scholarly journals Protocol optimization and histological analysis of in vitro plant regeneration of RB92579 and RB93509 sugarcane cultivars

2012 ◽  
Vol 43 (1) ◽  
pp. 49-54 ◽  
Author(s):  
Roberson Dibax ◽  
Giovana Bomfim de Alcantara ◽  
Marília Pereira Machado ◽  
João Carlos Bespalhok Filho ◽  
Ricardo Augusto de Oliveira

The objectives of this study were to establish appropriate conditions for obtaining plant regeneration and acclimatization of the 'RB92579' and 'RB93509' sugarcane cultivars and to elucidate the shoots origin through histological analysis. For both cultivars, obtaining shoots showed better results with the culture of explants on a callus induction medium containing 2.0mg L-1 2,4-dichlorophenoxyacetic acid, followed by cultivation on a shoot induction medium containing 0.1mg L-1 kinetin and 0.2mg L-1 benzilaminopurine. The MS medium without growth regulators proved to be appropriate for elongation and rooting of shoots and the use of the composed substrate of vermiculite + MS salts was effective for acclimatization. Histological analysis revealed that the origin of the shoots in both cultivars occurred through indirect organogenesis.

Agronomy ◽  
2020 ◽  
Vol 10 (6) ◽  
pp. 839
Author(s):  
Dorota Weigt ◽  
Idzi Siatkowski ◽  
Magdalena Magaj ◽  
Agnieszka Tomkowiak ◽  
Jerzy Nawracała

Ionic liquids are novel compounds with unique chemical and physical properties. They can be received based on synthetic auxins like 2,4-dichlorophenoxyacetic acid or dicamba, which are commonly used hormones in microspore embryogenesis. Nevertheless, ionic liquids have not been adapted in plant in vitro culture thus far. Therefore, we studied the impact of ionic liquids on the ability to undergo microspore embryogenesis in anther cultures of wheat. Two embryogenic and two recalcitrant genotypes were used for this study. Ten combinations of ionic liquids and 2,4-dichlorophenoxyacetic acid were added to the induction medium. In most cases, they stimulated induction of microspore embryogenesis and green plant regeneration more than a control medium supplemented with only 2,4-dichlorophenoxyacetic acid. Two treatments were the most favorable, resulting in over two times greater efficiency of microspore embryogenesis induction in comparison to the control. The effect of breaking down the genotype recalcitrance (manifested by green plant formation) was observed under the influence of 5 ionic liquids treatments. Summing up, ionic liquids had a positive impact on microspore embryogenesis induction and green plant regeneration, increasing the efficiency of these phenomena in both embryogenic and recalcitrant genotypes. Herbicidal ionic liquids can be successfully used in in vitro cultures.


2008 ◽  
Vol 21 (1) ◽  
pp. 43-48
Author(s):  
S. M. H. Kabir ◽  
M. S. Ali ◽  
M. K. Islam

The Experiment was conducted to establish an efficient plant regeneration protocol from hypocotyl sections of soybean. Callus initiation, shoot and root development were observed by using different concentrations and combinations of growth regulators. The best result for callus induction was observed in MS medium supplemented with 1.5 mg/l Kinetin and 2.0 mg/l NAA. The calli were transferred to shoot induction medium. The best shoot induction occurred in the medium containing 3.0 mg/l BAP and 0.5 mg/l NAA. The elongated shoots developed roots on MS medium supplemented with different IBA concentrations where 1.5 mg/l IBA was the best for root development. Plantlets with a well developed root system were transplanted in plastic container with a soil mixture of cowdung and fine sand. Plantlet survival rate was 70%. Through this experiment, a general suitable regeneration protocol from hypocotyls of soybean has been developed which can potentially be used for micropropagation and future transformation research in soybean.DOI: http://dx.doi.org/10.3329/bjpbg.v21i1.17049


HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 461d-461
Author(s):  
Richard L. Bell ◽  
Ralph Scorza ◽  
Chinnathambi Srinivasan

An efficient regeneration/transformation system was developed for `Beurre Bosc' pear. Young leaves were harvested from in vitro shoots proliferated on a medium containing MS basal salts and 5 BAP, 0.5 μM IBA, and 0.6M3. Shoot regeneration was optimized using a modification of the medium of Chevreau and Leblay (1993). Explants were cultured on shoot induction medium contained 10 μM TDZ and 1 μM IBA for 4 weeks in the dark, and then transfered to a similar, but auxinless, regeneration medium until shoots developed, usually after an additional 4 to 8 weeks. Leaf tissues were transformed by co-cultivation for 3 days with Agrobacterium tumefaciens EHA101 carrying a pGA482 plasmid containing NPTII, GUS, and rolC genes, followed by cultivation on SIM containing 300 mg/L timentin. Putative transgenic plants were selected on shoot induction medium containing 80mg/L kanamycin, and multiplied on shoot proliferation medium. Four clones were confirmed as transgenic using the GUS histochemical assay and Southern blots for the NPTII and rolC genes. Plants of each clone have been rooted and successfully transfered to the greenhouse for further analysis of gene expression.


HortScience ◽  
2001 ◽  
Vol 36 (6) ◽  
pp. 1102-1106 ◽  
Author(s):  
V.R. Bommineni ◽  
H. Mathews ◽  
S.B. Samuel ◽  
M. Kramer ◽  
D.R. Wagner

Improved in vitro clonal propagation methods are valuable tools for nurseries and growers, and are essential for manipulation and improvement of tree fruit germplasm using the tools and techniques of biotechnology. We have developed a rapid shoot multiplication procedure for clonal propagation of apple, Malus ×domestica cv. Gale Gala and pear, Pyrus communis L. cv. Bartlett. Rapid clonal multiplication was achieved after the following series of steps: pre-conditioning of micropropagated shoots, sectioning pre-treated stems into thin slices, placing slices onto shoot induction medium and incubating directly under cool-white fluorescent lights or after a brief dark incubation. Multiple induction of shoots recovered from stem slice explants within three weeks of culture. A maximum of 37% of cultured apple stem slices, and 97% of pear stem slices, showed induction of shoots. More shoots were recovered on phytagel solidified shoot induction medium than on agar. Cultured stem slices of both apple and pear showed maximum recovery of shoots from shoot induction medium supplemented with thidiazuron (TDZ) compared to medium supplemented with BAP and kinetin. Under ideal conditions, pear stems generated four times the shoots as the same quantity or length of apple shoots. Micropropagated shoots were rooted and transferred to the greenhouse and field nursery for further evaluation. Chemical names used: N-phenyl-N′-1,2,3-thidiazol-5-ylurea (thidiazuron or TDZ); 6-benzylaminopurine (BAP).


2018 ◽  
Vol 22 (2) ◽  
pp. 66
Author(s):  
RINDANG - DWIYANI ◽  
HESTIN - YUSWATI ◽  
UTAMI -

ABSTRACT  The objective of the study was to obtain the best 2,4-D concentration on callus induction of the banana flowers in banana propagation using indirect organogenesis method. Kesuna, local banana cultivar obtained from Sembung Gede, Tabanan was used as explant material. Callus induction was performed using 2,4-Dichlorophenoxyacetic acid with concentration of 0; 0.5; 1.0; 1.5 and 2.0 ppm. Each treatment was represented by 3 bottles and each bottle was planted with 3 explants, so each treatment was represented by 9 explants of banana flowers. The results showed that the concentration of 2.0 ppm 2.4-D induced callus with the fastest time and gave the highest percentage of the explants producing callus. The calluses were subsequently subcultured into regeneration medium using 0.5 mg/L Benzylaminopurine (BAP) and 0.005 mg/L Napthaleneaceticacid (NAA). The calluses were subsequently sub-cultured into a regeneration medium using 0.5 ppm (BAP) and 0.005 ppm Naphthalene acetic acid (NAA) to induce shoots and roots and performed plantlets.   Keywords: 2,4-Dichlorophenoxyacetic acid, banana’s flowers, callus


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).


2013 ◽  
Vol 153 ◽  
pp. 109-116 ◽  
Author(s):  
R. Collado ◽  
N. Veitía ◽  
I. Bermúdez-Caraballoso ◽  
L.R. García ◽  
D. Torres ◽  
...  

New Forests ◽  
2011 ◽  
Vol 43 (2) ◽  
pp. 143-154 ◽  
Author(s):  
Qingbin Jiang ◽  
Yong Zhang ◽  
Chonglu Zhong ◽  
Bingshan Zeng ◽  
Didier Bogusz ◽  
...  

Plants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 199
Author(s):  
Milica D. Bogdanović ◽  
Katarina B. Ćuković ◽  
Angelina R. Subotić ◽  
Milan B. Dragićević ◽  
Ana D. Simonović ◽  
...  

Somatic embryogenesis (SE) is a developmental process during which plant somatic cells, under suitable conditions, produce embryogenic cells that develop into somatic embryos (se). SE is the most important method for plant propagation in vitro, having both fundamental and applicative significance. SE can be induced from different tissues and organs, but when se are used as explants, the process is recognized as secondary or cyclic SE. We induced secondary SE in Centaurium erythraea by application of 2,4-dichlorophenoxyacetic acid (2,4-D) and N-(2-chloro-4-pyridyl)-N′-phenylurea (CPPU). A medium containing 0.1 mgL−1 2,4-D and 0.25 mgL−1 CPPU was optimal in terms of the number of primary SE explants forming se, the number of well-developed se per explant, and morphological appearance of the obtained se. These concentrations allowed SE to progress through three cycles, whereas at higher concentrations of 0.2 mgL−1 2,4-D and 0.5 mgL−1 CPPU, only two cycles were achieved. Histological analysis revealed that secondary se are formed both directly and indirectly. Secondary SE readily germinated and converted into plantlets. Induction of cyclic SE contributes to the conservation efforts of this endangered medicinal plant and expands the spectrum of in vitro developmental pathways described in centaury—an emerging model in developmental biology.


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