scholarly journals In Vitro Effects of Different Combinations of Phytohormones on Callus Induction from Different Explants of Cabbage Brassica Oleracea Var. Capitata L. Seedlings

2021 ◽  
pp. 3476-3486
Author(s):  
Alaa. M. Hasan ◽  
Ekhlas. A.J. ElKaaby ◽  
Rakad. M.Kh. AL-Jumaily
Keyword(s):  
Brassica Oleracea ◽  
Callus Induction ◽  
Basal Medium ◽  
Culture Conditions ◽  
Control Treatment ◽  
Fresh Weight ◽  
Superior Result ◽  
Significant Difference ◽  
In Vitro Effects

    The leading purpose of this work is the development of efficient culture conditions to induce calli from cabbage (Brassica oleracea var. capitata L.) under in vitro conditions. The mature seeds were surface sterilized with combinations of different concentrations of ethanol and NaOCl in different time durations and  were germinated on MS basal medium. The results revealed that the best sterilization method of cabbage seeds was by using 70% ethanol for one minute, followed by 15 min in 2% (NaOCl). Seedlings were used as donor sources for hypocotyls, cotyledon leaves, true leaves, and shoot tip explants. These explants were cultured on different combinations of cytokinins (TDZ, BAP, Ad) and auxins (IAA, NAA, 2, 4-D) then implanted in Murashige and Skoog (MS) media. 4 weeks after culturing, a significant difference was found among the explants in response to plant hormones. The maximum percentage of callus induction (100%) was using the combinations of 1 BAP + 1 2, 4-D, 1 BAP + 1 NAA, and 1 BAP + 2 2,4-D mg. l-1. In addition, explants responses varied and the hypocotyls showed a superior result (85.71 %) as compared to other explants. For callus fresh weight, the combination of 0.22 TDZ + 79.9 Ad mg. l-1    had a significant effect, causing the highest fresh weight (0.2745g), while control treatment gave the lowest mean of 0.0066 g. Data showed that cotyledon explants were significantly superior in giving highest callus fresh weight with the mean of 0.1723 g. On the other hand, hypocotyl explants gave the lowest mean, reaching 0.1542 g.

scholarly journals Effect of sucrose and mannitol on Ajmalicine production from leaves induced callus of Catharanthus roseus L. G. Don in vitro

2014 ◽  
Vol 8 (2) ◽  
pp. 27-34
Author(s):  
Emad H. Jassim ◽  
Sami K. M. Ameen
Keyword(s):  
Catharanthus Roseus ◽  
Callus Induction ◽  
Control Treatment ◽  
Ms Medium ◽  
Fresh Weight ◽  
And Control ◽  
Different Levels

An experiment on the effect of sucrose and mannitol on leave induced callus of Catharanthus roseus was conducted from February 2011 to May 2012. Callus induction was achieved by culturing leaves of the plant on MS medium supplemented with 0.5 mg /L 2,4-D and 1mg / L Kin, The best medium to maintain callus was on MS medium modified with 0.5 mg /L 2,4-D and 1.5 mg / L Kin. when different levels were added to MS medium for each Mannitol 0, 6000 ,8000 ,10000 mg /L and Sucrose 40, 60 ,80, 100 g /L in split experimental and control treatment was MS medium supplemented with 30 g/L sucrose. The results showed that medium supplemented with 100 gL of sucrose gave the highest quantity of Ajmalicine 32.27 µg/100 mg fresh weight of callus,as well as medium supplemented with 8000 mgL of Mannitol gave the highest value of Ajmalicine 120.19 µg/100 mg fresh weight of callus. The concentrations of Ajmalicine, derived from the leaves of the plants grown in pot, were lowest than the concentrations produced by the callus grown in vitro it was 0.047 µg/100 fresh weight of the leaves.

scholarly journals (282) In Vitro Indirect Organogenesis of Leucocoryne purpurea, an Ornamental Geophyte Species Endemic to Chile

HortScience ◽  
2005 ◽  
Vol 40 (4) ◽  
pp. 1051B-1051
Author(s):  
Luis Humberto Escobar Torres ◽  
Eduardo Alejandro Olate Muñoz ◽  
Miguel Jordan ◽  
Marlene Gebauer
Keyword(s):  
Callus Induction ◽  
Basal Medium ◽  
Average Frequency ◽  
Dichlorophenoxyacetic Acid ◽  
Fresh Weight ◽  
Root Tips ◽  
Indirect Organogenesis ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Different Sources

Callus induction (CI) and later shoot induction (SI) were studied in Leucocoryne purpurea, a native and endemic Chilean geophyte species. Basal leaf portions (BL), bulb basal plate (BP), and root tips (RT) from in vitro plants were used as explants. Treatments for CI included all three explants and media containing different sources and concentrations of auxins and cytokinins as plant growth regulators (PGRs). Plant material was initiated on MS basal medium (Murashige and Skoog, 1962), supplemented with vitamins, 30 g·L-1 sucrose, 6.0 g·L-1 agar and pH adjusted to 5.7 before autoclaving. The experiments were carried on a growth chamber at 24 ± 1.5 °C. CI cultures were maintained in darkness for 16 weeks, and SI for 12 weeks in a 16-hour photoperiod. BL and RT explants did not respond to any of the CI treatments. BP explants cultured on MS basal medium without PGRs also did not produce any callus. The average frequency of callus induction for BP was 78% and the average fresh weight of callus was 10.06 g/explant after 16 weeks of culture. Best treatment for CI was BP cultured on 4.52 μm 2,4-dichlorophenoxyacetic acid (2,4-D) in combination with 0.45 μm 6-benzyladenine (BA), when they were compared to 2,4-D alone or picloram as auxin source. After 16 weeks of culture, calli were transferred to SI medium, supplemented with three different concentrations of thidiazuron (TDZ), either intact or subdivided (150 mg/explant). SI treatments had a greater and significant response when the callus came from a CI medium containing auxin and cytokinin combined, in comparison to those coming from a CI medium containing auxins only.

scholarly journals In vitro growth and indoleacetic acid production by Mesorhizobium loti SEMIA806 and SEMIA816 under the influence of copper ions

2017 ◽  
Vol 8 (2) ◽  
Author(s):  
Jéssica Dutra Vieira ◽  
Paulo Roberto Diniz Da Silva ◽  
Valdir Marcos Stefenon
Keyword(s):  
Contaminated Soils ◽  
Acid Production ◽  
Indoleacetic Acid ◽  
Copper Ions ◽  
Control Culture ◽  
Control Treatment ◽  
Symbiotic Interaction ◽  
Mesorhizobium Loti ◽  
Significant Difference

The indoleacetic acid produced by symbiotic bacteria is an important phytohormone signaling microbe-plant interaction, being therefore essential for rhizoremediation. In this study, the effect of different concentrations of copper ions on the bacterial growth and indoleacetic acid production was investigated in two strains of Mesorhizobium loti in in vitro conditions, aiming to determine critical concentrations of this heavy metal for rhizoremediation of contaminated soils using this bacterium. The experiment consisted on a control culture without copper and three treatments supplemented with 10 mg.L-1, 20 mg.L-1 or 50 mg.L-1 of CuSO4. For both strains, the growth stopped after 48h and no significant difference was observed across treatments. The production of indoleacetic acid by the control treatment without copper was significantly higher in comparison to the copper- containing treatments. Mesorhizobium loti SEMIA806 and SEMIA816 are resistant to up to 50 mg.L-1 of CuSO4 in the culture medium, presenting effective growth. The synthesis of indoleacetic acid was strongly reduced but not excluded by ions copper in the medium. So, it is expected that environmental copper found in the soil up to the concentration of 50 mg.L-1 will not preclude the symbiotic interaction between M. loti and leguminous host plant in rhizoremediation enterprises.

scholarly journals In vitro assessment of wheat tolerance to drought

Genetika ◽  
2005 ◽  
Vol 37 (2) ◽  
pp. 165-171 ◽  
Author(s):  
Vladislava Galovic ◽  
Zorana Kotaranin ◽  
Srbislav Dencic
Keyword(s):  
Drought Stress ◽  
Nutrient Medium ◽  
Callus Tissue ◽  
Geographic Origin ◽  
Control Group ◽  
Zygotic Embryos ◽  
Fresh Weight ◽  
Effects Of Drought ◽  
In Vitro Effects

Analyzed in this paper were the in vitro effects of drought stress in 13 genotypes of winter wheat, one genotype of spring wheat, and three Triticale genotypes of different geographic origin. Callus tissue was induced from immature zygotic embryos (10-15 days after pollination) on a modified MS nutrient medium. After two weeks, callus tissue was transplanted onto the same medium enriched with 5% high-molecular polyethylene glycol (PEG 6000), which was used as the stress agent to produce the effect of drought chemically. A control group of calluses was grown on an identical medium but without PEG. After four weeks of growing calluses on these mediums, we assessed callus mass survival ability of the genotypes before the transplantation as well as percentage reduction of callus fresh weight after the transplantation onto the nutrient medium with 5% PEG. Statistically significant differences were found among the genotypes in their response to the induced stress. The best survival ability before the transplantation was found in the genotype Mexicol20 (83%), while the lowest was recorded in Slavija (11.3%). Culture growing under stress conditions significantly reduced callus fresh weight in all of the genotypes. The lowest decrease of the callus mass relative to control was recorded in Rozofskaja (14.4%) and the highest in Miranovska (58.4%), indicating the genotypes' tolerance levels towards drought stress.

326 EVALUATION OF CULTURE DURATION AND PARTIAL CUMULUS CELL REMOVAL DURING CANINE OOCYTE MATURATION

2006 ◽  
Vol 18 (2) ◽  
pp. 270
Author(s):  
C. Hanna ◽  
C. Long ◽  
M. Westhusin ◽  
D. Kraemer
Keyword(s):  
In Vitro Maturation ◽  
Calf Serum ◽  
Cumulus Cell ◽  
Germinal Vesicle ◽  
Cumulus Cells ◽  
Culture Conditions ◽  
Germinal Vesicle Breakdown ◽  
Significant Difference ◽  
Culture Duration

The objectives of this study were to determine whether the percentage of canine oocytes that resume meiosis during in vitro maturation could be increased by either increasing culture duration or by removing approximately one-half of the cumulus cells 24 h after oocytes were placed into culture. Canine female reproductive tracts were collected from a local clinic and ovaries were minced in warm TL-HEPES. Oocytes with a consistently dark ooplasm and at least two layers of cumulus cells were selected, cultured in a basic canine oocyte in vitro maturation medium consisting of TCM-199 with Earl's salts, 2.92 mM Ca-lactate, 20 mM pyruvic acid, 4.43 mM HEPES, 10% fetal calf serum, 1% Penicillin/Streptomycin (GibcoBRL, Grand Island, NY, USA), and 5 μg/mL porcine somatotropin, and incubated at 38.5°C in 5% CO2 in humidified air. Treatment groups were randomly assigned and oocytes were cultured for 60, 84, or 132 h (Basic). From each of these groups, one-half of the oocytes were pipetted through a fine bore pipette to partially remove the cumulus cells 24 h after the start of culture (Basic–1/2). At the end of culture, all oocytes were denuded and the nuclear status was observed with Hoechst 33342 under ultraviolet fluorescence. All data were analyzed by ANOVA with P < 0.05. Since the canine oocyte is ovulated at the germinal vesicle (GV) stage of meiosis and requires up to five days to mature in the oviduct, it was hypothesized that an increased culture time would allow for more oocytes to undergo nuclear maturation to metaphase II (MII). It was also hypothesized that partial removal of cumulus cells would decrease the cumulus cell component in the ooplasm that sustains meiotic arrest, allowing for more oocytes to resume meiosis (RM = germinal vesicle breakdown to MII). Results within each treatment group indicate that there is no significant difference between culture duration and the percent of oocytes that mature to MII. Additionally, there was no significance in the percent of oocytes that resumed meiosis after partial cumulus cell removal. Taken together, these data suggest that neither treatment is effective in canine in vitro maturation systems, given the current maturation culture conditions. Table 1. Nuclear status* of oocytes for three time periods with or without partial cumulus cell removal

Development of prepubertal goat oocytes after their in vitro maturation and chemical activation

Zygote ◽  
2020 ◽  
Vol 28 (6) ◽  
pp. 447-452
Author(s):  
Seungbum Hong ◽  
Binoy S. Vettical ◽  
Nisar Ahmad Wani
Keyword(s):  
Culture Media ◽  
Chemical Activation ◽  
Control Treatment ◽  
Developmental Potential ◽  
Embryo Culture Medium ◽  
Significant Difference ◽  
Maturation Rate ◽  
Further Development ◽  
Oviductal Fluid

SummaryExperiments were conducted to study in vitro maturation of prepubertal goat oocytes and their developmental potential after chemical activation. In Experiment 1, cumulus–oocytes complexes collected from the ovaries of prepubertal goats slaughtered at a local abattoir were matured in vitro in TCM-199-based medium supplemented with 10 µg/ml luteinizing hormone (LH) (treatment 1) or 10 µg/ml LH + 0.1 mM l-cysteine (treatment 2). In Experiment 2, mature oocytes were activated with either 5 µM ionomycin or 7% ethanol. After 18 h, some oocytes were randomly fixed and stained to evaluate their chromatin status, while others were cultured in embryo culture medium to study their further development. In Experiment 3, oocytes activated with 5 µM ionomycin were cultured for 7 days in one of the four different culture media [Charles Rosenkrans medium (CR-1), TCM-199, potassium simplex optimization medium (KSOM) and synthetic oviductal fluid (SOF)] to study their developmental potential. The maturation rate in control, treatment 1, and treatment 2 media did not differ from each other (P > 0.05). However, the lowest degeneration of oocytes was observed in treatment 3 (P < 0.05) when compared with the other two groups. The proportion of activated oocytes was higher, while non-activated oocytes were lower in ionomycin group when compared with the group activated with ethanol (P < 0.05). The proportions of oocytes cleaved were 65.7, 56.8, 61.0 and 54.4% in CR-1, TCM-199, KSOM and SOF medium, respectively, with no significant difference. However, further development of cleaved oocytes was better in KSOM followed by SOF.

IN VITRO EFFECTS OF LATS AND TSH ON PHOSPHODIESTERASE ACTIVITY IN HUMAN THYROID HOMOGENATES

1971 ◽  
Vol 67 (2) ◽  
pp. 209-215 ◽  
Author(s):  
K. Miyai ◽  
N. Amino ◽  
M. Azukizawa ◽  
Y. Kumahara
Keyword(s):  
Enzyme Activity ◽  
Adenosine Monophosphate ◽  
Human Thyroid ◽  
Phosphodiesterase Activity ◽  
Significant Difference ◽  
In Vitro Effects ◽  
Long Acting ◽  
Bovine Tsh

ABSTRACT The in vitro effects of long-acting thyroid stimulator (LATS) and thyrotrophin (TSH) on phosphodiesterase activity in human thyroid homogenates were studied. The enzyme activity was estimated by the destruction of 3H-cyclic 3′,5′-adenosine monophosphate. Bovine TSH had no effect on the enzyme activity. Five experiments were carried out using different batches of LATS-IgG and normal IgG which were preincubated with thyroid homogenates or not. There was no significant difference between the enzyme activity with LATS-IgG and that with normal IgG, while the activity was mostly inhibited by theophylline. These data are not consistent with the hypothesis that LATS acts as an antibody to phosphodiesterase.

scholarly journals Influence of pesticide-treated seeds on survival of Mesorhizobium sp. Cicer, symbiotic efficiency and yield in chickpea

2012 ◽  
Vol 48 (No. 1) ◽  
pp. 37-43 ◽  
Author(s):  
Kunal ◽  
P. Sharma
Keyword(s):  
Grain Yield ◽  
Seed Treatment ◽  
Fungal Pathogens ◽  
Winter Season ◽  
Control Treatment ◽  
Symbiotic Efficiency ◽  
Effective Measure ◽  
Significant Difference ◽  
Uninoculated Control

Chemical seed protectants are used to reduce the adverse effects of seedling fungal pathogens or insect attack on legume pastures and crops. Chickpea seeds are also frequently treated with Mesorhizobium sp. Cicer inoculant to promote effective symbiotic nitrogen fixation (SNF), which seems to be a cost effective measure. The population of viable Mesorhizobium sp. Cicer on seeds of chickpea declined with time of storage (4&deg;C) in pesticide treated and untreated chickpea seeds in vitro. A significant reduction in chickpea rhizobia was observed in seed treatment with Captan followed by Endosulfan and Chlorpyrifos. In a field experiment during the winter season 2006&ndash;2008, no difference in the emergence count of chickpea plants was observed. Treatments inoculated with Mesorhizobium sp. Cicer alone or along with Captan, Chlorpyrifos or Endosulfan showed improved plant growth and symbiotic parameters (plant height, nodulation, leghaemoglobin content, and nitrogen content) in comparison with the uninoculated control treatment. Significantly higher grain yield (9.6%) was observed in the treatment inoculated with Mesorhizobium sp. Cicer alone as compared to the uninoculated control. A non-significant difference in grain yield among treatments where Mesorhizobium sp. Cicer along with a mixture of fungicide and insecticides was applied was observed in contrast to the Mesorhizobium sp. Cicer treatment. In conclusion, the recommended rates of fungicide and insecticides as seed treatment were not detrimental to chickpea-Mesorhizobium sp. Cicer symbiosis, hence they can be safely used to obtain higher productivity. &nbsp;

scholarly journals PEMBENTUKAN KALUS DAN EMBRIO SOMATIK KAKAO MENGGUNAKAN THIDIAZURON MELALUI SATU TAHAP INDUKSI KALUS

2020 ◽  
Vol 20 (4) ◽  
pp. 179 ◽  
Author(s):  
NUR AJIJAH
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Growth ◽  
Callus Induction ◽  
Theobroma Cacao ◽  
Somatic Embryos ◽  
Agricultural Research ◽  
Basal Medium ◽  
Significant Difference ◽  
Four Levels ◽  
Theobroma Cacao L

<p>ABSTRAK</p><p><br />Embriogenesis somatik kakao (Theobroma cacao L.) telah banyak<br />dilaporkan  dengan  penggunaan  zat  pengatur  tumbuh  (ZPT)  yang<br />bervariasi. Penggunaan thidiazuron untuk menginduksi embriogenesis<br />somatik kakao telah dilaporkan melalui dua tahap induksi kalus. Penelitian<br />ini bertujuan untuk mengevaluasi efektivitas thidiazuron menginduksi<br />embriogenesis somatik kakao melalui satu tahap induksi kalus. Penelitian<br />dilaksanakan di Laboratorium Kultur Jaringan Unit Pengembangan Benih<br />Unggul, Badan Litbang Pertanian, Bogor. Empat taraf thidiazuron (0; 2,5;<br />5,0; dan 10 µg/l) dikombinasikan dengan 2,4-D 2 mg/l<br />digunakan untuk<br />menginduksi kalus dan embrio somatik 3 klon kakao (TSH858, Sca6, dan<br />ICS13) menggunakan eksplan mahkota bunga dan staminoid. Media dasar<br />DKW tanpa ZPT digunakan sebagai kontrol. Penelitian disusun dalam<br />rancangan lingkungan acak lengkap dengan lima ulangan. Setiap unit<br />percobaan terdiri dari sepuluh eksplan. Peubah yang diamati meliputi<br />persentase pembentukan kalus umur 2 dan 4 minggu, penampakan visual<br />kalus, persentase eksplan membentuk embrio somatik, dan jumlah embrio<br />somatik per eksplan umur 10 dan 14 minggu. Kalus terbentuk pada media<br />dengan penambahan hanya 2,4-D atau 2,4-D + thidiazuron, namun embrio<br />somatik hanya terbentuk pada media dengan penambahan 2,4-D +<br />thidiazuron. Pembentukan kalus dan embrio somatik sangat dipengaruhi<br />oleh tipe eksplan dan genotipe. Klon Sca6 lebih responsif dibandingkan<br />TSH858 dan ICS13 dan eksplan staminoid lebih responsif dibandingkan<br />mahkota bunga. Hasil studi ini menunjukkan terdapat pengaruh interaksi<br />yang kuat antara ZPT, genotipe, dan tipe eksplan terhadap pembentukan<br />kalus dan embrio somatik kakao serta tidak terdapat perbedaan hasil yang<br />nyata antara pembentukan embrio somatik melalui satu dan dua tahap<br />induksi kalus.<br />Kata kunci: Theobroma cacao L., genotipe, eksplan, zat pengatur tumbuh</p><p>ABSTRACT</p><p><br />Somatic embryogenesis of cacao (Theobroma cacao L.) has been<br />widely reported with varied of plant growth regulators (PGR) used. The<br />use of thidiazuron in inducing somatic embryogenesis of cacao has been<br />reported through a two-step callus induction. The study aimed to evaluate<br />the effectiveness of thidiazuron in inducing somatic embryogenesis of<br />cacao through a one-step of callus induction. The study was conducted at<br />the tissue culture laboratory of Agricultural Seed Development Unit,<br />Indonesian Agency for Agricultural Research and Development, Bogor.<br />Four levels of thidiazuron (0; 2.5; 5.0; and 10 µg/l) in combination with 2<br />mg/l  2,4-D  were  used  for  inducing  callogenesis  and  somatic<br />embryogenesis of three cacao clones (TSH858, Sca6, and ICS13) using<br />petals and staminoids explants. DKW basal medium without PGR was<br />used as a control. The result showed that callus were formed on medium<br />containing only 2,4-D or 2,4-D + thidiazuron, while embryos were only<br />formed on medium containing 2,4-D + thidiazuron. The formation of<br />callus and somatic embryos were highly affected by explant types and<br />genotypes. Sca6 clone was more responsive than TSH858 and ICS13 and<br />staminoids were more responsive than petals. The results of this study<br />revealed that there was a strong interaction between the PGRs, genotypes,<br />and explant types on the formation of cacao callus and somatic embryos.<br />Results of this study also showed no significant difference between the<br />formation of somatic embryos through one and two steps of callus<br />induction.<br />Keywords: Theobroma cacao L., genotypes, explants, plant growth<br />regulators</p>

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