scholarly journals Optimization Studies of Culture Media for In-Vitro Clonal Micropropagation of New Grape Varieties

2021 ◽  
Author(s):  
Abdulmalik Batukaev ◽  
Eliza Sobralieva ◽  
Diana Palaeva

This article describes the effect of Gautheret, White, Heller and Murashige & Skoog mineral salts during in-vitro clonal micropropagation of new grape varieties. The optimal mineral compositions of the culture medium that support the in-vitro regeneration of isolated grape explants were identified. The grapes that were studied were the Bart and Augustine varieties. Primary grape explants were cultivated for 30 days in a non-transplanted culture. Increased regenerative activity was observed in the Murashige & Skoog and White media. Increased haemogenesis occurred and shoots regenerated. The addition of cytokinin 6-BAP to the medium for obtaining aseptic culture led to an increase in the frequency of shoot-bud production by 5 to 6 times, depending on the type of medium. Combining 6-BAP with the auxin NAA provided an additional increase in the frequency of shoot-bud production, but to a lesser extent. Adding growth regulators to the culture medium also reduced the frequency of explant necrosis. Keywords: grapes, mineral salts, culture medium, microclonal propagation, in-vitro, cytokinins, auxins

2018 ◽  
Vol 12 (2) ◽  
pp. 117
Author(s):  
Cecília Moreira Serafim ◽  
Arlene Santisteban Campos ◽  
Priscila Bezerra Dos Santos Melo ◽  
Ana Cecília Ribeiro de Castro ◽  
Ana Cristina Portugal Pinto de Carvalho

Faced with the demand for plants potted for their foliage, Anthurium maricense is seen as a viable option. However, most of the studies on obtaining micropropagated plantlets are for A. andraeanum, with nothing yet reported for A. maricense. The aim of this study therefore, was to evaluate the effect of four cytokinins in different concentrations, on the in vitro induction of shoots from nodal segments of A. maricense. The experimental design was completely randomised in a 4 x 4 factorial scheme, with four cytokinins (BAP, ZEA, CIN and 2iP) and 4 concentrations (0, 2.22, 4.44 and 6.66 μM), for a total of 16 treatments, with 6 replications of five test tubes, and using one nodal segment. Cultures were kept in a growth room at 25 ± 2°C, a photoperiod of 16 h and a light intensity of 30 μmolm-2 s-1 for 60 days. After this period, the number of shoots formed per node was evaluated. The addition of a cytokinin to the culture medium was determinant for the in vitro regeneration of shoots in A. maricense. The greatest estimated number of shoot formations in A. maricense were obtained in the culture media containing ZEA (3.87) and BAP (3.30), both at concentration of 6.66 μM.


2021 ◽  
Vol 4 (46) ◽  
pp. 17-17
Author(s):  
Alexander Saakian ◽  
◽  

Abstract The aim of this study is to develop and improve methods of in vitro propagation of representatives of Dactylorhiza: D.baltica , D. fuchsii. For the study, we used protocorms obtained by the asymbiotic germination of seed during 90 days. It has been established that half-strength of Murashige and Skoog (1962) medium (½ MS) supplemented with 1-2 mg/l 6-Benzylaminopurine(6-BAP), potato puree (20g/l), and charcoal (1g/l) effectively influenced the development of protocorms, and seedlings formation in the studied species. The result of the study showed that the survival rate of protocorms was high in all experimental culture media, but in D. fuchsii it was better at a concentration 2mg/l of 6-BAP (95.4%), while in D. baltica it was high at 1mg/l (87.0%). The highest percentage of multiple protocorms (68%) and the formation of new secondary protocorms in D. fuchsii (5,5±0,3 units) were observed on a culture medium containing 2 mg/l 6-BAP. The highest percent of rooting of D. fuchsii protosoms (78%) and length of roots (0.9cm) observed in ½ MS medium without growth regulators. During the development of D. baltica protosoms, the culture medium of ½ MS containing 1 mg/l 6-BAP had the best effect on the number of roots (1.8±0.1root/protosom), while the medium supplemented with 2mg/l of 6-BAP contributed to the formation of a larger number of new secondary protocorms (3,2±0,1protocorm/unit). During the subsequent cultivation of protosoms of D. baltica on a culture medium containing 1 mg/l it was observed an increase in the height of shoots (4,8±0,3 см), and the length of roots (2,2±0,1 см), wherein the number of newly formed protocorms was higher by 30% on the medium supplemented with 2 mg/l 6-BAP. Keywords: DACTYLORHIZA BALTICA, DACTYLORHIZA FUCHSII, IN VITRO, PROTOCORMS, ORGANIC ADDITIVES


2005 ◽  
Vol 11 (2) ◽  
pp. 105-108
Author(s):  
P. Szarvas ◽  
A. Zsila-André ◽  
Z. Kováts ◽  
M. G. Fári

We conducted experiments for developing an in vitro micropropagation protocol starting from meristems of Rudbeckia hirta L seedlings. We pre-soaked the seeds in sterile ion-exchanged water for 17 hours, and then achieved surface disinfection in two separate steps. First, we used concentrated household sodium-hypochloride solution for 20 minutes and, also for 20 minutes, we applied hydrogen peroxide of 10%, which was followed by washing with sterile ion-exchanged water three times. For the propagation of seedling meristems, the combination of half-strenght solid Murashige and Skoog (1962) culture medium containing 10 mg/1 of kinetin or 2 mg/I of kinetin + 0.1 mg/1 of 2iP proved to be the most suitable. The average number of shoot-buds developed from the seedling axillary meristem in the best culture media varied between 5 and 17. Without separating them, we inoculated the shoot-bud clusters on MS culture medium containing 2 mg/1 of IAA. After four weeks of incubation we obtained elongated shoots which we separated and inoculated into a new culture medium and we obtained elongated roots. The rooted plants were gradually acclimatised in the cultivation room, potted and carried to a greenhouse, and then planted in open field for subsequent observation. By adopting this method, our laboratory started the micropropagation of the superior and/or elite genotypes of the Rudbeckia hirta L. being of special value in respect of breeding.


2019 ◽  
Vol 41 ◽  
pp. e47105
Author(s):  
Marcos Vinícius Marques Pinheiro ◽  
Daniele Cristina Fontana ◽  
Jullie dos Santos ◽  
Matheus Milani Pretto ◽  
Gabrieli Cristina Vitalli de Azevedo ◽  
...  

In vitro propagation increases the supply and commercialisation of products of interest. For this, optimising the growing conditions and the composition of the culture medium is crucial to benefit the full development of the plants. Thus, the objective was to evaluate the in vitro propagation of Aloysia triphylla on different culture media, with varying agar and sucrose concentrations. The experiment was conducted as a completely randomised design, 3×3×3 factorial scheme, with three culture media (MS, JADS and WPM), three sucrose concentrations (8, 10 and 12 g L-1) and three agar concentrations (15, 30 and 45 g L-1), with three replicates each and experimental units composed of one plant per replicate. After 25 days of cultivation, the fresh and dry mass of the plants, numbers of leaves, numbers of nodes, plant lengths, numbers of oxidised leaves, hyperhydricity and acclimatization percentages were evaluated. The WPM medium resulted in a reduced fresh mass, reflecting in the low hyperhydricity observed in the explants, and favoured the acclimatization of the plants. Thus, the WPM medium with sucrose (15 g L-1) and agar (12 g L-1) is recommended as the medium most suitable for the in vitro regeneration of Aloysia triphylla.


2019 ◽  
Vol 9 (4) ◽  
pp. 676-678
Author(s):  
I. D. Borodulina ◽  
A. Trufanova ◽  
G. Shevchenko ◽  
G. Sokolova ◽  
T. Plaksina

Micropropagation of Chrysanthemum is an alternative to the traditional method of reproduction. Thanks to this method, the Chrysanthemum reproduction time is reduced to 3-4 months. For clonal micropropagation, sterile microshoots of Chrysanthemums of the varieties Snow White, Stranger, and Baltica White were used. At the stage of the micropropagation, the Murashige-Skoog (MS) medium with the full and half composition of mineral salts and growth regulators (kinetin, 6-benzylaminopurine, β-indolylacetic acid) were used. A universal culture medium for clonal micropropagation of all varieties of Chrysanthemum and optimal mediums, taking into account variety-specificity were established. It was noted that under in vitro conditions, high proliferative activity was observed in Neznakomka variety.


2020 ◽  
Vol 25 ◽  
pp. 05003
Author(s):  
Saule Kazybayeva ◽  
Irina Kovalchuk ◽  
Timur Turdiyev ◽  
Shokan Kulshanov ◽  
Laura Azhitayeva

The article shows the improvement of the process of initiation into in vitro the culture and the clonal micropropagation of grape varieties. The optimal culture media for the initiation and cloning of grapes in vitro have been selected. During initiation on Murashige and Skoog, agar medium with ½ or ¾ concentration of macroand micronutrients and hormones (0.5 mg/l BAP and 0.1 mg/l IBA) is optimal. For micropropagation is suitable MS medium modified by some elements of mineral nutrition: 825 mg/l NH4NO3, 166 mg/l CaCl2, 15 mg/l ferrum chelate; best hormonal composition depends on variety: a) 0.5-1 mg/l BAP and 0.1-0.5 mg/l IBA; b) 0.5 mg/l 2-iP and 0.5 mg/l GA3.


2020 ◽  
Vol 224 ◽  
pp. 04001
Author(s):  
A Sh Tevfik ◽  
N. A. Yegorova

Thymus vulgaris L. is one of the widely known spicy aromatic and medicinal plants. Thyme plant material is widely used in medicine, cooking and perfumery. To increase the efficiency of breeding and seed production, it is necessary to develop biotechnological techniques, in particular, clonal micropropagation. The aim of the research is to optimize the composition of culture media for the main stages of propagation in vitro and to select adaptation ex vitro conditions for the development of Thymus vulgaris. clonal micropropagation. The article presents the results of studies of explant morphometric parameters cultivated on 20 variants of culture media at firstsecond stages of micropropagation. It was found that the optimal culture medium at the introduction stage is MS medium with 1.0 mg/l Kin and 1.0 mg/l GA3, on which, on average, 2.2 microshoots per explant with a length of 1.9 cm were obtained. Both high vitrification rate of microshoots and formation of small shoots (0.6-0.9 cm) were observed on media supplemented with BAP or TDZ. The most effective culture medium at the proper propagation stage is MS with 1.0 mg/l Kin, on which 4.6 shoots per explant and the multiplication index 12.8 were obtained. It is advisable to root microshoots at the 3rd stage of micropropagation on MS culture medium supplemented with 1.0 mg/l IBA or 1.0 mg/l IAA. It has been shown that it is possible to obtain high plant survival rate (89.5%) during adaptation ex vitro, using a substrate consisting of peat and perlite (1:1).


2006 ◽  
pp. 53-59
Author(s):  
Pál Szarvas ◽  
Anikó Zsila-André ◽  
Zoltán Kovács ◽  
Miklós Gábor Fári

We conducted experiments for developing an in vitro micropropagation protocol starting from meristems of Rudbeckia hirta L seedlings. We pre-soaked the seeds in sterile ion-exchanged water for 17 hours, and then achieved surface disinfection in two separate steps. First, we used concentrated household sodium-hypochloride solution for 20 minutes and, also for 20 minutes, we applied hydrogen peroxide of 10%, which was followed by washing with sterile ion-exchanged water three times. For the propagation of seedling meristems, the combination of half-strength solid Murashige and Skoog (1962) culture medium containing 10 mg/l of kinetin and 2 mg/l of kinetin + 0.1 mg/l of 2iP proved to be the most suitable. The average number of shoot-buds developed from the seedling axillary meristem in the best culture media varied between 5 and 17. Without separating them, we inoculated the shoot-bud clusters on MS culture medium containing 2 mg/l of IAA. After four weeks of incubation, we obtained elongated shoots, which we separated and inoculated into a new culture medium and from which we obtained elongated roots. The rooted plants were gradually acclimatised in the cultivation room, potted and carried to a greenhouse, and then planted in open field for subsequent observation. By adopting this method, our laboratory started the micropropagation of the superior and/or elite genotypes of the Rudbeckia hirta L. being of special value in respectt to breeding.


Author(s):  
E. N. Raeva-Bogoslovskaya ◽  
O. I. Molkanova

In vitro culture conditions were optimized for representatives of the genus Amelanchier Medik. at the stages of micropropagation and rooting. A significant effect of the mineral and hormonal compositions of culture media on the morphogenetic potential of the cultivars of serviceberry has been established. The use at the stage of micropropagation of the MS culture medium with addition 1,0 mg / L 6-benzylaminopurine (6-BAP) promoted the active microshoot regeneration of the studied genotypes. For the induction of rhizogenesis the type of auxin as IB A at a concentration of 1,0 mg / L was used.


Author(s):  
RK Niroula ◽  
BP Sah ◽  
HP Bimb ◽  
S Nayak

This study was under taken to elucidate the effect of genotypes and media compositions on callus induction from mature rice seeds. Three different callus induction media, designated as A (N6 mineral salts + N6 vitamins, 2 mg/l each + myoinositol, 100 mg/l + 2,4-D, 2.5 mg/l + kinetin, 0.5mg/l + AgNO3, 10 mg/l + maltose, 50 gm/l); B (MS organic salts + N6 mineral salts + NAA, 4 mg/l + kinetin, 2 mg/l + AgNO3, 5mg/l and sucrose, 60 gm/l); and C (B media without AgNO3), and six rice genotypes viz. Jumlimarshi, Tilki, Jethobudo, Manshara, Masuli and Pahenle were evaluated. The modified N6 medium supplemented with 2, 4-D, 2.5 mg/l and AgNO3, 10 mg/l exhibited better performance in callus induction. Among genotypes, callus induction frequency was higher (100%) in Masuli, Tilki and Jumlimarshi regardless of media tested. The positive effect of AgNO3 was only observed in medium A for quality callus induction and subsequent plant regeneration. The genotype Tilki performed better regarding plant regeneration (27.77%). Therefore, it is suggested that application of medium A is advantageous to accomplish overall efficiency of callus induction and plant regeneration from seeds of various rice genotypes. Key words: 2, 4-D; AgNO3; dehulled rice; in vitro; regeneration J. Inst. Agric. Anim. Sci. 26:21-26 (2005)


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