Encapsulation of upland cotton, Gossypium hirsutum L., axillary buds as artificial seeds
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摘要:目的
研究利用陆地棉Gossypium hirsutum L.营养芽制作人工种子的方法，为陆地棉人工种子技术开发应用提供一条新途径。
方法以陆地棉营养芽为外植体，利用海藻酸钠+MSB包埋外植体，CaCl2溶液凝固制作人工胚乳珠，再利用较高浓度海藻酸钠附加不同浓度的甲基纤维素包埋人工胚乳珠，用CaCl2溶液固化获得人工种皮，研究陆地棉营养芽人工种皮内添加抑菌剂后在有菌基质上的萌发效果。
结果制作人工种子的最佳外植体长度为5 mm; 最佳胚乳珠包埋方案为w为3%的海藻酸钠+MSB+0.5 mg·L-1 NAA+0.5 mg·L-1 IBA，在100 mmol·L-1 CaCl2中固化20 min; 人工种皮最佳配方为w为4%海藻酸钠+w为1%甲基纤维素+w为1.0%苯甲酸钠+0.02~0.04 mg·L-1青霉素+0.5 mg·L-1多效唑，在200 mmol·L-1 CaCl2固化20 min。该人工种子的无菌萌发率为95.0%，生根率为35.8%，可以在有菌人工基质中萌发成苗。
结论获得一条利用陆地棉营养芽制作人工种子的新途径，该技术制作的人工种子在有菌环境下有较高的萌发率并能成苗。
Abstract:ObjectiveTo provide a new manufacturing method for the artificial seeds of upland cotton, Gossypium hirsutum L., using axillary buds.
MethodThe axillary buds of upland cotton were used as explants. The explants were embedded by embedding solutions which were composed of different concentrations of sodium alginate and MSB after sterilization, and then immersed in different concentrations of CaCl2 solutions to solidify and obtain artificial endosperm beads. The artificial seed coat was obtained on the outer synthetic endosperm beads which were immersed in the solution composed of high concentration of sodium alginate and methyl cellulose, and then immersed in CaCl2 liquid. After added the bacteriostatic agent to the artificial seed coat, the seed germination experiment was done in nonsterile substrate.
ResultThe axillary buds could be made into artificial seeds after disinfection treatment, and the optimal length of explant was 5 mm. The optimal embedding program was found to be the combination of 3 % sodium alginate+MSB+0.5 mg·L-1 NAA+0.5 mg·L-1 IBA, and solidified in 100 mmol·L-1 CaCl2 for 20 min. The optimal combination of artificial seed coat was the solution of 4 % sodium alginate+1% methyl cellulose+1.0% sodium benzoate+0.02-0.04 mg·L-1 penicillin+0.5 mg·L-1 paclobutrazol, and solidified in 200 mmol·L-1 CaCl2 for 20 min. The germination rate of synthetic seed was 95.0 %, the rooting rate was 35.8 %.The artificial seed could germinate and grow into seedling in nonsterile substrate.
ConclusionA new asexual reproduction technology making artificial seed is obtained using upland cotton axillary bud, and the artificial seed can be converted into seedling in nonsterile substrate.
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Keywords:
- Gossypium hirsutum /
- artificial endosperm /
- sodium alginate /
- paclobutrazol /
- axillary bud
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图 1 处理前后的陆地棉营养芽
a:植株上的营养芽; b:处理后的营养芽。
Figure 1. The axillary buds of Gossypium hirsutum before and after processing
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图 2 陆地棉营养芽人工种子发育成苗
a:制备好的人工种子; b:人工种子在MS培养基上萌发; c:人工种子3周开始生根; d: 4周后形成人工种子萌发小苗。
Figure 2. Plantlet regeneration from encapsulated axillary buds of Gossypium hirsutum
表 1 海藻酸钠和CaCl2浓度对人工种子成型及萌发影响1)
Table 1 Effects of different concentrations of sodium alginate and CaCl2 on germination of encapsulated axillary buds
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表 2 人工胚乳中激素对种子萌发生根的影响1)
Table 2 Effects of different growth regulators on synthetic seed germination and root formation
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表 3 营养芽长度对人工种子萌发成活的影响1)
Table 3 Effect of nutrition bud length on the survival of artificial seed germination
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表 4 海藻酸钠与甲基纤维素混合种皮对种子失水率的影响1)
Table 4 Effects of different concentrations of sodium alginate and methyl cellulose on water loss rates
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表 5 人工种子种皮添加抑菌剂对种子污染和生根的影响1)
Table 5 Effects of bacteriostatic agents added into artificial seed coat on pollution and root of seed
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