scholarly journals Genetic transformation of Eucalyptus camaldulensis by agrobalistic method

2013 ◽  
Vol 37 (3) ◽  
pp. 419-429 ◽  
Author(s):  
Evânia Galvão Mendonça ◽  
Vanessa Cristina Stein ◽  
Flávia Pereira Balieiro ◽  
Carolina Delfin Fernandes Lima ◽  
Breno Régis Santos ◽  
...  
Keyword(s):  
Genetic Transformation ◽  
Transient Expression ◽  
Culture Media ◽  
Tissue Injury ◽  
Callus Formation ◽  
Nitrogen Concentration ◽  
Eucalyptus Camaldulensis ◽  
Ms Medium ◽  
Gus Gene

Eucalyptus stands in the setting of worldwide forestry due to its adaptability, rapid growth, production of high-quality and low cost of wood pulp fibers. The eucalyptus convetional breeding is impaired mainlly by the long life cycle making the genetic transformation systems an important tool for this purpose. However, this system requires in vitro eficient protocols for plant induction, regeneration and seletion, that allow to obtain transgenic plants from the transformed cell groups. The aim of this work was to evaluate the callus formation and to optimize the leaves and callus genetic transformation protocol by using the Agrobacterium tumefaciens system. Concerning callus formation, two different culture media were evaluated: MS medium supplemented with auxin, cytokinin (M1) and the MS medium with reduced nitrogen concentration and supplemented with auxin, cytokinin coconut water (M2). To establish the leave genetic transformation, those were exposed to agrobiolistics technique (gene gun), to tissue injury, and A. tumesfasciens EHA 105 contening the vetor pCambia 3301 (35S::GUS::NOS), for gene transference and to establish the callus transformation thoses were exposed only to A. tumefasciens. For both experiments, the influence of different infection periods was evaluated. The M2 medium provided the best values for callus sizea and fresh and dry weight. The leaves genetic transformation using the agrobiolistics technique was effective, the gus gene transient expression could be observed. No significant differences were obtained in the infection periods (4, 6 and 8 minutes). The callus genetic transformation with A. tumefaciens also promotend the gus gene transient expression on the callus co-cultiveted for 15 e 30 minutes. The transformed callus was transfered to a regeneration and selection medium and transformed plants were obtained.

scholarly journals Lippia graveolens (Lamiales: Verbenaceae) and Oryganum vulgare (Lamiales: Lamiaceae) obtained by means of the in vitro propagation technique

2021 ◽  
Author(s):  
María A. Aguilar Morales ◽  
Armandina De la Cruz Olvera ◽  
E. Archundia-Garduño ◽  
Rosy G. Cruz Monterrosa ◽  
Mayra Díaz-Ramírez ◽  
...  
Keyword(s):  
Activated Carbon ◽  
Culture Media ◽  
Callus Formation ◽  
Block Design ◽  
Leaf Explants ◽  
Basal Medium ◽  
Culture Technique ◽  
Ms Medium ◽  
Antioxidant Agents

Objective: The objective of this study was to establish the method of propagation of Oryganum vulgare and Lippia graveolens employing a plant tissue culture technique that decreased the phenolization percentages and increased the multiplication coefficients. Design/ methodology/ approach: The in vitro germination percentage was evaluated in both MS and MS medium + activated carbon. Microcuttings (small shoots) of both species were established in base medium added with different antioxidant agents to decrease the phenolization of explants; the treatments were arranged in a completely randomized block  design. For the propagation phase, a completely randomized factorial design was used, where the auxin/cytokinin phytoregulators, type of explants (axillary buds and leaves), and the species (Lippia graveolens and Oryganum vulgare)  were considered as factors. Results: maximum germination (63.3% ±12.5) was obtained on day 15 ​​in both culture media for L. graveolens and O. vulgare. The use of antioxidant agents mainly activated carbon, increased the in vitro establishment and activation of vegetative buds in both species by up to 90%. There were significant differences in the variables evaluated regarding the treatments, the explant, and the species in the multiplication phase. The combination 1.0/ 0.5 mg L-1 BA/AIB induces callus formation for both species. When used as leaf explants, callus formation was potentiated. Study Limitations / Implications: The results presented are advances from a long-term experiment. Findings/conclusions: The germination of L. graveolens seeds can be achieved in MS medium after 15 days. Microcuttings of both L. graveolens and O. vulgare were successfully established in MS basal medium enriched with 1 g L-1 charcoal that showed low oxidation percentages and induced up to 90% the production of shoots in the explants. The mixture of 1.0/0.5 mg L-1 BA/AIB induces callus formation for both species; when this medium is in contact with leaves as an explant, its formation is potentiated, achieving diameters up to 15 mm. In order to achieve the induction of shoots and roots, buds should be established in MS medium enriched with 0.5 mg L-1 IBA for both species; this mixture encreased the multiplication coefficients

scholarly journals IN VITRO SHOOT REGENERATION FROM WESTERN SOAPBERRY

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 697c-697
Author(s):  
Mahmoud B. Arif ◽  
Houchang Khatamian
Keyword(s):  
Nicotinic Acid ◽  
Shoot Regeneration ◽  
Culture Media ◽  
Callus Formation ◽  
Ms Medium ◽  
Full Strength ◽  
Basal Media ◽  
In Vitro Shoot Regeneration

Surface sterilized stem nodal sections of western soapberry (Sapindus drummondii Hook. & Arn.) were cultured on Murashige and Skoog (MS) medium. The basal media consisted of one half and full strength MS medium each supplemented with the following (mg-1): Nicotinic acid 0.5, pyridoxine Hcl 0.5, Glycine 2.0, myo-inositol 100, sucrose 30,000 and agar 8000. Each medium also was supplemented with either 0, 0.01, 0.1, 1.0 and 10 mg/l Thidiazuron (TZD) or 0, 0.5, 2.0 and 5 mg/l 6-Benzyladenine (BA). The pH of all media was adjusted to 5.8 ± 0.1. The culture media were autoclaved at 120°C at 1.5 Kgcm-1 pressure for 15 min. The highest percentage of nodal sections resulting in shoot regeneration occurred on 1/2 MS with TZD at 0.01 mg/l and MS medium containing 0.5 mg/l of BA Increasing the TZD concentration above 0.1 mg/l resulted in callus formation on cut surfaces.

scholarly journals Somaclonal variation on in vitro callus culture potato cultivars

2004 ◽  
Vol 22 (2) ◽  
pp. 300-304 ◽  
Author(s):  
Patricia N. Bordallo ◽  
Derly H. Silva ◽  
José Maria ◽  
Cosme D. Cruz ◽  
Elizabeth P. Fontes
Keyword(s):  
Somaclonal Variation ◽  
Culture Media ◽  
Callus Formation ◽  
Potato Cultivars ◽  
Synthetic Seed ◽  
Arbitrary Sequence ◽  
Stem Explants ◽  
Factors Affecting ◽  
High Level

Synthetic seeds can be an alternative for those species in which botanical seeds are not viable. One of the major problems of in vitro plant cultivation is the high level of somaclonal variation. The most common factors affecting somaclonal variation are genotype, explant source, in vitro period and cultivation conditions in which the culture is established. In this work, calli were induced using leaf and stem explants of the commercial potato cultivars Achat, Baraka, Baronesa, Bintje, and Contenda in MS culture media supplemented with 1.65 mM of picloram and 11.5 mM of 2,4-D. Seventy and 90 days after induction, DNA samples of 40 calli were compared concerning the effects of the two explant (leaf and stem) and two growth regulator sources on five potatoes cultivars. A total of 20 arbitrary sequence primers were evaluated. The RAPD pattern generated by these primers suggested a high percentage of polymorphic fragments among the five genotypes, indicating a high level of genetic variation among cultivars. Cultivar Baronesa showed the highest number of polymorphic fragments for all treatments. The cultivar Contenda showed the smallest somaclonal variation, for most of the treatments, except for the treatment which consisted of stem explants, picloram (1.65 mM) application, and a 70-day period of callus formation. 'Contenda' is, therefore, the most suitable cultivar for synthetic seed production.

scholarly journals In vitro Regeneration of Dalbergia sissoo Roxb. and the Potential for Genetic Transformation

2012 ◽  
Vol 40 (2) ◽  
pp. 140 ◽  
Author(s):  
Hafiz Mamoon REHMAN ◽  
Iqrar Ahmad RANA ◽  
Siddra IJAZ ◽  
Ghulam MUSTAFA ◽  
Faiz Ahmad JOYIA ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Genetic Transformation ◽  
Callus Induction ◽  
In Vitro Regeneration ◽  
Induction Medium ◽  
Native Forest ◽  
Ms Medium ◽  
Dalbergia Sissoo ◽  
Callus Induction Medium

Dalbergia sissoo Roxb. ex DC. (Sissoo) is a native forest tree species in Pakistan. Many ecological and economical uses are associated with this premier timber species, but dieback disease is of major concern. The objective of this study was to develop a protocol for in vitro regeneration of Sissoo that could serve as target material for genetic transformation, in order to improve this species. Callus formation and plantlet regeneration was achieved by culturing cotyledons, immature seeds, and mature embryos on a modified Murashige and Skoog (1962) (MS) medium supplemented with plant growth regulators. Callus induction medium containing 2.71 ?M 2, 4-dichlorophenoxyacetic acid (2,4-D) and 0.93 ?M kinetin produced better callus on all explants tested compared to other treatments, such as 8.88 ?M 6-benzylaminopurine (BA) and 2.69 ?M ?-naphthalene acetic acid (NAA), or 2.71 ?M 2, 4-D and 2.69 ?M NAA. Shoot regeneration was best on MS medium containing 1.4 ?M NAA and 8.88 ?M BA compared to other treatments, such as 1.4 ?M NAA and 9.9 ?M kinetin, or 2.86 ?M indole-3-acetic acid and 8.88 ?M BA. Murashige and Skoog medium containing 1.4 NAA ?M and 8.88 ?M BA was better in general for regeneration regardless of callus induction medium and the type of explant used. Rooting was best on half-strength MS medium with 7.35 ?M indole-3-butyric acid. Regenerated plantlets were acclimatized for plantation in the field. Preliminary genetic transformation potential of D. sissoo was evaluated by particle bombardment of callus explants with a pUbiGus vector. The bombarded tissue showed transient Gus activity 1week after bombardment. Transformation of this woody tree is possible provided excellent regeneration protocols. The best combination for regeneration explained in this study is one of such protocols.

scholarly journals Cell suspension culture and plant regeneration of a Brazilian plantain, cultivar Terra

2008 ◽  
Vol 43 (10) ◽  
pp. 1325-1330 ◽  
Author(s):  
Lucymeire Souza Morais-Lino ◽  
Janay Almeida dos Santos-Serejo ◽  
Sebastião de Oliveira e Silva ◽  
José Raniere Ferreira de Santana ◽  
Adilson Kenji Kobayashi
Keyword(s):  
Plant Regeneration ◽  
Suspension Culture ◽  
Cell Suspension ◽  
Cell Suspension Culture ◽  
Somatic Embryos ◽  
Culture Media ◽  
Ms Medium ◽  
Regenerated Plants ◽  
Male Flowers

The objective of this study was to establish cell suspension culture and plant regeneration via somatic embryogenesis of a Brazilian plantain, cultivar Terra Maranhão, AAB. Immature male flowers were used as explant source for generating highly embryogenic cultures 45 days after inoculation, which were used for establishment of cell suspension culture and multiplication of secondary somatic embryos. Five semisolid culture media were tested for differentiation, maturation, somatic embryos germination and for plant regeneration. An average of 558 plants per one milliliter of 5% SCV (settled cell volume) were regenerated in the MS medium, with 11.4 µM indolacetic acid and 2.2 µM 6-benzylaminopurine. Regenerated plants showed a normal development, and no visible somaclonal variation was observed in vitro. It is possible to regenerate plants from cell suspensions of plantain banana cultivar Terra using MS medium supplemented with 11.4 µM of IAA and 2.2 µM of BAP.

scholarly journals Induction of callus and adventitious shoots on epicotyl and hypocotyl segments of cumaru (Dipteryx odorata)

2018 ◽  
Vol 9 (3) ◽  
pp. 475-480
Author(s):  
Paulo Tarso Barbosa Sampaio ◽  
Lyana Silva Jardim ◽  
Ariel Dotto Blind ◽  
Flavio Mauro Souza Bruno
Keyword(s):  
Native Species ◽  
Callus Formation ◽  
Adventitious Shoots ◽  
Ms Medium ◽  
Adventitious Buds ◽  
Clonal Multiplication ◽  
Benefit Ratio ◽  
Dipteryx Odorata ◽  
Hypocotyl Segments

Somatic embryogenesis from callus induced in epicotyl and hypocotyl segments can be viable native species in order to better -benefit ratio costs, and rates of clonal multiplication. In this sense, two trials were established to induce callus and adventitious buds on hypocotyl and epicotyl segments of cumaru bean seedlings germinated in vitro in different concentrations and combinations of growth regulators. At first, we used the MS medium supplementwith ANA (0.0, 1.5 mg.L-1) and TDZ (0.0, 4.0 and 8.0 mg.L-1) distributed in factorial 2 x 3 x 2 (x auxin cytokinin x explant) with eight replications. In the second, it was used the WPM medium supplemented with BAP (2.0 mg L-1) and plus 2,4-D (2.0 and 4.0 mg L-1) in a factorial 2 x 2 (auxin x explant) with 15 repetitions each. They were evaluating callus formation and the average number of adventitious shoots during the period of 90 days. The results indicated that the highest average for callus formation was observed when the explants were subjected to concentrations of 8.0 mg L-1 TDZ combined with 1.5 mg L-1 ANA in MS medium. For the formation of buds, the WPM medium plus 2.0 mg L-1 2,4-D in the second experiment, induced higher number of shoots, being significant the use of auxin, and its interaction with the type of explant.

scholarly journals Sterilisasi dan Induksi Daun Muda Durian (Durio zibethinus) Dalam Medium MS Dengan Penambahan Kinetin dan IAA Secara In Vitro

2021 ◽  
Vol 1 (1) ◽  
pp. 34-38
Author(s):  
Gatot Supangkat ◽  
Innaka Ageng Rineksane ◽  
Kurniawati Pamuji
Keyword(s):  
Vitamin C ◽  
Callus Formation ◽  
Tween 20 ◽  
Second Phase ◽  
Induction Phase ◽  
Ms Medium ◽  
Sterilization Method ◽  
Central Java ◽  
Durio Zibethinus

A research  to study the sterilization   method  and application   of Kinetin  and IAA to induce the Durian  young  leaf (Durio zibethinus) in MS  medium   was conducted in Balai Benih Induk Hortikultura in Salaman  Magelang  district  of Central  Java  started  on September  until December 2003. The Laboratory experiment   was arranged  in two phases,  which were  the optimation  phase of sterilization   and  induction   phase.  At  the  first  phase,  the  sterilization method  used  was  the modification   of Mulya  (2001) method.  The modification   use of sterilant,  vitamin  C antioxidant, Alcohol  70 %, Benlate, Agrept,  Tween-20  and Betadine  were done to obtain  effectiveness   of the sterilization.  Explants  planted  then in MS medium  for two weeks. Contamination   time, percentage of contamination   and viabilitas  (percentage of living explants)  were observed  then.  At the second phase,  the treatments were arranged  in a 3 x 3 factorial  completely   randomized   design  (CRD)  to observed  the influence  of Kinetin  and IAA combination.   The concentration   of Kinetin  observed were 2, 4, and 6 mg/I, where  as the IAA concentration   were 0.5,  1.0, and  1.5 mg/I. All treatments were  repeated  three  times,  with three samples  on each  replication.   The percentage   of browning explants, percentage  of contaminated   explants,  site of  contamination   and percentage of explants live were observed  at the end of incubation. The results  showed that sterilization  of Durian young leaves explants  with 1  g/l deterjent  for 15 minutes  then by 2 g/l Benlate  and Agrept  for 10 minutes,  then by 1  g/200 mg Vitamin C, then by Alcohol  70 % for 1  minute, then by 20% Clorox,  then by 2 drip of Tween-20  for 10 minute and then by Betadine  decreased  the contamination down to 50 %, and this kind of sterilization  was relatively better than  the other  kinds.  Application   of growth  regulators   were  not  able  to induce  explants growth,  but stimulated  callus formation  at the cutting surface though,  in the application  of Kinetin 4 mg/1 + IAA 0,5 mg/I, Kinetin 4 mg/1 + IAA  1,5 mg/1, Kinetin  6 mg/I+  IAA 0,5  mg/1 and Kinetin 6 mg/l+IAA   1,0 mg/I.

scholarly journals INDUKSI KALUS PASAK BUMI (Eurycoma longifolia Jack) MELALUI EKSPLAN DAUN DAN PETIOL

2016 ◽  
Vol 6 (1) ◽  
pp. 33
Author(s):  
ROSMAINA ROSMAINA ◽  
ZULFAHMI ZULFAHMI ◽  
PROBO SUTEJO ◽  
ULFIATUN ULFIATUN ◽  
MAISUPRATINA MAISUPRATINA
Keyword(s):  
Slow Growth ◽  
Callus Formation ◽  
Germination Percentage ◽  
Ms Medium ◽  
Petiole Explants ◽  
Eurycoma Longifolia ◽  
Yellow Color ◽  
Eurycoma Longifolia Jack ◽  
Short Time

One of the problem of Eurycoma longifolia Jack propagation was low germination percentage due to recalcitrant seeds and slow growth of seedling from cutting propagation. To overcome this problem is required propagation of Eurycoma longifolia via in vitro culture. The objective of this research was to know the effect of Auxin (2,4-D and NAA) and Cytokines (BAP and Kinetin)  on Eurycoma longifolia callus induction via leaf and petiole explants. In this study, we used plant growth regulator of 2,4 D, NAA, BAP and Kinetin in several levels.  The observed variables were appearing callus time, callus color and callus texture. The results of this study showed that MS medium supplemented with 1 ppm NAA+ 1 ppm BAP was able to induce callus formation in leaf explant for 6 months after culture. While MS medium supplemented with 1 ppm 2,4-D, 1 ppm BAP, combination of 2,4-D and Kinetin and combination of 2,4-D and BAP can induce callus formation from petiole. All the callus formation has yellow color and yellow brown color. The petiole explant that is grown in MS medium supplemented with 1 ppm BAP induced of callus in short time (18 days after culture).

scholarly journals Adjustment of Mineral Elements in the Culture Medium for the Micropropagation of Three Vriesea Bromeliads from the Brazilian Atlantic Forest: The Importance of Calcium

HortScience ◽  
2009 ◽  
Vol 44 (1) ◽  
pp. 106-112 ◽  
Author(s):  
Alice Noemí Aranda-Peres ◽  
Lázaro Eustáquio Pereira Peres ◽  
Edson Namita Higashi ◽  
Adriana Pinheiro Martinelli
Keyword(s):  
New Media ◽  
Mineral Composition ◽  
Culture Media ◽  
Dry Weight ◽  
Defined Medium ◽  
Ms Medium ◽  
Media Formulation ◽  
Half Strength

Many different species of Bromeliaceae are endangered and their conservation requires specific knowledge of their growth habits and propagation. In vitro culture of bromeliads is an important method for efficient clonal propagation and in vitro seed germination can be used to maintain genetic variability. The present work aims to evaluate the in vitro growth and nutrient concentration in leaves of the epiphyte bromeliads Vriesea friburguensis Mez, Vriesea hieroglyphica (Carrière) E. Morren, and Vriesea unilateralis Mez, which exhibit slow rates of growth in vivo and in vitro. Initially, we compared the endogenous mineral composition of bromeliad plantlets grown in half-strength Murashige and Skoog (MS) medium and the mineral composition considered adequate in the literature. This approach suggested that calcium (Ca) is a critical nutrient and this was considered for new media formulation. Three new culture media were defined in which the main changes to half-strength MS medium were an increase in Ca, magnesium, sulfur, copper, and chloride and a decrease in iron, maintaining the nitrate:ammonium rate at ≈2:1. The main difference among the three new media formulated was Ca concentration, which varied from 1.5 mm in half-strength MS to 3.0, 6.0, and 12 mm in M2, M3, and M4 media, respectively. Consistently, all three species exhibited significantly higher fresh and dry weight on M4, the newly defined medium with the highest level of Ca (12 mm). Leaf nitrogen, potassium, zinc, magnesium, and boron concentrations increased as Ca concentration in the medium increased from 1.5 to 12 mm.

scholarly journals Callus Induction in Baru (Dipteryx alata Vog.) Explants

2018 ◽  
Vol 47 (2) ◽  
pp. 538-543
Author(s):  
Rodrigo Kelson S. REZENDE ◽  
Ana Maria N. SCOTON ◽  
Maílson V. JESUS ◽  
Zeva V. PEREIRA ◽  
Fernanda PINTO
Keyword(s):  
Ascorbic Acid ◽  
Callus Induction ◽  
Callus Formation ◽  
Leaf Explants ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Alternative Technique ◽  
Propagation Methods ◽  
Dipteryx Alata

Baru (Dipteryx alata Vog.) is a species with great economic and environmental potential; it has popular acceptance, besides being a very productive species. Alternative propagation methods are important for species maintenance and exploration. Thus, micropropagation emerged as an alternative technique, providing genetic stability and the production of a large number of seedlings. The aim of the present investigation was to develop a callus induction protocol for in vitro baru explants. The tested explants were nodal, internodal and foliar segments. The explants were disinfected for 30 seconds in 70% alcohol (v/v) and 2 minutes in sodium hypochlorite (1.25% active chlorine). This was followed by triple washing. The inoculation was carried out in test tubes containing 15 mL MS medium (30 g L-1 sucrose, 6 g L-1 agar and 100 mg L-1 ascorbic acid) supplemented with 2.0 mg L-1 naphthalene acetic acid (NAA). The solution also contained 0.0, 2.5 or 5.0 mg L-1 of 6-benzylaminopurine (BAP) with the pH adjusted to 5.8. In the incubation phase, the explants were cultured for seven days in the dark and then subjected to a photoperiod of 16 hours (43 µmol m-2 s-1) at 25 ± 2 °C. The treatments were studied with 2.5, 5.0, 7.5 or 10.0 mg L-1 BAP additions to the MS. Callus formation, contamination and oxidation evaluations were undertaken. The results obtained when using 2.0 mg L-1 NAA concluded that such a treatment should be used to induce callogenesis from nodal explants, while for the tested baru leaf explants, the best results for callus formation were given by the combination of 2.0 mg L-1 NAA with 2.5 mg L-1 of BAP to.

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