Plant Productions

Effect of Kind and Plant Growth Regulator Composition on Micropropagation of Three Begonia Species

Fateme Hosseini; Nasrin Moshtaghi; Ahmad Sharifi; Abdolreza Bagheri; Hasan Marashi; Fateme Keykha Akhar

Volume 44, Issue 1 , June 2021, , Pages 25-36

https://doi.org/10.22055/ppd.2019.27321.1660

Abstract

AbstractBackground and ObjectivesIntroduction: Begonia is one of the most important ornamental plants around the world. Begonia species are grown for their attractive leaves and flowers ... Read More AbstractBackground and ObjectivesIntroduction: Begonia is one of the most important ornamental plants around the world. Begonia species are grown for their attractive leaves and flowers in indoor and outdoor conditions. The tissue culture technique is an alternative method for the propagation of begonia species. It can overcome the difficulties of vegetative propagation and lead to high-quality and uniform planting material under disease-free conditions irrespective of the season and weather. In begonia tissue culture, different species showed diverse responses to the medium composition, especially plant growth regulators. Also, lack of information on large scale micropropagation of begonia in Iran highlights research on optimization of begonia species propagation on in vitro culture condition. Thus, in this study the effect of plant growth regulators on shoot regeneration and proliferation of three begonia species (B. elatior‌‌, B. soli-mutata, and B. tiger), the impact of Auxin and sucrose on rooting of B. elatior‌‌and diferent pot substrate on acclimation were survived. Materials and MethodsIn this experiment, petioles of three begonia species (B. soli-mutata, B. elatior, and B. tiger) were used as explant sources. So explants were washed under tab water for 30 mins and sterilized with 1.5% sodium hypochlorite for 15 mins. After sterilization, thin Cell Layers (TCL) with 2 mm thickness were prepared from petiols and cultured in MS medium with different concentrations (0.2, 1, 2 mg/L) of kinetin (kin) or thidiazuron (TDZ) in combination with NAA (0, 0.2 mg/L). Next, the effect of auxin type (1 mg/L of IAA, IBA, and NAA) and sucrose concentration (30 and 40 g/l) on rooting of B. elatior plantlets were determined on in vitro condition. For plantlet acclimation, sand, CocoPeat perlite mixture (1:1 ratio), and Peat moss as pot substrate were evaluated. Results The results showed the shoot regeneration response of begonia species to the plant growth regulators of the medium, and there was a significant difference between them in all of the species adding the Auxin to the medium increased shoot regeneration. In B. elatior and B. tiger lacking of NAA to the medium caused no shoot regeneration. Maximum shoot regeneration (100%) was achieved in B. soli-mutata species in medium containing 1 or 2 mg/L of kin and 0.2 mg/l NAA, in B. elatior at 1 or 2 mg/L of TDZ in combination with 0.2 mg/l NAA and in B. tiger at 2 mg/L of both plant growth regulator (TDZ and Kin) and 0.2 mg/l NAA. DiscussionFor rooting B. elatior, MS medium containing 1 mg/L IAA plus 30 g/L sucrose was the best and favorite acclimation was acheived in cocopeat perlite mixture and peat moss pot substrate.

Effect of Medium, Iron-Chelating Agent and Plant Growth Regulator on Micropropagation of Pomegranate (Punica granatum L.)

Tayyabeh Eshaghi Sanayi; Mohammad Zare Mehrjerdi; Ahmad Sharifi

Volume 43, Issue 2 , September 2020, , Pages 309-322

https://doi.org/10.22055/ppd.2019.27232.1655

Abstract

Abstract Background and ObjectivesPomegranate (Punica granatum L.) is a fruit tree that originated from Iran and has been cultivated since ancient times. Classical propagation ... Read More Abstract Background and ObjectivesPomegranate (Punica granatum L.) is a fruit tree that originated from Iran and has been cultivated since ancient times. Classical propagation methods of pomegranate like stem cutting and air-layering have seasonal timing and a higher risk of pathogens contamination. Micropropagation by tissue culture could be considered a method for production of steady and pathogen free plants in this species. However, factors such as lack of rooting and browning due to the oxidation of phenolic components under invitro culture can limit its micropropagation efficiency. To overcome these problems, the effects of various factors, including media, iron-chelating agent, and plant growth regulators, were examined on the pomegranate propagation through in vitro conditions. Materials and MethodsTo obtain sterile explants, stem cuttings of the pomegranate cultivar Shishe Cap Ferdos were treated with different disinfection treatments. Then, the effect of DKW, MS and WPM basal media and two types of iron chelating agents (EDDHA and EDTA) were evaluated to decrease the secretion of phenolic compound from explant and subsequent browning. In the next step, the effect of plant growth regulators treatments, including 1-2 mg/l of BA in combination with 0.2 mg/l of various auxins (IAA, NAA or IBA) on shoot development and proliferation was studied. After that, rooting of regenerated plantlets was explored at WPM medium supplemented with 0.5 or 1 mg/l IAA, IBA or NAA. Finally, pomegranate plantlets were transferred to different soil mixtures, including eat moss and perlite (2:1), cocopeat and perlite (2:1), peat moss and vermiculite (2:1) and cocopeat and vermiculite (2: 1) for acclimation. ResultsThe results indicated that sterilization with 2% sodium hypochlorite for 15 minutes followed by washing with 1g/l carbendazim for 10 minutes, and finally culturing the explants in the medium containing 1 g/l carbendazim had the best effect on reducing explant contamination. Among different basal media and iron chelating agents, WPM resulted in the lowest explant browning. Also, EDDHA was more efficient compared to the EDTA so that explant browning in WPM media containing Fe-EDDHA reduced by 55.8% compared to DKW containing Fe-EDTA. In addition, WPM basal medium supplemented with 2 mg/l BA and 0.2 mg/l IAA induced higher shoot proliferation in node explant of pomegranate. For rooting treatments, after 45 days, WPM supplemented with 1 mg/l IAA induced higher rooting percentage, root length and number in pomegranate shoots. Among different acclimatizing treatments, plantlets that grew on peat moss and perlite (2: 1) had the highest leaf number and stem height. DiscussionOur results suggested that the medium composition and chelating agent affected pomegranate explants browning. EDDHA appears to have a positive effect on reducing browning by chelating copper, which is a phenol oxidase cofactor. Also, EDDHA was more efficient compared to the EDTA.

Quantification of Taxol and 10-Deacetyl Baccatin III in the Leaf and Cell Suspension Cultures of Two Taxus L. Species

H. Ahadi; M.H. Mirjalili; M. Farzaneh; H. Rezadoost

Volume 41, Issue 3 , December 2018, , Pages 95-104

https://doi.org/10.22055/ppd.2018.20838.1440

Abstract

Abstract Background and Objectives Taxanes are a family of diterpene alkaloids which is currently known as the most important anticancer compounds to suppress and inhibit cell growth, ... Read More Abstract Background and Objectives Taxanes are a family of diterpene alkaloids which is currently known as the most important anticancer compounds to suppress and inhibit cell growth, differentiation and proliferation in indefinitely known cancer cell lines. The most well recognized member of the taxane family is taxol (paclitaxel), a minor cytotoxic component derived originally from yew bark (Taxus brevifolia). 10-Deacetylbaccatin III (10-DAB), which is a homolog of taxol, can be extracted at high yields from the needles of Taxus baccata and is currently considered the most appropriate precursor for semi-synthesizing taxol and its analogue, taxotere (docetaxel). The natural source of both compounds is the inner bark and needles of several Taxus species, especially T. baccata and T. brevifolia. Plant cell culture is an environmentally sustainable source of taxol and offers several advantages as it is not subjected to weather, season or contamination. The aim of the present study was to evaluate taxol and 10-DAB content of two Taxus species and their ability in production of both taxanes through cell culture. Materials and Methods The plant materials of T. baccata and T. berevifolia were collected from Shahid Beheshti University (35° 48' N, 51° 23' E at an altitude of 1785 m) and botanical garden of Faculty of Agriculture, University of Tehran, Karaj (35° 48' N, 50° 59' E at an altitude of 1238 m), respectively. Callus culture of both species was initiated from internodal segments on B5 medium supplemented with plant growth reaulators. Cell suspension culture was established on the medium supplemented with 2.0 mg L−1 naphtalene acetic acid (NAA) and 0.1 mg L−1 benzyl amino purine (BAP). Cell viability was determined by Evan’s blue staining test. After 21 days, the amount of taxol and 10-DAB was analysed by HPLC. Results The results showed that mother stocks of T. baccata and T. brevifolia contain 45 and 27 µg/g DW of taxol and 40 and 4 µg/g DW of 10-DAB, respectively. Cell growth was estimated by measuring fresh weight (FW) and dry weight (DW). Results showed that the maximum growth was reached after 21 days of cultivation. Cell cultures of T. baccata and T. brevifolia produced 540 and 20 µg/l taxol and 1 and 110 µg/l of 10-DAB, respectively. Taxol production in T. baccata cell culture was 26-fold higher than in T. berevifolia cell culture. Discussion As far as our literature survey could ascertain, growth and taxol production in T. baccata cell cultures have previously been reported. Although, T. baccata cells are more potent for the production of taxol, T. brevifolia cell culture can be considered for the production of 10-DAB, a homolog of taxol, as an appropriate precursor for semi-synthesizing taxol and its analogue, taxotere (docetaxel). However, it is essential to study and quantify the effect of selected key medium components on growth as well as product accumulation and strike a balance between the two to enhance the yield and productivity.

Utilization of Indirect Regeneration to Induce Micropropagation of Fritillaria imperialis L. on in vitro Culture

Mojtaba Rahimi; Mohammad Hosein Daneshvar

Volume 39, Issue 4 , March 2017, , Pages 31-42

https://doi.org/10.22055/ppd.2017.12066

Abstract

Background and Objectives Iran is home of one of the most delightful and graceful flower of Fritillaria in the world. Its habitation is commonly expanded on the vast areas: growing ... Read More Background and Objectives Iran is home of one of the most delightful and graceful flower of Fritillaria in the world. Its habitation is commonly expanded on the vast areas: growing in beds, borders, backgrounds and rock gardens. Besides, it is also suitable for naturalizing and excellent for cut flowers. Fritillaria imperialis as a medicinal plant has pharmaceutical purpose in which possesses a health beneficial essence that used in production of mucus and cough medicine. In addition, extractions of plant tissues contain certain of properties which affect to reduce blood pressure and blood sugar. Unfortunately, the habitation of this ornamental plant is under threat and majority of its habitation already being to ruin due to human activities and urbanization. The propagation of Fritillaria through offsets (bulblets) is prolonging approach which takes approximately 3-4 years. Whereas, using tissue culture technique for flower production is required less time probably within a few months and economically has a huge advantage due to low cost and a numerous plant production in a short time. Materials and methods This research was carried out in four experiments: callus formation, bulblet production, root formation and acclimatization. In the present study, a factorial experiment was assigned as completed randomized design (CRD) with three replications. In the first experiment, leaf primordia and scale explants were cultured on MS medium supplemented with different concentrations of NAA and IBA in order to produce callus formation. In the second experiment, the calli were cultured on organogenesis medium which were contained MS medium supplement, with kin. and TDZ, after 6 and 12 weeks. In the third experiment, the bulblets were transferred into rooting media, which were contained MS medium supplemented with different concentrations of NAA (0, 0.5, 1.0 and 2.0 mg/l). In the fourth experiment, after washing the bulblet whit warm distilled water (40ᵒC) that contain 2.0 mg/l benlate fungicide, they then were planted into the pots containing sterilized perlite, cocopeat and perlite along with cocopeat. Results In the first experiment, the results demonstrated that the highest weight of callus (3.29g) was sustained in scale explants on basal MS medium which was supplemented with 0.5mg/l NAA along with 0.5mg/l IBA. In second experiment, the result indicated that the numerous of bulblets were developed on MS medium supplemented with 0.5mg/l TDZ along with 0.5mg/l kin. Which of these, the callus scale were produced on MS medium supplemented with 0.5mg/l NAA. In the third experiment, in order to retain rooting the bulblets, they were cultured on MS medium supplemented with different concentration of NAA. The result elucidated that the large quantity of roots (7.7) were obtained from leaf primordia on MS medium supplemented with 1.0mg/l NAA. In the fourth experiment, rooted bulblets were washed with warm distilled water for 5-10min, and then they were transferred into pots with perlite, cocopeat and perlite + cocopeat. Discussion The results revealed information that the rooted bulblets from scale growing on cocopeat substrate had retained the most diameter (21.64mm) as compared to other treatments. The finding results under this investigation strongly suggested that using micropropagation it is definitely a possible trend to abolish the barriers which caused to deteriorate of Fritillaria imperiallis.

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Keywords = In vitro culture

Effect of Kind and Plant Growth Regulator Composition on Micropropagation of Three Begonia Species

Abstract

Effect of Medium, Iron-Chelating Agent and Plant Growth Regulator on Micropropagation of Pomegranate (Punica granatum L.)

Abstract

Quantification of Taxol and 10-Deacetyl Baccatin III in the Leaf and Cell Suspension Cultures of Two Taxus L. Species

Abstract

Utilization of Indirect Regeneration to Induce Micropropagation of Fritillaria imperialis L. on in vitro Culture

Abstract