scholarly journals An efficient in vitro regeneration system via callus from hypocotyl and root of Iris laevigata

2020 ◽  
Author(s):  
Fengyi Li ◽  
Lijuan Fan ◽  
Haijing Fu ◽  
Yujia Liu ◽  
Ling Wang
Keyword(s):  
In Vitro Regeneration ◽  
Genetic Material ◽  
Reproductive Capacity ◽  
Penicillin G ◽  
Induction Medium ◽  
Multiplication Rate ◽  
Regeneration System ◽  
Root Explants ◽  
2,4 Dichlorophenoxyacetic Acid

Abstract Background Iris laevigata is an ornamental plant with strong cold resistance. However, its low reproductive capacity limits its landscape applications. The I. laevigata wild genetic resources also need to be protected. In order to develop an effective regeneration system, the optimum agar concentration for the induction medium was determined. Two explants (hypocotyl and root) were then cultured on medium containing different concentrations of plant growth regulator (PGR). In addition, three antibiotics were evaluated for controlling endophyte contamination. Results The highest induction rate (75.00%) was obtained from hypocotyl explants on Murashige and Skoog salt mixture (MS) medium containing 6-benzylaminopurine (6-BA) 0.5 mg L-1 + 2,4-Dichlorophenoxyacetic acid (2,4-D) 1.0 mg L-1 + 1-naphthylacetic acid (NAA) 0.4 mg L-1. The medium containing 6-BA 0.5 mg L-1 + 2,4-D 0.5 mg L-1 + NAA 0.2 mg L-1 achieved the greatest multiplication rate (73.33%). Media containing indole-3-butyric acid (IBA) 0.5 mg L-1 + 6-BA 1.5 mg L-1 + NAA 1.0 mg L-1 achieved the highest differentiation rate (39.72%) for hypocotyl induced calli. Medium containing 6-BA 2.0 mg L-1 + NAA 0.4 mg L-1 + kinetin (KT) 1.0 mg L-1 resulted in the highest differentiation rate (49.52%) for root induced calli. One hundred mg L-1 penicillin G resulted in the optimal rate for reducing endophyte contamination. Conclusions The research determined the optimum PGR concentrations for inducting and multiplying I. laevigata calli from hypocotyl and root explants and a satisfactory means for control endophyte contamination. This research will result in the efficient and reliable reproduction of I. laevigata for landscape applications, genetic development of new Iris varieties, and preservation of the wild genetic material.

scholarly journals 117 An In Vitro Regeneration System for Basil (Ocimum basilicum L.)

HortScience ◽  
1999 ◽  
Vol 34 (3) ◽  
pp. 461E-461
Author(s):  
Winthrop B. Phippen ◽  
James E. Simon
Keyword(s):  
In Vitro Regeneration ◽  
Morphological Characteristics ◽  
Leaf Explants ◽  
Basal Medium ◽  
Induction Medium ◽  
Shoot Induction ◽  
Regeneration System ◽  
Transformation Protocol ◽  
Regeneration Procedure

A plant regeneration protocol was successfully developed for basil (O. basilicum L.). Explants from 1-month-old seedlings yielded the highest frequency of regeneration of shoots (37%) with an average number of 3.6 shoots per explant. Calli and shoot induction were initiated on Murashige and Skoog (MS) basal medium supplemented with thidiazuron (TDZ) (4 mg/L) for ≈30 days. Shoot induction and development was achieved by refreshing the induction medium once after 14 days. The most morphogenetically responsive explants were basal leaf explants from the first fully expanded true leafs of greenhouse-grown basil seedlings. Developing shoots were then rooted on MS media in the dark without TDZ. Within 20 days, rooted plantlets were transferred and acclimatized under greenhouse conditions where they developed normal morphological characteristics. This is the first report of a successful in vitro regeneration system for basil through primary callus. The establishment of a reliable regeneration procedure is critical when developing a transformation protocol for enhancing the production of basil for insect and disease resistance and improved essential oil constituents.

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 Optimization of Callogenesis/Caulogenesis Induction Protocol in Saffron Plant (Crocus sativus L.) Using Response Surface Methodology

2021 ◽  
Vol 12 (4) ◽  
pp. 4731-4746
Keyword(s):  
Callus Formation ◽  
In Vitro Regeneration ◽  
Shoot Formation ◽  
Crocus Sativus ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Regeneration System ◽  
Indole Butyric Acid ◽  
2,4 Dichlorophenoxyacetic Acid

The Crocus sativus, an endangered medicinal and aromatic plant in Morocco, has a low propagation rate in natural conditions and, therefore, an efficient method for in vitro propagation is required. This study investigated the effects of various hormones on the induction of callogenesis and callogenesis in C. sativus corms using the Box-Behnken experimental design. The best shoot formation was obtained with Murashige and Skoog fortified with 3 mg/L 6-Benzylaminopurine. On the other hand, callus formation was obtained with 3 mg/L 1-Naphthaleneacetic Acid or 3 mg/L 2,4-Dichlorophenoxyacetic Acid. However, a combination of 3 mg/L 6-Benzylaminopurine, 1.056 mg/L Indole Butyric Acid, and 3 mg/L 2,4-Dichlorophenoxyacetic Acid allows 50% caulogenesis and 60% callogenesis. The in vitro regeneration system could be utilized for both conservation and largescale multiplication of Crocus sativus corms.

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.

Establishment of in vitro regeneration system for Acaciella angustissima (Timbe) a shrubby plant endemic of México for the production of phenolic compounds

2016 ◽  
Vol 86 ◽  
pp. 49-57 ◽  
Author(s):  
Jannette Alonso-Herrada ◽  
Félix Rico-Reséndiz ◽  
Juan Campos-Guillén ◽  
Ramón G. Guevara-González ◽  
Irineo Torres-Pacheco ◽  
...  
Keyword(s):  
Phenolic Compounds ◽  
In Vitro Regeneration ◽  
Regeneration System

scholarly journals In vitro Regeneration of Dalbergia sissoo Roxb. and the Potential for Genetic Transformation

2012 ◽  
Vol 40 (2) ◽  
pp. 140 ◽  
Author(s):  
Hafiz Mamoon REHMAN ◽  
Iqrar Ahmad RANA ◽  
Siddra IJAZ ◽  
Ghulam MUSTAFA ◽  
Faiz Ahmad JOYIA ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Genetic Transformation ◽  
Callus Induction ◽  
In Vitro Regeneration ◽  
Induction Medium ◽  
Native Forest ◽  
Ms Medium ◽  
Dalbergia Sissoo ◽  
Callus Induction Medium

Dalbergia sissoo Roxb. ex DC. (Sissoo) is a native forest tree species in Pakistan. Many ecological and economical uses are associated with this premier timber species, but dieback disease is of major concern. The objective of this study was to develop a protocol for in vitro regeneration of Sissoo that could serve as target material for genetic transformation, in order to improve this species. Callus formation and plantlet regeneration was achieved by culturing cotyledons, immature seeds, and mature embryos on a modified Murashige and Skoog (1962) (MS) medium supplemented with plant growth regulators. Callus induction medium containing 2.71 ?M 2, 4-dichlorophenoxyacetic acid (2,4-D) and 0.93 ?M kinetin produced better callus on all explants tested compared to other treatments, such as 8.88 ?M 6-benzylaminopurine (BA) and 2.69 ?M ?-naphthalene acetic acid (NAA), or 2.71 ?M 2, 4-D and 2.69 ?M NAA. Shoot regeneration was best on MS medium containing 1.4 ?M NAA and 8.88 ?M BA compared to other treatments, such as 1.4 ?M NAA and 9.9 ?M kinetin, or 2.86 ?M indole-3-acetic acid and 8.88 ?M BA. Murashige and Skoog medium containing 1.4 NAA ?M and 8.88 ?M BA was better in general for regeneration regardless of callus induction medium and the type of explant used. Rooting was best on half-strength MS medium with 7.35 ?M indole-3-butyric acid. Regenerated plantlets were acclimatized for plantation in the field. Preliminary genetic transformation potential of D. sissoo was evaluated by particle bombardment of callus explants with a pUbiGus vector. The bombarded tissue showed transient Gus activity 1week after bombardment. Transformation of this woody tree is possible provided excellent regeneration protocols. The best combination for regeneration explained in this study is one of such protocols.

scholarly journals Assessment of In-vitro culture through Nodal explants of Dendrocalamus hamiltonii

2021 ◽  
Vol 2 (1) ◽  
pp. 130-133
Author(s):  
Abha Jha ◽  
◽  
Sunila Das ◽  
Keyword(s):  
Experimental Study ◽  
Growth Regulators ◽  
In Vitro Regeneration ◽  
Shoot Multiplication ◽  
Growth Hormones ◽  
Nodal Explants ◽  
Bud Break ◽  
Multiplication Rate ◽  
Dendrocalamus Hamiltonii

The present experimental study was aimed to overcome the traditional methods of propagation that limit the number of propagules by in-vitro regeneration through nodal explants of Dendrocalamus hamiltonii with a comparative study of growth regulators during the shooting and rooting process. Dendrocalamus hamiltonii is distributed from the Himalayas (Nepal) to the northern part of Burma. Collection of explants was done from different selected sites of CPTs. There was the use of HgCl2 and Ca (OCl)2 as sterilizing agents in different concentrations and its effect was visualized during the sprouting stage. Culm explants were cultured in a bottle containing White media (Wm) supplemented with BA and Kinetin for sprouting and IAA, IBA, NAA for rooting. There is also the use of IAA+IBA+NAA in combined form as a supplementary solution 0.1% HgCl2 treatment for 20-minute results into77.80% aseptic buds and 72% bud -break. Among the used growth-hormones, BA with concentration 0.25mg/l and 0.50mg/l respectively were appropriate for shoot-multiplication rate, 4.01±0.3 and 4.3±0.4 were ideal observation incorporation with BA (1.00mg/l) and BA (1.50mg/l) respectively. Maximum sprouting rate14.77±3.37with application of BA (2.00mg/l) and maximum shoot length4.3±0.4 is observed at BA (1.50mg/l). The applications of rooting hormone IAA+IBA+NAA in the concentration of 1.0 mg/l results in 72.5±0.3(rooting) and 11.1±0.3 (av. No. of the root).

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