Kim Jong Youn, Adhikari Prakash Babu, Ahn Chang Ho, Kim Dong Hwi, Chang Kim Young, Han Jung Yeon, Kondeti Subramanyam, Choi Yong Eui
Department of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University, Chuncheon, Republic of Korea.
Department of Herbal Crop Research, National Institute of Horticulture and Herbal Science, Rural Development Administration, Eumseong, Republic of Korea.
J Ginseng Res. 2019 Jan;43(1):38-48. doi: 10.1016/j.jgr.2017.08.002. Epub 2017 Aug 11.
Interspecific ginseng hybrid, × () has vigorous growth and produces larger roots than its parents. However, F progenies are complete male sterile. Plant tissue culture technology can circumvent the issue and propagate the hybrid.
Murashige and Skoog (MS) medium with different concentrations (0, 2, 4, and 6 mg/L) of 2,4-dichlorophenoxyacetic acid (2,4-D) was used for callus induction and somatic embryogenesis (SE). The embryos, after culturing on GA supplemented medium, were transferred to hormone free ½ Schenk and Hildebrandt (SH) medium. The developed taproots with dormant buds were treated with GA to break the bud dormancy, and transferred to soil. Hybrid plants were verified by random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analyses and by LC-IT-TOF-MS.
We conducted a comparative study of somatic embryogenesis (SE) in and its parents, and attempted to establish the soil transfer of propagated tap roots. The explants showed higher rate of embryogenesis (56% at 2 mg/L 2,4-D concentration) as well as higher number of embryos per explants (7 at the same 2,4-D concentration) compared to its either parents. The germinated embryos, after culturing on GA supplemented medium, were transferred to hormone free ½ SH medium to support the continued growth and kept until nutrient depletion induced senescence (NuDIS) of leaf defoliation occurred (4 months). By that time, thickened tap roots with well-developed lateral roots and dormant buds were obtained. All tap roots pretreated with 20 mg/L GA for at least a week produced new shoots after soil transfer. We selected the discriminatory RAPD and ISSR markers to find the interspecific ginseng hybrid among its parents. The F hybrid () contained species specific 2 ginsenosides (ginsenoside Rf in and pseudoginsenosides F in ), and higher amount of other ginsenosides than its parents.
Micropropagation of interspecific hybrid ginseng can give an opportunity for continuous production of plants.
种间人参杂种×（）生长旺盛，其根比亲本大。然而，F代完全雄性不育。植物组织培养技术可以解决这个问题并繁殖杂种。
使用含有不同浓度（0、2、4和6 mg/L）2,4-二氯苯氧乙酸（2,4-D）的Murashige和Skoog（MS）培养基进行愈伤组织诱导和体细胞胚胎发生（SE）。在添加GA的培养基上培养后的胚，转移到无激素的1/2 Schenk和Hildebrandt（SH）培养基上。将带有休眠芽的发育主根用GA处理以打破芽休眠，然后转移到土壤中。通过随机扩增多态性DNA（RAPD）和简单重复序列间（ISSR）分析以及液相色谱-离子阱-飞行时间质谱（LC-IT-TOF-MS）对杂种植物进行验证。
我们对和其亲本的体细胞胚胎发生（SE）进行了比较研究，并尝试建立繁殖的主根向土壤的转移。与任何一个亲本相比，外植体显示出更高的胚胎发生率（在2 mg/L 2,4-D浓度下约为56%）以及每个外植体更高的胚数（在相同的2,4-D浓度下约为7个）。在添加GA的培养基上培养后的萌发胚，转移到无激素的1/2 SH培养基上以支持其持续生长，并保持到叶片落叶的营养耗尽诱导衰老（NuDIS）发生（4个月）。到那时，获得了具有发达侧根和休眠芽的加粗主根。所有用20 mg/L GA预处理至少一周的主根在转移到土壤后都产生了新芽。我们选择了具有鉴别性的RAPD和ISSR标记来在其亲本中找到种间人参杂种。F杂种（）含有物种特异性的2种人参皂苷（中的人参皂苷Rf和中的假人参皂苷F），并且其他人参皂苷的含量比其亲本高。
种间杂种人参的微繁殖可为植物的持续生产提供机会。
