scholarly journals IN VITRO PROPAGATION OF VIRGINIA PINE FROM COTYLEDONS

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1101H-1102
Author(s):  
Carole H. Saravitz ◽  
Frank A. Blazich ◽  
Henry V. Amerson

Adventitious shoots developed on cotyledons of Virginia pine (Pinus virginiana Mill.) excised from seeds germinated for 3, 6, or 9 days and cultured on media containing 0.5 to 10 mg/liter benzyladenine (BA). Shoot regeneration was greatest (46 shoots per embryo) on cotyledons from seeds germinated for 6 days and placed on medium containing 10 mg/liter BA. Shoots were excised and elongated on medium lacking BA. Following elongation, shoots were placed on media containing 0 to 40 mg/liter indolebutyric acid (IBA) for 14 days followed by transfer to the same medium lacking auxin. Without IBA treatment, percent rooting was 3% and increased to 50% for concentrations of 5 to 40 mg/liter. Rooted shoots averaged 2.0 roots per shoot without auxin treatment, 3.3 roots when treated with 5 mg/liter IBA and root number increased linearly with increased IBA concentration up to 40 mg/liter (4.5 roots). Plant lets were transferred to growing medium and acclimated successfully to greenhouse conditions.

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1101h-1102
Author(s):  
Carole H. Saravitz ◽  
Frank A. Blazich ◽  
Henry V. Amerson

Adventitious shoots developed on cotyledons of Virginia pine (Pinus virginiana Mill.) excised from seeds germinated for 3, 6, or 9 days and cultured on media containing 0.5 to 10 mg/liter benzyladenine (BA). Shoot regeneration was greatest (46 shoots per embryo) on cotyledons from seeds germinated for 6 days and placed on medium containing 10 mg/liter BA. Shoots were excised and elongated on medium lacking BA. Following elongation, shoots were placed on media containing 0 to 40 mg/liter indolebutyric acid (IBA) for 14 days followed by transfer to the same medium lacking auxin. Without IBA treatment, percent rooting was 3% and increased to 50% for concentrations of 5 to 40 mg/liter. Rooted shoots averaged 2.0 roots per shoot without auxin treatment, 3.3 roots when treated with 5 mg/liter IBA and root number increased linearly with increased IBA concentration up to 40 mg/liter (4.5 roots). Plant lets were transferred to growing medium and acclimated successfully to greenhouse conditions.


1991 ◽  
Vol 116 (2) ◽  
pp. 362-365 ◽  
Author(s):  
Carole H. Saravitz ◽  
Frank A. Blazich ◽  
Henry V. Amerson

Adventitious shoots developed on cotyledons of Virginia pine (Pinus virginiana Mill.) excised from seeds subjected to H2O2 treatment for 3, 6, or 9 days and cultured on media containing 0.5 to 10 mg BA/liter. Shoot regeneration was greatest (42 shoots per embryo) on cotyledons from seeds treated with H2O2 for 6 days and placed on medium containing BA at 10 mg·liter-1. Excised shoots elongated on medium lacking BA. Following elongation, shoots were placed on media containing IBA at 0 to 40 mg·liter-1 for 14 days followed by transfer to the same medium lacking auxin. Without IBA treatment, rooting was 3%, and increased to 50% for 5 to 40 mg·liter-1. Rooted shoots averaged 2.0 roots per shoot without auxin incorporation, 3.3 roots when treated with 5 mg IBA/liter, and the number of roots increased linearly with increased IBA concentration up to 40 mg·liter-1 (4.5 roots). Plantlets were transferred to growing medium and acclimated successfully to greenhouse conditions. Chemical names used: N- (phenylmethyl)-1 H- purine-6-amine (BA), 1 H- indole-3-butyric acid CBA).


1987 ◽  
Vol 65 (1) ◽  
pp. 72-75 ◽  
Author(s):  
J. Y. Peron ◽  
E. Regnier

A method for rapid micropropagation of sea kale (Crambe maritima L.) was developed. Petiole explants placed in vitro on a medium containing 0.5 mg/L indoleacetic acid (IAA), 6.0 mg/L kinetin, and 1.5 mg/L benzylaminopurine developed callus within 15 days and shoots within 28 days. Nearly four adventitious shoots could be developed within 3 weeks by placing the initial shoot on media without IAA. To develop roots, the shoots were then transferred to the basal medium containing 0.1 to 1.0 mg/L indolbutyric or α-naphthaleneacetic acid. Rooted plantlets were obtained within 2 or 3 weeks. After an acclimatization period of 6 weeks in a greenhouse in unsterilized medium, the plantlets could be set outdoors.


HortScience ◽  
2017 ◽  
Vol 52 (7) ◽  
pp. 996-999 ◽  
Author(s):  
Carlos Alberto Lecona-Guzmán ◽  
Sheila Reyes-Zambrano ◽  
Felipe Alonso Barredo-Pool ◽  
Miguel Abud-Archila ◽  
Joaquín Adolfo Montes-Molina ◽  
...  

Factors such as slow growth, low rates of sexual and asexual reproduction, and viability of seeds among others limit the massive propagation of Agave americana L. by conventional methods. In this study, callus induction and shoot proliferation was determined in A. americana using Murashige and Skoog (MS) medium supplemented with dicholorophenoxyacetic acid (2,4-D) and 6-benzyl adenine (BA). Meristematic tissue was used as the explants, and were placed on MS medium supplemented with 30.0 g·L−1 sucrose with 0.11, 0.18, or 0.45 μm 2,4-D and 11.0, 22.0, 38.2, 44.0, 58.7, or 73.3 μm BA. Treatments were implemented according to factorial experimental design 3 × 6. After 1 month, the number of explants with callus was determined, whereas the numbers of shoots per explant were monitored after 4, 16, 20, and 36 weeks. The maximum percent of explants with callus was obtained with 0.11 μm 2,4-D and 58.7 and 73.3 μm BA, whereas the maximum numbers of shoots per explant (71) were obtained with 0.11 μm 2,4-D and 73.3 μm BA. The effect of different concentrations of indolebutyric acid (IBA) in the rooting of shoots was evaluated. There were no significant effects of IBA on the number of roots, root length, and axillary roots. Plantlets were acclimatized in the glasshouse and they did not show any phenotypic alteration. This is a highly efficient protocol for the in vitro propagation of A. americana via indirect organogenesis.


Author(s):  
Lani Irvin ◽  
Yarelia Zavala Ortiz ◽  
Kamila Rivera Rivera ◽  
Brajesh Nanda Vaidya ◽  
Samantha H Sherman ◽  
...  

We report the development of in vitro propagation protocols through adventitious shoot induction pathway for a rare and medicinal Scutellaria havanensis. In vitro propagation studies using nodal explants showed MS medium supplemented with 10µM 6-Benzylaminopurine induced highest number of adventitious shoots in a time dependent manner. A ten - day incubation was optimum for shoot bud induction as longer exposures resulted in hyperhydricity of the explants and shoots induced. We also report preliminary evidence of Agrobacterium tumefaciens EHA105 - mediated gene transfer transiently expressing of green fluorescent protein in this species. Transformation studies exhibited amenability of various explant tissues, internode being the most receptive. As the plant has medicinal value, research was carried out to evaluate its potential antioxidant capacity and the efficacy of methanolic leaf extracts in curbing the viability of human colorectal cancer cell line HCT116. Comparative total polyphenol, and flavonoid content measurement of fresh and air dried leaf extract revealed that the fresh leaf extracts contain higher total polyphenol and flavonoid content. The HCT 116 cell viability assessed by colorimetric assay using a 3-(4, 5-dimethyl-thiazol-2-yl)-2, 5-diphenyltetrazolium bromide, showed a steady growth inhibition after 24 hours of incubation. Scanning Electron Microscopy of leaf surface revealed high density of glandular and non-glandular trichomes. This research provides basis for the conservation of this rare plant and future phytochemical screening and clinical research.


2020 ◽  
Vol 8 (10) ◽  
pp. 237-244
Author(s):  
Duru Maduabuchi ◽  
◽  
Mbata Ikechukwu ◽  
Osikwe Keziah ◽  
Ukaoma Adamma ◽  
...  

The study investigated an in vitro propagation of Manihot esculenta Crantz in a substituted substrate regime. The aim was to proffer and affordable alternative to the expensive high tech media formulations usually employed in tissue culture protocol. The experiment was conducted on laboratory bench, using standard tissue culture and micropropagation methods under aseptic conditions. The morphogenesis effect of the substrate was determined based on the integer number of explants’ callus and adventitious shoot regeneration. Results showed that MS + Agar, supported embryogenic callus formation with 38% viability, NH4NO3 + KH2PO4 + Agar, supported same with 29%. MS + 2, 4-D + BAP +Agar supported shoot establishment with 32%. While NH4NO3 + KH2PO4 + Zea mays extracts + Agar, did same with 43.26%. MS + Soil, supported callugenesis with 27% viability while NH4NO3 + KH2PO4 + Soil supported the callus establishment with 25%. MS + 2,4 - D + BAP + Soil, supported shoot establishment with 38.41% viability while NH4NO3 + KH2PO4 + Zea mays Extracts + Soil supported same with 36%. The application of crude Zea mays seedling extracts can serve as potent alternative to the synthetic 2, 4 – D and BAP, in in vitro somatic cell morphogenesis. NH4NO3 + KH2 + PO4 can substitute for the MS salt in the same protocol. Loamy top soil can be a good alternative to agar powder as gelling agent in cassava somatic cell embryogenesis and shoot regeneration. Keywords: Ammonium nitrate, Potassium biphosphate, MS salt, axillary meristem, morphogenesis.


HortScience ◽  
2000 ◽  
Vol 35 (4) ◽  
pp. 749-750 ◽  
Author(s):  
Michael E. Compton

Organic competence of different explant sizes and locations on watermelon seedlings was determined by calculating the percentage of cotyledon explants that produced adventitious shoots. About 52% (214/412) of explants prepared from the proximal region of cotyledons formed shoots, whereas only ≈6% (24/411) of distal explants did so. Shoot formation was limited to the proximal end of basal explants but was not restricted to any specific region on distal ones. The percentage of explants that produced harvestable shoots was greater from basal halves than basal quarters in `Sweet Gem', `Crimson Sweet', and `Minilee', but explant size did not affect adventitious shoot regeneration of `Yellow Doll', resulting in significant interaction between cultivar and explant size. This study indicates that cultivars that respond poorly to in vitro procedures may have fewer cells competent for shoot regeneration, requiring special care during explant preparation.


HortScience ◽  
2006 ◽  
Vol 41 (1) ◽  
pp. 193-201 ◽  
Author(s):  
Ana Carolina Espinosa ◽  
Paula M. Pijut ◽  
Charles H. Michler

A complete regeneration protocol was developed for Prunus serotina Ehrh., an important hardwood species for timber and sawlog production in the central and eastern United States. Nodal sections were cultured on Murashige and Skoog (MS) medium supplemented with 4.44 μm 6-benzylaminopurine (BA), 0.49 μm indole-3-butyric acid (IBA), and 0.29 μm gibberellic acid (GA3). In vitro leaf explants of three genotypes were placed on woody plant medium (WPM) supplemented with 0, 2.27, 4.54, or 6.81 μm thidiazuron (TDZ) in combination with 0, 0.54, 1.07, or 5.37 μm naphthaleneacetic acid (NAA), and on WPM supplemented with 0, 4.44, 8.88, or 13.32 μm BA in combination with 0, 0.54, 1.07, or 5.37 μm NAA. Cultures were maintained either in continuous darkness for 5 weeks, or in the dark for 3 weeks and then transferred to a 16-hour photoperiod. TDZ and the genotype had a significant effect on the number of shoots regenerated. The maximum mean number of shoots regenerated per explant (5.05 ± 1.14) was obtained with 2.27 μm TDZ plus 0.54 μm NAA with the 3-week dark period then light treatment. The highest percent shoot regeneration (38.3) and mean number of shoots (4.13 ± 0.97) was obtained with 6.81 μm TDZ plus 1.07 μm NAA. The highest rooting (27%) of adventitious shoots and number of roots per shoot (2.3 ± 0.2) was obtained with 2.5 μm IBA when shoots were maintained for 7 days in the dark on rooting medium before transfer to a 16-hour photoperiod. The highest rooting (70%) of nodal explant-derived stock cultures and number of roots per shoot (2.7 ± 0.9) was also obtained with 2.5 μm IBA, but when shoots were maintained for 4 days in the dark before transfer to a 16-hour photoperiod. In total, 86% of the plantlets survived acclimatization to the greenhouse and 100% survival after overwintering in cold-storage.


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