李志瑛 穆治华 肖勇 范海阔

摘  要：本文對几个影响矮种椰子成熟胚的愈伤组织诱导及褐化程度的因素进行研究。以海南优良矮种椰子‘文椰2号和‘文椰4号11月龄的成熟胚为材料，研究不同的基因型、凝固剂、激素浓度和接种方式对愈伤组织的诱导率和外植体褐化程度的影响。结果显示：以‘文椰2号成熟胚为材料，愈伤率最高个体为H1;接种至PhytagelTM-P8169凝固剂的愈伤率最高，为71.43%;接种至Gelrite-G1910凝固剂的褐化率最低，为40.00%;接种至添加110 μmol/L 2，4-D的培养基的愈伤率最高，为63%，褐化率最低，为46.67%;剥取胚芽接种的愈伤率最高，为48.72%，其褐化率与完整胚接种的无显著性差异。以‘文椰4号成熟胚为材料，愈伤率最高个体为X1;接种至Agar-A8190凝固剂的愈伤率和褐化率最高，分别为46.15%和30.77%;接种至Gelrite-G1910凝固剂的褐化率最低，为16.67%;接种至添加600 μmol/L 2，4-D的培养基愈伤率最高，为40.00%，但与110 μmol/L 2，4-D无显著性差异;接种至110 μmol/L 2，4-D的褐化率最低，为50.00%;纵切接种获得愈伤率最高，为40.00%;完整胚接种褐化率最低，为33.33%。结果表明：基因型、凝固剂、激素浓度和外植体接种方式对愈伤组织的诱导率均有显著影响。‘文椰2号愈伤组织诱导的最佳2，4-D浓度为110 μmol/L，最佳凝固剂为PhytagelTM-P8169，最佳接种方法为剥取胚芽接种。‘文椰4号愈伤组织诱导的最佳2，4-D浓度为110 μmol/L，最佳凝固剂为Agar-A8190，最佳接种方法为胚纵切接种。

关键词：椰子;愈伤组织;组织培养

Abstract： Tissue culture technique is an efficient way to speed-up coconut multiplication. In this study， the factors limiting callus induction efficiency and the method to control browning rate were studied. Improvement of callus induction efficiency of coconut and the theoretical basis for an optimized condition of induction were presented. The 11-month-old mature embryos of ‘Wenye 2 and ‘Wenye 4， two elite dwarf coconut cultivars， were used as the explant materials. Experiments was conducted to evaluate the influence of genotype， optimum hormone concentration， gelling agent and inoculation method on callus induction and explant browning. When using the embryos from ‘Wenye 2 as the explant， a higher callus induction rate （83.33%） was from H1. The highest number of calluses were observed in the medium with 4 g/L PhytagelTM-P8169 （71.43%） and 110 μmol/L 2，4-D （63.00%） respectively， while rest treatments showed no significant difference. The highest rate of callus induction （48.72%） was observed in the treatment using plumule isolation techniques. As to the browning rate， the lowest were found in the medium containing 3 g/L Gelrite-G1910 （40.00%） and 110 μmol/L 2，4-D （46.67%） and in the plumule isolation treatment （66.67%）. When using the embryos from ‘Wenye 4 as the explant， the improved treatments for callus induction of coconut were genetics X1 （66.67%）， 6 g/L Agar-A8190 （46.15%）， 600 μmol/L 2，4-D （40.00%） and longitudinal cutting （58.33%）， but the difference was not significant between 600 μmol/L 2，4-D and 110 μmol/L 2，4-D. As to the browning， the lowest browning rate was showed in the medium containing 3 g/L Gelrite-G1910 （16.67%） and 110 μmol/L 2，4-D （50.00%） and in the induction method of embryo （33.33%）. This study explored that genotype of parent plants， the concentration of 2，4-D， gelling agent and condition of induction also had significant effects on the rate of callus induction and ‘Wenye 2 was better as the explant than ‘Wenye 4. For the callus induction of ‘Wenye 2， the suitable hormone concentration， gelling agent and inoculation were 110 μmol/L 2，4-D， PhytagelTM-P8169 and inducing plumule. For ‘Wenye 4， 110 μmol/L 2，4-D， Agar-A8190 and longitudinal embryo cutting techniques were the best for callus induction.

Keywords： coconut; callus; tissue culture

椰子（Cocos nucifera L.）为棕榈科椰子属多年生乔木，是典型的热带木本油料作物和食品作物，也是热带沿海生态林建设和园林绿化的重要植物之一，主要分布在我国海南地区。……