scholarly journals In vivo polyploidy induction of Phalaenopsis amabilis in a bubble bioreactor system using colchicine

2021 ◽  
Vol 27 (2) ◽  
pp. 204-212
Author(s):  
Mohsen Mohammadi ◽  
Behzad Kaviani ◽  
Shahram Sedaghathoor
Keyword(s):  
Genetic Improvement ◽  
Ornamental Plants ◽  
Guard Cells ◽  
Antimitotic Agent ◽  
Hoagland Solution ◽  
Stomata Size ◽  
Half Strength ◽  
Bioreactor System

Abstract Phalaenopsis amabilis Blume var. grandiflora Bateman is economically important as cut and pot flower. Polyploidy is considered as a valuable tool in improvement and evolution of ornamental plants. Protocorm-like bodies (PLBs) of P. amabilis were cultured on Murashige and Skoog medium containing 0.20 mg L-1 IBA together with 2.00 mg L-1 KIN and 1.00 g L-1 activated charcoal and grown for a period of five months. Fully-developed plantlets from in vitro grown PLBs were immersed in a bubble reactor filled with half-strength Hoagland solution containing the antimitotic agent colchicine (0.05%, 0.10% and 0.15%, w/v) for 72 h with a few drops (1 mL of 0.1%) of octylphenoxypolyethoxyethanol or Nonidet (P-40) as a surfactant. Plantlets were aerated to prevent hypoxia. Colchicine-treated and untreated plantlets were transferred to pots for a period of 60 days. Tetraploidy was successfully induced by 0.15% colchicine. Polyploidy levels were firstly detected using flow cytometry and then confirmed by cytological and morphological observations. The chromosome number was 2n = 2x = 38 in diploids and 2n = 4x = 76 in tetraploid. Incubation of plantlets in liquid medium containing 0.15% colchicine induced the maximum recovered tetraploids with minimum frequency of survival (50%). The tetraploid plants were more compact and exhibited round and thick leaves with darker green color than diploids. Stomata size in tetraploids were larger with less density than diploids. Chloroplast number in guard cells of tetraploids was about two times more than that of control. These results indicate that induction of tetraploids are a reliable and powerful tool for generation of novel phenotypes with ornamental and horticultural value for genetic improvement and breeding. Produced tetraploids in current study have potential in the ornamental/floriculture trade.

scholarly journals Sand Particle Size and Phosphorus Amount Affect Rhizophagus irregularis Spore Production Using In Vitro Propagated Spore as a Starter Inoculum in Rhizosphere of Maize (Zea mays) Plantlets

2021 ◽  
Vol 7 (10) ◽  
pp. 846
Author(s):  
Pulak Maitra ◽  
Jubair Al-Rashid ◽  
Nirmal Chandra Barman ◽  
Md. Niuz Morshed Khan ◽  
Dipa Mandal ◽  
...  
Keyword(s):  
Particle Size ◽  
Zea Mays ◽  
Am Fungi ◽  
Spore Production ◽  
Sand Particle ◽  
Monoxenic Culture ◽  
Hoagland Solution ◽  
Half Strength

Microbial inoculants, particularly arbuscular mycorrhizal (AM) fungi, have great potential for sustainable crop management. In this study, monoxenic culture of indigenous R. irregularis was developed and used as a tool to determine the minimum phosphorus (P) level for maximum spore production under the in vitro conditions. This type of starter AM fungal inoculum was then applied to an in vivo substrate-based mass-cultivation system. Spore production, colonization rate, and plant growth were examined in maize (Zea mays L.) plant inoculated with the monoxenic culture of R. irregularis in sand graded by particle size with varying P levels in nutrient treatments. In the in vitro culture, the growth medium supplemented with 20 µM P generated the maximum number of spores (400 spores/mL media) of R. irregularis. In the in vivo system, the highest sporulation (≈500 spores g−1 sand) occurred when we added a half-strength Hoagland solution (20 µM P) in the sand with particle size between 500 µm and 710 µm and omitted P after seven weeks. However, the highest colonization occurred when we added a half-strength Hoagland solution in the sand with particle sizes between 710 µm and 1000 µm and omitted P after seven weeks. This study suggests that substrate particle size and P reduction and regulation might have a strong influence on the maximization of sporulation and colonization of R. irregularis in sand substrate-based culture.

scholarly journals In vitro multiplication, micromorphological studies and ex vitro rooting of Hybanthus enneaspermus (L.) F. Muell. – a rare medicinal plant

2018 ◽  
Vol 77 (1) ◽  
pp. 80-87 ◽  
Author(s):  
Mahipal S. Shekhawat ◽  
M. Manokari
Keyword(s):  
Medicinal Plant ◽  
Large Scale ◽  
Nodal Segments ◽  
Ms Medium ◽  
Ex Vitro ◽  
Culture Bottle ◽  
Hybanthus Enneaspermus ◽  
Half Strength

AbstractHybanthus enneaspermusis a rare medicinal plant. We defined a protocol for micropropagation,ex vitrorooting of cloned shoots and their acclimatization. Surface-sterilized nodal segments were cultured on Murashige and Skoog (MS) medium with different concentrations of 6-benzylaminopurine (BAP) and kinetin (Kin). Medium supplemented with 1.5 mg L−1BAP was found optimum for shoot induction from the explants and 6.4±0.69 shoots were regenerated from each node with 97% response. Shoots were further proliferated maximally (228±10.3 shoots per culture bottle with 7.5±0.43 cm length) on MS medium augmented with 1.0 mg L−1each of BAP and Kin within 4–5 weeks. The shoots were rootedin vitroon half strength MS medium containing 2.0 mg L−1indole-3 butyric acid (IBA). The cloned shoots were pulse-treated with 300 mg L–1 of IBA and cultured on soilrite® in a greenhouse. About 96% of the IBA-pulsed shoots rootedex vitroin soilrite®, each shoot producing 12.5±0.54 roots with 5.1±0.62 cm length. Theex vitrorooted plantlets showed a better rate of survival (92%) in a field study thanin vitrorooted plantlets (86%). A comparative foliar micromorphological study ofH. enneaspermuswas conducted to understand the micromorphological changes during plant developmental processes fromin vitrotoin vivoconditions in terms of variations in stomata, vein structures and spacing, and trichomes. This is the first report onex vitrorooting inH. enneaspermusand the protocol can be exploited for conservation and large-scale propagation of this rare and medicinally important plant.

scholarly journals Adjustment of Mineral Elements in the Culture Medium for the Micropropagation of Three Vriesea Bromeliads from the Brazilian Atlantic Forest: The Importance of Calcium

HortScience ◽  
2009 ◽  
Vol 44 (1) ◽  
pp. 106-112 ◽  
Author(s):  
Alice Noemí Aranda-Peres ◽  
Lázaro Eustáquio Pereira Peres ◽  
Edson Namita Higashi ◽  
Adriana Pinheiro Martinelli
Keyword(s):  
New Media ◽  
Mineral Composition ◽  
Culture Media ◽  
Dry Weight ◽  
Defined Medium ◽  
Ms Medium ◽  
Media Formulation ◽  
Half Strength

Many different species of Bromeliaceae are endangered and their conservation requires specific knowledge of their growth habits and propagation. In vitro culture of bromeliads is an important method for efficient clonal propagation and in vitro seed germination can be used to maintain genetic variability. The present work aims to evaluate the in vitro growth and nutrient concentration in leaves of the epiphyte bromeliads Vriesea friburguensis Mez, Vriesea hieroglyphica (Carrière) E. Morren, and Vriesea unilateralis Mez, which exhibit slow rates of growth in vivo and in vitro. Initially, we compared the endogenous mineral composition of bromeliad plantlets grown in half-strength Murashige and Skoog (MS) medium and the mineral composition considered adequate in the literature. This approach suggested that calcium (Ca) is a critical nutrient and this was considered for new media formulation. Three new culture media were defined in which the main changes to half-strength MS medium were an increase in Ca, magnesium, sulfur, copper, and chloride and a decrease in iron, maintaining the nitrate:ammonium rate at ≈2:1. The main difference among the three new media formulated was Ca concentration, which varied from 1.5 mm in half-strength MS to 3.0, 6.0, and 12 mm in M2, M3, and M4 media, respectively. Consistently, all three species exhibited significantly higher fresh and dry weight on M4, the newly defined medium with the highest level of Ca (12 mm). Leaf nitrogen, potassium, zinc, magnesium, and boron concentrations increased as Ca concentration in the medium increased from 1.5 to 12 mm.

scholarly journals Phenotypic aberrations during micropropagation of Soymida febrifuga (Roxb.) Adr. Juss

2014 ◽  
Vol 6 (1) ◽  
pp. 99-104 ◽  
Author(s):  
Kishore Kumar CHIRUVELLA ◽  
Arifullah MOHAMMED ◽  
Rama Gopal GHANTA
Keyword(s):  
Seed Viability ◽  
Ms Medium ◽  
Stem Cuttings ◽  
Endemic Plant ◽  
Seedling Mortality ◽  
Root Regeneration ◽  
Half Strength ◽  
Shoot Necrosis

Like most of the medicinal plants Soymida febrifuga (Meliaceae) possess significance for its valuable secondary metabolites. Multiplication of this endemic plant is limited by difficulty in rooting of stem cuttings, high seedling mortality rates and low seed viability period. Hence efficient protocols for in vitro mass propagation has been established from field grown and aseptic seedlings explants. Strikingly, we observed aberrant structures such as vitrified shoots, faciated shoots, albino shoots as well shoot necrosis during its micropropagation. These phenotypic maladies were observed during organogenesis and rooting. Compared to other abnormalities, shoot necrosis nonetheless was frequent and pronounced leading to plant death. Shoots when subjected to rooting also displayed necrosis which was controlled by transferring to MS medium containing various concentrations and combinations of calcium levels, activated charcoal, glucose, fructose and auxins. Microshoots initiated roots on half strength MS medium with IBA and IAA individually or in combination within two weeks. MS half strength solid medium supplemented with CAN (556 mg l–1), CAP (1.0 mg l–1), IAA (2.0 mg l–1) and IBA (2.0 mg l–1) in combination was found to be more efficient in showing high frequency (95%) of root regeneration. Rooted plantlets were successfully hardened and 70-85% of regenerated plants were successfully acclimatized to natural environment. In vitro derived plantlets were morphologically similar to in vivo plants.

IN VIVO STUDIES ON RED CELLS AND PLATELETS STORED IN HALFSTRENGTH CITRATE

1987 ◽  
Author(s):  
C Prewse ◽  
K Bell ◽  
B Griffin
Keyword(s):  
Factor Viii ◽  
Coagulation Factor ◽  
Red Cells ◽  
In Vivo Studies ◽  
Dramatic Improvement ◽  
Half Strength ◽  
Survival Studies ◽  
Cellular Components

We have previously shown that donation of blood into anticoagulants containing half the normal amount of citrate results in a dramatic improvement in the stability of coagulation factor VIII and has no adverse effect on the in vitro qualities of red cells or platelets during storage. To confirm the viability of stored cellular components we are now performing autologous survival studies in healthy volunteers using radiolabelled cells from red cells and platelets stored for 35 and 5 days respectively. Results to date indicate a 24 hour survival of 80% for red cells stored at a haematocrit of 0.70 for 35 days. Infusion of Ill-In oxine labelled platelets after storage for 5 days in full or half-strength citrate gave recoveries of 40% and survivals of 7 days. These encouraging results suggest use of halfstrength citrate may be a route to increasing factor VIII supply without any additional donor recruitment. Further in vitro studies have also been performed on cellular components and reveal adequate in vitro quality for half-strength citrate blood held at room temperature for 20 hours prior to component preparation.

scholarly journals Improved micropropagation and foliar micromorphological studies in Turnera ulmifolia L. – An important medicinal plant

2018 ◽  
Vol 30 (2) ◽  
pp. 283-294 ◽  
Author(s):  
Mani Manokari ◽  
Mahipal S. Shekhawat
Keyword(s):  
Shoot Proliferation ◽  
Nodal Segments ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Half Strength ◽  
Turnera Ulmifolia ◽  
Semi Solid ◽  
Number Of Shoots

Abstract The present study reports an efficient in vitro propagation system for Turnera ulmifolia using nodal segments as explants. Turnera ulmifolia (Passifloraceae) is an important garden plant with multipotent medicinal values. Effective shoot proliferation was achieved on agar gelled MS medium (Murashige and Skoog, 1962). The maximum number of shoots (8.3 ± 0.57) per initial explant was obtained on MS medium supplemented with 8.88 mM of 6-benzylaminopurine (BAP) and 0.54 mM of α-naphthalene acetic acid (NAA). The highest number of shoots (59.5 ± 2.10) proliferated on semi-solid MS medium (with agar) augmented with 2.22 mM of BAP and 2.32 mM of kinetin (Kin) along with 0.54 mM of NAA. Longer (4-5 cm) and healthy shoots were rooted (12.0 ± 0.10 roots per shoot) on half-strength MS medium fortified with 9.84 mM of indole-3 butyric acid (IBA). The in vitro regenerated plantlets were hardened in the greenhouse and transferred to the field. Significant developmental changes were observed in the foliar micromorphology of in vitro raised plantlets when these were transferred to the field. The stomatal index was gradually reduced (26.72 to 21.25) in the leaves from in vitro to field environments. But, vein-islets and veinlet terminations (13.4 and 7.6) were increased (39.7 and 18.4) respectively from in vitro to in vivo grown plants. Simple, unicellular, less frequent and underdeveloped trichomes were observed with the leaves of in vitro plants but fully developed trichomes recorded in the field transferred plants. The study could help in understanding the response and adaptation of tissue culture raised plantlets towards changed environmental conditions.

scholarly journals A Review on Haploid and Double Haploids in Ornamental Plants

2021 ◽  
Vol 2 (3) ◽  
pp. 1-7
Author(s):  
Rakesh Kumar Meena ◽  
Keyword(s):  
Production System ◽  
Ornamental Plants ◽  
Cultivated Plants ◽  
Dh Population ◽  
New Varieties ◽  
Haploid Species ◽  
Short Time ◽  
Ornamental Species

Sporophyte plants with many gametophytic chromosomes are called haploid plants. These plants can be produced naturally or through in vitro or in vivo induction techniques. Double haploid (DH) can be obtained by doubling the number of haploid chromosomes spontaneously or artificially. They are homozygous, and this homozygosity will be realized in the life cycle of a generation using the DH production system. This production system is used to correct heterosis. Easy to interact with the DH population. DH can be used as parental inbreds of new varieties or self-pollinated plants or cross-pollinated plants. Haploids can be used to isolate mutants, especially if the mutant allele is not diploid. If the haploid is transformed immediately after the chromosome is copied, the plant can be obtained step by step. By combining biotechnological means with conventional methods, the important goal of improving cultivated plants can be achieved in a short time. This article analyzes the various developments in the field of haploid species related to economically important ornamental species.

scholarly journals Validation of a novel perfusion bioreactor system in cancer research

2020 ◽  
Vol 74 (3) ◽  
pp. 187-196
Author(s):  
Jasmina Stojkovska ◽  
Jovana Zvicer ◽  
Milena Milivojevic ◽  
Isidora Petrovic ◽  
Milena Stevanovic ◽  
...  
Keyword(s):  
Cancer Research ◽  
Drug Screening ◽  
Cancer Drug ◽  
Perfusion Bioreactor ◽  
Animal Testing ◽  
Anti Cancer ◽  
Medium Flow ◽  
Bioreactor System

Development of drugs is a complex, time- and cost-consuming process due to the lack of standardized and reliable characterization techniques and models. Traditionally, drug screening is based on in vitro analysis using two-dimensional (2D) cell cultures followed by in vivo animal testing. Unfortunately, application of the obtained results to humans in about 90 % of cases fails. Therefore, it is important to develop and improve cell-based systems that can mimic the in vivo-like conditions to provide more reliable results. In this paper, we present development and validation of a novel, user-friendly perfusion bioreactor system for single use aimed for cancer research, drug screening, anti-cancer drug response studies, biomaterial characterization, and tissue engineering. Simple design of the perfusion bioreactor provides direct medium flow at physiological velocities (100?250 ?m s-1) through samples of different sizes and shapes. Biocompatibility of the bioreactor was confirmed in short term cultivation studies of cervical carcinoma SiHa cells immobilized in alginate microfibers under continuous medium flow. The results have shown preserved cell viability indicating that the perfusion bioreactor in conjunction with alginate hydrogels as cell carriers could be potentially used as a tool for controlled anti-cancer drug screening in a 3D environment.

scholarly journals Effect of Drupe size Grading on in Vivo and in Vitro Germination and its Dormancy Mechanism of Teak (Tectona grandis Linn. F)

2021 ◽  
Vol 17 (4) ◽  
pp. 673-683
Author(s):  
Poomaruthai Masilamani ◽  
Venugopal Rajanbabu ◽  
Subramanian Venkatesan
Keyword(s):  
Tectona Grandis ◽  
In Vitro Germination ◽  
Seed Filling ◽  
Large Size ◽  
True Seed ◽  
Physiological Dormancy ◽  
Half Strength ◽  
Filling Capacity

The effect of drupe size on in vivo and in vitro germination and its dormancy mechanism in teak have been studied in this paper. Teak drupes were size graded to five categories namely very large with more than 15 mm, large with 13 to <15 mm, medium with 11 to <13 mm, small with 9 to <11 mm and very small with less than 9 mm of diameter. Under in vivo pot culture condition very low germination percent of drupes was observed. It was observed that very large size drupe had highest 100 drupe weight, 100 true seed weight and seed filling capacity, but it was negatively correlated with in vivo germination of teak drupes. When true seeds isolated from the drupes were grown under in vitro condition in half strength MS medium the germination percent was significantly increased up to 54.1. True seeds obtained from small size graded drupes recorded highest germination per cent in MS media. This showed that the drupe size did not play major role in the germination and early growth in teak. Very small size graded drupes have low seed filling capacity, and it leads to decrease the germination percent. Poor germination of drupes showed the presence of mechanical and physiological dormancy in drupes. Similarly, the presence of morphological and embryo dormancy in teak may limit the true seed germination potential to 54.1 per cent. In addition, in vitro germination of true seeds will be a promising approach to obtain large number of saplings in teak.

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