摘要
沉香是国际上极负盛名的药用和香料资源之一。在亚洲许多国家,沉香产业既是传统行业也是发展迅猛的新兴产业。沉香优良种质的大规模应用对沉香产业的可持续发展具有重大意义。因此优良沉香种质资源是近年沉香产业关注的重点和热点,但一些优良种质的品质特性无法通过有性繁殖遗传,大规模繁育存在技术障碍。植物组织培养技术在品种优良性状保持、种子资源保存、快速繁殖等方面具有非常突出的优越性,被广泛应用于中药资源保护和中药产业发展。为建立工业化生产的沉香广谱再生体系,本文以白木香无菌苗和成龄植株的枝条为材料从外植体消毒、丛生芽诱导和生根培养三方面进行组织培养研究。结果表明,通过0.1%多菌灵溶液浸泡3 min,流动水冲洗3 h处理,对采摘自大田的白木香枝条有较好的除菌效果。0.1%升汞溶液消毒6~8 min,可有效保持外植体的成活率。在培养基中适当添加灭菌剂也能有效控制材料染菌。无菌苗茎段丛生芽诱导较易,大田枝条的丛生芽诱导较难。WPM+20 g/L蔗糖+0.5 mg/L 6-BA+0.2 mg/L NAA(G_(4))有利于无菌苗的丛生芽诱导;1/2 MS+25 g蔗糖+2 mg/L 6-BA+0.01 mg/L NAA(I_(3)),1/4 MS+20 g/L蔗糖+2 mg/L 6-BA+0.5 mg/L NAA(J_(2))和WPM+20 g蔗糖+1 mg/L 6-BA+0.01 mg/L NAA(K_(2))有利于成龄植株枝条外植体的丛生芽诱导,将膨大的丛生芽团切割后转接于不含植物生长调节剂的WPM培养基中可促进丛生芽伸长。WPM培养基中添加10~20 g/L蔗糖,并加入0.1~0.5 mg/L NAA可诱导无菌苗和茎段外植体的新芽生根。本研究以沉香2种外植体进行组织培养,成功获得再生植株,为沉香优良种质的无性繁殖提供技术参考。
Agarwood,Aquilaria sinensis(Lour.)Gilg,is one of the most famous medicinal and spice resources in the world.In many Asian countries,the agarwood industry is both a traditional and rapidly developing emerging industry.As we all know,the large-scale application of high-quality agarwood germplasm resources is of great significance for the sustainable development of the agarwood industry.Therefore,high quality agarwood germplasm resources have been the focus and hotspot of the agarwood industry in recent years.However,the characteristics of some high quality germplasm cannot be inherited through sexual reproduction,and the technical barriers for large-scale breeding exist in now.Tissue culture has outstanding advantages in aspects of maintaining excellent traits of varieties,preservation of seed resources,and rapid propagation,which is widely used in Chinese medicine resources conservation and traditional Chinese medicine industry development.In order to establish a broad-spectrum agarwood regeneration system for in-dustrial production,aseptic seedlings and mature plant branches were used to plant regeneration techniques research from explant disinfection,cluster bud induction and rooting culture in this study.The results showed that the tender stems picked in the field were effectively sterilized by soaked in 0.1%carbendazim solution for 3 min and rinsed with flowing water for 3 h.The survival rate of explants could be maintained by disinfecting with 0.1%mercury liter solution for 6~8 min.In addition,appropriate addition of sterilizer into the culture medium could also control material contami-nation effectively.The stems of aseptic seedlings were easier to induce new buds,while the stems of branch were more difficult to induce secondary shoots as explants.The multiple shoots of aseptic seedlings could induced in WPM medium supplemented with 0.5 mg/L 6-BA,0.2 mg/L NAA and 20 g/L sucrose(G_(4)).The three advisable multiplication media for branches were 1/2 MS+25 g/L sucrose+2 mg/L 6-BA+0.01 mg/L NAA(I_(3)),1/4 MS+20 g/L sucrose+2 mg/L 6-BA+0.5 mg/L NAA(J_(2))and WPM+20 g/L sucrose+1 mg/L 6-BA+0.01 mg/L NAA(K_(2)).Then the enlarged clump bud clusters were cut and transferred to WPM medium without plant growth additives which helped grow buds to become rootless shoots.New shoots could be induced to root in the WPM medium with adding 20-30 g/L sucrose 0.1-0.2 mg/L NAA.In this paper,two explants of agarwood were used for tissue culture and successfully obtained regenerated plants,which provided a technical reference for the asexual reproduction of high-quality agarwood germplasm.
作者
张燕
孟慧
吕菲菲
魏建和
樊小红
何欣
袁白雪
陈波
杨云
ZHANG Yan;MENG Hui;LYU Feifei;WEI Jianhe;FAN Xiaohong;HE Xin;YUAN Baixue;CHEN Bo;YANG Yun(Hainan Branch Institute of Medicinal Plant,Chinese Academy of Medical Sciences&Peking Union Medical College/Hainan Provincial Key Laboratory of Resources Conservation and Development of Southern Medicine/Key Laboratory of State Admini-stration of Traditional Chinese Medicine for Agarwood Sustainable Utilization,Haikou,Hainan 570311,China;Institute of Me-dicinal Plant Development,Chinese Academy of Medical Science&Peking Union Medical College/Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine,Ministry of Education/National Engineering Laboratory for Breeding of Endangered Medicinal Materials,Beijing 100193,China)
出处
《热带作物学报》
CSCD
北大核心
2022年第7期1467-1476,共10页
Chinese Journal of Tropical Crops
基金
国家重点研发计划项目(No.2018YFC1706401)。
关键词
白木香
组织培养
外植体消毒
丛生芽诱导
生根培养
Aquilaria sinensis(Lour.)Gilg
in vitro culture
explant disinfection
multiple bud induction
rooting culture
