The characteristics of the thornless evergreen cultivar multiplication through micro-cloning
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LOZINSCHII, Mariana; CHIORCHINA, Nina. The characteristics of the thornless evergreen cultivar multiplication through micro-cloning. In: Conservation of plant diversity. Ediția 4-a, 28-30 septembrie 2015, Chișinău. Chișinău: Gradina Botanica (Institut), 2015, p. 75. ISBN 978-9975-3036-8-2.
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Conservation of plant diversity
Ediția 4-a, 2015
Simpozionul "Conservation of plant diversity"
Chișinău, Moldova, 28-30 septembrie 2015

The characteristics of the thornless evergreen cultivar multiplication through micro-cloning

Pag. 75-75

Lozinschii Mariana, Chiorchina Nina
Botanical Garden (Institute) of the Academy of Sciences of Moldova
Disponibil în IBN: 15 mai 2019

Thornless, in vitro, medium, culture, multiplication


 Blackberries are native to Asia, Europe, North America and South America. Blackberries have been used in Europe for over 2000 years in pharmaceutical and food industries. Blackberries, often called "berries", are a diverse group of species and hybrids of the Rubus genus. They are members of the Rosaceae family, subfamily Rosoideae. Rubus lacinatus with its Thornless evergreen representative is a European cultivar from North America; its branches can reach up to 6 m in length. With leaves pinnately veined, with white flowers with shades of pink, having shiny round black fruit weighted 4.5 g, it has a particular flavor. This shrub can be grown for ornamental purposes having a decorative appearance due to its vigorous evergreen leaves.         In vitro tissue culture is considered to be the most effective method for multiplication of blackberries [1]. In result, we have a material with a strong resistance to diseases, ability of quick multiplication, a healthy material with the same genetic lines. Before placing it into the in vitro culture, an important step was to do the sterilization of the plant material taken from the donor plant which is an essential contribution to the initial stage of the in vitro culture. The best period of time for all types of explants while treating them with diacid is 7 minutes. The inoculation of different types of meristems led to the start of the morphogenesis and organogenesis processes. The medium created for inoculation was MS 100 % modified. The period between passing is approximately one month, when the plant material increases considerably (10 times) in this time limit.         The following medium has a positive influence on the Thornless evergreen cultivar. It was noted that on medium with naphthaleneacetic acid (NAA) 0.1 mg / l supplement, a broader development of the vegetative material is observed after 12 to 15 days. At this time, the shoot length is 5 cm. In comparison with other varieties of shrubs, the Thornless evergreen cultivar has a better observed rootedness. Rootedness was stimulated by filling the nutrient medium with indolle-3-byric acid (IBA) and NAA, the formation of roots was visible after 10-12 days of cultivation. On medium with IBA are developing roots and is observed a white callus, but on mediums with NAA is developing radicular system and the plant in full. To optimize the mediums through  micropropagation of this cultivar we used medium MS 50 % with 30 g commercial sugar, liquid with  NAA in concentration of  0.1, 0.2 mg/ l, on this medium shoot grows 12 cm in 30 days and takes roots in 14 days. The cytokinins 6-benzyl aminoppurine (BAP) used in different concentrations 0.1, 0.3, 0.5, 0.7 had an important role in the initiation of morphogenesis. The most effective multiplication medium for the Thornless evergreen is the MS 100% supplemented with BAP (0.5 mg / l). In that medium, for a period of over 50-55 days, a quantity of 19 to 20 adventitious shoots with a length of 7 cm was obtained.   During the ex vitro acclimatization process, the substrate in which we plant is taken in great consideration. For the ex vitro transfer, the multiplied plantlets were taken from culture tubes, their base has been washed with hot water in order to remove the culture medium and the base roots were slightly cut to better stimulate the rooting process. The plantlets, which previously passed through a weak pink solution of KMnO4, are transferred into a solid substrate. The most appropriate and optimal substrate for plant growth and development was composed only of commercial peat with a pH of 5.8-6.5. Plants with substrate are planted in palettes composed of cells provided with holes. These cells are filled with a mixture of substrate in advance. The plant material is planted in the palettes for the ex vitro process, the palettes are enwrapped in transparent sheets to maintain humidity. For all cultivars it is very important the substrate to be as drained as possible, otherwise the percentage of plant survival is minimal. This culture has a growth rate of acclimatization 89-90% in winter months, but 95-99% – in summer months.         As a result of the investigations it has been established that the tissue culture for the Thornless evergreen cultivar was successfully used, being improved with time, this allowing to obtain perfectly healthy plants and high efficiency of seedlings. Nowadays, this method has turned into a modern technique for rapid multiplication of different species and cultivars including the one described above.