1)；福建农林大学科技发展基金项目（KF2015085） |
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| Establishment and optimization of tissue culture system of Fokienia hodginsii |
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ZHOU Chengcheng,YU Jianghong,CHEN Lingyan,et al
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| Abstract: |
| 【Objective】This study aimed to provide technical support for establishment of Fokienia hodginsii tissue culture system and explore optimal phytohormones of different explants in regeneration.【Method】Taking F. hodginsii leaf and shoot as explants,0.1% HgCl2 (5,8,10,12,15 min) was used to study the disinfection effects of different explants after soaking with detergent for 20 min and 75% alcohol for 30 s.Taking F. hodginsii leaf and shoot as materials,using MS medium as basic medium,the effects of NAA and 2,4-D with different concentrations on callus induction from different explants were analyzed,and the orthogonal experiment was used to study the influence of NAA,2,4-D and 6-BA ratios.Taking F. hodginsii calluses as materials, using MS medium as basic medium,the orthogonal experiment was used to study the influence of NAA,2,4-D,6-BA and KT ratios on subculture and proliferation of callus.Taking F. hodginsii shoot with bud as materials,using MS medium as basic medium,the effects of treatments with 6-BA at different concentrations on bud induction and germination were studied.Taking F. hodginsii differentiated seedlings as materials,using 1/2MS medium as basic medium,the orthogonal experiment was used to study the influence of 6-BA,2,4-D and KT ratios on subculture and growth of differentiated seedlings.Taking F. hodginsii rootless seedlings as materials,the propagation of rootless seedlings was also studied.【Result】（1） The optimal prescription of leaf disinfection was 75% alcohol treatment for 30 s and 0.1% HgCl2 treatm for 10 minutes with disinfection rate of 16.67%.The shoot prescription was 75% alcohol treatment for 30 s and 0.1% HgCl2 treatment for 15 min with disinfection rate of 23.33%.（2）The optimal phytohormones of leaf callus were MS medium,NAA 2.0 mg/L,2,4-D 1.0 mg/L and 6-BA 1.0 mg/L with induction rate of 81.67%, and the white calluses grew well.The shoot prescriptions were MS medium,6-BA 2.0 mg/L,2,4-D 1.0 mg/L and KT 0.5 mg/L with induction rate of 78.33%,and the green calluses grew well.（3）The optimal phytohormones of callus proliferation were MS medium,6-BA 2.0 mg/L,2,4-D 1.0 mg/L and KT 0.5 mg/L with proliferation weight of 1.984 g and browning rate of 23.33%.（4）The optimal phytohormones of bud induction and germination were MS medium and 6-BA 2.0 mg/L with average number of bud induction of 2.05,and average number of bud germination and differentiation of 1.70.（5） The optimal phytohormones of differentiated seedling growth were 1/2MS medium,KT 1.0 mg/L,2,4-D 0.5 mg/L with average length of 2.241 cm,no browning,propagation survival rate of 95.50% and propagation coefficient of 3.18.【Conclusion】Bud shoots were best explant for bud proliferation and regeneration pathway of F. hodginsii and leaves were the best explant for the callus regeneration pathway,which also optimized the tissue culture system. |
| Key words: Fokienia hodginsii callus regeneration in vitro induction and proliferation |
