The use of artificial light sources such as light-emitting diodes (LEDs) has become a prerequisite in tissue culture studies to obtain morphogenetic enhancements on in vitro plants. This technology ...is essential for developmental enhancements in the growing plant cultures due to its light quality and intensity greatly influencing the in vitro growing explants at a cellular level. The current study investigates the effects of different light-emitting diode (LED) spectra on the growth of apical buds of Ficus carica var. Black Jack. Ficus carica, commonly known as figs is rich in vitamins, minerals, and phytochemicals capable of treating microbial infections and gastric, inflammatory, and cardiac disorders. Apical buds of Ficus carica var. Black Jack, presented morphogenetic changes when grown under six different LED spectra. The highest multiple shoots (1.80 per growing explant) and healthy growing cultures were observed under the blue + red LED spectrum. Wound-induced callus formation was observed on apical buds grown under green LED spectrum and discolouration of the growing shoots were observed on the cultures grown under far-red LED spectrum. Multiple shoots obtained from the blue + red LED treatment were rooted using 8 µM indole-3-acetic acid (IAA), and the rooted plantlets were successfully acclimatised. Compared with the other monochromatic LEDs, blue + red proved to be significantly better for producing excellent plant morphogeny. It is apparent that blue and red LED is the most suitable spectra for the healthy development of plants. The findings have confirmed that the combination of blue + red LED can potentially be used for enhancing growth yields of medicinally and commercially important plants.
•First report that demonstrates in-vitro protocorm-like-body (PLB) cultures habituation towards illumination source.•Discovery of several anticancer bioactive compounds in PLB.•High intensity ...(16.9 μmol/s) green light supplementation promotes the accumulation of total phenolic content.•PLB serves as excellent source for the acquisition of anticancer phytochemicals due to its high proliferation capacity.
Gas-chromatography-mass-spectrometry revealed the presence of various bioactive compounds with anticancer properties in protocorm-like-body (PLB) cultures of a Dendrobium hybrid orchid (Dendrobium Enopi x Dendrobium Pink Lady). Pre-illumination of red fluorescent light lessened the stimulating effects of light-emitting diodes (LEDs) on secondary metabolites production among in vitro PLB cultures, possibly due to habituation. The highest flavonoid content of 16.79 μmol/ g of fresh weight (FW) was achieved under blue-red (1:1) LED for PLBs pre-treated with white LED for more than 3 subculture cycles. Phenolics content significantly reduced as PLBs pre-cultured under red fluorescent light for 2 subculture cycles were exposed to LED illuminations, where far red LED resulted in the lowest total phenolic content (18.85 μmol/ g FW). High intensity green LED (16.9 μmol/s) enhanced the accumulation of phenolics while amino acids such as L-leucine, glycine and proline exhibited no significant stimulating effect for secondary metabolites production.
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Plant tissue culture involves the use of explants obtained from plants to induce organogenesis with the help of plant growth regulators (PGRs). Micropropagation techniques provide a ...faster and economical solution to the limitations associated with traditional methods of plant cultivation. The present study focuses on the multiple shoot induction and proliferation of Ficus carica var. Black Jack. Factors that influence the growth of in vitro multiple shoots on the apical buds, which include growth media and PGRs, were investigated in this study. Different concentrations of cytokinins like 6-benzylaminopurine (BAP), Thidiazuron (TDZ), and Kinetin (Kin) were used on woody plant medium (WPM) for the optimization of media for multiple shoot induction and proliferation.
Apical buds of Ficus carica var. Black Jack growing in WPM supplemented with BAP produced the healthiest plantlets, with the highest number of multiple shoots. The most efficient medium composition which produced the highest number of multiple shoots (37.8) per growing explant was WPM supplemented with 20 µM BAP. Proliferated multiple shoots were efficiently rooted using WPM + 20 µM BAP + 8 µM indole-3-acetic acid (IAA). This optimized medium composition significantly enhanced the production of multiple, disease-free plantlets using single apical bud explants of Ficus carica var. Black Jack.
In the present study the observations indicate that WPM supplemented with 20 µM BAP is the best-suited medium for organogenesis and multiple shoot culture of Ficus carica var. Black Jack, and this technique can be potentially applied for commercialization of the plant.
How to cite: Parab AR, Chew BL, Yeow LC, et al. Organogenesis on apical buds in common fig (Ficus carica) var. Black Jack. Electron J Biotechnol 2021;54. https://doi.org/10.1016/j.ejbt.2020.01.010
Clitoria ternatea
L., known as the butterfly pea plant, is from the Fabaceae family and is commonly used in traditional medicine or as a brain tonic in Asia. The root extracts of
C. ternatea
have ...been previously studied for its neuroprotective properties linked to memory enhancement of the mammalian brain, indirectly delaying the occurrence of neurodegenerative diseases. The current study aims to establish in vitro adventitious root cultures of
C. ternatea
via cotyledon explants and to identify the phytoconstituents of the induced adventitious roots. In this study, the highest percentage of induction (95.24%) was achieved in half-strength MS medium supplemented with 2.50 mg/L NAA (1-naphthaleneacetic acid) after 6 weeks of culture. Results also indicated that the supplementation of 1.50 mg/L NAA induced the highest number of adventitious roots (12.86 ± 2.14), while 2.50 mg/L 4-Cl-IAA (4-chloroindole-3-acetic acid) yielded higher adventitious root length (0.75 ± 0.13 cm) in comparison with the other auxin treatments. Microscopic analysis indicated thicker in vitro root morphology for all NAA treatments in comparison with the roots of in vitro seedling. Ethanolic extract of in vitro-generated adventitious roots showed potential anti-acetylcholinesterase activities (IC
50
= 1519.89 ± 0.18 μg/mL) albeit being lower than the control (IC
50
= 70.37 ± 0.01 μg/mL), while gas chromatography–mass spectrometry analysis identified the presence of pentacyclic triterpenes, phytosterols and fatty acids in the adventitious root culture that are linked to the mammalian neuroprotective properties of
C. ternatea.
The current investigation reports on the potential of
C. ternatea
root cultures being further utilized to produce valuable metabolites linked to the mammalian memory enhancement.
Gac fruit ('Momordica cochinchinensis' Spreng) is an exploitable perennial dioecious fruit found commonly in Southeast Asian countries (SA). Gac fruit has been traditionally used in folk medicine due ...to its high medicinal and nutritive value. Generally, the aril of the fruit is used as a red colourant for cuisines and supplement for healthier vision. Gac fruit is highly rich in lycopene and beta- carotene, fatty acids, vitamin E, polyphenols like phenolic acids, flavonoids and trypsin inhibitors. These compounds are linked with many important bioactivities such as provitamin A, antioxidant, antimicrobial, antiulcer and anticancer activities. This paper reviews the findings on nutritional properties, phytochemical composition and the biological activities of Gac fruit. The review has also briefly discussed the suitable conditions for the storage and utilisation of the fruit in food, beverage, nutraceutical, pharmaceutical and cosmeceutical industries.
Different designs of the plant tissue culture vessel, such as size, material, and shape, may alter its microenvironment atmosphere. The present study was conducted on protocorm-like bodies (PLBs) of
...Sabin Blue orchid to determine the development of PLBs on plastic and glass culture vessels of different sizes. PLBs were cultured in half-strength Murashige and Skoog (MS) medium with the same initial weight of 0.5 g in 10 replicates. The growth index of the PLBs was calculated after 11 weeks to study their growth in every vessel; additionally, biochemical analysis was performed to determine carbohydrate content, proline concentration, and photosynthesis pigments in the PLBs. Scanning electron microscopy (SEM) was performed to study stomata development on PLBs in each vessel, and histological analyses were conducted to study the cell structure. Overall, the PLBs cultured in a large 470 ml plastic vessel showed successful growth with a high growth index, high carbohydrate content, low-stress condition, and high chlorophyll content. SEM confirmed that the presence of trichome and rhizoid in PLBs cultured in the 470 ml plastic vessel. Histological analysis showed the formation of the shoot on the PLBs and the presence of starch granules. Thus, the use of plastic as a culture vessel provides a good impact for culturing PLBs and has low cost.
Background
Clonal propagation is one of the attributes of plant tissue culture. Therefore, analysis of genetic stability among the in vitro cultured plants is a crucial step. It helps to signify the ...clonal propagation of the micropropagated plants. Regenerated
Ficus carica
var. Black Jack plantlets were established using woody plant medium supplemented with 20 μM 6-Benzylaminopurine and 8 μM Indole-3-acetic acid under different light treatments such as normal fluorescent white light (60 μmol m
−2
s
−1
), and four different LED spectra, white (400–700 nm), blue (440 nm), red (660 nm) and blue + red (440 nm + 660 nm). Genetic stability analysis was performed on the in vitro and ex vitro plants of
Ficus carica
var. Black Jack.
Methods and results
Ten primers of each, ISSR and DAMD molecular markers, were used to assess the genetic stability of the eight samples of
Ficus carica
var. Black Jack. ISSR markers showed 97.87% of monomorphism whereas DAMD markers showed 100% monomorphism. Polymorphism of 2.13% was observed for the UBC840 ISSR–DNA primer which was negated under the genetic similarity index analysis for the eight samples. The findings of this study revealed that ISSR and DAMD markers are efficient in determining the polymorphism and monomorphism percentage among the in vitro and ex vitro samples of
Ficus carica
var. Black Jack.
Conclusion
Monomorphism of 100% obtained using DAMD markers and more than 95% of monomorphism obtained using ISSR markers indicate that the regenerated plants are significantly genetically stable. These molecular markers can be used to test the genetic stability of in vitro regenerated plants. It is recommended that genetic stability analysis should be performed for long-term maintenance of such micropropagated plants.
•Exogenous ascorbic acid does not guarantee improvement of growth recovery in cryopreserved PLBs.•Exogenous ascorbic acid and cryopreservation stages has a significant interaction effect to total ...soluble protein content, SOD and APX activities.•Changes in total protein content, SOD and CAT activities do not correlated with growth recovery of cryopreserved PLBs.•High APX activities are associated with growth recovery of cryopreserved explants.•Genetic stability of regenerants recovered from cryopreserved Aranda PLBs was confirmed based on RAPD and SCoT molecular markers.
A droplet-vitrification cryopreservation protocol has been successfully developed for Aranda Broga Blue orchid hybrid using protocorm-like bodies (PLBs). However, maximum growth regeneration percentage was recorded at 5% only based on previous report. Thus, to improve growth recovery of cryopreserved PLBs, cryopreservation stages were supplemented with ascorbic acid, tested at 50, 100 and 150 mg/L. However, results demonstrated that exogenous ascorbic acid was not favorable in regeneration of cryopreserved explants (maximum value of 1.67 % with 50 mg/L ascorbic acid supplementation). Total soluble protein and various antioxidant enzyme activities such as catalase (CAT), superoxide dismutase (SOD) and ascorbate peroxidase (APX) were evaluated after each cryopreservation stages in conjunction with the application of exogenous ascorbic acid. Addition of antioxidant must be carefully evaluated and its application may not guarantee successful growth recovery. RAPD and SCoT molecular analysis confirmed the genetic stability of regenerated cryopreserved PLBs as no polymorphism was detected compared to control PLBs culture.
Ficus carica L. is a common fig that is an incredibly nutritional fruit, well-known for its medicinal and economic values. This study aims to establish an efficient protocol for the mass propagation ...of fig plantlets (Ficus carica L.) for the cultivar ‘Violette de Solliès’. Surface-sterilized shoot-tip explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of cytokinins (6-benzylaminopurine, BAP; thidiazuron, TDZ; kinetin, Kn; and zeatin, Zea). Induced shoots were rooted on Woody Plant Medium (WPM) with different concentrations of auxins (naphthalene-acetic acid, NAA; indole-3-acetic acid, IAA; and indole-3-butyric acid, IBA). Rooted explants were acclimatized in eight different soil substrates prior to cultivation in a commercial plot. The propagated plantlets were analyzed for genetic stability and clonal fidelity using RAPD and SCoT molecular markers, whereas scanning electron microscopy (SEM) was performed to observe the stomata morphology of post-acclimatized plants. MS media supplemented with 5.0 mg/L BAP was the optimal treatment for multiple shoot induction (15.20 ± 1.03 shoots), whereas the highest percentage of rooting (93.33%) was achieved in WPM supplemented with 3.0 mg/L IBA. Plantlets were successfully acclimatized in biochar soil with a survival rate of 100%. RAPD and SCoT analysis showed no polymorphism occurrences across six subculture cycles, whereas observations via SEM indicated normal stomata structures on the leaves of acclimatized plantlets. This study documents an efficient micropropagation protocol for Ficus carica cv. ‘Violette de Solliès’ for the production of uniformed and true-to-type plant stocks suitable for commercial propagation.
This study investigated conserving an endangered terrestrial jewel orchid
using in vitro grown axillary buds. Excised segments of axillary buds (4-5 mm in length) were precultured on a modified ...Murashige and Skoog (MS) medium supplemented with 0.2 M sucrose for 24 h and osmoprotected in a loading solution for 20 min. Then, axillary buds were dehydrated in plant vitrification solution 2 (PVS2) for 10 min at 0 °C and incubated in liquid nitrogen for 1 h. Subsequently, axillary buds were rewarmed rapidly by dilution solution and transferred to a growth recovery medium supplemented with 0.05 µM melatonin, which led to an improved survival chance (16.67%) for cryopreserved
. The osmotic stress and the overproduction of reactive oxygen species (ROS) during cryopreservation stages may result in cryoinjuries and poor survival as increased levels of proline (5.51 µmol/g), catalase (85.64 U/g), peroxidase (565.37 U/g), and ascorbate peroxidase activities (12.19 U/g) were detected after dehydration, preculture, rewarming, and loading stage, respectively. Results obtained from this study indicate that further experimental designs which apply different PVS and exogenous antioxidants are needed for improved survival and regrowth of
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