Micropropagation of berry crops for creation of germplasm cryobanks / Micropropagação de fruteiras para criação de criobancos de germoplasma

One of the main stages of cryopreservation of meristematic tissues in vegetative plants is a clonal micropropagation, which includes isolating the explants of the raw material in vitro and optimizing the culture medium for micropropagation. As the result of our studies, the optimal periods for in vitro micropropagation are: first - isolation of explants from initiated shoots of dormant buds (blackcurrants and raspberries) in January-March; the second - from actively growing shoots (blackcurrants and raspberries) in May-June, from the formed mustache (strawberry) in July-August. The optimal drugs for sterilization of raspberry explants are: a) 0.1% HgCl2 (6 min), then 3% H2O2 (15 min); b) chlorine-containing bleach «Domestos¼ in the dilution of H2O 1:9 (10 min). For blackcurrant: a) 0.1% HgCl2 (5 min) in combination with 0.1% fungicide "Topaz" (30 min); b) 0.1% HgCl2 (5 min) in combination with the treatment with KMnO4 (30 min); c) "Domestos" in the dilution of H2O 1:5 (20 min). For strawberry: a) 0.1% HgCl2 (6 min) followed by treatment with 3% H2O2 10 (min); b) 1% deochlor (7 min), 3% H2O2 (10 min); c) "Domestos" in the dilution of H2O 1:5 (8 min) with subsequent treatment 0,1% HgCl2 -7 min, then 0,20 mg/l КМnO4 - 30 min. Optimal compositions of culture media for micropropagation of blackcurrant - Murashige and Skoog (MS) medium with 0.5 mg L-1 BAP, 0.5 mg L-1 GA3, 0.1 mg L-1 IBA and 20 g L-1 glucose. For raspberry -MS medium with 0.5 mg L-1 BAP, 0.1 mg L-1 IBA, 10 mg L-1 iron chelate and 30 g L-1 sucrose. For strawberry - MS medium with 0.3 mg L-1 BAP, 0.01 mg L-1 IBA, 0.2 mg L-1 GA3, 10 mg L-1 iron chelate and 30 g L-1 sucrose. Based on these studies, the cryobank was created, which include the germplasm of in vitro meristematic tissues in 66 cultivars, hybrids and wild-growing forms of blackcurrant, raspberry and strawberry. Therefore, the aim of the research was to obtain aseptic plants, clonal micropropagation and the creation of a cryogenic collection of germplasm based on the developed technology.

Uma das principais etapas da criopreservação de tecidos meristemáticos em plantas vegetais é a micropropagação clonal, que inclui o isolamento dos explantes da matéria-prima in vitro e a otimização do meio de cultura para micropropagação. Como resultado deste estudo, os períodos ideais para micropropagação in vitro são: primeiro - isolamento de explantes de brotos iniciados de gemas dormentes (groselhas e framboesas) em janeiro a março; o segundo - de brotos em crescimento ativo (groselhas e framboesas) em maio a junho, do bigode formado (morango) em julho a agosto. As drogas ideais para esterilização de explantes de framboesa são: a) HgCl2 0,1% (6 min), depois H2O2 3% (15 min); b) lixívia contendo cloro "Domestos" na diluição de H2O 1:9 (10 min). Para groselha preta: a) HgCl2 a 0,1% (5 min) em combinação com fungicida "Topaz" a 0,1% (30 min); b) HgCl2 0,1% (5 min) em combinação com o tratamento com KMnO4 (30 min); c) "Domestos" na diluição de H2O 1:5 (20 min). Para morango: a) 0,1% HgCl2 (6 min) seguido de tratamento com 3% H2O2 10 (min); b) dicloro a 1% (7 min), H2O2 a 3% (10 min); c) "Domestos" na diluição de H2O 1:5 (8 min) com tratamento posterior 0,1% HgCl2 - 7 min, depois 0,20 mg/l КМnO4 - 30 min. Composições ótimas de meios de cultura para micropropagação de groselha negra - meio Murashige e Skoog (MS) com 0,5 mg L-1 BAP, 0,5 mg L-1 GA3, 0,1 mg L-1 IBA e 20 g L-1 de glicose. Para meio framboesa - MS com 0,5 mg L-1 de BAP, 0,1 mg L-1 de AIB, 10 mg L-1 de quelato de ferro e 30 g L-1 de sacarose. Para morango - meio MS com 0,3 mg L-1 BAP, 0,01 mg L-1 IBA, 0,2 mg L-1 GA3, 10 mg L-1 quelato de ferro e 30 g L-1 sacarose. Com base nesses estudos, um criobanco foi criado incluindo o germoplasma de tecidos meristemáticos in vitro em 66 cultivares, híbridos e formas silvestres de groselha, framboesa e morango. Portanto, o objetivo da pesquisa foi a obtenção de plantas assépticas, micropropagação clonal e a criação de uma coleção criogênica de germoplasma com base na tecnologia desenvolvida.

Micropropagation of berry crops for creation of germplasm cryobanks.

One of the main stages of cryopreservation of meristematic tissues in vegetative plants is a clonal micropropagation, which includes isolating the explants of the raw material in vitro and optimizing the culture medium for micropropagation. As the result of our studies, the optimal periods for in vitro micropropagation are: first - isolation of explants from initiated shoots of dormant buds (blackcurrants and raspberries) in January-March; the second - from actively growing shoots (blackcurrants and raspberries) in May-June, from the formed mustache (strawberry) in July-August. The optimal drugs for sterilization of raspberry explants are: a) 0.1% HgCl2 (6 min), then 3% H2O2 (15 min); b) chlorine-containing bleach «Domestos¼ in the dilution of H2O 1:9 (10 min). For blackcurrant: a) 0.1% HgCl2 (5 min) in combination with 0.1% fungicide "Topaz" (30 min); b) 0.1% HgCl2 (5 min) in combination with the treatment with KMnO4 (30 min); c) "Domestos" in the dilution of H2O 1:5 (20 min). For strawberry: a) 0.1% HgCl2 (6 min) followed by treatment with 3% H2O2 10 (min); b) 1% deochlor (7 min), 3% H2O2 (10 min); c) "Domestos" in the dilution of H2O 1:5 (8 min) with subsequent treatment 0,1% HgCl2 -7 min, then 0,20 mg/l КМnO4 - 30 min. Optimal compositions of culture media for micropropagation of blackcurrant - Murashige and Skoog (MS) medium with 0.5 mg L-1 BAP, 0.5 mg L-1 GA3, 0.1 mg L-1 IBA and 20 g L-1 glucose. For raspberry -MS medium with 0.5 mg L-1 BAP, 0.1 mg L-1 IBA, 10 mg L-1 iron chelate and 30 g L-1 sucrose. For strawberry - MS medium with 0.3 mg L-1 BAP, 0.01 mg L-1 IBA, 0.2 mg L-1 GA3, 10 mg L-1 iron chelate and 30 g L-1 sucrose. Based on these studies, the cryobank was created, which include the germplasm of in vitro meristematic tissues in 66 cultivars, hybrids and wild-growing forms of blackcurrant, raspberry and strawberry. Therefore, the aim of the research was to obtain aseptic plants, clonal micropropagation and the creation of a cryogenic collection of germplasm based on the developed technology.