Biotechnology and Tissue culture
Somaye Amini; Seyed Mahdi Ziaratnia; Nasim Adibpour
Abstract
Purpose: The objective of this study was to enhance the MS-based medium to maximize cell biomass obtained from saffron corm within a liquid culture system. Research Method: The initial experiment employed a factorial design to examine three key factors influencing cell biomass: total nitrogen concentrations ...
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Purpose: The objective of this study was to enhance the MS-based medium to maximize cell biomass obtained from saffron corm within a liquid culture system. Research Method: The initial experiment employed a factorial design to examine three key factors influencing cell biomass: total nitrogen concentrations (20, 40, and 60 mM), ammonium/nitrate ratios (100:0, 75:25, 50:50, 25:75, and 0:100), and sucrose concentrations (3, 6 and 9%). In the second experiment, we evaluated the effect of a basal liquid MS medium enriched with L-glutamine, L-cysteine, PVP, chitosan, and KH2PO4 on cell biomass. The optimized medium from Experiment 1 was further enhanced based on the results obtained from Experiment 2 in order to increase cell biomass. Its effects were then compared to those of the standard MS medium by measuring fresh weight of cells over a six-week period in Experiment 3. Findings: Results from the initial experiment demonstrated a significant increase in cell biomass (3.37 g) when using 3% sucrose with nitrate instead of ammonium. Lowering the nitrogen concentration from 60 mM to 40 mM significantly improved cell growth. Additionally, both PVP and KH2PO4 contributed to increased saffron cell fresh weight in the second experiment. However, it was noted that even at low concentrations, chitosan application significantly enhanced cell death. The findings from the third experiment revealed that the modified MS culture medium, combined with potassium phosphate, significantly enhanced cell biomass growth compared to the standard MS medium. Research Limitations: No limitations were identified during this study. Originality/Value: This study evaluated key factors affecting cell biomass in saffron, but future research should explore the production of saffron metabolites under these influences.
Biotechnology and Tissue culture
Maryam Dehestani-Ardakani; Mohsen Karimi Dorche; Maryam Rahmati
Abstract
Purpose: Caladium bicolor is highly valued as both a landscape and indoor plant, primarily for its decorative appeal stemming from its diverse leaf shapes and vibrant, multicolored foliage. LED (light-emitting diode) lighting serves as a cost-efficient and potent means of promoting plant growth and development. ...
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Purpose: Caladium bicolor is highly valued as both a landscape and indoor plant, primarily for its decorative appeal stemming from its diverse leaf shapes and vibrant, multicolored foliage. LED (light-emitting diode) lighting serves as a cost-efficient and potent means of promoting plant growth and development. The impact of different LED lights was investigated on callus induction, regeneration, and plantlet growth of two cultivars of Caladium bicolor (‘White’ and ‘Red’). Research Method: Leaf explants were cultured on Murashige and Skoog (MS) medium supplemented with 1.5 mg L-1 IBA and 1 mg L-1 BA and moved to racks equipped with various LED lighting (100% red lights (R), 100% blue lights (B), 50% blue + 50% red lights (B+R), and 100% white fluorescent lamps (W)). Findings: Results showed W light was the best for maximum callus induction, leaf number, and plantlet height in both cultivars. Red + blue LED light spectrum motivated proliferation percentage of callus in both cultivars as compared to other light spectra. Conservation of ‘White’ caladium plantlets in R and B light spectra resulted in no hyperhydric micro shoot formation incidences. When examining various growth characteristics, it was evident that the B+R light spectrum of ‘Red’ caladium showed the best performance, while the B light spectrum in both cultivars had the least favorable outcomes compared to all other light spectra. Research limitations: There was no limitation. Originality/Value: Our findings offer a deeper understanding of how the quality of LED light impacts the in vitro propagation of caladium, potentially enhancing the cultivation of these plantlets through specific spectral exposure.
Biotechnology and Tissue culture
Habibeh Behnam; Azim Ghasemnezhad; Mahdi Alizadeh; Alireza Sadeghi Mahonak
Abstract
Purpose: The objective of the present study was to examine the impact of explant type and varying concentrations of 2,4-Dichlorophenoxyacetic acid and 6-Benzyladenine growth regulators on the San Pedro cactus callus morphological and biochemical characteristics. Research method: Four types of explants ...
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Purpose: The objective of the present study was to examine the impact of explant type and varying concentrations of 2,4-Dichlorophenoxyacetic acid and 6-Benzyladenine growth regulators on the San Pedro cactus callus morphological and biochemical characteristics. Research method: Four types of explants were used i.e. explants with areola, without areola, with truncated areola, and with central tissue. Additionally, five combinations of BA and 2,4-D, were tested (0 mg/L BA + 2 mg/L 2,4-D, 2 mg/L BA + 2 mg/L 2,4-D, 3 mg/L BA + 3 mg/L 2,4-D, 4 mg/L BA + 4 mg/L 2,4-D, 0 mg/L BA + 0 mg/L 2,4-D). Findings: The results indicated that callus formation induced in all treatments 6 days after inoculation. There were significant differences in growth parameters, including fresh weight, volume, moisture, tissue firmness, total phenols, total flavonoids and antioxidant activity of the callus (P < 0.01) and dry weight of callus (P < 0.05). Explants holding a segment of central tissue, yielded the least favorable results in most of experimental treatments, and the application of 2,4-D in the absence of BA had an inhibitory and toxic effect on the San Pedro cactus explants. Research limitations: No limitations were found. Originality/Value: Specifically, use of 2 mg/L BA + 2 mg/L 2,4-D and explants with areola resulted in callus with higher fresh weight, volume and total flavonoids, as well as good tissue integrity and firmness. The reported results are a valuable resource for future research related to cell tissue culture and the elicitation of secondary metabolites in Echinopsis spp.
Biotechnology and Tissue culture
Mohammad Musavi Ahmadabadi; Nima Ahmadi; Maryam Dehestani-Ardakani
Abstract
Purpose: To investigate the effects of Putrescine and Indole-3-Butyric Acid (IBA) on the adventitious rooting of micro-cuttings and semi-hardwood cutting of Rosa damascena, this study was conducted under both in vitro and in vivo conditions. Research Method: The rooting of micro-cuttings was induced ...
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Purpose: To investigate the effects of Putrescine and Indole-3-Butyric Acid (IBA) on the adventitious rooting of micro-cuttings and semi-hardwood cutting of Rosa damascena, this study was conducted under both in vitro and in vivo conditions. Research Method: The rooting of micro-cuttings was induced on the basal MS medium supplemented with five concentrations (0, 0.25, 0.5, 1 and 2 mg/L) of IBA and putrescine. In vivo experiment, putrescine and IBA at five concentrations (0, 0.25, 0.5, 1 and 2 g/L) were applied on semi-hardwood damask cuttings, while a downward wounding was created by a sharp blade on the bases of cutting as another treatment. Findings: Data showed significant variations in the root number and root length for in vitro and in vivo cuttings treated with different concentrations of putrescine and IBA. The obtained results revealed that presence of putrescine and IBA in both conditions enhanced root formation, as significantly improved the number of roots and root length in each explant. Under in vitro conditions, the maximum root length and root number were observed on the MS medium supplemented with 1 mg/l IBA+1 mg/l putrescine. Research limitations: No limitations were found. Originality/Value: The present study highlighted the role of putrescine and IBA in the adventitious rooting of R. damascena, under both in vitro and in vivo situations.
Biotechnology and Tissue culture
Somaye Amini; Seyed Mahdi Ziaratnia; Ghadir Rajabzadeh
Abstract
Purpose: This study aims to explore the potential of in vitro culture as a method for scaling up the production of saffron based medicinal compounds, the most expensive spice renowned. Emphasis is placed on the critical role of friable callus (FC) formation as a prerequisite for successful suspension ...
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Purpose: This study aims to explore the potential of in vitro culture as a method for scaling up the production of saffron based medicinal compounds, the most expensive spice renowned. Emphasis is placed on the critical role of friable callus (FC) formation as a prerequisite for successful suspension culture. Research method: The research primarily investigates FC formation, focusing on the impact of varying strengths of Murashige and Skoog (MS) medium as well as combinations of NAA or 2,4-D and BA or Kin on compact callus. Subsequently, the study involves supplementing the MS medium with different concentrations of 2,4-D, kin, zeatin, glutamine, sucrose, and nitrogen to establish a cell suspension culture. Findings: The highest FC yield was achieved on a solid medium containing 2,4-D (1 mg l-1)+Kin (0.2 mg l-1), resulting in a fresh weight (FW) of 0.413 g. Furthermore, MS combined with 2,4-D (1 mg l-1)+Kin (0.2 mg l-1)+glutamine (10 mg l-1), as well as MS+2,4-D (0.5 mg l-1)+zeatin (0.3 mg l-1)+glutamine (10 mg l-1), demonstrated the highest FW under suspension conditions. The study also identified that 30 g l-1 sucrose and 30 µM were optimal for inducing maximum FW. Research limitations: Cell biomass is influenced by several factors that should to be optimized. Originality/Value: This research concludes that a cell suspension system holds promise for rapidly generating sufficient cell biomass to produce valuable secondary metabolites within a limited timeframe and space. Notably, the system successfully increased biomass from 0.2 to 1.2 g, underscoring its potential for efficient saffron-based product development.
Biotechnology and Tissue culture
Naser Askari
Abstract
Purpose: Lily is one of the most economically ornamental plants and tissue culture plays a vital role in accelerating mass propagation of lily. In lily tissue culture, the production of bigger bulblets is highly desirable. The objective of the present investigation is to examine the impact of gibberellin ...
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Purpose: Lily is one of the most economically ornamental plants and tissue culture plays a vital role in accelerating mass propagation of lily. In lily tissue culture, the production of bigger bulblets is highly desirable. The objective of the present investigation is to examine the impact of gibberellin on the in vitro growth of lily bulblets, administered at two distinct time intervals. Research method: In the present investigation, various concentrations of gibberellins (0, 0.1, and 1 µM) were employed at two distinct time points: the commencement of the culture period and the fifth week of culture period. After 11 weeks the fresh weight of bulblets, the number of bulblets, the fresh weight of leaves, the fresh weight of roots and the fresh weight of scale explant were scored and analyzed. Findings: The application of 1 µM gibberellin during bulblet induction yielded noteworthy outcomes, including a substantial 91% increase in the fresh weight of the bulblets, a significant 38% augmentation in the fresh weight of the leaves, as well as a 40% increase in the fresh weight of the roots. Research limitations: The quantification of endogenous phytohormones in lily scale explants was deemed unfeasible. Originality/value: The development of lily bulblets experienced a notable enhancement while the medium was supplemented with gibberellin in bulblet induction stage.
Biotechnology and Tissue culture
Mariem Lotfi
Abstract
Purpose: In Tunisia, Pyrus communis ‘Arbi’ is broadly imperiled by Erwinia amylovora. The breeding of resistant rootstocks is an effectual control strategy for disease management. Therefore, a sound protocol for micropropagation and for the extensive production of high-quality plantlets was ...
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Purpose: In Tunisia, Pyrus communis ‘Arbi’ is broadly imperiled by Erwinia amylovora. The breeding of resistant rootstocks is an effectual control strategy for disease management. Therefore, a sound protocol for micropropagation and for the extensive production of high-quality plantlets was developed. Research method: Three groups of LED treatments were carried out: (1) 100% blue (B) LED, (2) 100% red (R) LED and (3) 50% B + 50% R (=BR) LED. Stock plants were micropropagtaed on modified Murashige and Skoog (MS) medium with half concentration of NH4NO3 and KNO3. Findings: Throughout the propagation stage, red LED displayed important advantages: it produced optimal shoot height and leaf surface. The least leaf area was obtained with fluorescent light. The blue / red combination yielded considerable amelioration. Shoot weight/callus weight was maximal, along with shoot number and shoot length. The root formation of in vitro grown pear plantlets was greatly influenced by the various light types and by the incorporation of a new root-promoting substance, phenyl acetic acid (PAA). When combining red light and PAA, 89 % of rooting was observed in pear plantlets. The acclimatized pear vitroplants achieved rapid growth without morphological anomalies. Research limitations: In order to improve the survival rates of the acclimatized vitroplants, the acclimatization stage needs to be further studied. Originality/Value: The study compared the impact of different combinations of monochromatic blue and red LED lights and phenyl acetic acid against that of fluorescent light during the micropropagation, rooting and acclimation of a resistant pear (Pyrus communis L., cv. Arbi).
Biotechnology and Tissue culture
Mohammad Matani Borkheyli; Seied Mehdi Miri; Amrollah Nabigol
Abstract
Purpose: Micropropagation of GF677 rootstock, the most widely clonal rootstock used in peach orchards, is an important method for large-scale production of disease-free plants. In this research, effects of media, plant growth regulators and carbohydrates in order to optimize the efficient micropropagation ...
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Purpose: Micropropagation of GF677 rootstock, the most widely clonal rootstock used in peach orchards, is an important method for large-scale production of disease-free plants. In this research, effects of media, plant growth regulators and carbohydrates in order to optimize the efficient micropropagation protocol of GF677 rootstock has been investigated. Research method: In vitro shoots were multiplicated on MS, WPM and DKW media supplemented with 0, 0.5, 1, 2 mg L-1 BA. Proliferated shoots were rooted on MS, WPM and DKW media containing 0, 0.5, 1, 2 mg L-1 IBA. In another experiment, the effect of carbohydrate type was investigated. Findings: High shoot number and node number as well as shoot fresh weight were achieved with shoot tips when cultured on MS medium supplemented with 1 or 2 mg L-1 BA. The highest percentage of rooted shoots was obtained on MS or WPM media supplemented with 1 or 2 mg L-1 IBA. Maximum root number was regenerated on WPM medium containing 1 mg L-1 IBA. Sorbitol was found to be more effective carbon source on shoot multiplication than sucrose, while the highest average of root number and root length were observed in the medium containing sorbitol and sucrose medium, respectively. Survival rate during the acclimatization in the greenhouse was 67%. Limitations: Plant acclimatization needs to be studied for commercial production. Originality/Value: This protocol has proven useful for micropropagation of GF677 rootstock.
Biotechnology and Tissue culture
Shahin Jahangirzadeh Khiavi; Koorosh Falakro; Sanam Safaei Chaeikar
Abstract
Purpose: A significant number of genetic resources of Camellia sinensis and its allied genotypes have been collected and preserved in Iran TRC. Information about them is mostly based on morphological data. Research method: PCR-RFLP technique and morphological characters were used for the identification ...
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Purpose: A significant number of genetic resources of Camellia sinensis and its allied genotypes have been collected and preserved in Iran TRC. Information about them is mostly based on morphological data. Research method: PCR-RFLP technique and morphological characters were used for the identification of organelle DNA (cpDNA) diversity in 25 tea genotypes. Twenty-one qualitative and quantitative characteristics were evaluated. Findings: A pair-wise similarity among the samples ranged from 0.14 to 0.66 based on morphological data. The dendrogram was designed, and samples were grouped into three main clusters at 0.38 similarity. Using three universal primer pairs which introduced for chloroplast amplified about 4070bp of cpDNA, following the digestion of fragments with three restriction endonucleases (HinfI, AluI and PstI) and the result of this method was introduced six haplotypes. The most significant and widespread haplotype was H2 (frequency ≈ 28%). All of the detected mutations were insertion-deletions and they ranged from 30 to 60 bp. The calculated total cpDNA diversity in populations (hT), a major portion of it was within populations were (hS) and genetic differentiation among populations (GST) were 0.43, 0.17 and 0.61, respectively. It should have been noted that the calculated GST was low and no structure could be identified. Limitations: Applying allied species and using more potent markers such as cpSSR and sequencing can lead to more accurate results. Originality/Value: The results of this study indicate that the PCR-RFLP method and morphological characters are applicable in the identification of tea genotypes and cultivars. In studying Camellia genus phylogeny, the polymorphism in cpDNA has to be considered carefully.
Biotechnology and Tissue culture
Mariem Lotfi; Chokri Bayoudh; Afifa Majdoub; Messaoud Mars
Abstract
Purpose: In Tunisia, pear cultivars are widely threatened by the attack of fire blight disease. Cultivation of tolerant cultivars is an effective control strategy for disease control. For this purpose, a reliable protocol was established for micropropagation of local Pyrus communis and Pyrus syriaca ...
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Purpose: In Tunisia, pear cultivars are widely threatened by the attack of fire blight disease. Cultivation of tolerant cultivars is an effective control strategy for disease control. For this purpose, a reliable protocol was established for micropropagation of local Pyrus communis and Pyrus syriaca L. and for large-scale production of high-quality plantlets. Research method: Using apical explants, different media and hormones were tested to establish a micropropagation procedure for local Tunisian Pyrus communis cultivars ‘Arbi’, ʻMaltiʼ, ʻMahdia 6ʼ and ʻMoknine 10ʼ and for Pyrus syriaca. Disinfection with 4% HgCl2 treatment for 20 minutes showed the highest percentage of plant survival. Successful initiation of the cultures was achieved on MS basal medium supplemented with 0.25 mg L-1 BA. Findings: During the proliferation stage, optimal shoot multiplication was obtained on MS medium with a half concentration of NH4NO3 and KNO3 supplemented with 0.1 mg L-1 IBA and 2 mg L-1 BA, but for maximum shoot length the BA concentration needed to be lowered to 1 mg L-1. A rooting rate of 100% and the highest root length and root number were attained on Cheng medium supplemented with 1.0 mg L-1 IBA. Pear vitroplants were successfully acclimatized on S2 substrate, composed by peat moss. Research limitations: Vitroplants acclimatization step needs to be well studied for the improvement of theacclimatized vitroplant survival rates by reducing the symptoms of crown rot. Originality/Value: This efficient optimized in vitro protocol will be successfully applied for large multiplication of high quality of Tunisian Pyrus vitroplants and cultivars.
Biotechnology and Tissue culture
Chadha Ayed; Chokri Bayoudh; Awatef Rhimi; Najla Mezghani; Faouzi Haouala; Bouthaina AL Mohandes Dridi
Abstract
Purpose: Tunisian garlic is widely threatened by the attack of several viruses genus. For this purpose, a reliable protocol was established for rapid in vitro propagation of local garlic (Allium sativum L.) cultivars for large-scale production of virus-free plants and high quality bulblets. ...
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Purpose: Tunisian garlic is widely threatened by the attack of several viruses genus. For this purpose, a reliable protocol was established for rapid in vitro propagation of local garlic (Allium sativum L.) cultivars for large-scale production of virus-free plants and high quality bulblets. Research method: Well disinfected shoot-tips of 1 mm were used as explants and cultivated on MS basal solid media enriched with various growth regulators: 6-Benzylaminopurine, α-Naphthaleneacetic acid, Kinetin, Indole-3-butyric acid and 2-isopentenyladenine for assessment of shoot formation, shoot proliferation and bulb formation. Findings: Among the different phytohormone concentrations and combinations, MS basal medium without any growth regulators (M0) was found optimal for shoot-tip initiation (96% explants development) and plantlets elongation (56.26 mm). For shoot proliferation, the M1 culture medium containing 1 mg L-1 BAP and 0.25 mg L-1 NAA was the best, giving a multiplication rate of 1.7 plantlets/explant. Shoots on M0 culture medium formed bulblets earlier. Multiple bulblets per explants were obtained on medium M22 containing 2 mg L-1 Kin and 0.1 mg L-1 NAA. Separated bulblets were transferred individually on bulbification media. Non-dividable bulblet was developed in various sizes. Research limitations: Bulblet acclimatization step needs to be well studied for high quality cloves production. Originality/value: This efficient optimized in vitro protocol will be successfully applied for large multiplication of virus-free garlic cultivars.
Biotechnology and Tissue culture
Soghra Rezaie; Maryam Dehestani-Ardakani; Kazem Kamali
Abstract
Purpose: Due to pharmaceutical value of Stevia plant (Stevia rebudiana Bertoni.), this study was done to introduce a new protocol for rapid mass propagation of itthrough tissue culture. Research Method: In MS medium shoot proliferation of stevia by six concentrations of BA (0, 0.1, 0.2, 0.3, 0.4, 0.5 ...
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Purpose: Due to pharmaceutical value of Stevia plant (Stevia rebudiana Bertoni.), this study was done to introduce a new protocol for rapid mass propagation of itthrough tissue culture. Research Method: In MS medium shoot proliferation of stevia by six concentrations of BA (0, 0.1, 0.2, 0.3, 0.4, 0.5 mg·l-1) and root induction by four concentrations of IBA (0, 0.025, 0.05, 0.1 mg·l-1) was investigated.Rooting of cuttings was done both in vitro and ex vivo conditions. Findings: According to the results, the most number of stems obtained in MS medium containing 0.3 mg l-1 BA. The highest length of stems obtained in MS medium without BA and the most number of leaves observed in MS medium supplemented with 0.4 mg·l-1 BA. In in vitro situation, the most number and length of roots obtained in MS medium without IBA. The most number of rooted cuttings was obtained in IBA solution after 72 hours and about 70% of rooted cuttings were healthy. Research limitations: It had no limitation to report. Originality/value: In conclusion, it seems that the potential of producing root and shoots in stevia plant is extremely high, so its proliferation is possible using low concentrations of plant growth regulators in in vitro culture.
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