scholarly journals Optimization of nutrient media for clonal micropropagation of the rootstock grape variety ‘Kober 5BB’

2020 ◽  
pp. 298-303
Author(s):  
Игорь Владимирович Гавриленко ◽  
Юлия Сергеевна Матяш ◽  
Анжела Владимировна Гавриленко ◽  
Дмитрий Александрович Шанин ◽  
Ирина Александровна Павлова ◽  
...  

Сорт винограда Кобер 5 ББ (Берландиери x Рипариа Кобер 5ББ) - один из основных подвоев, используемых в питомниководстве для получения привитых саженцев, поэтому в настоящее время крайне актуально создание маточников данного подвоя посадочным материалом категории «оригинальный». Это объясняет необходимость проведения исследований, связанных с оптимизацией условий культивирования сорта Кобер 5ББ, для повышения эффективности массового клонального микроразмножения с сохранением его генетической однородности и стабильности. Целью исследования являлась оптимизация и подбор питательных сред для клонального микроразмножения сорта-подвоя Кобер 5 ББ на этапе тиражирования (микрочеренкования). Материалом для исследований служили растения in vitro сорта подвоя Кобер 5ББ, свободные от основной патогенной инфекции (по результатам тестирования). Исследования проводили на средах: МS; WPM; DKW; PG (контроль). В качестве регуляторов роста использовали GA (гиббереллиновая кислота) в концентрациях: 0,2; 0,6; 1; 1,4 мг/л в сочетании с NAA (α-нафтилуксусная кислота) 0,05 мг/л. Показано, что растения на среде WPM, содержащей NAA-0,05 мг/л, по биометрическим показателям превосходили развившиеся на среде PG с аналогичным гормональным составом. Проведенные исследования по оптимизации среды культивирования для ускорения ростовых процессов позволили по результатам биометрических показателей выделить блок сред с основой WPM для размножения сорта-подвоя Кобер 5 ББ на этапе микрочеренкования. После проведения дополнительных исследований с расширенной выборкой среду WPM можно будет рекомендовать для клонального микроразмножения винограда на этапе микрочеренкования. The grape variety ‘Kober 5 BB’ (‘Berlandieri x Riparia Kober 5BB’) is one of the main rootstocks used in rootstock-growing farming to obtain grafted seedlings. Currently it is a hot issue to create nurseries for the rootstock grapevine with planting material of the "original" category. This explains the need for research related to optimization of cultivation conditions of the variety ‘Kober 5BB’ to increase the efficiency of mass clonal micropropagation while retaining its genetic homogeny and stability. The aim of the study was to optimize and select nutrient media for clonal micropropagation of the rootstock variety ‘Kober 5 BB’ at the stage of tiraging (micropropagation by cutting). The material of research was the in vitro plants of ‘Kober 5BB’ rootstock variety, free from basic pathogenic infection (according to the test results). The studies were carried out on media: MS; WPM; DKW; PG (control). Gibberellic acid (GA) was used as a growth regulator at concentrations: 0.2; 0.6; 1; 1.4 mg/l in combination with NAA (α-naphthyl acetic acid) 0.05 mg/l. Plants on WPM medium containing NAA-0.05 mg/l in biometric parameters were superior to those grown on PG medium with a similar hormonal composition. According to the results of biometric parameters the studies on optimization the culture medium for accelerating growth processes made it possible to isolate a group of media with a WPM base for propagation of the rootstock variety ‘Kober 5 BB’ at the stage of microcutting. After additional studies with expanded selection, the WPM medium can be recommended for clonal micropropagation of grapes at the stage of microcutting.

2014 ◽  
Vol 962-965 ◽  
pp. 681-690 ◽  
Author(s):  
Dmitriy Nikolaevich Zontikov ◽  
Svetlana Zontikova ◽  
Roman Sergeev ◽  
Alex Shurgin ◽  
M. Sirotina

The stages of getting the planting material for laying plantations of the triploid aspen (Populustremula L.) are illustrated in this work. The source material selected from the genetic fund “The gigantic aspens” is characterized. The peculiarities of using the method of clonal micropropagation when getting the planting material, the usage of a rootstock as a source of donor explants for the introduction into the culture in vitro, the influence of a kind of nutrient media and growth regulators NAA and BAP on the growth and the development of microsprouts. The distinctions in the speed of the growth of the culture in vitro of diploid and triploid clones have been analyzed. The adaptation of the plants-regenerants to the soil-conditions.


2021 ◽  
Vol 12 (3) ◽  
Author(s):  
Y Kolomiiets ◽  
◽  
A Skuba ◽  

The study presents the results of obtaining regenerated plants of asparagus from seeds. Surface sterilizing the seeds by 0,75% sodium hypochlorite for 30 min is effective, during this obtained 83% viable sterile plants. The Murashige and Skoog medium supplemented with 6‑benzylaminopurine (2 mg/L), inositol (100 mg/L) and thiamine (0,4 mg/L) was found to be the best for seed germination. The expediency of using kinetin (1 mg/L) as a growth regulator to obtain a homogeneous plant material was established. The reproduction coefficient was 6,0. Only 11% of the explants formed callus. For the selection needs and production of somaclonal variants, the use of the culture medium with indole-3-acetic acid (0,2 mg/L) and 6‑benzylaminopurine (1 mg/L) is justified. In this condition reproduction coefficient was 3,7, and the level of different intensity callusogenesis was 59%. The rooting of obtained plants was performed in Murashige and Skoog medium supplemented with a half dose of macro- and micronutrients and growth regulators. Rooting frequency was up to 63%. The knowledge of hormonal requirements helps to promote isolated tissue and cells technologies of asparagus with purpose of rapid propagation and obtaining healthy, high-quality planting material.


2021 ◽  
Vol 12 (2) ◽  
pp. 71-80
Author(s):  
S. Yu. Bilous ◽  
O. O. Oliinyk ◽  
O. O. Hunko

The peculiarities of obtaining planting material of rare representatives Drosera spatulate L. and Drosera aliciae L. using microclonal propagation in order to preserve and cultivate them in ex vitro conditions were studied. The method of sterilization of D. spatulate and D. aliciae explants with 80-90% obtaining aseptic material has been developed. The influence of different sterilization options on the development of microshoots has been studied. The best mode of sterilization is 0,1% solution of AgNO3 and 12.5% solution of H2O2. The features of organogenesis and regeneration of the whole organism from cultivated tissues and organs of Drosera L. was investigated. The effect of exogenous growth regulators at different stages of plant morphogenesis in vitro is shown. Improved conditions of rhizogenesis in vitro. It was found experimentally that MS nutrient media with the addition of 2 g∙l-1 PVP is optimal at the stage of introduction into culture in vitro D. spatulate and D. aliciae. The regeneration of microshoots of D. spatulate and D. aliciae depending on the type of explant and the composition of nutrient media was studied. Morphogenesis was most effective on nutrient media with the addition of 0.25 mg∙l-1 kinetin and on the hormone free MS media. Such cultivation conditions provided 100% regeneration of plants with a reproduction rate of 1:8. Studying the effect of cytokinins on the microclonal reproduction of D. spatulate and D. aliciae, it was found that the development and induction of multiple shoot formation in vitro is best performed on hormone free MS media. To induce the formation of the root system, it is necessary to add into MS nutrient media 0.5 mg∙l-1 IBA. According to the results of the research, a method of microclonal propagation was developed by cutting stem culture, which made it possible to obtain genetically stable, disease-free regenerating plants of D. spatulate and D. aliciae with an optimally formed root system and vegetative mass. The obtained homogeneous planting material can be used in floriculture, creation of terrariums, for pharmacological purposes and for the purpose of introduction. Keywords: Drosera L., microclonal reproduction, morphogenes.


2022 ◽  
Vol 51 (4) ◽  
pp. 712-722
Author(s):  
Elena Kulikova ◽  
Sergey Makarov ◽  
Irina Kuznetsova ◽  
Anton Chudetsky

Introduction. The demand for honeysuckle berries and planting material is growing. Clonal micropropagation is the most effective method for industrial plantations. The research objective was to study the effect of cytokinins and auxins on Russian and Canadian honeysuckle microshoots and roots. Study objects and methods. The study featured regenerated honeysuckle (Lonicera edulis Turcz.) of three Russian cultivars (Bakcharsky Velikan, Doch Velikana, Yugana) and two Canadian cultivars (Boreal Beauty, Boreal Beast). The experiment focused on the effect of sterilizing agents and sterilization time on the viability of honeysuckle explants at the stage of culture introduction in vitro. The effect of the growth regulator Cytodef in the QL nutrient medium on organogenesis was studied at the stage of micropropagation proper, the effect of auxin IBA on plant root formation – at the stage of rooting in vitro. Results and discussion. The greatest viability of honeysuckle explants (80–94%) was registered in the samples affected by Lizoformin 3000 (5%) and silver nitrate (0.2%) as sterilizing agents with a sterilization time of 10 min at the stage of in vitro culture introduction. The biggest quantity (8.8 pcs.) and total length (40.1 cm) of microshoots were observed when the content of cytokinin Cytodef in the culture medium QL was 0.3 mg/L at the stage micropropagation proper. The Boreal Beast cultivar had the largest total length of shoots (29.0 cm). The biggest quantity (5.5 pcs.) and total length (30.8 cm) of roots resulted from 0.5 mg/L of auxin IBA at the stage of rooting in vitro. Coconut substrate produced the highest survival rate (92–99%) at the stage of adaptation to non-sterile conditions in vivo, with the greatest number of leaves (8.1–10.2 pcs.) observed in Canadian cultivars. Conclusion. Cytodef and IBA proved to be effective growth-regulating substances for microplants of Russian and Canadian honeysuckle cultivars in vitro, which makes them promising for berry plantations.


2021 ◽  
Vol 937 (4) ◽  
pp. 042093
Author(s):  
O V Yakimova ◽  
N A Yegorova

Abstract Melissa officinalis L. is a perennial herbaceous essential and medicinal plant widely used in pharmacology, perfumery and cosmetics, as well as in alcoholic beverage and food industries. The low content of essential oil in lemon balm raw material determines the selection work aimed at creating high-oil cultivars. The use of clonal micropropagation method in vitro will increase the efficiency of this process and accelerate promising breeding samples multiplication. The aim of our research was to study the influence of cultivation conditions and cultivar on the M. officinalis in vitro rhizogenesis and ex vitro adaptation. It was found that the maximum frequency of shoot rooting (up to 93.3%) in cultivars ‘Citronella’ and ‘Sobornaya’ was on MS culture medium supplemented with 0.5 mg/l NAA. The number of roots was 10.1 and 13.6 pcs. per shoot, respectively. The highest rates of root formation for the cv. ‘Crimchanka’ was found on a culture medium supplemented with 1.0 mg/l IAA (8.7 roots per shoot). The mixture of peat, sand and perlite (2:1:2) as a substrate provided up to 93% of adapted ex vitro lemon balm microplants. The presented studies were used to develop a technique for clonal micropropagation of M. officinalis.


2021 ◽  
pp. 301-308
Author(s):  
Olesya Nikolayevna Mazko ◽  
Lyudmila Ivanovna Tikhomirova ◽  
Lyudmila Vladimirovna Shcherbakova ◽  
Natal'ya Grigor'yevna Bazarnova ◽  
Dmitriy Alekseyevich Karpitsky

The aim of this study was to evaluate the effect of 6-benzylaminopurine (BAP) separately and in interaction with auxins on the change in the qualitative and quantitative composition of flavonoids in the raw materials of regenerating plants Iris sibirica L. Cambridge grade in comparison with aeroponic and intact raw materials using the method of high-performance liquid chromatography. Raw materials of I. sibirica Cambridge variety obtained in vitro culture had a richer qualitative composition of flavonoids than intact plants. The dependence of the accumulation of flavonoids on the concentration of 6-benzylaminopurine in nutrient media was noted. The presence of 13 compounds was observed in extracts of 70% ethyl alcohol from regenerating plants grown at the lowest concentration of BAP (1.0 µM) within the experiment. In quantitative terms, the flavonoid apigenin was maximally determined on a medium with BAP 1 µM, and kaempferol - on media with BAP 5.0 µM, supplemented with auxins. For a medium with 7.5 µM BAP, the lowest variety of compounds was observed (9) and the lowest kaempferol content. Auxins influenced the synthesis of flavonoids. The amount of flavonoids in all variants of the experiment increased by an average of 13% in the presence of auxins. The stages of the technological process of obtaining raw materials I. sibirica Cambridge variety on the basis of clonal micropropagation and cultivation in aeroponics conditions allowed to obtain raw materials that do not contain heavy and toxic metals, are not infected with pathogens and pests. With 1 m2 of useful area of aeroponics for 1 year, it is possible to collect 5 times more raw materials than with field cultivation. According to the qualitative composition of phenolic compounds, aeroponic raw materials are identical to intact plants.


Author(s):  
Sergey Makarov ◽  
Sergey Rodin ◽  
Irina Kuznetsova ◽  
Anton Chudetsky ◽  
Svetlana Tsaregradskaya

Introduction. Forest berry plants are popular on the food market and in pharmacy for their high nutritional and medicinal value. Plantations of forest berry plants can proliferate on unused lands, including depleted peatlands. Clonal micropropagation is the most effective method for obtaining large quantities of high quality planting material. Light-emitting diodes are highly effective for clonal micropropagation. The research objective was to study the effect of different spectral ranges on the process of root formation of forest berry plants in vitro. Study objects and methods. The research featured regenerant plants of half-highbush blueberry, arctic bramble, American cranberry, European cranberry, lingonberry, and Kamchatka bilberry of different cultivars. A set of experiments made it possible to study the effect of lighting type on the growth and development of the root system of forest berry plants in vitro using white fluorescent lamps, white spectrum LED lamps, and LED lamps with a combination of white, red, and blue spectra at the in vitro rooting stage of clonal micropropagation. Results and its discussion. The largest number (3.4–14.6 pcs.) and the maximum total length (10.0–156.9 cm) of roots were observed under LED lamps with a combination of white, red, and blue spectra. The effect was by 1.1–2.8 and 2.0–4.5 times higher than in the case of white-spectrum LED lamps, and by 2.3–7.0 and 3.3–14.9 times than in the case of fluorescent lamps. Variety and shape proved to have no significant effect on biometric indicators. Conclusion. LED lamps had a positive effect on the process of rhizogenesis of forest berry plants during clonal micropropagation. They appeared to be more effective than fluorescent lamps. The combination of white, blue, and red spectra increased the biometric parameters of plants at the stage of in vitro rooting.


2021 ◽  
Vol 34 ◽  
pp. 03002
Author(s):  
Viktor Klimenko ◽  
Ekaterina Lushchay ◽  
Valeryi Zlenko

In vitro experimental plants obtained by clonal micropropagation of 9 grape somaclones of 5 original forms were the material for cytogenetic research. A biological microscope XSP-146TP was used for cytogenetic analysis. 823 cases of deviation from diploidy were observed in total. Significant tissue ploidy was observed in the meristematic tissue of in vitro plants of grape somaclones obtained by colchicine treatment of proembryogenic cells of various varieties. The significant direct correlation was found between the frequency of polyploidy in meristem tissues of in vitro plants and the number of chloroplasts in the stomata of grape somaclones. The reverse correlation was found between the frequency of polyploidy and the number of stomata on the leaf area. Somaclone No. 72, obtained as a result of regeneration from colchicinated proembryogenic cells of the Ruta grape variety and identified as a tetraploid (2n = 4x = 76), is recommended for use in the polyploid creation program.


2021 ◽  
Vol 51 (1) ◽  
pp. 67-76
Author(s):  
Sergey Makarov ◽  
Irina Kuznetsova ◽  
Mikhail Upadyshev ◽  
Sergey Rodin ◽  
Anton Chudetsky

Introduction. The last decade saw a considerable increase in the demand for European cranberry planting material (Oxyccocus palustris Pers.) among consumers of non-timber forest products. Cranberry possesses high nutritional and medicinal value. Cultivars and hybrids of European cranberry prove extremely productive for plantation growth using the method of clonal micropropagation with revitalized planting material. Study objects and methods. The research featured European cranberry plants of the Dar Kostromy cultivar and its hybrid form 1-15-635. The study focused on the effect of various medications and growth regulators on the biometric profile of European cranberry and its adaptation to non-sterile conditions at all stages of in vivo clonal micropropagation. Results and discussion. During the introduction stage, the highest viability belonged to the explants treated with AgNO3 (95–96%) and Lizoformin 3000 (5%) as the main sterilizing solutions at a 10-min exposure and a 5% solution of Ecosterilizer (1:1) at a 20-min exposure (90–95%). During the micropropagation proper, the number, average length, and total growth of shoots increased as the concentration of cytokinin 2ip in the WPM 1/4 nutrient medium rose from 1.0 to 5.0 mg/L. At the stage of in vitro rooting, the maximal number, average length, and total growth of roots in regenerated plants for both cultivars were observed when Kornerost 5.0 mg/L was added to the WPM 1/4 nutrient medium. At the stage of adaptation to in vivo conditions, Micogel 0.2 mg/L contributed to the highest survival rate (94–100%). Conclusion. During clonal micropropagation in vitro, the biometric profile of European cranberry (Oxyccocus palustris Pers.) and its survival rate under non-sterile conditions in vivo proved to depend on various growth-regulating substances and their concentrations.


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