scholarly journals 112 The Influence of Plant Growth Regulators and Light Levels on Shoot Morphogenesis from Leaf Explants of Highbush Blueberry

HortScience ◽  
1999 ◽  
Vol 34 (3) ◽  
pp. 460F-461
Author(s):  
Xiaoling Cao ◽  
F.A. Hammerschlag
Keyword(s):  
Light Intensity ◽  
Shoot Regeneration ◽  
High Efficiency ◽  
Leaf Explants ◽  
Vaccinium Corymbosum ◽  
Zeatin Riboside ◽  
Highbush Blueberry ◽  
Shoot Cultures ◽  
Regeneration Medium

As part of a program to develop transgenic highbush blueberry (Vaccinium corymbosum L.) cultivars, studies were conducted to determine optimum conditions for high efficiency shoot regeneration from leaf explants of in vitro-propagated shoot cultures. The effect of either thidiazuron at 1 or 5 μM, or zeatin riboside at 20 μM, and two lit levels (18 ± 5 or 55 ± 5 μmol·m-2·s-1) on shoot organogenesis were investigated. With the exception of `Bluecrop', which did not regenerate shoots, maximum shoot regeneration of 13, 12.7, 12.6 and 4.6 shoots per explant for cultivars Duke, Georgiagem, Sierra, and Jersey, respectively, occured on regeneration medium with zeatin riboside and under a light intensity of 55 μmol·m-2·s-1. Whereas `Duke' regenerated equally well on regeneration medium with either zeatin riboside or 5 μM thidiazuron, regeneration frequencies for `Georgiagem' and `Sierra' were significantly higher on zeatin riboside. A light intensity of 55 μmol·m-2·s-1 significantly increased regeneration of cultivars Duke, Jersey, and Sierra on zeatin riboside, but inhibited regeneration of Duke on 5 μM thidazuron.

scholarly journals 307 Growth Regulator Pretreatments Significantly Enhance the Efficiency of Shoot Organogenesis from Leaf Explants of Highbush Blueberry Cultivar Bluecrop

HortScience ◽  
2000 ◽  
Vol 35 (3) ◽  
pp. 445A-445
Author(s):  
X. Cao ◽  
F. Hammerschlag
Keyword(s):  
Shoot Regeneration ◽  
Growth Regulators ◽  
High Efficiency ◽  
Shoot Organogenesis ◽  
Leaf Explants ◽  
Vaccinium Corymbosum ◽  
Zeatin Riboside ◽  
Highbush Blueberry ◽  
Shoot Cultures ◽  
Regeneration Medium

As part of a program to develop transgenic highbush blueberry (Vaccinium corymbosum L.) cultivars, studies were conducted to determine optimum conditions for high-efficiency shoot regeneration from leaf explants of in vitro propagated, commercially important, tissue culture-recalcitrant `Bluecrop' shoot cultures. The effects of pretreatments, growth regulators, and age of explant source on shoot organogenesis were investigated. A maximum of 98% shoot regeneration and 10 shoots regenerating per leaf explant occurred when explants of 2-week-old shoot cultures were incubated in the dark (for a total of 14 days) on pretreatment medium #1 containing 2.6 μM NAA and 5 μM TDZ for 4 days, next on pretreatment medium #2 containing 2.6 μM NAA and 7 μM zeatin riboside for 3 days, then on regeneration medium containing 1 μM TDZ for 6 weeks, and last on medium without growth regulators for 10 days. No shoot regeneration occurred if explants were incubated without exposure to pretreatments before incubation on regeneration medium. There were no significant differences in percentage of regeneration or the number of shoots regenerating per explant from leaf explants derived from either 1-, 2-, or 3-week-old shoot cultures. Shoot production per explant on 1 μM TDZ was about three times that on either 0.5 μM TDZ or 20 μM zeatin riboside, and nine times that on 5 μM TDZ.

scholarly journals Improved Shoot Organogenesis from Leaf Explants of Highbush Blueberry

HortScience ◽  
2000 ◽  
Vol 35 (5) ◽  
pp. 945-947 ◽  
Author(s):  
X. Cao ◽  
F.A. Hammerschlag
Keyword(s):  
High Efficiency ◽  
Shoot Organogenesis ◽  
Leaf Explants ◽  
Vaccinium Corymbosum ◽  
Zeatin Riboside ◽  
Photon Flux ◽  
Highbush Blueberry ◽  
Shoot Cultures ◽  
Regeneration Medium ◽  
Number Of Shoots

As part of a program to develop transgenic highbush blueberry (Vaccinium corymbosum L.) cultivars, studies were conducted to determine optimum conditions for high efficiency shoot regeneration from leaf explants of shoots propagated in vitro. The effects on shoot organogenesis of age of explant source, length of dark treatment, the addition of either thidiazuron (TDZ) at 1 or 5 μm, or zeatin riboside at 20 μm to the regeneration medium, and a photosynthetic photon flux (PPF) of either 18 ± 5 or 55 ± 5 μmol·m–2·s–1 were investigated. A maximum of 13.0, 13.0, 12.6, and 4.6 shoots regenerating per explant for cultivars Duke, Georgiagem, Sierra, and Jersey, respectively, occurred on regeneration medium with zeatin riboside and under a PPF of 55 ± 5 μmol·m–2·s–1. `Duke' regenerated equally well on medium with either zeatin riboside or 1 μm TDZ, whereas the number of shoots per explant for `Georgiagem' and `Sierra' was significantly higher on zeatin riboside. Regeneration of `Duke', `Jersey', and `Sierra' on zeatin riboside was significantly better under a PPF of 55 ± 5 μmol·m–2·s–1 than under 18 ± 5 μmol·m–2·s–1, but the higher PPF inhibited regeneration of `Duke' on 5 μm TDZ. There were no significant differences in percentage of regeneration or the number of shoots per explant from leaf explants derived from either 1-, 2-, or 3-week-old shoot cultures, or when either 1 week or 2 weeks of darkness preceded light treatments. Chemical names used: 1-phenyl-3-(1,2,3-thiadiazol-5-yl)urea (thidiazuron, TDZ); 9-(-β-ribofuranosyl)-6-(4-hydroxy-3-methyl-but-2-enylamino)purine (zeatin riboside).

scholarly journals A Two-step Pretreatment Significantly Enhances Shoot Organogenesis from Leaf Explants of Highbush Blueberry cv. Bluecrop

HortScience ◽  
2002 ◽  
Vol 37 (5) ◽  
pp. 819-821 ◽  
Author(s):  
Xiaoling Cao ◽  
Freddi A. Hammerschlag ◽  
Larry Douglass
Keyword(s):  
Growth Regulators ◽  
Shoot Organogenesis ◽  
Leaf Explants ◽  
Vaccinium Corymbosum ◽  
Zeatin Riboside ◽  
Highbush Blueberry ◽  
Shoot Cultures ◽  
Regeneration Medium ◽  
Explant Source ◽  
Shoot Production

As part of a program to improve highbush blueberry (Vaccinium corymbosum L.) cultivars via tissue culture and genetic engineering, studies were conducted to determine optimum conditions for organogenesis from leaf explants of the previously recalcitrant cv. Bluecrop. The effects of a pretreatment, growth regulators, and age of explant source on shoot organogenesis were investigated. A maximum of 98% explants regenerated shoots with a mean of 11 shoots per leaf explant after 62 days when explants of 2-week-old shoot cultures were incubated on the following regime: pretreatment medium #1 containing 5 μm TDZ and 2.6 μm NAA for 4 days, pretreatment medium #2 containing 7 μm zeatin riboside and 2.6 μm NAA for 3 days, regeneration medium containing 1 μm TDZ for 6 weeks, and last on medium without growth regulators for 10 days. No shoot regeneration occurred if explants were incubated without exposure to pretreatment prior to incubation on regeneration medium. There were no significant differences in percentage of regeneration or the number of shoots regenerating per explant from leaf explants derived from either 1-, 2-, or 3-week-old shoot cultures. Shoot production per explant on regeneration medium containing 1 μm TDZ was about three times that on 0.5 μm TDZ or 20 μm zeatin riboside, and nine times that on 5 μm TDZ. Chemical names used: 1-phenyl-3-(1,2,3-thiadiazol-5-yl)urea (thidiazuron, TDZ); 9-(β-D-ribofuranosyl)-6-(4-hydroxy-3-methyl-but-2-enylamino)purine (zeatin riboside).

scholarly journals Development of an in vitro Callus Induction Protocol and Shoot Proliferation for Selected Jatropha (Jatropha curcas) Accessions

2020 ◽  
Vol 30 (1) ◽  
pp. 131-141
Author(s):  
Hundessa Fufa ◽  
Jiregna Daksa
Keyword(s):  
Shoot Regeneration ◽  
Callus Induction ◽  
Jatropha Curcas ◽  
Plant Tissue ◽  
Leaf Explants ◽  
Shoot Proliferation ◽  
Adventitious Shoot ◽  
Regeneration Medium ◽  
Adventitious Shoot Buds

The present study was undertaken to establish a protocol for in vitro callusing of three Jatropha accessions, namely Metema, Adami Tulu and Shewa Robit from leaf explants. The medium supplemented with combination of 4.44 μM BAP and 4.52 μM 2,4-D resulted in maximum percentage of callus (100%) formed for all accessions. The maximum shoot regeneration (66.67%) from callus with 10.13 number of shoot was obtained from Shewa Robit in MS medum fortified with TDZ (2.27 μM ) and IBA (0.49 μM ). The presence of TDZ in the shoot regeneration medium has greater influence on the induction of adventitious shoot buds, whereas MS supplemented with BAP alone and combination with IBA did not induce shoot regeneration from callus culture. The results obtained in the present study would facilitate the high callus induction and regeneration responses in Jatropha for its improvement using biotechnological tools. Plant Tissue Cult. & Biotech. 30(1): 131-141, 2020 (June)

scholarly journals Use of a Cytokinin Conjugate for Efficient Shoot Regeneration from Leaf Sections of Highbush Blueberry

HortScience ◽  
1992 ◽  
Vol 27 (10) ◽  
pp. 1127-1129 ◽  
Author(s):  
Lisa J. Rowland ◽  
Elizabeth L. Ogden
Keyword(s):  
Tissue Culture ◽  
Shoot Regeneration ◽  
Culture Medium ◽  
Vaccinium Corymbosum ◽  
Zeatin Riboside ◽  
Tissue Culture Medium ◽  
Highbush Blueberry ◽  
Leaf Section ◽  
Number Of Shoots

Conditions for improving the efficiency of shoot regeneration from leaf sections of highbush blueberry (Vaccinium corymbosum L.) were investigated. Effectiveness of tissue culture medium supplemented with the cytokinin conjugate zeatin riboside or the cytokinin zeatin at 10, 20, or 30 μm was compared with medium supplemented with the optimum 2iP concentration of 15 μm. Use of 20 μm zeatin riboside resulted in the most shoots per leaf section, » 6-fold higher than the number of shoots produced on 2iP medium. The number of shoots produced on medium supplemented with zeatin was not significantly higher than the number of shoots produced on 2iP medium. Consequently, we concluded that the cytokinin conjugate zeatin riboside was more effective than either of the free cytokinins, 2iP or zeatin, in promoting shoot regeneration from leaf sections of highbush blueberry. Chemical names used: 6-(y,y-dimethylallylamino)-purine (2iP); 6-(4-hydroxy-3-methyl-but-2-enylamino)purine (zeatin).

scholarly journals PCIB Enhances Regeneration of Ipomoea cordatotriloba Dennstedt Leaf Explants and Protoplasts

HortScience ◽  
1994 ◽  
Vol 29 (4) ◽  
pp. 327-328
Author(s):  
Ruth S. Kobayashi ◽  
John C. Bouwkamp ◽  
Stephen L. Sinden
Keyword(s):  
Abscisic Acid ◽  
Shoot Regeneration ◽  
Callus Induction ◽  
Leaf Explants ◽  
Dichlorophenoxyacetic Acid ◽  
Regeneration Medium ◽  
Ipomoea Cordatotriloba ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Chlorophenoxyisobutyric Acid

Leaf callus of Ipomoea cordatotriloba was initiated by culturing explants on Linsmaier and Skoog medium containing 3 g yeast extract/liter, 18.9 μm ABA, 2.3 μm 2,4-D, and 0.15 m sucrose. Calluses were transferred to Murashige and Skoog media containing 17.8 μm BA and 0, 1, 10, or 100 μm PCIB. The number of shoots from calluses grown on medium containing 10 μm PCIB increased significantly, and the percentage of calluses exhibiting shoot regeneration almost doubled compared to calluses grown on regeneration medium without PCIB. Protoplasts isolated from stem and petiole tissues of in vitro-grown plants were cultured in Kao and Michayluk 8p medium to the callus stage. Calluses (4-6 mm) were transferred to the callus induction and regeneration media used to regenerate leaf-explant callus. Of the protoplast-derived calluses cultured on media containing 10 or 100 μm PCIB, ≈13% and 18%, respectively, regenerated shoots after 2 months; none regenerated on the medium without PCIB. Chemical names used: abscisic acid (ABA); 2,4-dichlorophenoxyacetic acid (2,4-D); N6-benzyladenine (BA); α -p-chlorophenoxyisobutyric acid (PCIB).

scholarly journals Optimizing the concentrations of plant growth regulators for in vitro shoot cultures, callus induction and shoot regeneration from calluses of grapes

OENO One ◽  
2015 ◽  
Vol 49 (1) ◽  
pp. 37 ◽  
Author(s):  
Nadra Khan ◽  
Maqsood Ahmed ◽  
Ishfaq Hafiz ◽  
Nadeem Abbasi ◽  
Shaghef Ejaz ◽  
...  
Keyword(s):  
Shoot Regeneration ◽  
Growth Regulators ◽  
Callus Induction ◽  
Leaf Explants ◽  
Shoot Cultures ◽  
Ms Medium ◽  
Grape Cultivar ◽  
Half Strength ◽  
Number Of Shoots

<p style="text-align: justify;"><strong>Aim</strong>: To optimize the concentrations of growth regulators in the media for the proficient micropropagation of grapevine (<em>Vitis vinifera </em>L.) cv. King’s Ruby.</p><p style="text-align: justify;"><strong>Methods and results</strong>: Apical meristems of the grape cultivar were used to establish <em>in vitro</em> shoot cultures. Nodal explants, each containing an axillary bud, taken from <em>in vitro</em> grown shoots were inoculated in shoot proliferation medium, i.e., half strength Murashige and Skoog (MS) medium supplemented with benzyl aminopurine (BAP), kinetin, glycine and gibberellic acid (GA<sub>3</sub>). A higher number of shoots (5.33) with greater shoot length (2.75 cm) was produced in the medium supplemented with 1.0 mg L<sup>-1</sup> BAP and 0.1 mg L<sup>-1</sup> GA<sub>3</sub>. Calluses were induced from leaf explants taken from <em>in vitro</em> grown shoots. Callus induction was greater (73.00%) on the medium containing 2.0 mg L<sup>-1</sup> 2,4-dichlorophenoxyacetic acid (2,4-D), 0.3 mg L<sup>-1</sup> BAP and 0.2 mg L<sup>-1</sup> α-naphthaleneacetic acid (NAA). The maximum frequency of shoot regeneration (53.33%) was achieved on the medium supplemented with 1.5 mg L<sup>-1</sup> BAP and 0.5 mg L<sup>-1</sup> NAA, and the regenerated shoots successfully formed roots on growth regulator-free half strength MS medium.</p><p style="text-align: justify;"><strong>Conclusion</strong>: Optimizing the concentration of BAP and GA<sub>3</sub> and omitting the glycine and kinetin in the culture medium increased the number and length of shoots. Similarly, for inducing the callus of the leaf explants, taken from <em>in vitro</em> grown shoots, it is recommended to adjust the medium with the higher concentration of 2,4-D and lower concentrations of BAP. Moreover, the maximum number of shoots was regenerated on a medium supplemented with relatively high levels of both BAP and NAA (1.5 and 0.5 mg L<sup>-1</sup>, respectively). Finally, we suggest the half strength MS medium that is free from growth regulators for the root formation of the regenerated shoots.</p><p style="text-align: justify;"><strong>Significance and impact of the study</strong>: Optimizing the concentration of growth regulators is crucial for the efficient micropropagation of a grape cultivar. Knowing the specific balance between the growth regulators is necessary to establish <em>in vitro</em> shoot cultures, callus induction and shoot regeneration and, hence, to propagate disease-free true to type grape cultivars in a short time.</p>

scholarly journals Chromosome Doubling in Genetically Diverse Bilberry (Vaccinium myrtillus L.) Accessions and Evaluation of Tetraploids in Terms of Phenotype and Ability to Cross with Highbush Blueberry (V. corymbosum L.)

Agronomy ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 2584
Author(s):  
Malgorzata Podwyszynska ◽  
Katarzyna Mynett ◽  
Monika Markiewicz ◽  
Stanisław Pluta ◽  
Agnieszka Marasek-Ciolakowska
Keyword(s):  
Chromosome Doubling ◽  
Diploid Species ◽  
Vaccinium Corymbosum ◽  
Pcr Analysis ◽  
Highbush Blueberry ◽  
Shoot Cultures ◽  
Triploid Block ◽  
Interspecific Crossing ◽  
In Vitro Shoot Cultures

To expand the gene pool and introduce new traits to the tetraploid cultivars of Vaccinium corymbosum from wild diploid species V. myrtillus, it is necessary to double the chromosome number in diploid species in order to overcome a post zygotic crossing barrier and a strong triploid block, existing within the genus Vaccinium. Five genetically diverse bilberry genotypes were selected from 21 accessions taken from the breeding collection of the National Institute of Horticultural Research (Skierniewice, Poland) for this study. The bilberry genotypes were derived from the Polish locations of Bolimów Landscape Park, Budy Grabskie and forest complex Zwierzyniec (Łódź Province), and habitats in Norway. The selection of genotypes was made based on the analysis of amplified fragment length polymorphism (AFLP-PCR). Analysis of the Jaccard similarity indexes and the UPGMA method revealed that the examined accessions formed two main groups on the dendrogram. The first group consisted of accessions from Norway, while the second group agglomerated Polish accessions. A further two classes were distinguished in the Polish group: the first included accessions from Budy Grabskie and the second from Zwierzyniec, located ca. 9 km from Budy Grabskie. In order to obtain plant material for in vitro polyploidisation, in vitro shoot cultures of the selected accessions were initiated and multiplied. Both antimitotics used, colchicine and APM, induced tetraploids for all of the accessions. The obtained tetraploids were multiplied, rooted ex vitro and grown in a greenhouse and then in a field. The first flowering was observed in 1.5-year-old plants, either diploid or tetraploid. Diploids bloomed slightly earlier and more profusely than tetraploid plants. Compared to diploids, autotetraploids had significantly larger flowers by ca. 64% and larger pollen tetrads by ca. 35%. The germination capacity of pollen tetrads was high in tetraploids (87.8%), although slightly lower than in diploids (94.3%). After pollinating the flowers of three highbush blueberry cultivars with pollen from the bilberry tetraploid accession, J-4-4x, the plants formed fruits, some of which contained properly formed seeds. The effectiveness of interspecific crossing between V. corymbosum and tetraploid V. myrtillus, defined as the percentage of obtained seedlings in relation to the number of pollinated flowers, was highest (53.3%) in the blueberry ‘Liberty’, and lower in ‘Bluecrop’ and ‘Northland’, 14.8% and 10.0%, respectively. Before using the seedlings for further breeding, their hybridity will be confirmed by molecular markers and the phenotype will be evaluated.

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