scholarly journals IN VITRO ELIMINATION OF DAHLIA MOSAIC VIRUS BY USING MERISTEM CULTURE, ELECTROTHERAPY AND CHEMOTHERAPY

2020 ◽  
Vol 51 (2) ◽  
pp. 665-674
Author(s):  
Nerway & et al.
Keyword(s):  
Salicylic Acid ◽  
Mosaic Virus ◽  
Meristem Culture ◽  
Ms Medium ◽  
Virus Elimination ◽  
In Vitro Techniques ◽  
Dahlia Mosaic Virus ◽  
Time Courses ◽  
Das Elisa

This study was conducted to investigate the effect of different in vitro techniques on elimination of dahlia mosaic virus. Meristems of Dahlia mosaic virus (DMV) infected dahlias, with 0.2-0.3 mm in length, were cultured on Murashige and Skoog (MS) medium. Based on DAS-ELISA, 100% of the survived plantlets by meristem culture were virus-free. DMV infected stem segments with two axillary buds were treated at three different levels of electric currents (15, 25 and 35 mA) and two time courses (10 and 20 min) in an electrophoresis tank and cultured on MS medium. The treatment of 35 mA for 20 min was the best for viral elimination from diseased dahlias by 85% followed by 25 mA for 20 min by 70%. Acyclovir and salicylic acid were the antivirals were used to eliminate DMV from the infected dahlias. For chemotherapy, nodes were cultured on MS media supplemented with antivirals at 0, 10, 20, 30, 40 and 50 mg/l for 20 and 30 days. The highest concentration of acyclovir (50 mg/l) for 30 days were the best treatments as its highest effectiveness on elimination ability (90.67%) but gave only 49% for plantlet survival. Whereas 40 mg/l acyclovir for 30 days gave elimination ability (80%) and plantlet survival (89.67%). The therapy containing salicylic acid at 40 mg/l for 30 days was the best as its highest effectiveness on DMV elimination ability (100%). But the concentration of salicylic acid at 30 mg/l for 30 days was the best treatment (chemotherapy efficiency= 49.21) as well as DMV elimination ability (75.33%) and plant survival (65.33%). The in vitro developed dahlia plantlets were tested for DMV-freeness using DAS-ELISA. The using therapies (electrotherapy and chemotherapy) can play a good role in virus elimination because they need the minimum period of time to regenerate fully developed healthy plants as long as 2-3 months.

scholarly journals Shoot Regeneration Rates of Perennial Phlox are Dependant on Cultivar and Explant Type

HortScience ◽  
2004 ◽  
Vol 39 (6) ◽  
pp. 1373-1377 ◽  
Author(s):  
Margarita Fraga ◽  
Mertxe Alonso ◽  
Marisé Borja
Keyword(s):  
Adventitious Shoots ◽  
Leaf Explants ◽  
Naphthaleneacetic Acid ◽  
Meristem Culture ◽  
Ms Medium ◽  
Virus Elimination ◽  
Open Flower ◽  
Regeneration Rate ◽  
Shoot Initiation

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).

Field evaluation of dasheen mosaic virus-free cocoyam plants produced by in vitro techniques

1997 ◽  
Vol 68 (1-4) ◽  
pp. 37-47 ◽  
Author(s):  
R. Valverde ◽  
L. Gomez ◽  
F. Saborio ◽  
S. Torres ◽  
O. Arias ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Field Evaluation ◽  
In Vitro Techniques

scholarly journals IN VITRO REGENERATION OF ARUNDINA GRAMINIFOLIA (D. DON) HOCHR

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Sharone gladies E ◽  
Chithra Devi B. S
Keyword(s):  
In Vitro Regeneration ◽  
Basal Medium ◽  
Culture Technique ◽  
Ms Medium ◽  
Asymbiotic Seed Germination ◽  
In Vitro Techniques ◽  
Murashige And Skoog Medium ◽  
Arundina Graminifolia ◽  
Indole 3 Acetic Acid

We can see Orchids come in a wide variety of shapes, sizes, colours, and textures far beyond the human mind’s imagination. They emerge from seeds in nature, but in the absence of suitable hosts, they do not germinate in sufficient numbers. This problem was solved by using the tissue culture technique for its germination. One of the successful method used for mass propogation of orchid plantlets is in vitro techniques. Therefore, an initial analysis was conducted in order to establish an appropriate procedure for mass multiplication of Arundina graminifolia. MS (Murashige and Skoog) medium was found to be suitable for the asymbiotic seed germination of Arundina graminifolia. Direct protocorm like bodies were induced by using combinations and individual supplement of MS medium with IAA (Indole-3-acetic acid), IBA (Indole-3- butyric acid), BAP (6-Benzylaminopurine) and KIN (Kinetin). Hormone-free MS basal medium was found suitable for the conversion of PLBs (protocorm-like bodies) into complete plantlets

scholarly journals Detection of viruses of Bangladeshi and Japanese garlic and their elimination through root meristem culture

2017 ◽  
Vol 28 (2) ◽  
pp. 55-63 ◽  
Author(s):  
MS Haque ◽  
K Hattori
Keyword(s):  
Root Meristem ◽  
Onion Yellow Dwarf Virus ◽  
Meristem Culture ◽  
Free Medium ◽  
Rt Pcr ◽  
Virus Elimination ◽  
Root Meristems ◽  
Yield And Quality ◽  
Yellow Dwarf Virus

A number of viruses cause considerable yield loss and quality deterioration in garlic. Root meristems of virus infected plants are known to be free from detectable viruses. This potentiality could be exploited to obtain virus free clones at a high frequency by culturing excised root meristems in vitro. We have developed efficient methods of direct and somatic embryo derived shoot regeneration from root meristems of garlic. The objectives of this work were to detect viruses infecting Bangladeshi and Japanese garlic clones and find an easy and efficient method of eliminating the viruses for the improvement of both yield and quality of garlic. At first, we confirmed the presence of detectable viruses in three Bangladeshi and one Japanese clones. The clones were infected with four different types of viruses: Garlic viruses (GarVs), Onion yellow dwarf virus (OYDV), Leek yellow stripe virus (LYSV), and Garlic common latent virus (GCLV). To eliminate those viruses, as per our previous method, root meristems were cultured on MS medium supplemented with 1.0 µM NAA and 10.0 µM BA. Shoot primordia developed from the cultured explants within 1 month. The regenerated individual shoot buds (2-5 mm) were separated from the mother explants and transferred to growth regulators free medium. RT-PCR confirmed that the viruses present in the mother garlic plants were absent in the shoots found after two-step culture. The regenerated shoots were rooted on growth regulator free medium and transferred to pots. Results indicated that the plants remained free from LYSV. Virus elimination through root meristem culture emerged as an efficient novel technique for the eradication of multiple viruses as confirmed by RT-PCR in this study. This technique has the potential for the production and supply of virus free propagules (plants/bulblets) for the yield and quality improvement of garlic.Progressive Agriculture 28 (2): 55-63, 2017

scholarly journals Relative efficiency of positive selection and tissue culture for generating pathogen-free planting materials of yam (Dioscorea spp.)

2017 ◽  
Vol 53 (No. 1) ◽  
pp. 9-16 ◽  
Author(s):  
M. Balogun ◽  
N. Maroya ◽  
J. Augusto ◽  
A. Ajayi ◽  
L. Kumar ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Positive Selection ◽  
Mosaic Virus ◽  
Relative Efficiency ◽  
Meristem Culture ◽  
Important Species ◽  
In Vitro Plantlets ◽  
Seed System ◽  
Planting Materials

Yams are staples in West Africa. They are propagated from tubers in an informal seed system. This encourages a build-up of diseases, and necessitates the rapid development of a formal seed system where certified seeds are functional. Although few reports exist on the use of meristem culture to generate pathogen-free yam, the success rate for the most economically important species in the sub-region, Dioscorea rotundata, for the most prevalent viruses is inadequate. To generate pathogen-free yam planting materials, the relative efficiency of tissue culture and positive selection was compared. Twenty-one asymptomatic yam plants were positively selected from 8187 stands of five landraces. Five of these stands were tested virus-negative by multiplex polymerase chain reaction (PCR) for Yam mosaic virus (YMV), Yam mild mosaic virus (YMMV) and Cucumber mosaic virus (CMV), and by PCR for the genus Badnavirus (BV), giving 0.08% success. Single nodes of the positively selected stands were used to establish in vitro plantlets, which were screened onto bacteriological indexing medium. The same was done for meristem- and node-derived plantlets of the improved variety TDr 95/19158. Incidence of endophytes ranged from 18 to 32% in the nodal plantlets while it was 0% in the meristem-derived plantlets. The effect of meristem culture combined with thermotherapy on the virus infection status was determined using virus-tested, one week old in vitro plantlets of eight improved genotypes. These in vitro plantlets were incubated at 36 ± 0.5°C and 16 h photoperiod for 21 days, after which meristems were excised, regenerated into plantlets and re-tested for viruses. Seventy-three percent of the samples were recovered from YMV but the effect on BV was inconsistent. Positive selection can be used as a palliative in generating quality declared seed but meristem culture combined with thermotherapy is more efficient for generating certified seed tubers of yam.

scholarly journals First Report of Cucumber mosaic virus in Paeonia lactifera in France

Plant Disease ◽  
2010 ◽  
Vol 94 (6) ◽  
pp. 790-790 ◽  
Author(s):  
L. Cardin ◽  
J. P. Onesto ◽  
B. Moury
Keyword(s):  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Polyclonal Antibodies ◽  
Tobacco Rattle Virus ◽  
Meristem Culture ◽  
Cut Flowers ◽  
Leaf Extracts ◽  
First Report ◽  
The Family ◽  
Das Elisa

Chinese peony (Paeonia lactiflora Pall.), a hardy ornamental plant of the family Paeoniaceae cultivated in gardens and for cut flower production, is frequently infected by Tobacco rattle virus (TRV) in the field. The virus usually induces severe mosaic and chlorotic ringspot symptoms in the leaves, decreasing the commercial value of cut flowers. TRV is routinely detected by mechanical inoculation onto Nicotiana tabacum cv Xanthi, where it induces typical necrotic local ringspots in 3 to 7 days, followed by a reverse transcription (RT)-PCR test (2). In 2004, Xanthi test plants inoculated with sap extracts from 4 of 36 P. lactiflora cv. Odile plants grown in a field plot in the region of Hyères (southeast France) showed systemic mosaic symptoms in addition to the TRV-typical response. In each case, Cucumber mosaic virus (CMV) was detected by the reactions of a range of inoculated plants (1), the observation of 30 nm isometric particles in crude leaf extracts with the electron microscope, and by positive reactions in double antibody sandwich (DAS)-ELISAs with specific polyclonal antibodies. In double-immunodiffusion analysis, these isolates were shown to belong to the group II of CMV isolates (3). ELISA of the peony plants confirmed the presence of CMV and revealed two additional infected plants in the spring of 2006. Following isolation from local lesions on Vigna unguiculata and multiplication in Xanthi tobacco plants, one of the isolates was used to inoculate manually or with Myzus persicae aphids 10 CMV-free plants of P. lactiflora cv. Odile obtained from meristem culture. Three months postinoculation, only three of the aphid-inoculated plants were CMV positive by DAS-ELISA. No change was observed at 1 year postinoculation and no symptoms have been observed, even in CMV-infected plants. CMV appears to be latent in P. lactiflora, therefore detection of CMV before vegetative propagation of the plants is advised because of the risks of synergism for symptoms with other viruses such as TRV. To our knowledge this is the first report of CMV in peony. References: (1) L. Cardin et al. Plant Dis. 87:1263, 2003. (2) D. J. Robinson J. Virol. Methods 40:55, 1992. (3) M. J. Roossinck. J. Virol. 76:3382, 2002.

scholarly journals Influence of L-Tryptophan and salicylic acid on secondary metabolites production from leaves induced callus of Catharanthus roseus L.G.Don in vitro

2014 ◽  
Vol 8 (2) ◽  
pp. 35-43
Author(s):  
Emad H. Jassim ◽  
Sami K. M. Ameen
Keyword(s):  
Salicylic Acid ◽  
Secondary Metabolites ◽  
Catharanthus Roseus ◽  
Leaf Explants ◽  
Ms Medium ◽  
Plant Leaves ◽  
Fresh Weight ◽  
Metabolites Production ◽  
Different Levels

An experiment was conducted to steady the effect of L-Tryptophan and salicylic acid on callus induced on leaf explants of Catharanthus roseus. Callus induction was achieved by culturing true leaves of the plant on Murashige and Skoog (MS) medium supplemented with 0.5 mg /L 2,4-D and 1mg / L Kin. The best medium to maintain callus was MS medium supplemented with 0.5 mg /L 2,4-D and 1.5 mg / L Kin. Different levels added to MS medium for each L-Tryptophan 0,200,300or400 mg /L and salicylic acid 0, 0.5,1or1.5mg /L. The medium supplemented with 30 g/ L sucrose was used as a control. Results showed the medium supplemented with 200 mgL of L-Tryptophan gave the highest quantity of Vincristine reached 48.66 µg/100 fresh weight of callus. MS medium content at the concentration 1 mg L of salicylic acid gave the highest level of Vinblastine recording 50.98 µg/100 fresh weight of callus. While MS medium supplemented with 0.5 mg /L of salicylic acid gave the highest level of Serpentine 24.76 µg/100 fresh weight of callus. The concentrations of active compound derived from plant leaves were much less than the concentrations produced by the callus grown in vitro. The concentration of Serpentine was 0.059 while Vinblastine was 0.183 and the concentration of Vincristine was 0.064.

scholarly journals (288) Determination of Genetic Stability of In Vitro-derived Birch Plants

HortScience ◽  
2006 ◽  
Vol 41 (4) ◽  
pp. 1077C-1077
Author(s):  
Wenhao Dai ◽  
Victoria Magnusson ◽  
Andrea Swanberg
Keyword(s):  
Genetic Stability ◽  
Random Amplified Polymorphic Dna ◽  
Genetic Fidelity ◽  
Genetic Variations ◽  
Meristem Culture ◽  
In Vitro Techniques ◽  
Somaclonal Variations ◽  
Regenerated Plants ◽  
Leaf Tissues

Many woody plants, including some birch species, can be cloned using such in vitro techniques as pre-existing meristem culture, organogenesis, and embryogenesis. However, clonal fidelity of in vitro-derived plants is always a big concern because somaclonal variations may be induced during the entire in vitro process. To address this issue, we used random amplified polymorphic DNA (RAPD) markers to determine the genetic stability of in vitro-propagated plants of Betula platyphylla `Fargo'. Forty-two greenhouse-grown birch plants derived from a 10-year shoot tip culture (shoot-derived) and 42 in vitro plants regenerated from leaf tissues (regenerated) were randomly selected and evaluated for their genetic fidelity by RAPD. To date, 20 primers (C1-C20, Operon Technologies) were screened for all 84 plants. Only strong bands that are conservative were scored. Each primer generated a unique set of amplification products. Most of scoreable bands are ranged from 350 to 1800 bp. A total of 3696 fragments were amplified from 42 shoot-derived plants by all 20 primers with an average of 4.4 bands per primer, in which 6 primers produced polymorphic bands, indicating some genetic variations within shoot-derived plants. Nineteen out of 20 primers yielded 2772 clear and reproducible bands (an average of 3.47 per primer) from 42 regenerated plants with no significant variations being detected. Our preliminary results showed that in vitro regenerated plants are genetically uniform. However, a long-term tissue culture might result in a few genetic variations of birch species.

scholarly journals In vitro Plant Production through Apical Meristem Culture of Bitter Gourd (Momordica charantia L.)

1970 ◽  
Vol 16 (1) ◽  
pp. 31-36 ◽  
Author(s):  
AKMN Huda ◽  
B Sikdar ◽  
B Sikdar
Keyword(s):  
Apical Meristem ◽  
Shoot Elongation ◽  
Plant Production ◽  
Bitter Gourd ◽  
Meristem Culture ◽  
Ms Medium ◽  
Semisolid Medium ◽  
Shoot Initiation ◽  
In Vitro Plantlets

The growth of meristem was observed on semisolid MS medium supplemented with 0.05 mg/l Kn + 0.1 mg/l GA3. After three weeks, meristems were transferred to MS supplemented with BA, Kn, IBA, NAA and IAA singly or in combination for shoot elongation and root initiation. Among different treatments for shoot initiation with elongation were obtained in MS supplemented with 1.0 mg/l BA + 0.1 mg/l IBA + 0.3 mg/l GA3. On the other hand good rooting was observed when 0.5 mg/l IBA and 0.1 mg/l NAA were used to fortify MS semisolid medium. Ten weeks old in vitro plantlets were successfully planted in soil through gradual acclimation.Key words: Apical meristem, Bitter gourd, In vitro, Plant production DOI = 10.3329/ptcb.v16i1.1103Plant Tissue Cult. & Biotech. 16(1): 31-36, 2006 (June)

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