scholarly journals Genetic identity of mint cultivars after in vitro conservation, assessed by ISSR primers

2021 ◽  
Vol 937 (4) ◽  
pp. 042014
Author(s):  
M S Zagorskaya ◽  
S F Abdurashytov
Keyword(s):  
Genetic Stability ◽  
Culture Media ◽  
Callus Formation ◽  
In Vitro Conservation ◽  
Artificial Culture ◽  
Full Compliance ◽  
Conservation Genetic ◽  
Somaclonal Variability ◽  
Issr Primers

Abstract The species of the genus Mentha have been known since ancient times and have significant value in the pharmaceutical, cosmetic and food industries, as well as in medicine. For the widespread use of mint, including in a variety of breeding programs, and the preservation of genetic diversity, effective methods of maintaining cultivars and collection samples are required. Thanks to the development of biotechnological methods, in particular, the creation of slow-growing collections, are now actively used as an effective alternative to field collections. It is known that the cultivation of tissues and organs on artificial culture media can cause somaclonal variability. The purpose of this work is to study the effect of in vitro storage at 4-6°C without illumination after 3 and 4 in vitro conservation cycles on the genetic stability of three cultivars of mint Azhurnaya, Bergamotnaya and Zagrava using ISSR primers. 1 cycle: 1 year of in vitro conservation, microcutting and 2 subcultures of regrowth in a culture room. After conservation, the number of viable explants was 70.0-82.1%. Callus formation at the base of the shoots was not observed in any of the cultivars. After 3 and 4 cycles of in vitro conservation, genetic stability was assessed using 11 ISSR primers. It was found that all three mint genotypes showed full compliance (length and number of amplicons) with the profiles of control samples for all studied markers. It was also found that the markers used by ISSR are highly informative for mint cultivars.

scholarly journals Somaclonal variation on in vitro callus culture potato cultivars

2004 ◽  
Vol 22 (2) ◽  
pp. 300-304 ◽  
Author(s):  
Patricia N. Bordallo ◽  
Derly H. Silva ◽  
José Maria ◽  
Cosme D. Cruz ◽  
Elizabeth P. Fontes
Keyword(s):  
Somaclonal Variation ◽  
Culture Media ◽  
Callus Formation ◽  
Potato Cultivars ◽  
Synthetic Seed ◽  
Arbitrary Sequence ◽  
Stem Explants ◽  
Factors Affecting ◽  
High Level

Synthetic seeds can be an alternative for those species in which botanical seeds are not viable. One of the major problems of in vitro plant cultivation is the high level of somaclonal variation. The most common factors affecting somaclonal variation are genotype, explant source, in vitro period and cultivation conditions in which the culture is established. In this work, calli were induced using leaf and stem explants of the commercial potato cultivars Achat, Baraka, Baronesa, Bintje, and Contenda in MS culture media supplemented with 1.65 mM of picloram and 11.5 mM of 2,4-D. Seventy and 90 days after induction, DNA samples of 40 calli were compared concerning the effects of the two explant (leaf and stem) and two growth regulator sources on five potatoes cultivars. A total of 20 arbitrary sequence primers were evaluated. The RAPD pattern generated by these primers suggested a high percentage of polymorphic fragments among the five genotypes, indicating a high level of genetic variation among cultivars. Cultivar Baronesa showed the highest number of polymorphic fragments for all treatments. The cultivar Contenda showed the smallest somaclonal variation, for most of the treatments, except for the treatment which consisted of stem explants, picloram (1.65 mM) application, and a 70-day period of callus formation. 'Contenda' is, therefore, the most suitable cultivar for synthetic seed production.

Control of morphogenetic pathways in thin cell layers of tobacco by pH

1989 ◽  
Vol 67 (3) ◽  
pp. 650-654 ◽  
Author(s):  
A. Cousson ◽  
P. Toubart ◽  
K. Tran Thanh Van
Keyword(s):  
Butyric Acid ◽  
De Novo ◽  
Culture Media ◽  
Callus Formation ◽  
Medium Ph ◽  
Vegetative Buds ◽  
Cell Layers ◽  
Initial Medium ◽  
Vegetative Bud

Thin cell layer explants of tobacco were floated in vitro on the surface of liquid culture media. The initial exogenous concentrations of indolyl-3-butyric acid, and kinetin, the initial medium pH, and the explant density were varied. Various patterns of de novo and direct differentiation without any intermediate callus (flower, vegetative bud, root) as well as the absence of morphogenesis and callus formation without any subsequent organogenesis were separately controlled on 100% of the explants. On the same exogenous combination of glucose, indolyl-3-butyric acid, and kinetin, changes in initial medium pH changed the pattern of morphogenesis. For a given initial exogenous indolyl-3-butyric acid concentration, vegetative buds were obtained at either pH 6.1 or 7.8, whereas a mixture of flowers and vegetative buds was obtained at pH 6.8. Furthermore, changes in explant density changed the morphogenetic response. It is suggested that the effects of the initial medium pH and explant density on morphogenesis may be related partially to modifications of the physicochemical properties of the cell wall and (or) plasmalemma.

scholarly journals Micropropagation and in vitro conservation of Alcantarea nahoumii (Bromeliaceae), an endemic and endangered species of the Brazilian Atlantic Forest

2020 ◽  
Vol 42 ◽  
pp. e52940
Author(s):  
Simone Sacramento dos Santos Silva ◽  
Everton Hilo de Souza ◽  
Fernanda Vidigal Duarte Souza ◽  
Cristina Ferreira Nepomuceno ◽  
Maria Angélica Pereira de Carvalho Costa
Keyword(s):  
Atlantic Forest ◽  
Culture Media ◽  
In Vitro Conservation ◽  
Naphthaleneacetic Acid ◽  
High Survival ◽  
Multiplication Rate ◽  
Ms Medium ◽  
Mature Seeds ◽  
Stem Segments

Alcantarea nahoumii (Leme) J. R. Grant is a species native to the Atlantic Forest that stands out for ornamental purposes. The objective of this work was to evaluate the in vitro germination of A. nahoumii seeds and establish a micropropagation protocol for production of seedlings so as to minimize the effects of predatory extractivism and develop an in vitro conservation system. Mature seeds were disinfested, established in three culture media (MS, MS½ and MS⅓) and incubated at four temperatures (20, 25, 30 and 35ºC) in a germination chamber. In the micropropagation experiment, stem segments were introduced in MS medium supplemented with 0.5 μM of 1-naphthaleneacetic acid (NAA) and 0.0, 2.2, 4.4 and 6.6 μM of 6-benzylaminopurine (BAP). For the in vitro conservation, plantlets were established in MS or MS½ medium supplemented with 15 g L-1 or 30 g L-1 of sucrose. The plants were acclimated with commercial substrate. The highest seed germination percentages were promoted by temperature conditions of 20 and 25ºC, with MS culture medium. The highest multiplication rate of shoots was obtained from the treatment without addition of the growth regulator or when combined with 2.2 μM of BAP + 0.5 μM of NAA. The acclimation of the plants occurred with high survival rate. The species can be conserved in vitro under slow growth condition for 24 months when incubated in MS medium supplemented with 30 g L-1 of sucrose.

scholarly journals Effects of Temperature and pH on Fusarium oxysporum and Soybean Seedling Disease

Plant Disease ◽  
2019 ◽  
Vol 103 (12) ◽  
pp. 3234-3243
Author(s):  
David R. Cruz ◽  
Leonor F. S. Leandro ◽  
Gary P. Munkvold
Keyword(s):  
Fusarium Oxysporum ◽  
Root Rot ◽  
Radial Growth ◽  
Culture Media ◽  
Fungal Growth ◽  
Artificial Culture ◽  
Vitro Assay ◽  
Seedling Disease ◽  
Soybean Seedlings

Fusarium oxysporum (Fo) is an important pathogen that reduces soybean yield by causing seedling disease and root rot. This study assessed the effects of pH and temperature on Fo fungal growth and seedling disease. In an in vitro assay, 14 Fo isolates collected from symptomatic soybean roots across Iowa in 2007 were grown on artificial culture media at five pH levels (4, 5, 6, 7, and 8) and incubated at four temperatures (15, 20, 25, or 30°C). In a rolled-towel assay, soybean seeds from Fo-susceptible cultivar Jack were inoculated with a suspension of a pathogenic or a nonpathogenic Fo isolate; both isolates were previously designated for their relative aggressiveness in causing root rot at 25°C. The seeds were placed in rolled germination paper, and the rolls were incubated in all combinations of buffer solutions at four pH levels (4, 5, 6, and 7), and four temperatures (15, 20, 25, or 30°C). There was a significant interaction between temperature and pH (P < 0.05) for in vitro radial growth and root rot severity. Isolates showed the most in vitro radial growth after incubation at pH 6 and 25°C. For the rolled-towel assay, the pathogenic isolate caused the most severe root rot at pH 6 and 30°C. Gaussian regression analysis estimates for optimal conditions were pH 6.3 at 27.1°C for maximal fungal growth and pH 5.9 at 30°C for maximal root rot severity. These results indicate that optimal pH and temperature conditions are similar for Fo growth and disease in soybean seedlings and suggest that Fo may be a more important seedling pathogen when soybeans are planted under warm conditions in moderately acidic soils.

In vitro conservation strategy for endemic and endangered Himalayan liverwort Stephensoniella brevipedunculata Kashyap (Marchantiophyta)

2018 ◽  
Vol 4 (02) ◽  
pp. 81-87
Author(s):  
A. K. Asthana ◽  
S. D. Tewari ◽  
Vishwa Jyotsna Singh ◽  
Isha Pathak ◽  
Vinay Sahu
Keyword(s):  
Culture Media ◽  
In Vitro Conservation ◽  
Conservation Strategy ◽  
Healthy Population ◽  
Salt Mixture ◽  
Young Thalli ◽  
Half Strength ◽  
Axenic Cultures ◽  
First Time

During the present study an effort has been made to propagate (in-vitro) the endangered and endemic Himalayan liverwort Stephensoniella brevipedunculata Kashyap using different culture media under controlled Laboratory conditions. Axenic cultures of the taxon have been established using tubers as explants. Seven combinations of media with Full Strength Knop’s macronutrients; Half-strength Knop’s macronutrients; Half-strength Knop’s macronutrients + Vitamins; Halfstrength Knop’s macronutrients + 0.2 mg L-1 IBA + 0.1 mg L-1 BAP; Half-strength Knop’s macronutrients + 0.1 mg L-1 Kinetin + 0.1 mg L-1 2,4D; Half-strength Knop’s macronutrients + 0.1 mg L-1 IBA + 0.2 mg L-1 BAP and Hoagland no. 2 basal salt mixture were used for culture. The best growth was observed in the Hoagland no. 2 basal salt mixture medium, in which dichotomously branched young thalli were successfully formed. Subsequently healthy population of culture grown plants has been raised on soil in pots for the first time.

scholarly journals Low Cost Tissue Culture Technologies in Vegetables: A Review

2020 ◽  
pp. 66-78
Author(s):  
Reshav Naik ◽  
Anil Bhushan ◽  
R. K. Gupta ◽  
Anamika Walia ◽  
Anshika Gaur
Keyword(s):  
Tissue Culture ◽  
Culture Media ◽  
Callus Formation ◽  
Low Cost ◽  
Direct Regeneration ◽  
Clonal Variation ◽  
In Vitro Mutagenesis ◽  
Vegetable Crops ◽  
Brown Sugar

The demand of vegetable crops is increasing day by day due to changes in consumption patterns, so the need of the hour is to develop technologies that enhance the vegetable production at a rapid rate. Plant Tissue culture is one such remarkable biotechnological tool that has its application in vegetable propagation and improvement, disease elimination, herbicide resistance, salinity tolerance, incorporation of high nutrient content, genetically improved plants and conservation of endangered plant species and in the near future usage of this technology is going to increase further manifold. It is used for production of disease free quality planting material and development of varieties through direct regeneration, anther/ovule culture, somatic embryogenesis etc. or for creation of new variation (organogenesis via callus formation, soma-clonal variation and in vitro mutagenesis). In spite of being a very important and viable non-conventional biotechnological tool, high cost of production of seedlings in vitro remains a major impediment in popularization of this technology. High cost of producing seedlings is due to availability of limited resources, high recurrent costs of consumables for media and lack of awareness, which limits its application only to a few institutions and rich farmers especially in developing countries. Therefore, in order to make this technology a successful and viable option for the farmers, future thrust must be on cost reduction of in vitro seedlings. The components of tissue culture technology such as culture media components, glassware, lighting and water for media preparation can be replaced with low cost alternatives to reduce the overall cost of tissue culture. The usage of alternatives for gelling agent’s like isabgol (potato, tomato, cassava, turmeric, ginger), sago (potato, tomato, turmeric, ginger) cassava starch (potato, cassava, sweet potato) barley starch, phytagel etc. and for carbon sources like table sugar (potato, turmeric, ginger), jaggery, sugarcane juice, cube sugar (bittergord), brown sugar etc have already been documented worldwide. The present paper reviews the work done by researchers around the globe in developing various low cost alternative technologies with focus on vegetable crops.

scholarly journals Dimorphism in Neopseudocercosporella capsellae, an Emerging Pathogen Causing White Leaf Spot Disease of Brassicas

2021 ◽  
Vol 11 ◽  
Author(s):  
Niroshini Gunasinghe ◽  
Martin J. Barbetti ◽  
Ming Pei You ◽  
Prabuddha Dehigaspitiya ◽  
Stephen Neate
Keyword(s):  
Leaf Spot ◽  
Culture Media ◽  
Control Measures ◽  
Effective Control ◽  
Leaf Spot Disease ◽  
In Planta ◽  
Artificial Culture ◽  
White Leaf ◽  
Yeast Phase

White leaf spot pathogen: Neopseudocercosporella capsellae causes significant damage to many economically important Brassicaceae crops, including oilseed rape through foliar, stem, and pod lesions under cool and wet conditions. A lack of information on critical aspects of the pathogen’s life cycle limits the development of effective control measures. The presence of single-celled spores along with multi-celled conidia on cotyledons inoculated with multi-celled conidia suggested that the multi-celled conidia were able to form single-celled spores on the host surface. This study was designed to demonstrate N. capsellae morphological plasticity, which allows the shift between a yeast-like single-celled phase and the multi-celled hyphal phase. Separate experiments were designed to illustrate the pathogen’s morphological transformation to single-celled yeast phase from multi-celled hyphae or multi-celled macroconidia in-vitro and in-planta. Results confirmed the ability of N. capsellae to switch between two morphologies (septate hyphae and single-celled yeast phase) on a range of artificial culture media (in-vitro) or in-planta on the host surface before infection occurs. The hyphae-to-yeast transformation occurred through the production of two morphologically distinguishable blastospore (blastoconidia) types (meso-blastospores and micro-blastospores), and arthrospores (arthroconidia).

scholarly journals Creation of clary sage cultivar using cell engineering methods. 1. Obtaining of plant-regenerants in callus culture in vitro

2021 ◽  
Vol 1 (25) ◽  
pp. 98-112
Author(s):  
N.A. Yegorova ◽  
◽  
I.V. Stavtzeva ◽  
Keyword(s):  
Callus Formation ◽  
Leaf Shape ◽  
Flower Color ◽  
Callus Cultures ◽  
Maximum Frequency ◽  
Agricultural Plant ◽  
Morphogenic Callus ◽  
Somaclonal Variability ◽  
Callus Tissue Culture

To increase the efficiency in agricultural plant breeding, including clary sage – one of the main essential oil crops grown in Russia, it is necessary to use biotechnological methods. One of these techniques is based on the induction of somaclonal variability in the callus tissue culture. To develop it, it is necessary to optimize the conditions for obtaining plant-regenerants in vitro and their analysis. The aim of this work was to study the features of morphogenesis and regeneration of plants from callus cultures to develop cell technologies for creating an initial breeding material based on somaclonal variability in Salvia sclarea L. In the course of the research, we found that the optimal explants for obtaining morphogenic callus, from which shoots were regenerated, were segments of buds and stems with a node (isolated from seedlings in vitro). Cytological analysis of callus cultures revealed two types of morphogenesis – organogenesis (gemmogenesis) and somatic embryogenesis. The features of the morphogenic callus formation of six sage cultivars and samples during the long-term cultivation were studied. The maximum frequency of morphogenesis was noted in the 2nd passage (from 32.4 to 85.2 %, depending on the genotype). Then, to the 8–10th passage, this indicator decreased to 0.0–3.9 %.‘S-785’ and ‘Taigan’ cultivars showed the highest morphogenesis frequency (81.5–85.2 %) and duration of callus regeneration potential (up to the 10th passage). The analysis of callus cultures of six donor plants of ‘S-785’ cultivar helped us to reveal their heterogeneity in morphogenesis induction ability. The maximum frequency of morphogenic callus formation (76.3–91.5 % in the 2nd passage) and the duration of the morphogenic potential preservation (up to the 12th passage) were observed in plants No. 3 and 9, whereas in No. 2, regeneration with a frequency of 3.6–9.7 % was observed only during three passages. Analysis of plants obtained from calli showed their variability in morphology – up to 12.5 % of the samples had deviations compared to the initial cultivar ‘S-785’ in leaf shape, inflorescence structure, flower color, etc. Somaclonal changes in morphological and economically useful traits revealed in regenerants indicate that they are promising for use in sage breeding.

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