scholarly journals Precise evaluation of tissue culture-induced variation during optimisation of in vitro regeneration regime in barley

2020 ◽  
Vol 103 (1-2) ◽  
pp. 33-50 ◽  
Author(s):  
Renata Orłowska ◽  
Piotr Tomasz Bednarek
Keyword(s):  
Tissue Culture ◽  
In Vitro Regeneration ◽  
Precise Evaluation ◽  
Induced Variation

scholarly journals Barley somatic embryogenesis-an attempt to modify variation induced in tissue culture

2021 ◽  
Vol 28 (1) ◽  
Author(s):  
Renata Orłowska
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Taguchi Method ◽  
Culture Conditions ◽  
Dna Demethylation ◽  
Silver Ion ◽  
Donor Plant ◽  
In Vitro Tissue Culture ◽  
Induced Variation

Abstract Background Somatic embryogenesis is a phenomenon carried out in an environment that generates abiotic stress. Thus, regenerants may differ from the source of explants at the morphological, genetic, and epigenetic levels. The DNA changes may be the outcome of induction media ingredients (i.e., copper and silver ions) and their concentrations and time of in vitro cultures. Results This study optimised the level of copper and silver ion concentration in culture media parallel with the induction medium longevity step towards obtaining barley regenerants via somatic embryogenesis with a minimum or maximum level of tissue culture-induced differences between the donor plant and its regenerants. The optimisation process is based on tissue culture-induced variation evaluated via the metAFLP approach for regenerants derived under varying in vitro tissue culture conditions and exploited by the Taguchi method. In the optimisation and verification experiments, various copper and silver ion concentrations and the different number of days differentiated the tested trials concerning the tissue culture-induced variation level, DNA demethylation, and de novo methylation, including symmetric (CG, CHG) and asymmetric (CHH) DNA sequence contexts. Verification of optimised conditions towards obtaining regenerants with minimum and maximum variability compared to donor plants proved useful. The main changes that discriminate optimised conditions belonged to DNA demethylation events with particular stress on CHG context. Conclusions The combination of tissue culture-induced variation evaluated for eight experimental trials and implementation of the Taguchi method allowed the optimisation of the in vitro tissue culture conditions towards the minimum and maximum differences between a source of tissue explants (donor plant) and its regenerants from somatic embryos. The tissue culture-induced variation characteristic is mostly affected by demethylation with preferences towards CHG sequence context.

scholarly journals CURRENT STATUS OF TRANSGENIC COTTON-UTILITY OF GENOTYPE INDEPENDENT IN PLANTA APPROACHES FOR THE GENERATION OF TRANSGENICS

2020 ◽  
Vol 11 (2) ◽  
Author(s):  
Kesiraju Karthik
Keyword(s):  
Tissue Culture ◽  
In Vitro Regeneration ◽  
Crop Improvement ◽  
Transgenic Cotton ◽  
Current Status ◽  
In Planta ◽  
Biotic And Abiotic Stresses ◽  
Independent Manner ◽  
Gossypium Spp

Cotton (Gossypium spp.), is a mercantile crop plant is grown for its fluffy fiber and cotton seed oil in around 70 countries worldwide. Cotton is an economically important crop, shows erratic productivity under rain feed conditions; it is bogged down with many biotic and abiotic stresses. Due to lack of resistant germplasm, crop improvement through conventional breeding practices has been lagging. Genetic engineering offers numerous protocols to engineer plants to overcome stress. Biotechnological intervention for cotton improvement has begun three decades ago. The recalcitrance of cotton to tissue culture has been the major constraint for in vitro regeneration. Alternate methods that evade tissue culture regeneration steps have thus been envisaged. Till date there are very few standardized protocols that can be employed to develop transgenics in a genotype independent manner. Thus, genotype independent in planta transformation strategies have gained momentum in the present days, but reproducibility of reported protocols remains an amigna in many cases. In planta transformations holds prominence due to viability and ease in generation of transgenic cotton plants with in less time. This review focuses on grouping efforts made by different research groups in this senior. Several reports and standardizations have been focused that reports development of transgenic cotton.

scholarly journals In vitro Propagation and Ex vitro Rooting of Blueberry Plantlets

1970 ◽  
Vol 18 (2) ◽  
pp. 187-195 ◽  
Author(s):  
Zhao Guang-jie ◽  
Wang Zhan-bin ◽  
Wang Dan
Keyword(s):  
Tissue Culture ◽  
Key Words ◽  
Plant Tissue ◽  
In Vitro Propagation ◽  
Root Formation ◽  
Early Stage ◽  
In Vitro Regeneration ◽  
Basal Medium ◽  
Ex Vitro

Effects of different concentrations of 2-ip and IBA in WPM basal medium for Blomidon blueberry in vitro propagation and four different rooting agents at the early stage after transplantation showed that 15 mg/l of 2-ip is the best concentration to induce shoots. For optimum in vitro root formation 10 µM IBA was found to be best and four rooting agents for seedling transplantation according to their effects were No.2>, No.4>, No.3 >, water > and No. 1. Key words: Blomidon, Tissue culture, In vitro regeneration, Rooting agent D.O.I. 10.3329/ptcb.v18i2.3650 Plant Tissue Cult. & Biotech. 18(1): 187-195, 2008 (December)

CALLUS INDUCTION AND PLANT REGENERATION FROM IMMATURE EMBRYOS CULTURE OF TROPICAL MAIZE

2018 ◽  
Vol 17 (3) ◽  
pp. 359
Author(s):  
MARIA JOSÉ VILAÇA VASCONCELOS ◽  
MAURICIO SCHUSTERSCHITZ ANTUNES ◽  
MAURÍLIO FERNANDES DE OLIVEIRA ◽  
MAURÍCIO ANTÔNIO LOPES ◽  
JOSÉ EDSON FONTES FIGUEIREDO
Keyword(s):  
Tissue Culture ◽  
Zea Mays ◽  
Callus Induction ◽  
Culture Media ◽  
In Vitro Regeneration ◽  
Immature Embryos ◽  
Type I ◽  
Type Ii ◽  
Tropical Maize

ABSTRACT - The development of protocols to overcome the current limitations of callus induction and in vitro regeneration of highly recalcitrant tropical maize is crucial for plant genetic transformation. The ability of embryogenic callus (EC) formation of 46 tropical maize hybrids and 14 inbred lines was tested using N6 medium with the following changes: medium A (N6 + 15 μM dicamba + 25 mM L-proline + 88 mM AgNO3), medium B (N6 + 30 μM dicamba + 25 mM L-proline + 88 mM AgNO3), medium C (N6+ 30 μM dicamba + 6 mM L-proline), and medium D (N6+ 30 μM dicamba + 25 mM L-proline). Compact (Type I) and friable (Type II) callus were induced in the four media. Twenty genotypes produced callus in all media, and four genotypes (CO32, AG8012, CMS477BC4F2, and CMS-HGZ10) produced the highest number of callus (114, 134, 131, and 126 calli, respectively). All immature embryos of ten genotypes produced callus in at least one medium, while eight genotypes were highly recalcitrant, and they did not produce any callus. The frequencies of EC ranged from 0% to 38%, and the highest rate of EC was observed on medium B (0.40) with a total of 865 calli, and the lowest induction rate was obtained with medium D (0.29) with 555 calli (P= 0.05). From the seventy-two EC of 26 genotypes transferred to Murashige & Skoog regeneration medium, twenty-four (66.7%) differentiated into green plants which produced seeds in R0 and R1 generations, and twelve (33.3%) developed into albino plants. The results demonstrated that the problem of the recalcitrant genotypes can be, at least partially, overcome by using immature embryos as explants together with tissue culture media formulations.Keywords: Zea mays, immature embryos, somatic embryogenesis, tissue culture, type I and type II calli. INDUÇÃO DE CALO E REGENERAÇÃO DE PLANTAS DA CULTURA DE EMBRIÕES IMATUROS DE MILHO TROPICAL  RESUMO - O desenvolvimento de protocolos para superar as limitações atuais de indução de calos e regeneração in vitro de milho tropical altamente recalcitrante é crucial para a transformação genética de plantas. Foi testada a capacidade de formação de calos embriogênicos (CE) de 46 híbridos de milho tropicais e 14 linhagens cultivadas em Meio N6 com as seguintes alterações: Meio A (N6 + 15 μM dicamba + 25 mM L-prolina + 88 mM AgNO3), Meio B (N6 + 30 μM de dicamba + 25 mM de L-prolina + 88 μM de AgNO3), Meio C (N6 + 30 μM de dicamba + 6 mM de L-prolina) e Meio D (N6 + 30 μM de dicamba + 25 mM de L-prolina). Calos compactos (tipo I) e friáveis (tipo II) foram induzidos nos quatro meios de cultura. Vinte genótipos produziram calos em todos os meios, e quatro genótipos (CO32, AG8012, CMS477BC4F2 e CMS-HGZ10) produziram o maior número de calos (114, 134, 131 e 126 calos, respectivamente). Todos os embriões imaturos de dez genótipos produziram calos em pelo menos um meio, enquanto oito genótipos foram altamente recalcitrantes e não produziram nenhum calo. As frequências de CE variaram de 0% a 38%, e a maior taxa de CE foi observada no meio B (0,40), com um total de 865 calos, e a menor taxa de indução com o meio D (0,29), com 555 calos (P = 0,05). Dos setenta e dois CE de 26 genótipos transferidos para meio de regeneração Murashige & Skoog, vinte e quatro (66,7%) se diferenciaram em plantas verdes que produziram sementes nas gerações R0 e R1, e doze (33,3%) se desenvolveram em plantas albinas. O problema relacionado dos genótipos recalcitrantes de milho pode ser, pelo menos parcialmente, contornado com o uso de explantes de embriões imaturos juntamente com a formulação dos meios de cultura.Palavras-chave: Zea mays, embriões imaturos, embriogênese somática, cultura de tecidos, calos tipo I e tipo II.

Establishment and standardization of in vitro regeneration protocol in Nothapodytes nimmoniana Graham and evaluation of camptothecine (CPT) in tissue culture plants

2015 ◽  
Vol 21 (1) ◽  
pp. 1-7 ◽  
Author(s):  
Ugraiah Amilineni ◽  
Vinod Gangal ◽  
Ravikanth Gudasalamani ◽  
Nataraja N. Karaba ◽  
Uma Shaanker Ramanan
Keyword(s):  
Tissue Culture ◽  
In Vitro Regeneration ◽  
Nothapodytes Nimmoniana

scholarly journals PLANT TISSUE CULTURE: A TOOL TO CONSERVE BIODIVERSITY THROUGH RAPID IN VITRO REGENERATION OF DIVERSE PLANT SPECIES

2017 ◽  
pp. 132-139
Author(s):  
Nirali Vora
Keyword(s):  
Tissue Culture ◽  
Plant Species ◽  
Plant Tissue Culture ◽  
Plant Tissue ◽  
In Vitro Regeneration ◽  
Ornamental Plants ◽  
Cost Effective ◽  
Human Consumption ◽  
Natural Forests

Biodiversity is declining with the loss of natural forests across the world. Plant tissue culture is an important biotechnological tool to raise large number of plant species in short spa of time. However, commercial tissue culture laboratories are working on raising plantlets important for human consumption only; which mainly include fruit crops, ornamental plants, timber-yielding forest trees and medicinal plants. There is an urgent need of raising all the different plant species rapidly through tissue culture. Through cultivation of these high yielding and disease-free crops in the forests for consumption of all other species of fauna; conservation of biodiversity can be achieved. However, as tissue culture plantlets are costlier than conventionally raised plants, despite of its advantages its utility is limited. To reduce cost of an important fruit crop, banana during its in vitro regeneration, cost-effective alternatives are proposed.

Genetic control of somatic embryogenesis in alfalfa

Genome ◽  
1992 ◽  
Vol 35 (3) ◽  
pp. 474-477 ◽  
Author(s):  
G. A. Kielly ◽  
S. R. Bowley
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Medicago Sativa ◽  
Genetic Control ◽  
In Vitro Regeneration ◽  
Breeding Population ◽  
Medicago Sativa L

The genetic control of somatic embryogenesis in alfalfa (Medicago sativa L.) was studied using one nonembryogenic and three embryogenic genotypes: A70-34, a selection from 'Rangelander'; RA3, a selection from 'Regen-S'; and C2-4, a selection from a breeding population that had A70-34 in its pedigree. Crosses of embryogenic × embryogenic and embryogenic × nonembryogenic and S1 and BC1 testcrosses were evaluated for in vitro regeneration. Selfing reduced the expression of the trait. Somatic embryogenesis was dominant and explained by two loci. All three regenerating genotypes shared a common genetic system.Key words: alfalfa, Medicago sativa, somatic embryogenesis, tissue culture.

scholarly journals Understanding In Vitro Tissue Culture-Induced Variation Phenomenon in Microspore System

2021 ◽  
Vol 22 (14) ◽  
pp. 7546
Author(s):  
Piotr Tomasz Bednarek ◽  
Katarzyna Anna Pachota ◽  
Wioletta Monika Dynkowska ◽  
Joanna Machczyńska ◽  
Renata Orłowska
Keyword(s):  
Tissue Culture ◽  
Plant Regeneration ◽  
Signaling Pathways ◽  
In Vitro Tissue Culture ◽  
Dna Mutations ◽  
Complicated Process ◽  
Multiple Levels ◽  
Induced Variation ◽  
Cellular Organelles

In vitro tissue culture plant regeneration is a complicated process that requires stressful conditions affecting the cell functioning at multiple levels, including signaling pathways, transcriptome functioning, the interaction between cellular organelles (retro-, anterograde), compounds methylation, biochemical cycles, and DNA mutations. Unfortunately, the network linking all these aspects is not well understood, and the available knowledge is not systemized. Moreover, some aspects of the phenomenon are poorly studied. The present review attempts to present a broad range of aspects involved in the tissue culture-induced variation and hopefully would stimulate further investigations allowing a better understanding of the phenomenon and the cell functioning.

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 Hormonal Effect on Mandarin Orange (Citrus reticulata Blanco) Micro-propagation

2016 ◽  
Vol 4 (1) ◽  
pp. 33-36 ◽  
Author(s):  
Resham Babu Amgai ◽  
Hari Kumar Prasai ◽  
Yama Raj Pandey
Keyword(s):  
Tissue Culture ◽  
In Vitro Regeneration ◽  
Flower Bud ◽  
Shoot Bud ◽  
Hormonal Effect ◽  
Medium Level ◽  
Micro Propagation ◽  
Planting Materials ◽  
Mandarin Orange

Tissue culture is the best option to produce disease free seedling of the fruit crop rapidly. Micro-propagation and use of the in-vitro grafting (micro-grafting) is very helpful for production of virus free planting materials in mandarin. Different levels of the in-vitro hormone affect the success of callusing, shooting and plant regeneration in mandarin. Shoot bud, flower bud and in-vitro seedling epicotyl was used as explants to study the hormonal effect on mandarin micro-propagation. Similarly, 10 levels of BAP and IAA combination on MS media for mandarin tissue culture were used. Observation was done for 100 test tubes per treatment combination after 4, 8 and 12 weeks of culture. Data was arc sine transformed for analysis. Shooting from explants was significantly higher (71.72%) on medium level of the BAP (0.5 mg/L) and IAA (0.2 mg/L) using in-vitro seedling stem as explant, however, it was 27.91% for stem bud as explant. Stem bud showed higher level of callusing (6.15%, p<0.001) in mandarin orange. However, flower bud didn’t develop shoot in mandarin tissue culture. Increment of the in-vitro regeneration of the shooting and callusing was observed by the increment of the in-vitro incubation duration in mandarin orange tissue culture.

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