scholarly journals Effect of Different Cellulase and Pectinase Enzyme Treatments on Protoplast Isolation and Viability in Lilium ledebeourii Bioss.

2012 ◽  
Vol 40 (2) ◽  
pp. 123 ◽  
Author(s):  
Esmaeil CHAMANI ◽  
Seyyed Karim TAHAMI ◽  
Nasser ZARE ◽  
Rasool Asghari-ZAKARIA ◽  
Mehdi MOHEBODINI ◽  
...  
Keyword(s):  
Analysis Of Variance ◽  
Treatment Time ◽  
Leaf Explants ◽  
Protoplast Isolation ◽  
Combination Method ◽  
Interspecific Incompatibility ◽  
Protoplast Yield ◽  
Protoplast Preparation ◽  
Protoplast Production ◽  
Protoplast Viability

For overcoming interspecific incompatibility, protoplast combination method is a proper procedure for making a new plant withdesired traits. For this purpose, protoplast preparation is a first and important step. Hence, experiments were conducted to evaluatevarious combinations of cellulose, pectinase and their treatment times on protoplast production and protoplast viability in Liliumledebeourii Bioss. The results of experiment revealed that the protoplast yield was significantly affected by different treatment levels.Cellulase at 4% gave the highest numbers of protoplasts at 3.71×105 protoplast/g FW. Pectinase at 1% gave the highest numbers ofprotoplast. For treatment times, the highest yield of protoplast was with leaf explants treated for 24 h. Analysis of variance indicated thatconcentration, time and three-way interaction of cellulase, pectinase and time were significant at p<0.01. Cellulase at 4% and pectinase at0.2% for 24 h gave the highest viability. Interactions of cellulase × pectinase, cellulase × time, pectinase × time and cellulase × pectinase× treatment time were significant at P≤0.05 for protoplast number. The highest and lowest protoplast numbers were produced in mediacontaining 4% cellulase and 1% pectinase for 24 h (6.65×105 protoplast/g FW) and 1% cellulase and 0.2% pectinase for 12 h, respectively.It’s concluded that, the best treatment for isolation of Lilium protoplast was 4% cellulase and 1% pectinase for 24 h.

Effect of biotic factors on the isolation of Lupinus albus protoplasts

2003 ◽  
Vol 51 (1) ◽  
pp. 103 ◽  
Author(s):  
Anupam Sinha ◽  
Andrew C. Wetten ◽  
P. D. S. Caligari
Keyword(s):  
Lupinus Albus ◽  
Protoplast Isolation ◽  
Biotic Factors ◽  
Average Yield ◽  
Fresh Tissue ◽  
Progressive Decline ◽  
Protoplast Yield ◽  
Protoplast Production ◽  
Protoplast Viability

Several tissue types of Lupinus albus L. were investigated as sources for the isolation of protoplasts. Cotyledons from in vitro seedlings were found to yield the highest number of protoplasts compared with leaves, hypocotyls and roots. A combination of the protoplast isolation enzymes, cellulase and Pectolyase Y23, was capable of releasing the highest number of protoplasts compared with a combination of cellulase and Macerase. Protoplast yield increased with increasing cotyledon age but was accompanied by a progressive decline in protoplast viability. The optimal combination of protoplast yield and viability occurred when the protoplasts were isolated from 14- to 18-day-old cotyledons. The ratio between the volume of enzyme solution and the tissue biomass did not affect the protoplast production significantly. This is the first report of the isolation of protoplasts from a lupin cotyledon and, following the procedure described in this paper, an average yield of 1.2 × 106 protoplasts per gram of fresh tissue was obtainable.

Some features of protoplast isolation from leaf explants of Ficus carica plantlets cultured in vitro

2019 ◽  
Vol 305 ◽  
pp. S52-S53
Author(s):  
I. Mitrofanova ◽  
N. Lesnikova-Sedoshenko ◽  
O. Krivenko
Keyword(s):  
Leaf Explants ◽  
Protoplast Isolation ◽  
Ficus Carica

scholarly journals Protoplast production from napier grass and pearl millet triploid hybrids

2010 ◽  
Vol 34 (5) ◽  
pp. 1219-1223
Author(s):  
Ana Luiza de Oliveira Timbó ◽  
Lisete Chamma Davide ◽  
José Eduardo Brasil Pereira Pinto ◽  
Antônio Vander Pereira
Keyword(s):  
Pearl Millet ◽  
Treatment Time ◽  
Enzymatic Digestion ◽  
Napier Grass ◽  
Future Studies ◽  
Protoplast Production ◽  
Chromosomal Duplication

The objective of this work was to obtain protoplasts from napier grass and pearl millet triploid hybrids as a basis for future studies on chromosomal duplication. Explants were taken from mesophyll of in vitro- and in vivo-cultured plants or from calli of two triploid hybrids (H1 and H2), which were treated with enzymatic solutions containing different concentrations of cellulase R-10 (0.5, 1.0, 1.5 and 2.0%) with an additional 0.2% macerozyme and 0.1% driselase or 1.0% pectolyase Y-23 and 0.5% hemicellulase. Enzymatic digestion was monitored once every hour for five hours. Protoplasts were obtained from in vitro and in vivo leaflets of both triploid hybrids, and in vitro leaflets were the best explant sources. The quantity of produced protoplasts varied according to the hybrid, the enzymatic solution and the treatment time.

scholarly journals Effects of environmental preconditioning, donor tissue and isolation conditions on tomato (Lycopersicon esculentum Mill.) protoplast yield

2013 ◽  
Vol 48 (2) ◽  
pp. 113-120
Author(s):  
Elżbieta Kuźniak ◽  
Marzena Wielanek ◽  
Urszula Małolepsza ◽  
Henryk Urbaniak
Keyword(s):  
Bovine Serum Albumin ◽  
Protoplast Isolation ◽  
Isolation Procedure ◽  
Low Light ◽  
Cold Pretreatment ◽  
Isolation Medium ◽  
Donor Tissue ◽  
Protoplast Yield ◽  
Pretreatment Conditions

The effects of soil or <em>in vitro</em> grown plants, pretreatment conditions, donor tissue and isolation procedure on protoplast yield from cotyledons and leaves of tomato cv. 'Perkoz' and 'Zorza' were studied. The highest protoplast yield of 1.5 x 10<sup>7</sup>/g FW was obtained from leaves of <em>in vitro</em> grown plants. Low light intensity during donor plants <em>in vitro</em> culture and dark pretreatment were essential for successful protoplast isolation while cold pretreatment was not. Tissue preplasmolysis prior to transfer to enzyme mixture increased 4-fold the number of isolated protoplasts. Glycine and bovine serum albumin in the isolation medium did not significantly influence the protoplast yield.

scholarly journals EXPERIENCE IN THE TREATMENT OF CHILDREN WITH VASCULAR TUMORS OF THE MAXILLOFACIAL AREA

2018 ◽  
Vol 14 (4) ◽  
pp. 82-86 ◽  
Author(s):  
Ирина Замятина ◽  
Irina Zamyatina ◽  
Евгения Бимбас ◽  
Evgeniya Bimbas ◽  
Валентина Вольхина ◽  
...  
Keyword(s):  
Blood Flow ◽  
Young Children ◽  
High Efficiency ◽  
Treatment Time ◽  
Combined Treatment ◽  
Treatment Method ◽  
Linear Velocity ◽  
Combination Method ◽  
Vascular Tumors ◽  
Comparison Groups

Subject. The article shows the effectiveness of the author's method of treatment of young children with vascular tumors of the maxillofacial area. The variety of the clinical course of vascular tumors, their localization in the maxillofacial area create difficulties and limit the choice of treatment method. The author's method combines high efficiency, minimally invasiveness, reduction of rehabilitation time and minimal adverse reactions. Objective ― evaluation of the experience of treating vascular tumors of the maxillofacial region in children using a combination method - taking propranolol and hardening. Methodology. The results of the treatment of 120 children with vascular tumors of the maxillofacial area aged 2 months to 2 years have been evaluated. Compared the results in 3 groups of children who were treated by various methods: combined - taking propranolol and hardening (FG); propranolol (HS-1); sclerotherapy method (HS-2). The efficacy of treatment was assessed by changing the volume of formation, the diameter of the supply vessels and the values of the linear velocity of blood flow. Results. According to the results of our clinical study, a group of young children with vascular tumors of the maxillofacial area in cosmetically significant areas that received treatment by the author's method, has demonstrated its effectiveness. A significant amount of regression was achieved (2.5 times) compared with the comparison groups. According to the ultrasound, the reduction in the diameter of the supply vessels and the linear velocity of blood flow reached zero after 6 months after the start of treatment, faster than in the comparison groups Conclusions. The use of the combined treatment - taking propranolol and hardening - is effective in treating children with vascular tumors of the maxillofacial area, reduces the treatment time and reduces the risk and degree of complications.

scholarly journals A study of the treatment of high-salt chromium-containing wastewater by the photocatalysis-constructed wetland combination method

2019 ◽  
Vol 80 (10) ◽  
pp. 1956-1966 ◽  
Author(s):  
Dandan Li ◽  
Fengzhen Chen ◽  
Jianqiu Han
Keyword(s):  
Constructed Wetland ◽  
Treatment Time ◽  
Reduction Rate ◽  
Oxygen Demand ◽  
High Salt ◽  
Combination Method ◽  
Treated Wastewater ◽  
System A ◽  
Combination System

Abstract In this study, iron ore slag as the photocatalyst was introduced into a constructed wetland simulation system. A comparative experiment of the constructed wetland method and photocatalysis-constructed wetland combination method that treats the high-salt chromium-containing wastewater was carried out. The best hydraulic retention time (HRT) of the photocatalysis-constructed wetland combination system was studied. The effects of these two methods on biochemical oxygen demand (BOD5), chemical oxygen demand (COD) removal and Cr(VI) reduction rate of the high-salt chromium-containing wastewater were analysed after 14 periods. The results showed that under the optimal HRT of 4 hours, the COD and BOD5 of the wastewater reduced by 47% and 31%, and the reduction rate of Cr(VI) was 83% separately in the constructed wetland system. The COD and BOD5 of the wastewater reduced by 83% and 42%, and the reduction rate of Cr(VI) was 96% separately in the photocatalysis-constructed wetland combination method system. At the same time, the changes in plant parameters under these two systems were studied, and the results showed that the addition of photocatalyst and hydrogen peroxide to constructed wetlands did not affect the normal indicators of plant growth. The results showed that the photocatalysis-constructed wetland combination method not only reduced the treatment time greatly, but also improved the quality of the treated wastewater significantly.

scholarly journals CMV-RESISTANT CULTIVAR SELECTION BY ENZYME-LIKED IMMUNOSORBENT ASSAY AND PROTOPLAST ISOLATION METHOD IN CUCUMBER.

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 672a-672
Author(s):  
Kuen-Woo Park ◽  
Min-Jea Kim
Keyword(s):  
Cell Wall ◽  
Immunosorbent Assay ◽  
Wall Thickness ◽  
Protoplast Isolation ◽  
Elisa Test ◽  
Resistant Cultivar ◽  
Isolation Method ◽  
Resistant Cultivars ◽  
Protoplast Yield ◽  
Cultivar Selection

This experiment was carried out to select resistant cultivar to CMV in cucumber using Elisa-test and protoplast isolation method. Twenty domestic cultivars or lines and 8 European cultivars were ested for resistance by Elisa test. Among the domestic cultivars, DADAKI group was found to be susceptible and CHEONGJANG group resistant. Among all the cultivars and lines tested, a European cultivar, DALIBOR and Janghyungheukjinju Korean line were found to be highly resistant. When comparing for the protoplast yield depending upon the positions of seedlings (cotyledon, young leaf and hypocotyl), the highest protoplast yield could be obtained from cotyledons in macerozyme 0.5% + cellulose 2.0%. Protoplast yields in susceptible cultivars were higher than those from resistant cultivars. Differences in cell wall thickness between susceptible and resistant cultivars were observed.

scholarly journals PROTOPLAST ISOLATION AND CULTURE OF KENAF (HIBISCUS CANNABINUS L.)

HortScience ◽  
1994 ◽  
Vol 29 (7) ◽  
pp. 729e-729
Author(s):  
Donglong Liu ◽  
Nancy A. Reichert
Keyword(s):  
Cell Division ◽  
Liquid Medium ◽  
Hydrolytic Enzymes ◽  
Leaf Tissue ◽  
Protoplast Isolation ◽  
Hibiscus Cannabinus ◽  
Cultivar Differences ◽  
Fusion Frequency ◽  
Protoplast Yield ◽  
Acceptable Range

Protoplast isolation and culture protocols were developed for leaf tissue from 6 kenaf cultivars [Everglades 41 (E41), E71, Guatemala 4 (G4), G45, G51, and Tainung 1]. For protoplast isolation, the best combination of hydrolytic enzymes was cellulysin (1% w/v; Calbiochem) plus macerase (0.5% w/v; Calbiochem), with a 24 hour digestion at 30°C in the dark. Yields reached 7.2 (10)6 protoplasts/g leaf tissue. Protoplast viabilities ranged from 65% to 96%. Minor cultivar differences were observed related to protoplast yield, but all viability estimates were in an acceptable range. Greatest cell division frequencies and plating efficiencies were obtained when protoplasts were initially cultured in liquid medium at a density of 1.0 (10)5 protoplasts/ml. Electrofusion protocols were developed for kenaf protoplasts testing the range from 1200 to 3000 V/cm. A fusion voltage of 2000 V/cm yielded the highest fusion frequency and retained viability above 80%.

scholarly journals Isolation of Protoplasts from Nepenthes - A Plant Carnivore

2014 ◽  
Vol 24 (1) ◽  
pp. 93-100 ◽  
Author(s):  
Jaylen Sweat ◽  
Michael S. Bodri
Keyword(s):  
Cell Wall ◽  
Cell Division ◽  
Effective Treatment ◽  
Wild Species ◽  
Plant Tissue ◽  
Protoplast Isolation ◽  
Protoplast Yield ◽  
Viable Culture

Protoplasts were isolated from the lamina of greenhouse grown Nepenthes ampullaria and the hybrid N. ‘Rokko’ Exotica in order to develop a protocol for protoplast isolation suitable for wild species of Nepenthes. Various molarities utilizing mannitol or sorbitol and different enzyme mixtures and concentrations as well as incubation times were evaluated to maximize protoplast yield and viability.  The most effective treatment, a 4 hrs incubation at 40 rpm and 25°C in a solution consisting of 0.5 M sorbitol, 5% cellulase ‘Onozuka’ R-10, 0.5% macerozyme R-10, and 0.3% pectolyase Y-23, generated 4.35 × 106 protoplasts/ gfw of which 62.1% were viable.  Culture was attempted in respect of  regeneration of the cell wall, however, no cell division was observed. Plant Tissue Cult. & Biotech. 24(1): 93-100, 2014 (June) D. O. I. http://dx.doi.org/10.3329/ptcb.v24i1.19217

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