Establishing the in vitro culture of and micropropagating edible honeysuckle

T. I. Khoruzheva, S. Borovaya, N. G. Boginskaya
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Abstract

Edible honeysuckle is a popular fruit crop. Its therapeutic and health-promoting effects are attributed to a high content of bioactive compounds in the fruits. Unlike the traditional plant multiplication methods, the in vitro propagation allows scientists to obtain high-quality planting material of honeysuckle in a great quantity and within a short time. The research was carried out at the Laboratory of Breeding and Genetic Research on Field Crops of the Federal Scientific Center of Agricultural Biotechnology of the Far East named after A.K. Chaiki. Honeysuckle variety Podarok amurchanam created by the Far Eastern State Agrarian University was used as the research object. The research materials were sterilized according to the methodology of N.I. Vavilov All-Russian Institute of Plant Genetic Resources with some modifications. Several products were used as chemical agents for sterilization in the following sequence: a 5% solution of surfactants, fungicide Fundazol, EC (1 g/l), the bleaching agent ACE freshly diluted with distilled water in the proportion 1:9 (0.50% of NaOCl in the working solution), and 70% ethanol. The primary explants were cultured on an MS containing 20 g/l sucrose and 6 g/l agar (hereafter – MS) and supplemented with 6-benzylaminopurine (BA) at a concentration of 0.5 mg/l. The pH of the medium was adjusted to 5.7-5.8 using 1N КОН. The explants (microcuttings with one-two internodes) were subcultured on an MS supplemented with BA (0.5 mg/l). The morphometric parameters of the plants were measured on the 35th day of cultivation. The sterilization of the explants with Fundazol (1 g/l) and the ACE diluted with distilled water in the proportion 1:9 allowed us to obtain a high number of viable microclones (50%). The elimination of leaves from the honeysuckle microcuttings drastically decreased the survival rate and led to the death of the microclones in most cases (the mortality rate was 98.7 %). Subculturing the microcuttings on the MS supplemented with BA at a concentration of 0.5 mg/l facilitated the normal growth and development of the regenerated honeysuckle plants (the average reproduction rate was 4.65).
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建立食用金银花的离体培养和微繁殖技术
食用金银花是一种广受欢迎的水果作物。其治疗和促进健康的功效归功于果实中高浓度的生物活性化合物。与传统的植物繁殖方法不同,体外繁殖使科学家能够在短时间内获得大量高质量的金银花种植材料。这项研究是在以 A.K. Chaiki 命名的远东农业生物技术联邦科学中心的大田作物育种和遗传研究实验室进行的。研究对象是远东国立农业大学培育的金银花品种 Podarok amurchanam。研究材料按照 N.I. Vavilov 全俄植物遗传资源研究所的方法进行灭菌,并做了一些修改。灭菌化学制剂的使用顺序如下:5% 的表面活性剂溶液、杀菌剂 Fundazol, EC(1 克/升)、用蒸馏水按 1:9 比例稀释的漂白剂 ACE(工作溶液中含有 0.50%的 NaOCl)和 70%的乙醇。初级外植体在含有 20 克/升蔗糖和 6 克/升琼脂的 MS(以下简称 MS)上培养,并添加浓度为 0.5 毫克/升的 6-苄基氨基嘌呤(BA)。培养基的 pH 值用 1N КОН 调节到 5.7-5.8。将外植体(一至两个节间的微切片)移栽到添加了 BA(0.5 毫克/升)的 MS 培养基上。在培养的第 35 天测量植株的形态参数。用 Fundazol(1 克/升)对外植体进行消毒,再用蒸馏水按 1:9 的比例稀释 ACE,这样就能获得大量存活的微克隆(50%)。去除金银花微切片中的叶片会大大降低存活率,在大多数情况下会导致微克隆死亡(死亡率为 98.7%)。在添加了 0.5 毫克/升浓度 BA 的 MS 上对微切片进行分培,有利于再生金银花植株的正常生长和发育(平均繁殖率为 4.65)。
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