scholarly journals Integrating the Roles for Cytokinin and Auxin in De Novo Shoot Organogenesis: From Hormone Uptake to Signaling Outputs

2021 ◽  
Vol 22 (16) ◽  
pp. 8554
Author(s):  
Martin Raspor ◽  
Václav Motyka ◽  
Abdul Rasheed Kaleri ◽  
Slavica Ninković ◽  
Ljiljana Tubić ◽  
...  
Keyword(s):  
Shoot Regeneration ◽  
De Novo ◽  
Shoot Organogenesis ◽  
In Vitro Regeneration ◽  
Genetic Regulation ◽  
Auxin Signaling ◽  
Plant Tissues ◽  
Cultivated Plant ◽  
The Media

De novo shoot organogenesis (DNSO) is a procedure commonly used for the in vitro regeneration of shoots from a variety of plant tissues. Shoot regeneration occurs on nutrient media supplemented with the plant hormones cytokinin (CK) and auxin, which play essential roles in this process, and genes involved in their signaling cascades act as master regulators of the different phases of shoot regeneration. In the last 20 years, the genetic regulation of DNSO has been characterized in detail. However, as of today, the CK and auxin signaling events associated with shoot regeneration are often interpreted as a consequence of these hormones simply being present in the regeneration media, whereas the roles for their prior uptake and transport into the cultivated plant tissues are generally overlooked. Additionally, sucrose, commonly added to the regeneration media as a carbon source, plays a signaling role and has been recently shown to interact with CK and auxin and to affect the efficiency of shoot regeneration. In this review, we provide an integrative interpretation of the roles for CK and auxin in the process of DNSO, adding emphasis on their uptake from the regeneration media and their interaction with sucrose present in the media to their complex signaling outputs that mediate shoot regeneration.

Development of an efficient protocol for high frequency in vitro regeneration of a horticultural plant Primulina tamiana (B.L. Burtt) Mich. Möller & A. Webber

2014 ◽  
Vol 94 (7) ◽  
pp. 1281-1287 ◽  
Author(s):  
Priya Padmanabhan ◽  
Susan J. Murch ◽  
J. Alan Sullivan ◽  
Praveen K. Saxena
Keyword(s):  
Shoot Regeneration ◽  
High Frequency ◽  
De Novo ◽  
Shoot Organogenesis ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Root Induction ◽  
Horticultural Plant ◽  
Leaf Segments

Padmanabhan, P., Murch, S. J., Sullivan, J. A. and Saxena, P. K. 2014. Development of an efficient protocol for high frequency in vitro regeneration of a horticultural plant Primulina tamiana (B.L. Burtt) Mich. Möller & A. Webber. Can. J. Plant Sci. 94: 1281–1287. A rapid and efficient in vitro regeneration system was established for Primulina tamiana, an attractive ornamental plant of horticultural importance. The successful regeneration protocol employed induction of shoot organogenesis on leaf explants. Among the various plant growth regulators evaluated, thidiazuron (TDZ) proved to be the most effective in inducing rapid de novo shoot formation. The highest shoot regeneration frequency within the shortest time from leaf explants was obtained on medium enriched with 2.5 µM TDZ. Furthermore, leaf segments were found to be a more suitable explant for in vitro shoot regeneration compared with petiole explants. The abaxial side of the leaf segments, which was in contact with the culture medium, generated more shoots than the adaxial side. Scanning electron microscopic studies carried out at various stages of de novo shoot differentiation demonstrated that shoot organogenesis involved the formation of actively dividing regions from the epidermal and subepidermal layers of the explants. In vitro rooting of the regenerated shoots was attempted using two different auxins, IAA (3-indoleacetic acid) and 3-indolebutyric acid (IBA) (0, 0.1, 0.5, 1.0, 2.0, and 5.0 µM). The optimum concentration for root induction was found to be 5 µM IBA. Approximately, 98% of the regenerated plants survived transfer and acclimation to greenhouse conditions.

scholarly journals The effects of β-lactam antibiotics and hygromycin B on de novo shoot organogenesis in apple cv. Golden Delicious

2018 ◽  
Vol 70 (1) ◽  
pp. 179-190 ◽  
Author(s):  
Mariana Stanisic ◽  
Slavica Ninkovic ◽  
Jelena Savic ◽  
Tatjana Cosic ◽  
Nevena Mitic
Keyword(s):  
Shoot Regeneration ◽  
De Novo ◽  
Shoot Organogenesis ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Selection Procedure ◽  
Transformation Process ◽  
Hygromycin B ◽  
Golden Delicious

Since the genetic transformation of the apple is strongly genotype-dependent and generally inefficient, the evaluation of factors affecting shoot regeneration are crucial for the establishment of a successful transformation process. In this report, we evaluated the effects of the ?-lactam antibiotics meropenem and timentin on in vitro regeneration via de novo shoot organogenesis from leaf explants of apple cv. Golden Delicious, as well as on the growth of the Agrobacterium tumefaciens strain EHA 105, and compared them with the commonly used ?-lactam cefotaxime. Also, we report for the first time the effect of hygromycin B as a selective agent in the domesticated apple, as regards shoot regeneration and shoot multiplication efficiency. We observed that cefotaxime and timentin at concentrations higher than 100 mg L-1 were sufficient to prevent Agrobacterium growth during a two-week period, while meropenem exhibited an inhibitory effect on bacterial growth at all tested concentrations (25-150 mg L-1). Cefotaxime at a concentration of 300 mg L-1 increased the number of regenerated shoots per explant (9.39) in comparison with the control (7.67). In contrast to cefotaxime, meropenem and timentin caused a decrease in shoot regeneration efficiency, but larger and more developed shoots were obtained on meropenem (25-125 mg L-1) after the same period of cultivation. Hygromycin B at a concentration of 5 mg L-1 or higher completely inhibited shoot regeneration and induced explant tissue necrosis. Therefore, the selection procedure with a final concentration of 4 mg L-1 throughout organogenesis and 10 mg L-1 for further shoot growth and multiplication is recommended for an efficient transformation process in apple cv. Golden Delicious.

scholarly journals Natural Variation in Plant Pluripotency and Regeneration

Plants ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 1261
Author(s):  
Robin Lardon ◽  
Danny Geelen
Keyword(s):  
De Novo ◽  
Shoot Organogenesis ◽  
In Vitro Regeneration ◽  
Relative Role ◽  
Root Explants ◽  
Experimental Systems ◽  
Complex Process ◽  
Insight Into ◽  
Molecular Framework

Plant regeneration is essential for survival upon wounding and is, hence, considered to be a strong natural selective trait. The capacity of plant tissues to regenerate in vitro, however, varies substantially between and within species and depends on the applied incubation conditions. Insight into the genetic factors underlying this variation may help to improve numerous biotechnological applications that exploit in vitro regeneration. Here, we review the state of the art on the molecular framework of de novo shoot organogenesis from root explants in Arabidopsis, which is a complex process controlled by multiple quantitative trait loci of various effect sizes. Two types of factors are distinguished that contribute to natural regenerative variation: master regulators that are conserved in all experimental systems (e.g., WUSCHEL and related homeobox genes) and conditional regulators whose relative role depends on the explant and the incubation settings. We further elaborate on epigenetic variation and protocol variables that likely contribute to differential explant responsivity within species and conclude that in vitro shoot organogenesis occurs at the intersection between (epi) genetics, endogenous hormone levels, and environmental influences.

scholarly journals Transcriptome Analysis of Melocactus glaucescens (Cactaceae) Reveals Metabolic Changes During in vitro Shoot Organogenesis Induction

2021 ◽  
Vol 12 ◽  
Author(s):  
Gabriela Torres-Silva ◽  
Ludmila Nayara Freitas Correia ◽  
Diego Silva Batista ◽  
Andréa Dias Koehler ◽  
Sheila Vitória Resende ◽  
...  
Keyword(s):  
De Novo ◽  
Shoot Organogenesis ◽  
Average Length ◽  
Transcriptome Assembly ◽  
Expression Patterns ◽  
Genetic Regulation ◽  
Pairwise Comparison ◽  
Illumina Hiseq ◽  
Real Time Quantitative Pcr

Melocactus glaucescens is an endangered cactus highly valued for its ornamental properties. In vitro shoot production of this species provides a sustainable alternative to overharvesting from the wild; however, its propagation could be improved if the genetic regulation underlying its developmental processes were known. The present study generated de novo transcriptome data, describing in vitro shoot organogenesis induction in M. glaucescens. Total RNA was extracted from explants before (control) and after shoot organogenesis induction (treated). A total of 14,478 unigenes (average length, 520 bases) were obtained using Illumina HiSeq 3000 (Illumina Inc., San Diego, CA, USA) sequencing and transcriptome assembly. Filtering for differential expression yielded 2,058 unigenes. Pairwise comparison of treated vs. control genes revealed that 1,241 (60.3%) unigenes exhibited no significant change, 226 (11%) were downregulated, and 591 (28.7%) were upregulated. Based on database analysis, more transcription factor families and unigenes appeared to be upregulated in the treated samples than in controls. Expression of WOUND INDUCED DEDIFFERENTIATION 1 (WIND1) and CALMODULIN (CaM) genes, both of which were upregulated in treated samples, was further validated by real-time quantitative PCR (RT-qPCR). Differences in gene expression patterns between control and treated samples indicate substantial changes in the primary and secondary metabolism of M. glaucescens after the induction of shoot organogenesis. These results help to clarify the molecular genetics and functional genomic aspects underlying propagation in the Cactaceae family.

scholarly journals In vitro shoot regeneration through anther culture of Brassica spp.

1970 ◽  
Vol 35 (2) ◽  
pp. 331-341 ◽  
Author(s):  
MA Sayem ◽  
M Maniruzzaman ◽  
SS Siddique ◽  
M Al-Amin
Keyword(s):  
Plant Regeneration ◽  
Anther Culture ◽  
Shoot Regeneration ◽  
Callus Induction ◽  
Maximum Rate ◽  
Media Composition ◽  
Shoot Initiation ◽  
The Media

The experiment was conducted to investigate the performance of three different genotypes (BARI Sarisha-6, BARI Sarisha-8, and BARI Sarisha-11) in two different media viz., MS and B5 with different concentrations of phytohormone (2, 4-D) for callus induction from uninucleate stage anthers of Brassica and subsequent plant regeneration in MS media with different concentrations of phytohormone (BAP and NAA). Among the genotypes, BARI Sarisha-8 showed the best performance for all the parameters of callus induction. The performance of BARI Sarisha-6 was poor compared to others. Maximum rate of callus induction (%) was observed in MS + 0.5 mg/L 2, 4-D followed by B5 + 0.5 mg/L 2,4-D. The media combination MS + 1.0 mg/L BAP 0.3 mg/L 2,4-D showed the best performance for maintenance of calli. Significant variations were observed among the genotypes and media composition for shoot regeneration. Among the genotypes, BARI Sarisha-8 showed the best performance for shoot regeneration followed by BARJ Sarisha-l1. The genotype BARI Sarisha-8 produced higher percent of shoots/calli and required minimum days for shoot initiation. Higher percent calli without shoot were produced by the genotype BARI Sarisha-6. The media combination MS + 2.0 mg/L BAP + 0.5 mg/L NAA showed the best performance for shoot regeneration and required maximum days for shoot initiation. Keywords: Regeneration; BARI Sarisha-6; BARI Sarisha-8; BARI Sarisha-11; anther culture; phytohormone  DOI: 10.3329/bjar.v35i2.5896Bangladesh J. Agril. Res. 35(2) : 331-341, June 2010

scholarly journals In vitro regeneration and polyploidy induction in Pelargonium × hortorum L. H. Bailey

2009 ◽  
Vol 57 (5) ◽  
pp. 305-312
Author(s):  
Hana Vejsadová ◽  
Petra Kuchtová-Jadrná
Keyword(s):  
Flow Cytometry ◽  
Plant Growth ◽  
Shoot Regeneration ◽  
In Vitro Regeneration ◽  
Infiltration Method

The objective of this work was to induce in vitro shoot regeneration as influenced by plant growth re­gu­la­tors and ascertain an effective method of polyploidy induction using colchicine and oryzalin in two diploid cultivars ‘Gizela’ and ‘Black Velvet Scarlet F1’ of Pelargonium × hortorum L. H. Bailey. In both cultivars, benzyladenine (BA) significantly improved shoot regeneration compared to zeatin. The infiltration and the dip methods of explant treatment were used for polyploidy induction. Regenerants were analyzed using the flow cytometry (FCM). In ‘Gizela’ and ‘Black Velvet Scarlet F1’, 10 tetraploids on the level of 2n = 4x were found by the infiltration method. The tetraploidy was determined by the dip method in 4 shoots of ‘Gizela’ and 11 tetraploids were detected in ‘Black Velvet Scarlet F1’.

scholarly journals In vitro regeneration performance of Corchorus olitorius

1970 ◽  
Vol 8 (1) ◽  
pp. 1-6 ◽  
Author(s):  
M Hoque ◽  
KM Nasiruddin ◽  
GKMN Haque ◽  
GC Biswas
Keyword(s):  
Shoot Regeneration ◽  
Callus Induction ◽  
Root Formation ◽  
Plantlet Regeneration ◽  
Ms Medium ◽  
Corchorus Olitorius ◽  
The Media ◽  
Regenerated Plantlets

The experiment was conducted during May to December 2008 in the Biotechnology Laboratory of Bangladesh Agricultural University, Mymensingh to observe the callus induction, regeneration potentiality and to establish a suitable in vitro plantlet regeneration protocol of Corchorus olitorius. MS medium supplemented with different phytohormone concentrations and combinations were used to observe the callus induction, shoot regeneration and root formation ability of the cotyledon with attached petiole derived explant of three genotypes viz. O-9897, O-72 and OM-1. The highest callus induction (92.85%) was observed in O-9897 followed by O-72 (82.14%) in the MS media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA. Genotype O-9897 in MS media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA produced the highest percentage of shoot regenerants (83.33%) followed by O-72 (75.00%) in the media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA. The root formation from regenerants was the best on halfstrength of MS media supplemented with 0.6 mg/L IBA in genotype O-9897 (45.00%). The in vitro regenerated plantlets from the genotypes O-9897 could be established in the field. Therefore, the genotypes O-9897 of C. olitorius in MS media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA could be used for callus induction and shoot regeneration. Keywords: Regeneration; Phytohormone; Corchorus olitorius DOI: 10.3329/jbau.v8i1.6390J. Bangladesh Agril. Univ. 8(1): 1-6, 2010

scholarly journals DNA Methylation and Histone Modifications Regulate De Novo Shoot Regeneration in Arabidopsis by Modulating WUSCHEL Expression and Auxin Signaling

PLoS Genetics ◽  
2011 ◽  
Vol 7 (8) ◽  
pp. e1002243 ◽  
Author(s):  
Wei Li ◽  
Hui Liu ◽  
Zhi Juan Cheng ◽  
Ying Hua Su ◽  
Hua Nan Han ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Histone Modifications ◽  
Shoot Regeneration ◽  
De Novo ◽  
Auxin Signaling

scholarly journals In vitro Regeneration and Rapid Multiplication of Dendrobium bensoniae, an Indigenous Ornamental Orchid

2017 ◽  
Vol 14 (2) ◽  
pp. 24-31 ◽  
Author(s):  
S S Riva ◽  
A Islam ◽  
M E Hoque
Keyword(s):  
Shoot Regeneration ◽  
In Vitro Regeneration ◽  
Ms Medium ◽  
Shoot Induction ◽  
Rapid Multiplication ◽  
Number Of Shoots

An experiment was conducted on in vitro regeneration and multiplication of Dendrobium bensoniae. Different concentrations of BA and IBA alone or combination of both hormones were used as treatment for regeneration.  It was revealed that shoot regeneration from node was the best at 2.0 mg/l BA supplemented to MS medium. It gave better responses than all other concentrations and combinations of BA and BA+IBA, used in the present study. The highest number of shoots and leaves were found when 1.0 mg/l BA with 1.5 mg/l IBA was supplemented into MS medium.  For rooting, 0.5 mg/l BA with 1.0 mg/l IBA was found to be the most effective. The well-rooted plantlets were successfully acclimatized under 70-80% humidity and planted in pots and transferred to the shade house for establishment. Around 85% of plantlets survived in the field. From the present result, it may be recommended that MS medium supplemented with 2.0 mg/l BA may be used for rapid shoot induction and regeneration of D. bensoniae.The Agriculturists 2016; 14(2) 24-31

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