scholarly journals Creating a novel petal regeneration system for function identification of colour gene of grape hyacinth

Plant Methods ◽  
2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Qian Lou ◽  
Hongli Liu ◽  
Wen Luo ◽  
Kaili Chen ◽  
Yali Liu
Keyword(s):  
Transformation Method ◽  
Breeding Cycle ◽  
Regeneration System ◽  
Flower Bud ◽  
Juvenile Period ◽  
Colour Gene ◽  
Function Identification ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Grape Hyacinth

Abstract Background Grape hyacinth (Muscari spp.) is one of the most important ornamental bulbous plants. However, its lengthy juvenile period and time-consuming transformation approaches under the available protocols impedes the functional characterisation of its genes in flower tissues. In vitro flower organogenesis has long been used to hasten the breeding cycle of plants but has not been exploited for shortening the period of gene transformation and characterisation in flowers. Results A petal regeneration system was established for stable transformation and function identification of colour gene in grape hyacinth. By culturing on Murashige and Skoog medium (MS) with 0.45 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 8.88 μM 6-benzyladenine (6-BA), during the colour-changing period, the flower bud explants gave rise to regeneration petals in less than 3 months, instead of the 3 years required in field-grown plants. By combining this system with Agrobacterium-mediated transformation, a glucuronidase reporter gene (GUS) was delivered into grape hyacinth petals. Ultimately, 214 transgenic petals were regenerated from 24 resistant explants. PCR and GUS quantitative analyses confirmed that these putative transgenic petals have stably overexpressed GUS genes. Furthermore, an RNAi vector of the anthocyanidin 3-O-glucosyltransferase gene (MaGT) was integrated into grape hyacinth petals using the same strategy. Compared with the non-transgenic controls, reduced expression of the MaGT occurred in all transgenic petals, which caused pigmentation loss by repressing anthocyanin accumulation. Conclusion The Agrobacterium transformation method via petal organogenesis of grape hyacinth took only 3–4 months to implement, and was faster and easier to perform than other gene-overexpressing or -silencing techniques that are currently available.

scholarly journals High Frequency Plant Regeneration System from Transverse Thin Cell Layer Section of In vitro Derived ‘Nadia’ Ginger Microrhizome

2014 ◽  
Vol 6 (1) ◽  
pp. 85-91
Author(s):  
Dikash Singh THINGBAIJAM ◽  
Devi Sunitibala HUIDROM
Keyword(s):  
High Frequency ◽  
Somatic Embryos ◽  
Cell Layer ◽  
Sucrose Concentration ◽  
Dichlorophenoxyacetic Acid ◽  
Thin Cell Layer ◽  
Regeneration System ◽  
Transverse Thin Cell Layer ◽  
2,4 Dichlorophenoxyacetic Acid

An efficient and reproducible procedure is outlined for rapid in vitro multiplication of Zingiber officinale var. ‘Nadia’ through high frequency shoot proliferation from transverse thin cell layer (tTCL) sections of in vitro derived microrhizome. In vitro derived microrhizome of size 500 μm in thickness was used as initial explants for induction of somatic embryos. Among the different phytohormones tested, tTCL explants shows maximum calli proliferation in medium containing 2 mg/L 2,4-Dichlorophenoxyacetic acid (88.30±0.11%). Reduced concentration of 2,4 Dichlorophenoxyacetic acid was supplemented with different cytokinins for regeneration of callus. Among the different medium tested, optimum redifferentiation of somatic embryos were observed in medium containing 0.2 mg/L 2,4 Dichlorophenoxyacetic acid and 6.0 mg/L BAP (141.08±0.25). Clump of regenerated plantlets were further subculture and transfer into microrhizome inducing medium containing high sucrose concentration (8%). Plantlets with well developed microrhizome were successfully acclimatized and eventually transferred to the field. The application of studying embryo section for regeneration of plants might be useful alternative to ginger improvement programme. Histological analysis showed formation of somatic embryos and regenerated adventitious shoot.

scholarly journals A Review on In-vitro Haploid Production in Orchids

2021 ◽  
pp. 15-19
Author(s):  
Teena Jaswal ◽  
Saranjeet Kaur
Keyword(s):  
Breeding Cycle ◽  
Fertilization In Vitro ◽  
Juvenile Period ◽  
Haploid Production ◽  
Natural Breeding ◽  
Time Required ◽  
Pure Lines ◽  
Haploid Plants

The aim of writing this paper is to review production of haploids in orchids in vitro. Haploids possess half number of chromosomes and do not undergo fertilization. In vitro conditions provide necessary nutrients and conditions that are required for growth of haploid plantlets. The natural breeding cycle of orchids is very slow as well as unpredictable. To reduce this time, the technique of in vitro haploid production is used. In vitro conditions can decrease or shorten the time required for juvenile period in Orchids. Haploid plants are produced from in vitro haploid culture. This technique is useful to produce homozygous pure lines and to increase the yield of a plant.

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 324 An in Vitro Regeneration System for Mass Production of Daylily (Hemerocallis fulva L.)

HortScience ◽  
2000 ◽  
Vol 35 (3) ◽  
pp. 448A-448
Author(s):  
Johnny Carter ◽  
Seema Dhir
Keyword(s):  
Tissue Culture ◽  
Mass Production ◽  
Somatic Embryos ◽  
In Vitro Regeneration ◽  
Regeneration System ◽  
Flower Buds ◽  
Flower Bud ◽  
Explant Source ◽  
Bud Size

A plant regeneration protocol has been successfully developed to mass propagate daylilies. Experiments were conducted to determine source (BA, KN, and ZT) and concentration (0, 1.0, 2.0, and 3.0 mg/L) of cytokinins and sugars (glucose, surcose, and maltose) to be used in the medium. Studies were also conducted to determine the influence of flower bud size (5, 10, 15, and 20 mm) as explant source. Based on results from these studies a protocol for propagating daylilies was developed. The procedure involved using filament explants from daylily flower buds ranging in sizes from 5 to 10 mm. The filaments when cultured on MS+BAP (3.0 mg/L)+ IAA (0.5 mg/L) medium,formed globular somatic embryos in 4 weeks. Complete plants were regenerated within a period of 6 to 7 months. Upon acclimatization, 100% of the tissue culture generated raised plants survived under greenhouse conditions.

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.

Establishment of in Vitro regeneration system for Ricinus communis

2014 ◽  
Vol 39 (5) ◽  
pp. 484-488
Author(s):  
Chao XING ◽  
Lu JIN ◽  
Peng LIU ◽  
Ying RUAN ◽  
Chun-lin LIU
Keyword(s):  
Ricinus Communis ◽  
Regeneration System

Introduction of Eucommia ulmoides Oliv. to in vitro Culture

2020 ◽  
pp. 38-50
Author(s):  
A.V. Zhigunov ◽  
◽  
Q.T. Nguyen
Keyword(s):  
Herbal Medicines ◽  
Naphthaleneacetic Acid ◽  
Eucommia Ulmoides ◽  
Juvenile Period ◽  
Dioecious Species ◽  
Relict Species ◽  
Mass Reproduction ◽  
Special Value ◽  
The Republic

The increasing need for herbal medicines requires the study of not only biological resources of medical plants, but also methods for their reproduction. Of special value are the medicinal plants that have a long history of success in traditional medicine. One of such plants is Eucommia ulmoides Oliv., which belongs to a rare relict species growing in natural conditions, for the most part, in the undergrowth of humid subtropical forests in China, mainly in the middle course of the Yangtze river. E. ulmoides compares favorably with most subtropical plants owing to its significant frost resistance, which makes it possible to cultivate it outside the humid subtropics. It has been widely introduced in Krasnodar Krai and in the Republic of Adygea (Russia) since the mid-20th century and successfully adapted to various environmental conditions in the Northwest Caucasus. The increasing demand for E. ulmoides bark can only be satisfied by laying out industrial plantations. However, the difficulties encountered in the traditional seed reproduction of E. ulmoides (dioecious species, pollen low quality, parthenocarpy, prolonged seed dormancy, irregular fruiting, long juvenile period, etc.) make scientists turn to modern biotechnological methods of plant propagation. While considering cultivation of planting material, we should focus on highly efficient methods that ensure stable and mass reproduction of the plants under study. An important role is played here by in vitro plant regeneration. The effectiveness of biotechnology methods is due to a reduction in timing of obtaining a large number of vegetative progeny of plants difficult for propagation, as well saving of the area required for their cultivation. The conditions for producing an aseptic culture of E. ulmoides were chosen based on the results of the studies. The highest degree of sterilization of E. ulmoides shoot segments was achieved when the explants were sequentially immersed first in 70 % ethanol (30 s) and then in 0.1 % mercuric chloride solution (5 min). With such a sterilization procedure, 63.3 % of the studied cuttings were made sterile, and 56.7 % of them proved to be viable. The optimal composition of the nutrient medium for regeneration of E. ulmoides microshoots has been determined: MS medium complemented with 1 mg/L 6-Benzylaminopurine (BAP) + 0.2 mg/L 1-Naphthaleneacetic acid (NAA). The best media for explant rooting are the following: 2/3 MS + 1.5 mg/L NAA + 30 g sucrose + 7 g agar; 2/3 MS + 1 mg/L NAA + 0.4 mg/L IBA + 30 g sucrose + 7 g agar.

FACTORS AFFECTING THE REGENERATION OF PEPPER (CAPSICUM ANNUUM L.)

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1120G-1120
Author(s):  
J. L. Jacobs ◽  
C. T. Stephens
Keyword(s):  
Growth Hormone ◽  
Silver Nitrate ◽  
Callus Formation ◽  
Leaf Explants ◽  
Regeneration System ◽  
Factors Affecting ◽  
Nitrate Concentrations ◽  
Leaf Differentiation ◽  
Whole Plants

Several growth hormone combinations and silver nitrate concentrations were examined for their effect on regeneration of different pepper genotypes. Primary leaf explants from in vitro seedlings were cultured on a revised Murashige and Skoog medium supplemented with auxin, cytokinin and 1.6% glucose. Combinations of different concentrations of indole-3-acetic acid (IAA), 0-5 mg/l, and 6-benzylaminopurine (BAP), 0-5 mg/l, were tested to determine the most effective medium for shoot primordium formation. Experiments with IAA and BAP did not result in a specific growth hormone combination appropriate for regeneration of all genotypes tested. Of the silver nitrate concentrations tested, 10 mg/l resulted in the best shoot and leaf differentiation and reduced callus formation. Differences in organogenic response of individual genotypes were evaluated on a single regeneration medium. Whole plants were regenerated from 11 of 63 genotypes examined. Based on these experiments, a reproducible regeneration system for pepper was developed with a total of 500 plants regenerated to date.

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.

Characterization of the Reproductive Mode in Guayule In Vitro

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 483a-483
Author(s):  
Roy N. Keys ◽  
Dennis T. Ray ◽  
David A. Dierig
Keyword(s):  
Field Experiments ◽  
Reproductive Mode ◽  
Dichlorophenoxyacetic Acid ◽  
Initial Field ◽  
Breeding Lines ◽  
Reproductive Modes ◽  
Desert Southwest ◽  
2,4 Dichlorophenoxyacetic Acid

Guayule (Parthenium argentatum Gray, Asteraceae) is a latex-producing perennial desert shrub that is potentially of economic importance as an industrial crop for the desert Southwest. It is known to possess complex reproductive modes. Diploids are predominantly sexual and self-incompatible, while polyploids show a range of apomictic potential and self-compatibility. This paper describes the development of a relatively rapid and simple technique for characterizing reproductive modes of breeding lines of P. argentatum. Initial field experiments were based on an auxin test used successfully to characterize reproductive mode in the Poaceae. The application of 2,4-dichlorophenoxyacetic acid inhibited embryo formation in P. argentatum, but this was not the case with other auxins tested. Results of field experiments were ambiguous because: 1) the floral structure of P. argentatum is such that auxins might not have penetrated to the ovules, and 2) there was potential self-fertilization by pollen released within isolation bags. Therefore, in vitro culture of flower heads was tested because it provided much better control of environmental conditions, growth regulator application, and pollen release. Auxin alone, or in combination with gibberellic acid or kinetin, inhibited parthenogenesis in vitro. Embryo production did not vary using two substantially different nutrient media. In vitro flower head culture using a (Nitsch and Nitsch) liquid nutrient medium without growth regulators, enabled characterization of the reproductive mode of seven breeding lines, ranging from predominantly sexual to predominantly apomictic. The results of this technique were substantiated using RAPD analyzes of progeny arrays from controlled crosses.

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