scholarly journals Efficient Plant Regeneration via Indirect Organogenesis in Carnation (Dianthus Caryophyllus Semperflorens flore pleno) Cultivars

2022 ◽  
Vol 0 (0) ◽  
Author(s):  
Hamid Reza SABAGHI ◽  
Gholamreza SHARIFI-SIRCHI ◽  
Pejman AZADI ◽  
Mohammad Hossein AZIMI

ABSTRACT Callus induction and plant regeneration are important steps of in vitro plant breeding of ornamental plants. In this study, the effects of different combinations of plant growth regulators (PGRs), promoters, and minerals on callus induction and plant regeneration in different carnation cultivars were studied in a completely randomized design with three replications. For callus induction, 16 different combinations of 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BA), 1-naphthaleneacetic acid (NAA), and casein hydrolysate (CH) were studied using in vitro leaf explants. The Murashige and Skoog (MS) medium supplemented with 0.2 mg·dm-3 of 2,4-D and 200 mg·dm-3 of CH showed the highest frequency of callus induction. Among the cultivars, ‘Noblesse’ showed the highest rate of callus induction (91.67%). Regarding regeneration, BA, NAA, silver nitrate (AgNO3), and adenine hemisulfate (As) were used in ten different combinations. The ‘Cameron’, ‘Tabasco’, and ‘Noblesse’ cultivars with 95.24% regeneration percentage showed the highest rate of plant regeneration. Generally, in most cultivars, the highest regeneration rate and shoot number per explant were found in the MS medium supplemented with 3 mg·dm-3 of BA, 0.6 mg·dm-3 of NAA, 5 mg·dm-3 of AgNO3, and 40 mg·dm-3 of As. According to the results, the highest regeneration frequency was obtained when 40 mg·dm-3 of As was added to the medium. Finally, the flow cytometry analysis indicated that there were no significant differences between in vitro regenerated and control plants in terms of DNA ratios.

2019 ◽  
Vol 27 ◽  
pp. 89-99
Author(s):  
M Haque ◽  
SMS Islam

This study was carried out to improve an efficient protocol for in vitro callus induction and plant regeneration using Bangladeshi barley genotypes collected from BARI, Gazipur, Bangladesh. After sterilization embryos were separated carefully from mature seeds of six barley genotypes (BB-1, BB-2, BB-3, BB-4, BB-5 and BB-6) and cultured them in MS medium supplemented with various concentration and combination of PGRs for callus induction and regeneration. Out of six genotypes BB-6 showed highest (38.17%) callus induction in MS + 4.0 mg/l 2,4-D + 200 mg/l L-proline + 300 mg/l casein hydrolysate; whereas, BB-4 and BB-5 showed no callus induction in the same medium. For plant regeneration from embryogenic calli the same genotype (BB-6) also performed the best results (19.25%) in MS medium supplemented with 1.5 mg/l BAP + 30 g/l sucrose. Analysis of variance (ANOVA) showed highly significant differences among the media and the genotypes. J. bio-sci. 27: 89-99, 2019


2015 ◽  
Vol 804 ◽  
pp. 259-262
Author(s):  
Chonnikarn Khunchuay ◽  
Kanokporn Sompornpailin

The optimum ratios of auxin and cytokinin are necessary for callus induction and plant regeneration. This ratio is a key function involving in the promoting cell division and proliferation in tissue culture. The axillary buds of in vitro plantlets fromVetiveria nemoralisA. Camuscv. Roiet were used as explants for the callus induction experiment. These explants were cultured on Murashige & Skoog (MS) medium [1] supplemented with various combinations of auxins and cytokinins. Under this experimental study, the highest frequency of callus induction was found on MS medium supplemented with 2 mgL-1α-naphthalene acetic acid (NAA) and 1 mgL-12-furanylmethyl-1H-purine-6-amine (kinetin) (62.5%). On the other hand the combination of 2, 4-dichlorophenoxyacetic acid (2, 4-D) and 6-benzylaminopurine (BAP) was toxicity to this explants. All culturing explants were dead and no calli appearance. The calli derived from each medium were transferred into the same regeneration medium (MS with 1 mgL-1NAA and 2 mgL-1BAP). After culturing on regeneration medium, calli induced from the highest callus induction medium have shown high frequencies of regeneration and also shoot number per callus (58.33% and 7.1 shoots).


2020 ◽  
Vol 5 (1) ◽  
pp. 75-84
Author(s):  
Seied Mehdi Miri

AbstractThe present study describes a protocol for micro-propagation, callus induction, and shoot regeneration of ginger (Zingiber officinale). The rhizomes were surface-sterilized with ethanol (70%) for 45 s, sodium hypochlorite (2.5%) for 10 min, and mercuric chloride (0.1%) for 10 min. Multiple shoots were induced from sprouting bud explants cultured on Murashige and Skoog (MS) medium supplemented with 6-benzyladenine (BA) combined with kinetin (Kin). The maximum shoot number was obtained from MS medium containing 10 mg/l BA with a mean of 20.6 shoots per explant. The leaf explants were cultured on MS medium supplemented with indole-3-acetic acid (IAA), naphthaleneacetic acid (NAA), 2,4-dichlorophenoxy acetic acid (2,4-D), Dicamba, or BA for callus culture. Green-red compact calli were induced using 2,4-D, Dicamba or BA. Also, BA successfully induced plant regeneration. The multiplied shoots that were transferred to the rooting medium (½MS supplemented with 0, 1 and 2 mg/l IAA, indole-3-butyric acid (IBA) or NAA) showed development of roots (100%). The rooted plantlets were transferred to pots containing a 1:1 mixture of cocopeat and perlite, and acclimatization was successful, resulting in 85% survival of the plantlets in the greenhouse.


Horticulturae ◽  
2021 ◽  
Vol 7 (10) ◽  
pp. 407
Author(s):  
Yung-Ting Tsai ◽  
Kin-Ying To

Wedelia chinensis, belonging to the Asteraceae family, has been used in folk medicine in East and South Asia for the treatment of common inflammatory diseases and protection against liver toxicity. Previously, in vitro propagation through different tissue explants has been reported, including through nodal segments, axillary buds, and shoot tips, whereas leaf segments failed to proliferate. Here, we report on the in vitro propagation of W. chinensis by culturing young leaf explants in MS medium supplemented with 0.5 mg/L α-naphthaleneacetic acid (NAA), 0.75 mg/L thidiazuron (TDZ), 1 mg/L gibberellic acid (GA3), 3.75 mg/L adenine, 3% sucrose, and 0.8% agar at pH 5.8. Calli were observed in all explants derived from the youngest top two leaves, and the average percentage of shoot regeneration was 23% from three independent experiments. Then, several shoots were excised, transferred onto MS basal medium supplemented with 3% sucrose and 0.8% agar at pH 5.8, and cultured in a growth chamber for 1 to 2 months. Roots were easily induced. Finally, plantlets carrying shoots and roots were transferred into soil, and all of them grew healthily in a greenhouse. No morphological variation was observed between the regenerated plantlets and the donor wild-type plants. In addition, we also established root cultures of W. chinensis in culture medium (MS medium, 3 mg/L NAA, 3% sucrose, pH 5.8) with or without 0.8% agar. To the best of our knowledge, this is the first paper reporting plant regeneration from leaf explants in the herbal plant W. chinensis.


HortScience ◽  
2004 ◽  
Vol 39 (6) ◽  
pp. 1373-1377 ◽  
Author(s):  
Margarita Fraga ◽  
Mertxe Alonso ◽  
Marisé Borja

Meristem culture and/or thermotherapy were used for virus elimination from ornamental Phlox paniculata L. (`Blue Boy', `Orange perfection' and `Starfire') mother plants. Shoot tip, leaf, node and flower ovary explants collected from greenhouse-maintained virus free plants were cultured in vitro for shoot initiation. Adventitious shoot initiation was observed on Murashige and Skoog (MS) medium containing the cytokinin BA with or without the auxin NAA. The addition of 0.4 mg·L-1 thiamine, 0.4 mg·L-1 folic acid, and 40 mg·L-1 adenine sulfate to the MS medium did not improve the regeneration rate. Multiplication and rooting were genotype dependent. Blue Boy and Orange Perfection cultivars regenerated the maximum number of shoots from leaf explants. `Blue Boy' leaf explants from in vitro plants had a lower regeneration rate than explants from greenhouse plants. Cultivar `Starfire' had the highest shoot formation with open flower ovary explants and failed to regenerate from leaf explants. In vitro rooting of adventitious shoots in the presence of auxins (IAA, NAA, or IBA) with or without BA was less effective than ex vitro rooting. Chemical names used: 6-benzyladenine (BA); indole-acetic acid (IAA); indole-3-butyric acid (IBA); α-naphthaleneacetic acid (NAA).


2017 ◽  
Vol 15 (10) ◽  
pp. 701-710
Author(s):  
Piyaporn SAENSOUK ◽  
Surapon SAENSOUK ◽  
Phattaraporn PIMMUEN

An efficient and rapid protocol for the micropropagation of Globba schomburgkii Hook. f. via bulbil explants was investigated. The long divided and undivided bubils of G. schomburgkii Hook. f. were cultured on MS medium (Murashige and Skoog) that had either 3 mg/l benzyladenine (BA) or 0.5 mg/l naphthaleneacetic acid (NAA) added for 8 weeks. The results indicated that the long divided bulbils of G. schomburgkii Hook. f. showed a greater amount of plant regeneration than the undivided bulbils. Callus induction, as well as shoot and root formation, were observed when culturing microshoots of 1 cm in length on media (MS) that had Thidiazuron (TDZ) or NAA plus BA added at a range of concentrations for 8 weeks. The highest percentage of callus induction was 40 % when culturing the microshoots on MS medium supplemented with NAA and BA. The best result for shoot formation was achieved when culturing the microshoots on MS medium with TDZ added. The highest number of roots was obtained when culturing the microshoots on MS medium with NAA and BA added. The in vitro-derived plantlets of G. schomburgkii Hook. f. were transplanted to pots containing different types of potting mixture in a greenhouse. The survival rates were 80 % when G. schomburgkii Hook. f. was transplanted to sand.


Plants ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 67
Author(s):  
Hsiao-Hang Chung ◽  
Hui-Yao Ouyang

Strawberry, belonging to the Fragaria genus, is an important crop that produces popular fruits globally. F. vesca, known as wild strawberry, has great characteristics, such as a robust and powerful aroma, making it an important germplasm resource. The present study aims to establish an efficient regeneration method for the in vitro propagation of F. vesca. Firstly, leaf explants were used to induce callus formation on a Murashige and Skoog medium with combinations of cytokinins (thidiazuron (TDZ) and 6-benzylaminopurine (BA)) and auxin (2,4-dichlorophenoxyacetic acid (2,4-D)). Among them, 0.45–4.54 µM TDZ combined with 0.45–4.53 µM 2.4-D exhibited a high induction rate after 4 weeks of culturing. Different explants were examined for their ability to form a callus, and whole leaves on the medium containing 2.27 µM TDZ and 2.27 µM 2,4-D showed the highest callus induction rate at 100% after 2 weeks of culturing in darkness. The highest shoot regeneration ability through organogenesis from the callus was obtained at 0.44 µM BA. After 2 weeks of culturing, the shoot regeneration rate and shoot number per explant were 96% and 19.4 shoots on an average, respectively. Rooting of shoots was achieved using indole-3-butyric acid (IBA) or an α-naphthaleneacetic acid (NAA)-containing medium, and the resulting plantlets were acclimatized successfully and formed flowers eventually. In this report, we demonstrated that shoot organogenesis was derived from the meristematic cells of calli and by transferring the induced calli to a 0.44 µM BA medium, the regeneration period can be shortened greatly. A protocol for the efficient regeneration of wild strawberry was established, which might be useful for their large-scale propagation or obtaining transgenic plants in the future.


Author(s):  
Norizzah Jaafar SIDIK ◽  
Norhayati DAUD ◽  
Som Cit SINANG ◽  
Nurul Fazira OMAR

This study examined the efficiency of callus induction on optimum concentrations of NAA (a-naphthaleneacetic acid) and BAP (6-benzyladenine) from culturing stem and leaf explants of finger eggplant (Solanum sp.) and investigated the phytochemical constituents of callus tissue. Seeds were sterilized by using 3 and 5 % Clorox solution, which gave the highest number of survival seeds (100 %) and were grown in vitro plantlets. The highest frequency of callus induction (100.00 ± 0.00 %) was obtained from stems and leaf explants that were excised from in vitro plantlets. The stem explants cultured on MS medium consisted of 1.0 mg/L NAA + 1.0 mg/L BAP, giving the maximum mean callus fresh weight (0.14 ± 0.05 g). Meanwhile, the leaf explants cultured on MS medium consisted of  0.5 mg/L NAA + 2.0 mg/L BAP, generating the maximum mean callus fresh weight (0.48 ± 0.10 g). The highest frequency of callus induction (88.00 ± 1.60 %) was obtained in solidified MS medium supplemented with 0.5 mg/L NAA + 2.0 mg/L BAP, producing the maximum mean fresh weight of callus (1.54 ± 0.27 g) and dry weight (0.90 ± 0.01 g). The results of the Phytochemical screenings of callus and dried leaf extracts indicated the presence of alkaloids, flavonoids, terpenoids, and saponins.


2015 ◽  
Vol 25 (1) ◽  
pp. 1-12 ◽  
Author(s):  
Kee Hwa Bae ◽  
Eui Soo Yoon

Lychnis cognate Maxim and Lychnis fulgens Fish. Ex Spreng are two valued ornamental plants in Korea. Soaking of seeds in GA3 solution remarkably promoted germination up to 60%, but the control (0 mg/l) was not effective (> 5%). To select an adequate temperature for seed germination, seeds, previously soaked in a 1000 mg/l GA3 for 24 hrs, were incubated at 15, 20, 25, and 30°C. Seed germination of over 20% was obtained at 15, 20, and 25°C, but only 10% at 30°C. These results indicate that the seeds of L. cognate and L. fulgens are in a such dormant state that they hardly germinate even by dormancy breaker (GA3) and low (15 ? 25°C) temperature treatment. The highest callus induction was observed in the leaf explants of the seedlings on MS containing specific concentrations of 3.0 mg/l BA and 1.0 mg/l NAA. The adventitious shoot was formed < 90% of calli on 1/2 WPM medium. The height of in vitro propagated plantlet was no different media used for regeneration. This in vitro propagation protocol should be useful for conservation of endangered and ornamental plant.Plant Tissue Cult. & Biotech. 25(1): 1-12, 2015 (June)


2017 ◽  
Vol 4 (2) ◽  
pp. 52-56
Author(s):  
Mallika Devi T

In the present study the protocol for callus induction and regeneration in Azima tetracantha has been developed in culture medium. The young apical leaf explants were used for callus induction on MS medium containing BAP and NAA at 1.0 and 0.4mgl-1 respectively showed maximum callus induction (73%). The amount of callus responded for shoot formation (74%) was obtained in the MS medium containing BAP (1.5 mgl-1) and NAA (0.3mgl-1).The elongated shoots were rooted on half strength medium supplemented with IBA (1.5 mgl-1) and Kn (0.4 mgl-1) for shoots rooted. Regenerated plantlets were successfully acclimatized and hardened off inside the culture and then transferred to green house with better survival rate.


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