scholarly journals Application of Artificial Neural Network for Modeling and Studying In Vitro Genotype-Independent Shoot Regeneration in Wheat

2020 ◽  
Vol 10 (15) ◽  
pp. 5370 ◽  
Author(s):  
Mohsen Hesami ◽  
Jorge A. Condori-Apfata ◽  
Maria Valderrama Valencia ◽  
Mohsen Mohammadi
Keyword(s):  
Neural Network ◽  
Shoot Regeneration ◽  
In Vitro Regeneration ◽  
Generalized Regression Neural Network ◽  
Naphthaleneacetic Acid ◽  
Regeneration Frequency ◽  
Artificial Neural ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
In Vitro Shoot Regeneration

Optimizing in vitro shoot regeneration conditions in wheat is one of the important steps in successful micropropagation and gene transformation. Various factors such as genotypes, explants, and phytohormones affect in vitro regeneration of wheat, hindering the ability to tailor genotype-independent protocols. Novel computational approaches such as artificial neural networks (ANNs) can facilitate modeling and predicting outcomes of tissue culture experiments and thereby reduce large experimental treatments and combinations. In this study, generalized regression neural network (GRNN) were used to model and forecast in vitro shoot regeneration outcomes of wheat on the basis of 10 factors including genotypes, explants, and different concentrations of 6-benzylaminopurine (BAP), kinetin (Kin), 2,4-dichlorophenoxyacetic acid (2,4-D), indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), 1-naphthaleneacetic acid (NAA), zeatin, and CuSO4. In addition, GRNN was linked to a genetic algorithm (GA) to identify an optimized solution for maximum shoot regeneration. Results indicated that GRNN could accurately predict the shoot regeneration frequency in the validation set with a coefficient determination of 0.78. Sensitivity analysis demonstrated that shoot regeneration frequency was more sensitive to variables in the order of 2,4-D > explant > genotype < zeatin < NAA. Results of this study suggest that GRNN-GA can be used as a tool, besides experimental approaches, to develop and optimize in vitro genotype-independent regeneration protocols.

scholarly journals In Vitro Shoot Regeneration and Polyploid Induction of Rhododendron ‘Fragrantissimum Improved’

HortScience ◽  
2010 ◽  
Vol 45 (5) ◽  
pp. 801-804 ◽  
Author(s):  
Cary J. Hebert ◽  
Darren H. Touchell ◽  
Thomas G. Ranney ◽  
Anthony V. LeBude
Keyword(s):  
Shoot Regeneration ◽  
In Vitro Regeneration ◽  
Naphthaleneacetic Acid ◽  
Regenerative Capacity ◽  
Leaf Segments ◽  
Ornamental Traits ◽  
Hybrid Protocols ◽  
Mitotic Inhibitor ◽  
In Vitro Shoot Regeneration

Rhododendron L.‘Fragrantissimum Improved’ is an attractive cultivar with showy, fragrant flowers but has limited potential for breeding because it is a sterile wide hybrid. Protocols for in vitro regeneration and polyploid induction were developed for this cultivar as a means to potentially restore fertility and enhance ornamental traits. Combinations of thidiazuron (TDZ) at 0, 5, 10, 15, or 20 μM and 1-naphthaleneacetic acid (NAA) at 0, 2.5, 5, or 10 μM were used to induce shoot regeneration from leaves. Shoot regeneration was optimized (68% of leaf segments produced shoots) using 8.8 μM TDZ and 10 μM NAA. To induce polyploidy, regenerative callus was treated with 7.5, 15, 30, 60, or 90 μM of the mitotic inhibitor oryzalin for 1, 3, 5, 7, or 14 d in various combinations. Oryzalin significantly affected survival and shoot regenerative capacity. A percentage of homogenous, tetraploid shoots was recovered from treatments of 30 μM oryzalin for 1 (13%) or 3 (13%) days and 7.5 μM oryzalin for 7 (20%) or 14 (7%) days.

scholarly journals Response of Different Explants for Callus Induction in Cucumber

2021 ◽  
Vol 2 (5) ◽  
pp. 71-75
Author(s):  
Hasina Sultana ◽  
Lutfun Nahar ◽  
M. Mofazzal Hossain ◽  
Totan Kumar Ghosh ◽  
Md. Sanaullah Biswas
Keyword(s):  
Shoot Regeneration ◽  
Callus Induction ◽  
In Vitro Regeneration ◽  
Leaf Disc ◽  
Dry Weight ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Regenerate Shoot ◽  
2,4 Dichlorophenoxyacetic Acid

In vitro regeneration of cucumber is relatively difficult for genetic improvement. In this regard, different concentrations of growth regulators and three types of explants (cotyledon, hypocotyl and leaf disc) were investigated for their efficiency on callus induction potential. Among different explants explored for callus induction with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D), leaf disc responded earlier (4.67 days) and showed higher percentage of callus induction (91.50%) with 2 mg/l 2,4-D supplemented Murashige and Skoog (MS) media. The same concentration of 2,4-D resulted in the maximum callus fresh (0.56 g) and dry weight (0.39 g) from leaf disc explant. Then the callus was transferred to untreated, 2.0 mg/l BAP + 0.2 mg/l NAA + 1.0 mg/l Kn, 2.0 mg/l BAP + 1.0 mg/l NAA + 1.0 mg/l Kn and 2.0 mg/l BAP + 1.5 mg/l NAA + 1.0 mg/l Kn fortified MS medium. After transferring the callus of different explants to shoot regeneration media containing different concentrations of 6-benzylaminopurine (BAP), 1-naphthaleneacetic acid (NAA) and Kinetin (Kn), only cotyledon callus started to regenerate shoot. The combination of BAP (2 mg/l) + NAA (0.2 mg/l) + Kn (1 mg/l) showed highest shoot regeneration percentage (67.77%) and the maximum number of shoots (5.12) per explant were recorded in the treatment combination of 2 mg/l BAP + 0.2 mg/l NAA + 1 mg/l Kn. These results provided a basis for the optimization of the callus induction protocol of cucumber for genetic transformation.

In Vitro Shoot Regeneration from Callus Derived from Marubakaido Apple Rootstock under Different Aluminium Concentrations

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 460e-460 ◽  
Author(s):  
Marisa F. de Oliveira ◽  
Gerson R. de L. Fortes ◽  
João B. da Silva
Keyword(s):  
Shoot Regeneration ◽  
Dichlorophenoxyacetic Acid ◽  
Apple Rootstock ◽  
Under Five ◽  
Significant Difference ◽  
Previously Treated ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
In Vitro Shoot Regeneration

The aim of this work was to evaluate the organogenesis of Marubakaido apple rootstock under different aluminium concentratons. The explants were calli derived from apple internodes treated with either 2,4-dichlorophenoxyacetic acid or pichloram at 0.5 and 1.0 μM and under five different aluminium concentrations (0, 5, 10, 15, 20 mg/L). These calli were then treated with aluminium at 0, 5, 10, 15, and 20 mg/L. It was observed shoot regeneration only for those calli previously treated with pichloram. There were no significant difference among the aluminium concentrations.

scholarly journals In vitro shoot regeneration of boldo from leaf explants

2010 ◽  
Vol 40 (10) ◽  
pp. 2210-2213
Author(s):  
Monalize Salete Mota ◽  
Juliana de Magalhães Bandeira ◽  
Eugenia Jacira Bolacel Braga ◽  
Valmor João Bianchi ◽  
José Antonio Peters
Keyword(s):  
Shoot Regeneration ◽  
Leaf Explants ◽  
Shoot Development ◽  
High Potential ◽  
Naphthaleneacetic Acid ◽  
Regeneration System ◽  
Bud Induction ◽  
Molecular Studies ◽  
In Vitro Shoot Regeneration

A shoot regeneration system for Plectranthus neochilus was studied from leaf explants. Leaves developed under in vitro conditions were cultured on Wood Plant Medium supplemented with 0.2mg dm-3 α-naphthaleneacetic acid (NAA) and different 6-benzilaminopurine (BAP) or thidiazuron (TDZ) concentrations (0, 1.5, 3.0, 4.5 and 6.0mg dm-3). An increase in percentage of responsive explants (85.3%) and in the number of shoots developed per explant (3.2) was observed when the explants were treated with 5.3 and 4.7mg dm-3 BAP, respectively. The leaf explants cultured on media supplemented with TDZ became vitreous and did not form buds. The regeneration system used is efficient for boldo bud induction and shoot development, showing high potential for advanced cellular and molecular studies.

scholarly journals Factors Affecting the Regeneration, via Organogenesis, and the Selection of Transgenic Calli in the Peach Rootstock Hansen 536 (Prunus persica × Prunus amygdalus) to Express an RNAi Construct against PPV Virus

Plants ◽  
2019 ◽  
Vol 8 (6) ◽  
pp. 178 ◽  
Author(s):  
Sabbadini ◽  
Ricci ◽  
Limera ◽  
Baldoni ◽  
Capriotti ◽  
...  
Keyword(s):  
Shoot Regeneration ◽  
Prunus Persica ◽  
Fluorescent Protein ◽  
In Vitro Regeneration ◽  
Naphthaleneacetic Acid ◽  
Prunus Amygdalus ◽  
Indirect Organogenesis ◽  
Egfp Gene ◽  
Rnai Construct

Prunus spp. is one of the most recalcitrant fruit tree species in terms of in vitro regeneration and transformation, mostly when mature tissues are used as explants. The present study describes the in vitro regeneration via indirect organogenesis, and Agrobacterium tumefaciens-mediated transformation of the peach rootstock Hansen 536 (Prunus persica × Prunus amygdalus) through the use of meristematic bulks (MBs) as starting explants. Efficient adventitious shoot regeneration was obtained when Hansen 536 MBs were cultured on an optimized medium consisting of modified McCown Woody Plant medium (WPM) enriched with 4.4 M 6-Benzyladenine (BA), 0.1 M 1-Naphthaleneacetic acid (NAA) and 6.0 g L−1 plant agar S1000 (B&V). MB slices were used later as starting explants for Agrobacterium-mediated transformation to introduce an RNAi construct “ihp35S-PPV194” against PPV virus. Transgenic events were identified by both green fluorescent protein (GFP) screening and kanamycin selection at different concentrations (0, 17 or 42 M). GFP-fluorescent proliferating callus lines were selected and confirmed to stably express the ihp35S-PPV194::eGFP gene construct by molecular analysis. Although shoot regeneration from these transgenic calli has not been obtained yet, this represents one of the few examples of successful attempts in peach genetic transformation from somatic tissues, and also serves as a useful in vitro system for future gene functional analysis in peach.

scholarly journals Plant regeneration from in vitro leaves of four commercial Pyrus species

2008 ◽  
Vol 54 (No. 4) ◽  
pp. 140-148 ◽  
Author(s):  
H. Tang ◽  
Y. Luo ◽  
C. Liu
Keyword(s):  
Plant Regeneration ◽  
Shoot Regeneration ◽  
Pyrus Communis ◽  
Naphthaleneacetic Acid ◽  
Complete Medium ◽  
Regeneration Frequency ◽  
Regenerated Plants ◽  
Equivalent Conditions

An efficient shoot regeneration from in vitro leaf sections of <I>Pyrus communis</I> Bartlett, <I>P. pyrifolia</I> Shenbuzhi, <I>P. bretschneideri</I> Zaosu and <I>P. ussuriensis</I> Manyuanxiang was successfully developed for use in future transgenic studies. On the basis of regeneration frequency and average shoot numbers, optimal shoot regeneration was obtained on leaf sections of <I>P. communis</I> Bartlett when cultured on Murashige and Skoog complete medium containing 6.0 mg/l BA (6-benzyladenine) and 0.1 mg/l NAA (&alpha;-naphthaleneacetic acid), while Quoirin and Lepoivre complete medium supplemented with 1.0 mg/l TDZ [thidiazuron (N-phenyl-N<sup>1</sup>-1,2,3-thiadiazol-5-ylurea)] and 0.1 mg/l NAA was found best for <I>P. pyrifolia</I> Shenbuzhi, and Nitsch and Nitsch complete medium containing 3.0 mg/l TDZ and 0.1 mg/l NAA or 0.2 mg/l IAA was suitable for<I>P. bretschneideri</I> Zaosu or <I>P. ussuriensis</I> Manyuanxiang, respectively. After cutting the leaves into three sections perpendicular to the midrib and culturing under the equivalent conditions, regeneration occurred more frequently on basal sections than middle sections, and no shoots formed on apical sections. A ratio of NH<sup>+</sup><sub>4</sub>-N/NO<sup>-</sup><sub>3</sub>- N of 1:2~1:7 was found beneficial for shoot regeneration. 75.0–87.5% of proliferating shoots formed roots after 4 weeks of transfer to 1/4 strength of Murashige and Skoog complete medium supplemented with 2.5 mg/l IBA (indole-3-butryric acid) and 30.0 g/l sucrose. Regenerated plants were successfully established under greenhouse conditions.

In vitro shoot regeneration in Myracrodruon urundeuva Fr. All

2021 ◽  
Vol 51 ◽  
Author(s):  
Tecla dos Santos Silva ◽  
Rosembrando Sosthenes Leite Carvalho Filho ◽  
Priscila Tavares Fonseca ◽  
José Raniere Ferreira de Santana
Keyword(s):  
Shoot Regeneration ◽  
Callus Formation ◽  
Cotyledonary Node ◽  
Quadratic Equation ◽  
Naphthaleneacetic Acid ◽  
Nodal Segment ◽  
The Mean ◽  
In Vitro Shoot Regeneration ◽  
Number Of Shoots

ABSTRACT Myracrodruon urundeuva Fr. All. is a tree threatened with extinction, which has wood and medicinal potential. This study aimed to analyze the in vitro shoot regeneration in M. urundeuva, in order to increase the species multiplication. Two experiments were conducted: 1) concentrations of 6-benzylaminopurine (BAP) (0.0, 2.0, 4.0, 8.0 and 16.0 µM), in association with naphthaleneacetic acid (NAA) (0.0, 1.5 and 3.0 µM), in explants (cotyledon, hypocotyl and cotyledonary node); 2) concentrations of meta-topolin (mT) (0.0, 2.0, 4.0, 8.0, 16.0 and 32.0 µM) in explants (biaxillary, medial uniaxillary and apical basal nodal segment). The percentage of explants responsive to shoot regeneration, percentage of callus explants, number of shoots and shoot length were evaluated. In the first experiment, the shoot regeneration occurred only in explants of the cotyledonary node and hypocotyl type, with the highest responsiveness percentage (76.67 %) and number of shoots (1.97 and 1.63) obtained for the cotyledonary node in the presence of 3.0 µM of NAA in association with 2.0 (1.97 shoots/explant) and 4.0 µM (1.63 shoots/explant) of mT. In the second experiment, the resolution of the obtained quadratic equation indicates that the use of basal explant with 24.59 µM of mT added to the culture medium leads to the highest number of shoots (1.86). However, despite the mT having increased the mean number of shoots, all treatments containing this cytokinin showed callus formation. As a conclusion, it is possible to regenerate shoots in M. urundeuva from the cotyledonary node using BAP in association with NAA.

scholarly journals In Vitro Shoot Regeneration and Polyploid Induction from Leaves of Hypericum Species

HortScience ◽  
2009 ◽  
Vol 44 (7) ◽  
pp. 1957-1961 ◽  
Author(s):  
Elisabeth M. Meyer ◽  
Darren H. Touchell ◽  
Thomas G. Ranney
Keyword(s):  
Shoot Regeneration ◽  
Chromosome Doubling ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Shoot Induction ◽  
Bluish Green ◽  
Spontaneous Chromosome ◽  
Ornamental Traits ◽  
In Vitro Shoot Regeneration

Hypericum L. H2003-004-016 is a complex hybrid among Hypericum frondosum Michx., Hypericum galioides Lam., and Hypericum kalmianum L. and exhibits valuable ornamental characteristics, including compact habit, bluish green foliage, and showy flowers. Inducing polyploidy may further enhance the ornamental traits of this hybrid and provide new opportunities for hybridizing with other naturally occurring polyploid Hypericum sp. In this study, in vitro shoot regeneration and treatment of regenerative callus with the dinitroaniline herbicide oryzalin (3,5-dinitro-N4,N4-dipropylsufanilamide) were investigated as a means of inducing allopolyploidy. First, in vitro regeneration was optimized for callus and shoot induction by culture of leaf explants on medium supplemented with benzylamino purine (BA) or meta-topolin (mT) at 5, 10, or 15 μM in combination with indoleacetic acid (IAA) at 0, 1.25, 2.5, or 5 μM. Both BA and mT treatments successfully induced regenerative callus and shoots. Multiple regression analysis estimated maximum regenerative callus (94%) and shoot induction (18 shoots per explant) in medium supplemented with 5 μM BA and 3.75 μM IAA. In the second part of the study, exposure of regenerative callus to oryzalin at 0, 7.5, 15, 30, 60, or 90 μM for durations of 3, 6, or 9 d was investigated for polyploid induction. There was no survival for any of the calli in the 60- or 90-μM oryzalin treatments, but calli subjected to the other treatments exhibited some survival and polyploid induction. Duration had no effect on callus survival or ploidy level, but oryzalin concentration was a significant factor in both. The greatest percentage (44%) of polyploids was induced with 30 μM oryzalin. Spontaneous chromosome doubling was observed in 8% of control explants receiving no oryzalin treatment.

scholarly journals The effect of auxins in inducing organogenesis or somatic embryogenesis in mature sunflower zygotic embryo derived apex

2020 ◽  
Vol 48 (1) ◽  
pp. 150-161
Author(s):  
Adriana AURORI ◽  
Imola MOLNAR ◽  
Elena RAKOSY-TICAN
Keyword(s):  
Zygotic Embryo ◽  
In Vitro Regeneration ◽  
Mature Stage ◽  
Induction Medium ◽  
Naphthaleneacetic Acid ◽  
Axillary Shoots ◽  
Mature Embryos ◽  
Cytokinin Treatment ◽  
2,4 Dichlorophenoxyacetic Acid

Induction of shoots or of somatic embryos is the key step for gaining the morphogenetic potential in sunflower (Helianthus annuus L.), species known as recalcitrant to in vitro regeneration. In the immature zygotic embryo derived tissues or in other juvenile tissues resulted from seedlings, the acquisition of the competence for regeneration can be achieved directly by cytokinin treatment or by preconditioning the explants on cytokinin containing medium. In this paper is presented a new type of explant for sunflower in vitro culture, consisting of the apex with primordial leaves, resulted from ungerminated mature zygotic embryo, in which a specific morphogenetic response was triggered by the exogenously applied auxins. Among the auxins tested, indole-3-acetic acid, indole-3-butyric acid and 1-naphthaleneacetic acid are inducers of an organogenetic response, apical/axillary shoots and adventitious buds being regenerated while 2,4-dichlorophenoxyacetic acid, 3,6-dichloro-2-methoxybenzoic acid and 4-amino-3,5,6-trichloropicolinic acid led to somatic embryo formation. Among the auxins tested only 4-amino-3,5,6-trichloropicolinic acid sustains the embryos development up to mature stage. A high amount of sucrose (120 g L-1) supplied during the auxin treatment promotes the maturation of the embryos directly on the induction medium for all tested auxins with embryogenic effect. These findings show that regardless of the type of morphogenetic response aimed in sunflower meristematic tissues resulted from mature embryos, the presence of auxins is mandatory.

scholarly journals Improving In Vitro Plant Regeneration from Leaf and Petiole Explants of `Marion' Blackberry

HortScience ◽  
2004 ◽  
Vol 39 (2) ◽  
pp. 316-320 ◽  
Author(s):  
Rengong Meng ◽  
Tony H.H. Chen ◽  
Chad E. Finn ◽  
Yonghai Li
Keyword(s):  
Leaf Explants ◽  
Petri Dish ◽  
Shoot Formation ◽  
Photon Flux ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Petiole Explants ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
In Vitro Shoot Regeneration

Experiments focusing on plant growth regulators' concentrations and combinations, mineral salt formulations, and TDZ pretreatment formations were conducted to optimize in vitro shoot regeneration from leaf and petiole explants of `Marion' blackberry. Optimum shoot formation was obtained when stock plants were incubated in TDZ pretreatment medium for 3 weeks before culturing leaf explants on regeneration medium (Woody Plant Medium with 5 μm BA and 0.5 μm IBA) in darkness for 1 week before transfer to light photoperiod (16-hour photoperiod at photosynthetic photon flux of ≈50 μmol·m-2·s-1) at 23 °C ± 2 °C for 4 weeks. Under these conditions, ≈70% of leaf explants formed ≈40 shoots per petri dish that could be harvested and rooted to form plantlets. Chemical names used: N6-benzyladenine (BA); 2,4-dichlorophenoxyacetic acid (2,4-D); gibberellic acid (GA3); indole-3-acetic acid (IAA); indole-3-butyric acid (IBA); α-naphthaleneacetic acid (NAA); N-phenyl-N'-1,2,3-thidiazol-5-ylurea [thidiazuron (TDZ)].

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