scholarly journals Temporal Control of Morphogenic Factor Expression Determines Efficacy in Enhancing Regeneration

Plants ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 2271
Author(s):  
Juan H. Gonzalez ◽  
Joseph S. Taylor ◽  
Kelsey M. Reed ◽  
R. Clay Wright ◽  
Bastiaan O. R. Bargmann
Keyword(s):  
Shoot Regeneration ◽  
Auxin Signaling ◽  
Transient Transformation ◽  
Shoot Induction ◽  
Root Explants ◽  
Ectopic Overexpression ◽  
Factor Expression ◽  
Regeneration Efficiency ◽  
Signaling Activation ◽  
Induction Stage

Background: Regeneration of fertile plants from tissue culture is a critical bottleneck in the application of new plant breeding technologies. Ectopic overexpression of morphogenic factors is a promising workaround for this hurdle. Methods: Conditional overexpression of WUS and ARF5Δ was used to study the effect of timing the overexpression of these morphogenic factors during shoot regeneration from root explants in Arabidopsis. In addition, their effect on auxin-signaling activation was examined by visualization and cytometric quantification of the DR5:GFP auxin-signaling reporter in roots and protoplasts, respectively. Results: The induced expression of both WUS and ARF5Δ led to an activation of auxin signaling in roots. Activation of auxin signaling by WUS and ARF5Δ was further quantified by transient transformation of protoplasts. Ectopic overexpression of both WUS and ARF5Δ enhanced regeneration efficiency, but only during the shoot-induction stage of regeneration and not during the callus-induction stage. Conclusions: The overexpression of WUS and ARF5Δ both lead to activation of auxin signaling. Expression during the shoot-induction stage is critical for the enhancement of shoot regeneration by WUS and ARF5Δ.

Mutagen treatment affects the regeneration efficiency of the shoot-tip explant in Celosia cristate L. (Cockscomb)

2020 ◽  
Vol 80 (03) ◽  
Author(s):  
Rupesh S. Badere ◽  
Pallavi K. Rinkey
Keyword(s):  
Plant Growth Regulator ◽  
Gamma Ray ◽  
Multiple Shoot ◽  
Shoot Tip ◽  
Ms Medium ◽  
Shoot Induction ◽  
Ethyl Methane Sulphonate ◽  
Multiple Shoot Induction ◽  
Methane Sulphonate ◽  
Regeneration Efficiency

The shoot-tip explant harvested from ethyl methane sulphonate (EMS) and gamma ray (GR) mutagenized seedling was cultured over MS medium fortified with NAA and BAP for five generations to amplify the mutated sector. Mutagens reduced the regeneration efficiency of the explant and affected its plant growth regulator-dependence for multiple shoot induction. While the 12d-old shoot-tip from GR-treated seedling induced shoots with 0.5µM NAA+6.6µM BAP; that from EMS-treated seedling induced shoots with 8.8µM BAP. The present study establishes that the mutagens affect the regeneration process in the explant.

scholarly journals Efficient Protoplast Regeneration Protocol and CRISPR/Cas9-Mediated Editing of Glucosinolate Transporter (GTR) Genes in Rapeseed (Brassica napus L.)

2021 ◽  
Vol 12 ◽  
Author(s):  
Xueyuan Li ◽  
Sjur Sandgrind ◽  
Oliver Moss ◽  
Rui Guan ◽  
Emelie Ivarson ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Shoot Regeneration ◽  
Gene Editing ◽  
Protoplast Culture ◽  
Callus Formation ◽  
Protoplast Regeneration ◽  
Induction Medium ◽  
Shoot Induction ◽  
Brassica Napus L ◽  
High Concentrations

Difficulty in protoplast regeneration is a major obstacle to apply the CRISPR/Cas9 gene editing technique effectively in research and breeding of rapeseed (Brassica napus L.). The present study describes for the first time a rapid and efficient protocol for the isolation, regeneration and transfection of protoplasts of rapeseed cv. Kumily, and its application in gene editing. Protoplasts isolated from leaves of 3–4 weeks old were cultured in MI and MII liquid media for cell wall formation and cell division, followed by subculture on shoot induction medium and shoot regeneration medium for shoot production. Different basal media, types and combinations of plant growth regulators, and protoplast culture duration on each type of media were investigated in relation to protoplast regeneration. The results showed that relatively high concentrations of NAA (0.5 mg l−1) and 2,4-D (0.5 mg l−1) in the MI medium were essential for protoplasts to form cell walls and maintain cell divisions, and thereafter auxin should be reduced for callus formation and shoot induction. For shoot regeneration, relatively high concentrations of cytokinin were required, and among all the combinations tested, 2.2 mg l−1 TDZ in combination with auxin 0.5 mg l−1 NAA gave the best result with up to 45% shoot regeneration. Our results also showed the duration of protoplast culture on different media was critical, as longer culture durations would significantly reduce the shoot regeneration frequency. In addition, we have optimized the transfection protocol for rapeseed. Using this optimized protocol, we have successfully edited the BnGTR genes controlling glucosinolate transport in rapeseed with a high mutation frequency.

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

scholarly journals Silicon Promotes Adventitious Shoot Regeneration and Enhances Salinity Tolerance ofAjuga multifloraBunge by Altering Activity of Antioxidant Enzyme

2014 ◽  
Vol 2014 ◽  
pp. 1-10 ◽  
Author(s):  
Iyyakkannu Sivanesan ◽  
Byoung Ryong Jeong
Keyword(s):  
Salinity Tolerance ◽  
Shoot Regeneration ◽  
Microscopic Analysis ◽  
Adventitious Shoot ◽  
Induction Medium ◽  
Shoot Induction ◽  
Electron Microscopic ◽  
Sod Activity ◽  
Scanning Electron Microscopic Analysis ◽  
Shoot Induction Medium

We investigated the effect of Si concentration on shoot regeneration and salinity tolerance ofAjuga multiflora. Addition of Si to the shoot induction medium significantly increased the frequency of shoot induction. The average number of shoots regenerated per explant decreased on the medium containing NaCl alone, while there was less decrease when the shoot induction medium was supplemented with both NaCl and Si. The shoot induction percentage increased linearly with increasing concentration of Si in the NaCl containing medium. Addition of Si to the shoot induction medium significantly increased SOD, POD, APX, and CAT activity in regenerated shoot buds as compared with the control. The inclusion of Si to the NaCl containing medium significantly increased the SOD activity in leaves and roots, while it decreased POD, APX, and CAT activity in both organs. Scanning electron microscopic analysis showed that there are no distinct differences in the structure of stomata between the control and Si-treated plants. However, NaCl treatment significantly affected the structure and number of stomata as compared to the control. Wavelength dispersive X-ray analysis confirmed the high Si deposition in trichomes of plants grown in the Si containing medium but not in plants grown in the medium without Si.

Adventitious shoot regeneration from leaf, stem and root explants of commercial cultivars of Gentiana

1996 ◽  
Vol 15 (8) ◽  
pp. 578-581 ◽  
Author(s):  
Keizo Hosokawa ◽  
Masaru Nakano ◽  
Yayoi Oikawa ◽  
Saburo Yamamura
Keyword(s):  
Shoot Regeneration ◽  
Adventitious Shoot ◽  
Adventitious Shoot Regeneration ◽  
Root Explants ◽  
Commercial Cultivars

Adventitious Shoot Induction from Internode and Root Explants in a Semiparasitic Herb Monochasma savatieri Franch ex Maxim

2017 ◽  
Vol 36 (3) ◽  
pp. 799-804 ◽  
Author(s):  
Yueya Zhang ◽  
Yulu Chen ◽  
Xinhua Zhang ◽  
Jaime A. Teixeira da Silva ◽  
Guohua Ma
Keyword(s):  
Adventitious Shoot ◽  
Shoot Induction ◽  
Root Explants

scholarly journals In Vitro Culture of Heuchera villosa ‘Caramel’

HortScience ◽  
2017 ◽  
Vol 52 (4) ◽  
pp. 622-624 ◽  
Author(s):  
Hua Q. Zhao ◽  
Qing H. He ◽  
Li L. Song ◽  
Mei F. Hou ◽  
Zhi G. Zhang
Keyword(s):  
Acetic Acid ◽  
In Vitro Culture ◽  
Butyric Acid ◽  
Shoot Regeneration ◽  
Ms Medium ◽  
Shoot Induction ◽  
Induction Rate ◽  
Regenerated Plantlets ◽  
Indole 3 Acetic Acid

The procedure for Heuchera villosa ‘Caramel’ propagation was investigated, which involves shoot regeneration, rooting of regenerated shoots, and acclimation of regenerated plantlets. Petioles, as explants, were cultured on MS medium supplemented with 1-naphthylacetic acid (NAA), benzylaminopurine (BA), thidiazuron (TDZ) and callus formed on all media. Shoots were observed to proliferate from callus on media with BA and NAA, whereas no shoots regenerated on media with TDZ and NAA. On media containing 0.5 or 1.0 mg·L−1 BA in combination with NAA, the regenerated shoots showed severe hyperhydricity, whereas on media containing 0.1 mg·L−1 BA in combination with NAA, the regenerated shoots grew normally. The highest shoot induction rate, 90.6%, was obtained on media containing 0.1 mg·L−1 BA and 0.01 mg·L−1 NAA. The effects of indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), and NAA on rooting of H. villosa ‘Caramel’ was explored. The highest rooting rate (95%) was obtained on 1/2 MS medium containing 0.2 mg·L−1 NAA. In the subsequent acclimation experiments, about 85% of rooted plantlets survived and grew normally.

scholarly journals Indirect in vitro Regeneration of the Medicinal Plant, Aspilia africana, and Histological Assessment at Different Developmental Stages

2021 ◽  
Vol 12 ◽  
Author(s):  
Denis Okello ◽  
Sungyu Yang ◽  
Richard Komakech ◽  
Yuseong Chung ◽  
Endang Rahmat ◽  
...  
Keyword(s):  
Medicinal Plant ◽  
Shoot Regeneration ◽  
Callus Induction ◽  
Developmental Stages ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Regeneration Capacity ◽  
Root Explants ◽  
Indirect Regeneration

The medicinal plant, Aspilia africana, has been traditionally used in several African countries to treat many diseases such as tuberculosis, cough, inflammation, malaria, osteoporosis, and diabetes. In this study, we developed a protocol for in vitro propagation of A. africana using indirect shoot organogenesis from leaf and root explants of in vitro-grown seedlings and assessed the tissues at different developmental stages. The highest callus induction (91.9 ± 2.96%) from leaf explants was in the Murashige and Skoog (MS) medium augmented with 1.0 mg/L 6-Benzylaminopurine (BAP) and 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) while from root explants, the highest callus induction (92.6 ± 2.80%) was in the same plant tissue culture medium augmented with 0.5 mg/L BAP and 1.0 mg/L 2,4-D. The best shoot regeneration capacity from leaf-derived calli (i.e., 80.0 ± 6.23% regeneration percentage and 12.0 ± 6.23 shoots per callus) was obtained in medium augmented with 1.0 mg/L BAP and 0.05 mg/L α-Naphthaleneacetic acid (NAA); the best regeneration capacity for root-derived calli (i.e., 86.7 ± 6.24% shoot regeneration percentage and 14.7 ± 1.11 shoots per callus) was obtained in the MS medium augmented with 1.0 mg/L BAP, 0.05 mg/L NAA, and 0.1 mg/L Thidiazuron (TDZ). Regenerated plantlets developed a robust root system in 1/2 MS medium augmented with 0.1 mg/L NAA and had a survival rate of 93.6% at acclimatization. The in vitro regenerated stem tissue was fully differentiated, while the young leaf tissue consisted of largely unorganized and poorly differentiated cells with large intercellular airspaces typical of in vitro leaf tissues. Our study established a protocol for the indirect regeneration of A. africana and offers a basis for its domestication, large-scale multiplication, and germplasm preservation. To the best of our knowledge, this is the first study to develop an indirect regeneration protocol for A. africana and conduct anatomical assessment through the different stages of development from callus to a fully developed plantlet.

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