scholarly journals In vitro mutagenesis in anthurium induced by colchicine

2022 ◽  
Vol 28 (1) ◽  
Author(s):  
María Isabel López-Martínez ◽  
◽  
Alejandrina Robledo-Paz ◽  
Luis Antonio Flores-Hernández ◽  
Tarsicio Corona-Torres ◽  
...  
Keyword(s):  
Morphological Changes ◽  
Lethal Dose ◽  
Adventitious Shoots ◽  
Root Apex ◽  
In Vitro Mutagenesis ◽  
Root Explants ◽  
Regenerated Plants ◽  
New Varieties ◽  
Induced Mutagenesis

Developing new varieties of anthurium by hybridization can take 8-10 years; therefore, induced mutagenesis can be an alternative strategy to hybridization. The objective of this work was to induce mutations in A. andreanum by exposing explants obtained from vitroplants to colchicine. Explants of leaves, nodes and roots obtained from vitroplants were exposed to 0.1 % colchicine for 0, 2, 3 and 4 h. The mean lethal dose (LD50), survival, number of explants that generated callus, number of explants that formed shoots and the number of shoots per explant were evaluated. The karyotype of the presumed mutated regenerated plants was determined by the root apex squash technique. The leaves showed the highest sensitivity to cochicine. The survival of the root explants treated with colchicine was 100 %; 4 % of roots exposed for 2 and 3 h formed adventitious shoots (120 shoots). For nodes, the LD50 was found at 3.98 h; 76 and 56 % of the nodes cultivated for 2 and 3 h with colchicine formed adventitious shoots (4.4 and 3.6 shoots). The plants regenerated from the explants exposed to colchicine showed morphological changes. The chromosomal number of the regenerated vitroplants from the explants exposed for 2 and 3 h to colchicine was 2n = 29, while that of those obtained from the explants that remained on the colchicine for 4 h was 2n = 31. The sensitivity to colchicine was a function of the type of explant and the dose used. Colchicine caused the loss (monosomy) or gain of chromosomes (trisomy).

scholarly journals In Vitro Mutagenesis of Etlingera elatior by Gamma Ray Intermittent Irradiation

2018 ◽  
Vol 5 (3) ◽  
pp. 111-118 ◽  
Author(s):  
Elmi Irmayanti Azzahra ◽  
Syarifah Iis Aisyah ◽  
Diny Dinarti ◽  
Krisantini Krisantini
Keyword(s):  
Gamma Ray ◽  
Morphological Changes ◽  
Leaf Shape ◽  
Morphological Diversity ◽  
Mutation Breeding ◽  
Ornamental Plant ◽  
In Vitro Mutagenesis ◽  
Split Dose ◽  
Etlingera Elatior

Etlingera elatior is tropical ornamental plant commonly called “torch ginger” from Zingiberaceae family. Conventional breeding of E. elatior is limited by cross incompatibility, poor fruit set and low seed production.  In this study irradiation of E. elatior with Gamma ray performed to induce mutation. This study was aimed to increase morphological diversity and to obtain unique morphological characters to increase the aesthetic value of E. elatior as ornamental plants and cut flower. Two genotypes of E. elatior, red and white flowers, were tested. The LD20, LD35, and LD50 were determined following intermittent (split dose) Gamma irradiation with a two-hour gap between each gamma ray shot. Red genotype E. elatior explants were irradiated with dose of 3 + 3 Gy (LD20); 4 + 4 Gy (LD35) and 5 + 5 Gy (LD50); white genotype were irradiated with a dose of 2 + 2 Gy (LD20); 2.8 + 2.8 Gy (LD35) and 3.7 + 3.7 Gy (LD50). Non-irradiated explants were set as control.  The results of this study indicated that the increase in dose of gamma ray irradiation changed shoot length and number of leaves in the red genotype per explant as compared to control. Morphological changes occurred in leaf shape at 5 + 5 Gy and 3.7 + 3.7 Gy and formation of variegated leaves at 2.8 + 2.8 Gy and 5 + 5 Gy. Keywords: mutation breeding, ornamental plant, split irradiation dose, torch ginger, Zingiberaceae

scholarly journals Antigiardial Effect of Kramecyne in Experimental Giardiasis

2017 ◽  
Vol 2017 ◽  
pp. 1-7
Author(s):  
Leticia Eligio-García ◽  
Elida Pontifez-Pablo ◽  
Salúd Pérez-Gutiérrez ◽  
Enedina Jiménez-Cardoso
Keyword(s):  
Side Effects ◽  
High Efficiency ◽  
Morphological Changes ◽  
Lethal Dose ◽  
Expression Of Genes ◽  
Antigiardial Activity ◽  
Resistant Strains ◽  
Polymerase Chain

A variety of drugs are used in giardiasis treatment with different levels of efficiency, presence of side effects, and even formation of resistant strains, so that it is important to search new only-one-dose treatments with high efficiency and less side effects. Kramecyne, an anti-inflammatory compound isolated from methanolic extract ofKrameria cytisoides, does not present toxicity, even at doses of 5,000 mg/kg. The objective was to determine the antigiardial effect of kramecyne overGiardia intestinalis in vitroandin vivoand analyze the expression of genes ERK1, ERK2, and AK on kramecyne treated trophozoites by Real Time Polymerase Chain Reaction (RTPCR). The median lethal dose (LD50) was 40 μg/mL and no morphological changes were observed by staining with blue trypan and light microscopy; experimental gerbil infection was eliminated with 320 μg/Kg of weight. After treatment there were no differences between intestines from treated and untreated gerbils. Kramecyne did not have significant effect over ERK1 and AK, but there are differences in ERK2 expression (p=0.04). Results show antigiardial activity of kramecyne; however the mode of action is still unclear and the evaluation of ultrastructural damage and expressed proteins is an alternative of study to understand the action mechanism.

scholarly journals Regenerating Adventitious Shoots from in Vitro Culture of Liatris spicata (L.) Willd. Cotyledons

HortScience ◽  
1996 ◽  
Vol 31 (1) ◽  
pp. 154-155
Author(s):  
Dennis P. Stimart ◽  
John C. Mather
Keyword(s):  
In Vitro Culture ◽  
Adventitious Shoots ◽  
Adventitious Shoot ◽  
Shoot Formation ◽  
Ms Medium ◽  
Adventitious Shoot Formation ◽  
Regenerated Plants ◽  
Murashige And Skoog Medium ◽  
Homogeneous Chemical

Cotyledons from developing 6- to 8-week-old embryos of Liatris spicata (L.) Willd. (blazing star) were cultured on Murashige and Skoog medium containing 0, 0.4, 4.4, or 44.4 μm BA or 0, 0.2, 2.2, or 22.2 μm TDZ to induce adventitious shoot formation. The highest percentage of cotyledons forming the most shoots was on medium containing 2.2 μm TDZ. Cotyledon-derived callus cultured on medium containing 4.4 μm BA formed ≈16 times more adventitious shoots than on 2.2 μm TDZ. Adventitious shoots derived from cotyledons or callus produced roots when placed on MS medium containing 5.0 μm IBA. Regenerated plants that flowered in the field appeared homogeneous. Chemical names used: N6-benzyladenine (BA), thidiazuron (TDZ), indole-3-butyric acid (IBA).

scholarly journals In vitro morphogenetic responses from obligatory apomictic Taraxacum belorussicum Val. N. Tikhom seedlings explants

2019 ◽  
Vol 139 (3) ◽  
pp. 505-522 ◽  
Author(s):  
Adrianna Gałuszka ◽  
Maciej Gustab ◽  
Monika Tuleja
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Material ◽  
Molecular Mechanisms ◽  
Adventitious Shoots ◽  
Indirect Organogenesis ◽  
Regenerated Plants ◽  
Blue Solution ◽  
Meristem Explants ◽  
Scanning Electron

Abstract Taraxacum belorussicum Val. N. Tikhom, a poorly known and obligatory apomictic species, is an attractive plant material for studying the embryological, genetic and molecular mechanisms of apomixis. This work aims to obtain an efficient protocol for Taraxacum belorussicum regeneration. Four types of explants (cotyledons, hypocotyls, meristems and roots) that were taken from 2-weeks-old seedlings were used for in vitro cultures, and a fast and efficient protocol of T. belorussicum regeneration was obtained. Various ½ MS-based media containing IAA (5.71 µM), TDZ (4.54 µM) and PSK (100 nM) were chosen to assess the morphogenetic abilities of selected T. belorussicum explants. Studies on the role of PSK were done in three independent experiments, where the most significant factors were always light and darkness. All explants produced callus by the third day of culture and adventitious shoots after 7 days, although in an asynchronous indirect manner, and with different intensities for all explant types. The most preferred medium culture for hypocotyl, cotyledon and meristem explants was ½ MS + TDZ, and ½ MS + IAA + TDZ + PSK for roots which were the only explant sensitive to PSK. A short darkness pretreatment (8 days) in PSK medium was found suitable to enhance organogenesis. Secondary organogenesis was observed for regenerated plants on meristem explants from the ½ MS + IAA + TDZ + PSK medium. A weak somatic embryogenesis was observed for hypocotyl and cotyledon explants from ½ MS + IAA + TDZ and ½ MS + IAA + TDZ + PSK media. Histological and scanning electron microscope images (SEM) of T. belorussicum confirmed indirect organogenesis and somatic embryogenesis. Plant material treated with aniline blue solution revealed the presence of callose in the cell walls of cotyledon and hypocotyl explants. The presence of extracellular matrix (ECM) and heterogenic structure of callus was also verified by scanning electron microscopy and light microscopy, confirming the high morphogenetic ability of T. belorussicum.

scholarly journals Nematicidal Activity of the Essential Oil of Three Varieties of Tagetes minuta from Argentina

2017 ◽  
Vol 12 (5) ◽  
pp. 1934578X1701200
Author(s):  
Yamile Massuh ◽  
Angel Cruz-Estrada ◽  
Azucena González-Coloma ◽  
Marta S. Ojeda ◽  
Julio A. Zygadlo ◽  
...  
Keyword(s):  
Lethal Dose ◽  
Nematicidal Activity ◽  
Root Knot Nematode ◽  
Egg Hatching ◽  
Tomato Seedlings ◽  
Tagetes Minuta ◽  
In Vivo Tests ◽  
New Varieties

Essential oils composition of three Tagetes minuta varieties and a wild population (WP) from Argentina and their in vitro and in vivo nematicidal activity against root-knot nematode, Meloidogyne javanica, are described. All T. minuta EOs tested were very active against nematode juveniles (J2), but the strongest nematicidal effects were exhibited by the TmV3 variety oil, characterized by a high content of ( E)-ocimenone. High nematode egg hatching suppression (> 90%) was induced by TmV3 EO after five days of incubation. In vivo tests on tomato seedlings showed a significant reduction of infection rate of M. javanica J2 treated with TmV3 and WP oils at sub lethal dose. Therefore, EOs from chemically stable new varieties of T. minuta could be environmentally friendly nematicidal agents.

scholarly journals Comparative Rooting of Shoot Tips of Four Japanese Persimmon Cultivars vs. Shoots Regenerated from Roots Cultured In Vitro

HortScience ◽  
2000 ◽  
Vol 35 (5) ◽  
pp. 940-944 ◽  
Author(s):  
Takuya Tetsumura ◽  
Hisajiro Yukinaga
Keyword(s):  
Histological Study ◽  
Adventitious Shoots ◽  
Shoot Proliferation ◽  
Shoot Tips ◽  
Diospyros Kaki ◽  
Japanese Persimmon ◽  
Rooting Ability ◽  
Regenerated Plants ◽  
In Vitro Roots

When cultured in vitro, roots of four Japanese persimmon (Diospyros kaki L.) cultivars formed adventitious shoots on MS medium with 10 μm zeatin and 0.01 μm indole-3-acetic acid, although their organogenetic capacities varied. Histological study revealed that the origin of the adventitious shoots was the pericycle. The regenerated shoots grew well on the shoot proliferation medium (MS with 5 μm zeatin). Final rooting percentages of shoots regenerated from roots of three of the four cultivars were greater than those of shoots that originated from shoot tips and that had been subcultured >50 times. Shoots regenerated from `Jiro' roots rooted 10 days earlier, had more roots than those from shoot tips, and maintained higher rooting ability over ten subcultures. Rooted `Hiratanenashi' shoots regenerated from roots survived better after acclimatization than those from shoot tips. No obvious variants were observed either in vitro or in the field. The trees regenerated from roots flowered within 4 years. These findings suggest that partial rather than true rejuvenation was responsible for both the early flowering and the juvenile characteristics, i.e., the enhanced rooting ability, observed in the regenerated plants. Chemical name used: 6-(4-hydroxy-3-methylbut-2-enylamino) purine (zeatin).

scholarly journals Induction in vitro of adventitious morphogenesis in isolated tissues of the genus Rubus plants using cytokinins from adenine and diphenylurea groups

2020 ◽  
pp. 21-27
Author(s):  
N. V. Solovykh ◽  
M. B. Yankovskaya
Keyword(s):  
Adventitious Shoots ◽  
Leaf Explants ◽  
Red Raspberry ◽  
Regenerated Plants ◽  
Ability To Regenerate ◽  
Organogenesis In Vitro ◽  
Heavy Metal Salts ◽  
Cultivation In Vitro ◽  
Isolated Tissues

The eff ect of cytokinins from the adenine and diphenylurea groups on adventitious organogenesis in vitro in isolated tissues of the genus Rubus plants was studied. Leaf explants and callus of red raspberry of the Volnitsa variety, blackberry of the Chester Thornless variety and the Rubus odoratus species were cultivated in the dark at a temperature of +25 ±2 °C on Murashige and Skoog medium. The medium contained 0.5 mg/l of gibberellic acid (HA), 0.5 mg/l of indolylacetic acid (IAA), and 6-benzylaminopurine (6-BAP) at concentrations of 0, 1, 2 and 4 mg/l or thidiazuron (TDZ) at concentrations of 0.1, 0.2 and 0.4 mg/l. The number of explants that formed adventitious shoots and the number of shoots per explant were taken into account. It has been established that for the induction of adventitious morphogenesis from leaf explants and calluses of red raspberry and blackberry the use of 6-BAP is more eff ective, thydiazuron is more eff ective for Rubus odoratus. The optimal concentration for 6-BAP is 2 mg/l and 0.2 mg/l for TDZ. Exceeding these concentrations of cytokinins can cause shoot vitrifi cation. For blackberry, it is possible to increase the content of 6-BAP in the medium for the induction of morphogenesis to 4 mg/l. The unequal ability to regenerate adventitious shoots in diff erent genotypes was revealed. In optimal variants of the experiment, the maximum frequency of their formation ranged from 13.3 % in Rubus odoratus to 40.0 % in blackberry. The use of the established optimal concentrations of growth regulators made it possible to obtain regenerated plants from callus that underwent long-term cultivation (for 10 months) on artifi cial nutrient media during tissue selection for tolerance to heavy metal salts and pesticides. Despite the reduced morphogenetic potential of tissues undergoing prolonged cultivation in vitro, 3 red raspberry regenerant plants and 1 Rubus odoratus plant were obtained from callus selected for tolerance to cobalt chloride. 9 blackberry plants and 7 red raspberry plants were regenerated from the tissues selected for pesticide tolerance.

scholarly journals Mutation Induction of Dendrobium ‘Earsakul’ Using Sodium Azide

HortScience ◽  
2016 ◽  
Vol 51 (11) ◽  
pp. 1363-1370 ◽  
Author(s):  
Anakkaorn Wannajindaporn ◽  
Chitpan Kativat ◽  
Piyada Alisha Tantasawat
Keyword(s):  
Reverse Osmosis ◽  
Sodium Azide ◽  
Chromosome Numbers ◽  
Lethal Dose ◽  
Morphological Differentiation ◽  
Morphological Characters ◽  
Mutation Induction ◽  
In Vitro Mutagenesis ◽  
Reduced Height

Dendrobium ‘Earsakul’ is an important commercial orchid in Thailand. Breeding new Dendrobium varieties for improved quality and yield is crucial. The objectives of this research were to perform in vitro mutagenesis of Dendrobium ‘Earsakul’ protocorm-like bodies (PLBs) using sodium azide (NaN3) and to select and evaluate the putative mutants using morphological characters, molecular markers, and the cytological method. The percentages of mortality of PLBs increased as concentrations of NaN3 increased. At 2 weeks, the lethal dose 30 (LD30) and LD50 were obtained with 0.1 and 0.5 mm NaN3, respectively. These two NaN3 concentrations were used for in vitro mutagenesis with reverse osmosis water (ROW; control 1) and 0 mm NaN3 (control 2) as controls. After the plants were cultured for 6 months, morphological differentiation was observed in some putative mutants: reduced height, higher numbers of nodes, reduced node length, shorter and thicker leaves, and shorter and fewer roots, compared with controls. When genetic profiles of 24 putative mutants were compared with controls, altered DNA profiles were found in 20 of 24 putative mutants (83.33%). Sixty-three polymorphic bands were produced from a total of 181 bands (34.81%) amplified by 10 polymorphic intersimple sequence repeat (ISSR) primers. When genetic diversity and relatedness, which were evaluated by ISSR analysis, and morphological characters were compared, the two markers were found to be uncorrelated. ISSR had a higher mutant differentiation capability than the morphological characters, indicating its higher efficiency. The chromosome numbers were similar in putative mutants and controls (2n = 2x = 24), suggesting that neither of the concentrations of NaN3 had any effect on the chromosome numbers in this experiment. These results indicate that NaN3 can be used effectively to mutagenize Dendrobium ‘Earsakul’ PLBs, and ISSR is a powerful tool for the identification of mutants. Chemical name: sodium azide (NaN3); reverse osmosis water (ROW).

scholarly journals The Effect of BAP and IBA on In Vitro Root Cultures of Acehnese Pomelo (Citrus maxima (Burm.) Merr.)

2020 ◽  
Vol 3 (1) ◽  
pp. 38
Author(s):  
Ira Handayani ◽  
Laila Nazirah ◽  
Rd. Selvy Handayani
Keyword(s):  
Tissue Culture ◽  
Adventitious Shoots ◽  
Shoot Formation ◽  
Root Explants ◽  
Nutritional Values ◽  
Organ Formation ◽  
Plant Organ ◽  
Randomized Design ◽  
Citrus Maxima

Acehnese pomelo is considered as a potential fruit to be developed for its economic and nutritional values. However, there is a main problem in developing this fruit. Pomelo sometimes is grown from seeds, but nowadays the fruits are often found to be having lesser seeds or even seedless. Besides, it is quite challenging to grow this local pomelo not only due to the scarce of the seeds, but it also cannot be conventionally germinated. Therefore, we need to develop this fruit through tissue culture. The purpose of this study was to investigate the effect of growth regulators on germination of pomelo seeds in vitro and its organ formation from in vitro root explants seed-originated of pomelo. The study was conducted at Plant Tissue Culture Laboratory, Faculty of Agriculture, Malikussaleh University. In October 2018 to April 2019. The research was divided into two separate experiments: 1) in vitro germination of pomelo seeds and 2) in vitro organ formation from root explants from of pomelo seeds. The first experiment using Complete Randomized Design (CRD) with 10 replications. The only factor observed was the use of BAP (0 and 2 mg/l). The second experiment was the examination of plant organ formation from root explants of pomelo seeds. This experiment had 2 factors observed. The first factor was different concentrations of BAP (0, 1, and 2 mg/l). The second factor was the different concentration of IBA (0, 1.25, and 2.5 mg/l). The results indicated that the application of BAP influenced the germination of pomelo seeds. The application of 2 mg/l BAP attributed to slower growth of shoots and roots, but it demonstrated better number of shoots, roots and leaves compared to application of 0 mg/l BAP. Pamelo root explants can form adventitious shoots on media eventhough without growth regulator (BAP 0 mg/l + IBA 0 mg/l), but optimization still needs to be done in order to obtain the best concentration to initiate shoot formation.

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