Anther Culture of Blast-resistant Indica-japonica Hybrid, Yunyin
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摘要:
目的 花药培养在不同培养基成分构成的条件下，诱导率和成苗率等都存在显著差异。 通过优化培养基成分以及其他培养条件，提高云引籼粳杂交后代花药培养效率，为稻瘟病抗性品种云引的挖掘和利用奠定基础。 方法 以稻瘟病抗性粳稻品种云引作为母本，与常规籼稻品种粤晶丝苗进行杂交，F1代幼穗经过消毒后接种。根据不同基础培养基，碳源以及2,4-D质量设置了12种诱导培养基，经过比较出愈率、绿苗率、得苗率和花药培养力，进而确定最佳的培养条件。 结果 NB培养基的最高培养力仅为0.52%，而M8培养基的培养力0.90%～1.92%；以蔗糖为碳源的M8培养基早期出愈率可达到11.32%，添加1.0 mg·L−1 2,4-D和2.0 mg·L−1 2,4-D花药培养力相当，分别为1.89%和1.92%；而0.2 cm大小的愈伤组织进行分化时绿苗率最高，达到18.3%。 结论 云引籼粳交材料的最佳花药培养条件为：M8基础培养基，添加50 g·L−1蔗糖作为碳源和1.0～2.0 mg·L−1 2,4-D，当花药愈伤组织长至0.2 cm时转移至分化培养基分化。 Abstract:Objective To improve callus induction and seedling formation rates of the anther culture for an expedited and stable breeding program utilizing blast-resistant Indica-japonica hybrid rice Yunyin as a parent, an optimized medium and conditions for the auxiliary breeding technology in creating new materials and varieties were investigated. Method The young spikes of F1 hybrid of Yunyin and a conventional Indica rice Yuejingsimiao were disinfected and inoculated. From 12 formulations of base media, carbon sources, and 2,4-D concentrations as well as culture conditions, an optimal anther cultivation according to the rates of callus induction, green seedling, seedling yield, and anther cultivation performance was selected. Result The maximum anther cultivation performance using the NB medium was merely 0.52%, whereas, M8 significantly higher at 0.90%–1.92%. The M8 medium with sucrose for carbon source reached 11.32% on the performance at the early stage of callus induction. The use of 1.0 mg·L−1 or 2 mg·L−1 2,4-D in the medium did not differ significantly on the performance, i.e., 1.89% vs 1.92%. A peaked green seedling rate up to 18.3% was achieved when 0.2 cm-sized calluses were used for the differentiation. Conclusion The optimal culture conditions of Yunyin Indica-japonica hybrid anthers were to use calluses inducing on M8 medium with added 50 g·L−1 sucrose and 1–2 mg·L−1 2,4-D, then, when grew to 0.2 cm in size, transferring to a differentiation medium. -
Key words:
- anther culture /
- Indica-japonica hybrid /
- Yunyin /
- blast-resistant variety
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表 1 培养基成分
Table 1. Components in culture media
培养基编号
NO. of media基础培养基
Basic media碳源
Carbon source2,4-D质量浓度
Concentration of 2,4-D/（mg·L−1）NS05 NB 蔗糖 0.5 NS10 NB 蔗糖 1.0 NS20 NB 蔗糖 2.0 MS05 M8 蔗糖 0.5 MS10 M8 蔗糖 1.0 MS20 M8 蔗糖 2.0 NM05 NB 麦芽糖 0.5 NM10 NB 麦芽糖 1.0 NM20 NB 麦芽糖 2.0 MM05 M8 麦芽糖 0.5 MM10 M8 麦芽糖 1.0 MM20 M8 麦芽糖 2.0 
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表 2 不同培养基组合的花培力的比较
Table 2. Anther cultivation performances on media of varied formulations
培养基
Media基础培养基
Basic media出愈率
Rate of callus formation/%绿苗率
Rate of green seedling/%花药培养力
Anther culture ability/%NS05 NB 2.32 e N/A N/A NS10 NB 5.88 dc 8.64 d 0.51 d NS20 NB 7.34 bc 7.32 d 0.52 d MS05 M8 6.75 bc 19.6 b 1.32 b MS10 M8 13.32 a 14.2 c 1.89 a MS20 M8 16.75 a 11.5 c 1.92 a NM05 NB 1.35 f N/A N/A NM10 NB 1.32 f N/A N/A NM20 NB 3.28 e N/A N/A MM05 M8 3.75 e 24.3 a 0.90 c MM10 M8 6.36 c 21.3 ab 1.34 b MM20 M8 8.23 b 17.5 b 1.40 b 注：①同列数据后含不同字母者在5%水平上差异显著，表3同。②N/A表示样本过少而不被采用。
Note: ① Figures marked with same letter on a same column are not significantly different at 5% level. Same for Table 3. ② N/A indicates sample size is too low to be useful.
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表 3 不同诱导时间的出愈率比较
Table 3. Callus induction rates by varied induction time
培养基编号
NO. of media28 d出愈率
Rate of callus formation in 28 d/%42 d出愈率
Rate of callus formation in 42 d/%28 d出愈率/
42 d出愈率
Ration of callus formation (28 d versus 42 d) /%MS05 2.52 cd 4.25 d 59.29 MS10 7.25 b 9.35 b 77.54 MS20 11.32 a 14.50 a 78.07 MM05 1.45 d 3.25 d 44.62 MM10 3.22 c 5.35 cd 60.19 MM20 3.35 c 7.35 bc 45.58
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表 4 不同愈伤组织大小进行分化成苗得苗率的比较
Table 4. Seedling yields after differentiation using calluses of different sizes
(单位/%) 组别
Groups愈伤组织大小 Callus Size 0.1 cm 0.2 cm 0.4 cm 0.8 cm 全部 Total seedlings 23.2 63.2 85.5 83.6 绿苗 Green seedlings 4.1 18.3 13.3 5.35
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