Plant Productions - Articles List

Effect of phytohormone concentration and composition on the micropropagation efficiency of Stevia rebaudiana Bertoni

Atefeh Amini Neisiani; Abbas Saidi; Masoud Tohidfar

Volume 47, Issue 4 , January 2025, , Pages 581-594

https://doi.org/10.22055/ppd.2024.47365.2187

Abstract

Introduction Stevia rebaudiana Bertoni is a medicinal plant commercially used as a non-caloric sweetener for diabetic patients. Stevia seeds are small and infertile, and exhibit a ... Read More Introduction Stevia rebaudiana Bertoni is a medicinal plant commercially used as a non-caloric sweetener for diabetic patients. Stevia seeds are small and infertile, and exhibit a relatively low germination rate. Moreover, given the steviol glycoside content and morphological characteristics (such as leaf shape and color), cross-pollination in this plant, leads to a great deal of diversity among the plants grown from the seeds. Consequently, asexual propagation methods are vital for the effective cultivation of this plant. Micropropagation not only enhances growth and reproduction rates, it also preserves genetic resources. Furthermore, it allows for the creation of a genetically homogeneous population with high yields of steviol glycosides, which makes it a suitable method for the propagation of Stevia. The present study aims to evaluate the effects of various growth regulators on this plant, presenting an efficient protocol for commercial cultivation. Materials and Methods Stevia seed germination rates were calculated on MS medium at four and eight days post-sowing. Shoot induction was examined using combinations of kinetin (KIN), gibberellic acid (GA3), and benzylaminopurine (BAP). Subsequently, rooting experiments were conducted with three auxin hormones: naphthaleneacetic acid (NAA), indole-3-acetic acid (IAA), and indole-3-butyric acid (IBA). The acclimatization of the samples was conducted following successful rooting. Results and Discussion The germination rate of Stevia seeds was found to be 33.38%. The best shoot proliferation was achieved with 0.5 mg/L BAP and 2 mg/L KIN, producing the highest number of leaves and lateral branches. Optimal fresh weight (437.29 g) and shoot length (15.38 cm) were obtained in the treatment with 1.5 mg/L GA3 and 2 mg/L KIN. Significant negative correlations were observed at the 1% probability level between leaf number and shoot length (-0.61**), leaf number and fresh weight (-0.46**), lateral branch number and shoot length (-0.81**), and lateral branch number and fresh weight (-0.69**). Significant positive correlations were found between leaf and lateral branch numbers (0.95**) and between shoot length and fresh weight (0.98**). The longest root was observed in the treatment with 0.5 mg/L NAA, and the highest number of roots was found in the treatment with 1 mg/L IBA. The study reveal the effectiveness of specific combinations of growth regulators for the micropropagation of Stevia rebaudiana. The findings indicate that the hormonal treatments significantly affect various growth parameters, including leaf number, shoot length, fresh weight, and root development. The negative correlations between some traits suggest that optimizing one growth parameter might compromise. The positive correlations between other traits, such as leaf and lateral branch number, suggest that some growth attributes can be simultaneously enhanced. Conclusion The optimized protocol for the micropropagation of Stevia rebaudiana (0.5 mg/L BAP and 2 mg/L KIN for shoot proliferation, and 0.5 mg/L NAA for root induction) demonstrated efficient and economical performance using fewer and less diverse hormones compared to existing micropropagation protocols. Therefore, it can be considered an optimum protocol for the propagation of Stevia, potentially benefiting commercial cultivation and research applications.

Effect of proline nanocomposite coated with chitosan and moderate salinity stress on in vitro microtuberization of potato (Solanum tuberosum L.) cv. Agria

Roghayeh Ghasemi; alireza motallebiazar; Sima Panahirad; Amin Gahanian

Volume 47, Issue 1 , June 2024, , Pages 117-130

https://doi.org/10.22055/ppd.2023.43532.2097

Abstract

Introduction In recent years, the application of nanoparticles has been used successfully in plant tissue culture, which has a tremendous impact on the removal of microbial contamination ... Read More Introduction In recent years, the application of nanoparticles has been used successfully in plant tissue culture, which has a tremendous impact on the removal of microbial contamination of the explant through the use of these materials and also the positive role of these materials in callus formation, organogenesis, somatic embryogenesis, somaclonal variation and production of secondary metabolites. Proline is a proteogenic amino acid that accumulates as a useful solvent in plants under both stress and non-stress conditions. Recent discoveries indicate that proline plays an important role in plant growth and differentiation throughout the life cycle. Chitosan was found to enhance secondary metabolite production in cell suspensions and calli of various species applied in micropropagation, chitosan could improve plantlet quality in vitro, in consequence facilitating the subsequent acclimatization of plantlets to ex vitro conditions. The aim of this work was to investigate whether the in vitro application of nanocomposite and moderate salinity can improve microtuberization. Materials and Methods After the cultivation, the jars containing the explants were transferred to the growth chamber under absolute darkness and at a temperature of 18±2°C. The cultures were kept in these conditions for 4 months without sub culturing the samples for this purpose. MS medium containing 80 g of sucrose, containing proline at concentrations of 20 and 40 mg L-1, chitosan at a concentration of 240 mg L-1, proline 20 and chitosan 240 mg L-1, proline 40 and chitosan 240 mg L-1, 120 mg L-1, proline nanocomposite coated with chitosan 120 and 240 mg L-1 and control treatment, as well as non-salinity treatments with the same compounds and concentrations mentioned for salinity were cultured. Factorial experiment was conducted base on a completely randomized design with three replications on single nude explants of mentioned cultivar. Results and Discussion The results of this study show that moderate salinity has a significant effect on the first, second, third and last month of microtuberization. (in 1% level). The results of the comparison of mean microtuber yield showed that the highest yield was obtained when 240 mg of chitosan was treated with 20 mg / l of proline at moderate salinity with an average weight of 210 mg. Moreover, a concentration of 20 mg / l proline alone in moderate salinity was not sufficient to stimulate microtubers., but a concentration of 40 mg L-1 proline in mild salinity was satisfactory. According to the results of this study, it can be suggested that the use of chitosan compounds with proline and the use proline nanocomposite coated with chitosan with moderate salinity can be used to produce suitable microtuberization in the in vitro conditions of potatoes. In order to prove our results with the results of previous researchers, the results of chitosan application experiment on the growth and yield of miniature showed that application of 500 mg L-1 chitosan produced minituber weighing more than 1.5 g per seedling. Chitosan increased other traits including number of minituber per seedling, number of leaves per seedling and seedling length (Asghari-Zakaria et al., 2009). In the study of the effect of cobalt nanoparticles on the morphological characteristics of Santana cultivar in vitro, it was observed that the presence of cobalt nanoparticles caused the formation of microtubers faster, also the maximum size and function of microtubers from the combination of 2.5 mg. Cobalt nanoparticles with 80 g L-1 sucrose were obtained (Hamza, 2019). Conclusions In this experiment, proline nanocomposite coated with chitosan was successfully synthesized and according to the obtained results, it can be used in plant tissue culture and in vitro microtuberization. In the field of potato microtuberization, the use of nanocomposite showed a significant increase in the number of microtubers compared to the control, which showed a significant effect in the late stages of microtuberization. Also, the use of moderate salinity with treatment nanocomposite is effective for stimulating microtuberization, which increases microtubers yield. Based on the results of this experiment, a concentration of 120 mg L-1 proline nanocomposite coated with chitosan is a more suitable concentration for potato microtuberization.

Comparison of germination and primary growth of Ophrys schulzei Bornm. & Fleischm seedling in different culture media under in vitro condition

Frouzan Valadi; Jalal Khorshidi; Ali akbar Mozafari; Yavar Vafaee

Volume 46, Issue 1 , June 2023, , Pages 129-140

https://doi.org/10.22055/ppd.2023.42629.2068

Abstract

Introduction Ophrys schulzei Bornm. & Fleischm is an orchid species which its underground organ is harvested and has numerous applications in the food and pharmaceutical industries. ... Read More Introduction Ophrys schulzei Bornm. & Fleischm is an orchid species which its underground organ is harvested and has numerous applications in the food and pharmaceutical industries. The population of this orchid species in the natural habitat has been greatly reduced and is at risk of extinction due to the low germination percentage, required specific ecological conditions and also overharvesting. Therefore, it is necessary to evaluate different propagation methods in order to approach ex situ conservation, domestication and cultivation of this plant species. Vegetative propagation of orchids is difficult, time-consuming and expensive, and also a small number of propagules can be obtained from each mother plant. Sexual propagation in ex vitro condition is also difficult due to low endosperm of orchid seeds, undifferentiated embryo and need to symbiosis with mycorrhizal fungi. Therefore, it seems that in vitro seed germination is the most suitable method for producing orchid seedlings. The most important and crucial step in seedlings mass production is selecting appropriate growth media for in vitro cultivation. Therefore, in this research, the effect of different culture media on the germination and seedling growth of Ophrys schulzei was studied. Materials and Methods In this research, germination, protocorm formation and growth indices of Ophrys schulzei seedlings were evaluated in 5 different culture media including Malmgren, Orchimax, P723, and . At first, the seeds were sown in the mentioned culture media and transferred to the growth chamber with 25ºC and dark conditions. After germination, they were placed in a refrigerator at 4ºC for two weeks. At the end of two weeks, the culture containers were placed in the growth chamber with 25ºC and light conditions 16/8 h (light/dark). After 12 months from seed sowing, the mentioned factors were evaluated. This research was conducted as a completely randomized design with 5 replications at Research Center of Medicinal Plants Breeding and Development of University of Kurdistan, in 2019-2022. Results and Discussion The highest (25.26%) and lowest (0%) germination percentage were observed in Malmgren and Orchimax culture media, respectively. Protocorms were appeared earlier in the Malmgren medium than other media (19.33 days after seed sowing). Protocorms on medium were stayed in initial form, and no significant development was observed. Protomeristems were appeared earlier in Malmgren medium compared to P723 and (105.66 days after seed sowing). There was no significant difference between P723 and in terms of protomeristem appearance time (1/4 B5 148.33 and P723 159 days after seed sowing). Seedlings were formed only in P723, Malmgren and culture media. The highest protocorm weight (0.12 g) was belonged to the Malmgren culture medium, and the highest root length (21.44 mm), root weight (0.19 g), aerial weight (0.05 g), seedling length (45.98 mm) and seedling weight (0.37 g) were belonged to the seedlings grown in P723 medium. In terms of the number of roots and shoot length, no significant differences were observed between the three culture media. Conclusion In general, among the studied culture media, Malmgren and P723 were determined the most suitable culture media for production of Ophrys schulzei seedlings.

Micropropagation of Polianthes tuberosa L. Through Direct Organogenesis

Mohammad Hosein Daneshvar; Mehri Havil; Amin Lotfi Jalal Abadi

Volume 45, Issue 2 , September 2022, , Pages 277-286

https://doi.org/10.22055/ppd.2022.36928.1973

Abstract

IntroductionPolianthes tuberosa L. is one of the most important ornamental bulbous plants in tropical and subtropical regions that is cultivating in some regions of Iran. Due to the ... Read More IntroductionPolianthes tuberosa L. is one of the most important ornamental bulbous plants in tropical and subtropical regions that is cultivating in some regions of Iran. Due to the transmission of fungal and bacterial diseases, propagation by bulbs is not affordable. Tissue culture is a good way to produce uniform and disease-free seedling. Therefore, in vitro cultivation of tuberose is a suitable solution to commercial reproduction. Materials and MethodsTo investigate the best method of micropropagation of Polianthes tuberosa, this study was conducted in three experiments (proliferation, root formation and adaptation). The explants used in this study were lateral buds. Ethyl alcohol (70%) and dilute laundry bleach (5.25% active chlorine), benomyl fungicide and dishwashing liquid were used to eliminate surface contamination of the explants. In the first experiment, the effect of different plant growth regulators and MS medium (MS, ½ MS, ¼ MS) on shoot formation of lateral bud was investigated. Lateral bud explants were cultured in MS medium with various plant growth regulators including KIN (0.25, 0.5, 0.1, 0.2 and 0.4 mg / L), BAP (0.25 (0.5, 0.1 and 2.0 mg / l) and then the length and number of shoots were evaluated. After forming shoots in the propagation medium, they were transferred to a new environment to form roots.The root formation experiment included tree levels of NAA concentrations, (0.5, 0.75 and 1 mg/L) in combination with 0.2 mg/L IBA and control (without growth regulator). Adaptation experiment with 5 Substrate of bed including Peat Moss, Cocopeat, Sand, Peat Moss with Cocopeat (1: 1) and Sand with Cocopeat and Peat Moss (1: 1: 1) was conducted. Results and DiscussionThe results showed that there was a significant difference between various types of plant growth regulators (PGR) and MS culture media at different concentrations and no branch was produced without the use of PGR. In shoot prolifferation experiment, the maximum number of shoots (4.8) was obtained from lateral bud explants in MS medium with 0.25 mg / l BAP in combination with 0.25 mg / l KIN. The highest length of lateral shoot shoots (3.75 cm) was observed in MS medium with 0.25 mg / l KIN in combination with 0.25 mg / l BAP.The results of root formation experiment showed that the highest number of roots was observed in MS medium containing 0.5 mg / l NAA and 0.2 mg / l IBA and the lowest number of roots in control (without plant growth regulator). Treatment of 0.5 mg / l NAA with 0.2 mg / l IBA had the highest root length (1.22 cm). The lowest root length was also observed in the control (without plant growth regulator). In the adaptation test, no significant difference was observed between treatments in survival rate. ConclusionThe results of this study showed that the shoot organogenesis is controlled by the cytokinin with MS medium in different concentrations. It should be mentioned that cytokinin had an important role in the synthesis of RNA, stimulation of the production of protein and the activity of some enzymes. Also, the results showed that the use of cytokinin alone has a beneficial effect on shoot organogenesis.

Investigate the Possibility of Early Prediction of Sweet Cherry Graft Incompatibility by in vitro Techniques

Arezoo Jalali; Ebrahim Ganji Moghaddam; Ali Marjani

Volume 45, Issue 2 , September 2022, , Pages 287-298

https://doi.org/10.22055/ppd.2022.37508.1982

Abstract

IntroductionGraft compatibility is one of the most problems in fruit tree. Determination graft compatibility, it may last a long time but it was achieved in a short time by in vitro ... Read More IntroductionGraft compatibility is one of the most problems in fruit tree. Determination graft compatibility, it may last a long time but it was achieved in a short time by in vitro techniques. Plants increase their capacity antioxidant rapidly when stress occurs to increase resistance and tolerance to the conditions created. Since grafting in plants is also considered a form of stress, this way. It can be said that the plant's ability to disperse excess energy and neutralize free radicals is impaired, thus increasing the antioxidant capacity to increase the resistance to stress, so in incompatible grafts increases phenolic compounds and peroxidase to minimize oxidative damage compared to compatible grafts. This study Investigation the possibility of early determination of grafting (in) compatibility of sweet cherry by compare phenolic compound and peroxidase activity in vitro techniques. Materials and MethodsTwo factorial experiment was carried out on the base of completely randomized design with three replications. Treatments including the first factor, four levels of sweet cherry cultivars “Bing’’, “Takdaneh”, “Siyah Mashhad”, and “Adli” and the second factor, four levels of rootstocks Gisela-5, Gisela-6 (compatibility control), Mahaleb-168, and GF-305 (incompatibility control). Measurement of total phenolic compounds was determind according to the Folin-Ciocalteu method (Singleton et al., 1999). Antioxidant enzyme extraction was determined with the method of Mac Adam (1992) with some modifications. Analysis of variance was carried out by SAS software ver. 9.1 and mean comparison was conducted using LSD test. Results and DiscussionThe result showed that the highest and lowest percentage of grafting were “Bing” cultivar on Gisela-6 rootstock with an average of 64.21% and “Bing” cultivar on GF-305 rootstock of 1.1%. The highest number of leaves and longitudinal growth of scion was in “Bing” cultivar on Gisela-6. The highest number of roots and root lengths were in “Takdaneh” cultivar on Gisela-6 and “Bing” on Gisela-6. The highest amount of total phenol at the graft union was GF-305 and the lowest at the graft union was Gisela-5. The highest and lowest of peroxidase were at the graft union of “Bing” on-G5-305 and “Bing” and “Siyah mashhad” on Gisela. In general, the highest levels of total phenol and peroxidase at the graft union were higher on the GF-305 incompatible rootstock than Gisela-6 compatible rootstock. Therefore, our finding leads to the conclusion that phenolic compounds and peroxidase activity can be used to pre-screen for incompatible grafts. In general, the amount of total phenol in the grafts made on Gisela-6 (compatible control) and Gisela-5 rootstocks was less than GF-305 (incompatible control). GF-305 is incompatible with cherry cultivars. Phenolic compounds such as Catechins above the graft site can be used as a biochemical marker in the diagnosis of graft incompatibility (Baron et al., 2019). Peroxidase levels in incompatible grafts were higher than compatible grafts. It seems that the peroxidase enzyme can be used to predict incompatible grafts quickly. Preliminary analyzes on the role of peroxidase indicate that this enzyme is involved in the formation of cell wall constituents and in the next step reduces the flexibility of the cell wall by creating cross-linking between phenolic polymers of the cell wall (Cassab & Varner, 1987). ConclusionThe highest levels of total phenol and peroxidase at the graft union were higher on the GF-305 incompatible rootstock than Gisela-6 compatible rootstock. Therefore, the finding of this study leads to the conclusion that phenolic compounds (mostly) and peroxidase activity can be used to pre-screen for incompatible grafts

Optimization of in vitro Cultural Conditions of Haworthia (Haworthia Cooperi Baker)

Mahdiyeh Kharrazi; Maryam Moradian; Nasim Safari; Azadeh Khadem; Ahmad Sharifi

Volume 45, Issue 1 , May 2022, , Pages 53-66

https://doi.org/10.22055/ppd.2021.36744.1971

Abstract

IntroductionHaworthia cooperi is a type of ornamental succulent that is propagated by separating the offsets and leaf cuttings. Due to the small number of produced offsets by the mother ... Read More IntroductionHaworthia cooperi is a type of ornamental succulent that is propagated by separating the offsets and leaf cuttings. Due to the small number of produced offsets by the mother plant, the rate of propagation in this way is low, so micropropagation can be considered as an effective and efficient way to propagate this ornamental plant. Materials and MethodsFor this purpose, in the present study, the growth response of Haworthia cooperi plants to in vitro culture conditions in 1398 in the research laboratory of the Department of Ornamental Plants Biotechnology was investigated. This research was conducted in the form of three separate experiments based on a completely randomized design with three replications. In the first experiment, the effects of cytokinin type (BA and KIN) and concentration of cytokinin (0, 1, 2 and 3 mg/l) on leaf explant regeneration were evaluated. In the second experiment, the effects of culture medium type (MS and B5) and concentration of culture medium (0.5 and 1) on plantlet propagation were investigated. In the third experiment, plantlet rooting rate was evaluated under the influence of auxin type (NAA, IBA and IAA) and concentration (0, 0.5, 1 and 1.5 mg/l) of auxin. Results and DiscussionThe results showed that application of BA and KIN hormones increased the number of regenerated plantlets. The highest number of plantlets (5.50) was recorded in the MS medium containing 3 mg/l BA with 0.5 mg/l NAA. Plantlets cultured in the hormone-free culture medium had no regenerated symptoms. The number of plantlets increased with increasing the concentration of both hormones, but BA hormone in the culture medium had a greater effect on this factor. Lack of cytokinin hormone application in culture medium led to root production; however, the use of BA and KIN hormones in the culture medium reduced the number of produced roots by plantlets. The highest number of roots was recorded in hormone-free culture medium and the lowest number was recorded in the high concentration of BA hormone. The reduction in the number of roots in the culture medium containing BA was greater than the culture medium containing KIN. The of the second experiment revealed that use of MS culture medium showed better performance compared to B5 culture medium. Thus, the highest dry weight of plantlet and roots was obtained at full concentration of MS culture medium. The results of rooting stage showed that the presence of NAA hormone in the culture medium increased rooting in the plant. However, the produced roots in this culture medium were so thick that these roots are not suitable for plant growth in the acclimation stage. Therefore, considering the number and length of produced roots, the optimal culture medium for plantlet rooting is ½ MS culture medium containing 1 mg/l IAA hormone. Determining the best type of culture medium and optimizing it for regeneration, proliferation and rooting is one of the important factors in success in vitro culture conditions. Based on the results of the current study, application of MS medium containing 3 mg/l BA with 0.5 mg/l NAA was so effective for regeneration phase of this ornamental plant. Also, the efficiency of MS culture medium in the process of propagation of this plant is most likely related to the suitability of the components of this culture medium. In general, due to the presence of plant growth regulators such as NAA, IBA and IAA in the culture medium, rooting is induced in plantlets. In the present study, the use of IAA hormone at a concentration of 1 mg/l was a suitable hormonal combination for rooting in this ornamental plant. ConclusionBased on the results of the present study, the application of MS culture medium containing 3 mg/l BA with 0.5 mg/l NAA is recommended for regeneration and proliferation of tissue culture plantlets of Haworthia cooperi. It is also recommended to use ½ MS culture medium containing 1 mg/l of IAA hormone for rooting of tissue culture plantlets.

Optimizing Micro Propagation of Red Flesh Apple Genotype

Nooshin Kazemi; Mohammad Esmaiil Amiri; Maryam Jaffarkhani Kermani; Zahra sadat Hosseini

Volume 43, Issue 3 , November 2020, , Pages 337-348

https://doi.org/10.22055/ppd.2019.26409.1649

Abstract

Abstract Background and Objectives Red flesh apple belongs to Malus niedzwetzkyana of Rosaceae family. It has high levels of important phytochemicals like antioxidants, flavonoids ... Read More Abstract Background and Objectives Red flesh apple belongs to Malus niedzwetzkyana of Rosaceae family. It has high levels of important phytochemicals like antioxidants, flavonoids and anthocyanins in its cortex, which, in addition to attractiveness, is a source of benefit compound. The aim of the present investigation was to optimize efficient protocols for micro propagation of red flesh apple in order to use the results in future breeding strategies. Materials and Methods During the early spring, stem cuttings of growing red flesh apple trees were surface-sterilized in 70% ethanol and 50% sodium hypochlorite. Then, sterilized samples were placed in the test tubes containing MS medium supplemented with 2 µM BAP. At the proliferation stage, interactive effects of BAP (0, 2, 4, 6 mM) and GA3 (0, 3, 6 mM) on various aspects of proliferation (multiplication rate and vegetative growth characteristics such as number of produced shoots per explant, shoots length, leaf numbers, and the fresh and dry weight of explants) were investigated. At rooting stage, the effect of different concentrations of IBA or NAA (0, 1, 2, 3,4 µM) on the percentage of rooting, number of roots per explant and length of roots were compared. At acclimatization stage, the plantlets were transferred into pots with a volume of 200 ml containing sterile compound at the greenhouse condition. Results The results showed that there was a significant difference between the effects of various concentrations of BAP and GA3 on micro propagation of red flesh apple. The best result in proliferation (7.9 of axillary shoots after 8 months) was obtained from explants that were cultured in the medium containing 4 mM BAP and 3 mM GA3. At rooting stage, the maximum rooting percentage (85.2%) was obtained in the medium containing 3 mM IBA. In addition, rooted explants from IBA treatments were successfully acclimatized (100%) in the greenhouse, but rooted explants in NAA treatments were weak in acclimatized stage (40%). Discussion Measuring the growth rate is the major economic parameter for successful plant production at in vitro conditions, which is usually characterized by the number of axillary shoots, number of regenerated leaves and increase in the length of shoots and fresh and dry weight. We obtained the highest growth rate of red flesh apple in the culture medium containing 4 μM of BAP. The efficiency of BAP as an important cytokinin in apple proliferation has been reported in other studies as well. The results of the present investigation showed that highest rooting percentage was obtained from culture medium containing 3 μM of IBA. In addition, morphology of roots varied in callus formation and longitudinal growth and the samples rooted in IBA treatments had stronger roots than the rooted samples in NAA treatments. It seems that exposure to NAA instead of IBA stimulates indirect organogenesis and produced undesirable side effects on roots, such as callus formation, which can cause plantlets weakness during the period of acclimatization.

Evaluation of Yield and Advantages of Forage Legumes with Cereals Intercropping as Second Crop in Paddy Fields

mohammad rabiee; fatemeh farahdahr

Volume 43, Issue 3 , November 2020, , Pages 363-374

https://doi.org/10.22055/ppd.2019.27838.1689

Abstract

Abstract Background and Objectives One of the key strategies in sustainable agriculture is to restore diversification into agricultural ecosystems and its effective management. ... Read More Abstract Background and Objectives One of the key strategies in sustainable agriculture is to restore diversification into agricultural ecosystems and its effective management. The results of various studies have shown that the ecological advantage of intercropping is due to the efficient use of environmental resources. The purpose of mixed cultivation experiments, especially forage plants, is to increase the yield of forage in quantitative and qualitative terms, maintain the ecological balance and stabilize the system. Materials and Methods In order to compare the forage yield and determine the best planting ratio in mixed cropping of Triticale, Barley, Crimson clover (Trifolium incarnatum L.), and Vicia sativa, an experiment was conducted based on the complete randomized block design with sixteen treatments and three replications in paddy fields of Rice Research Institute of Iran (Rasht) during 2012-2014 for two years. Experimental treatments included pure planting of Triticale, Crimson clover, Vicia sativa, and Barley, planting mixtures of Triticale and Crimson clover with ratios of 70:30, 50:50 and 30:70, planting mixtures of Triticale and Vicia sativa with ratios of 70:30, 50:50 and 30:70, planting mixtures of Barley and Crimson clover with ratios of 70:30, 50:50 and 30:70 as well as planting mixtures of Barley and Vicia sativa with ratios of 70:30, 50:50 and 30:70. The studied traits included plant height, number of stems, number of tillers, number of plants per m2, fresh forage yield, dry forage yield as well as land equivalent ratio (LER) values for fresh and dry forages. The obtained data were analyzed by SAS software and mean comparisons were performed using LSD test. Results The results of analysis of the variance showed significant differences between the experimental treatments in most traits. Mean comparison showed that planting ratio of 30% Vicia sativa + 70% Triticale had maximum fresh and dry forage yields with averages of 48844 and 15269 kg. ha-1, respectively. Most treatments had LER of greater than one. Planting ratio of 70% Triticale and 30% Vicia sativa showed LER values of 1.31 and 1.54 for fresh and dry forage yields. The treatment is recommended for planting in paddy fields of Guilan area due to its relative advantage with respect to other planting ratios. Discussion Considering that the main objectives of crop forage production in rice paddy fields are the optimum use of paddy fields, sustainability of rice production, production of forage for livestock, and increasing farmers' incomes, the cultivation of all treatments tested as pure and mixed crops in rotation with rice has no problem with regard to the interference of harvesting of the forages with the time of cultivating the seedling rice.

Effect of Different Plant Growth Regulators, Explant Types and Light on Callogenesis Traits of Medicinal Plant Nettle (Urtica dioica L.)

Abdolkarim Zarei; Fatemeh Salajeghe; Mojahed Kamalizadeh

Volume 43, Issue 2 , September 2020, , Pages 255-266

https://doi.org/10.22055/ppd.2019.27548.1670

Abstract

Abstract Background and ObjectivesNettle (Urtica dioica L.) is considered a valuable plant with high level of various beneficial phytochemical, which has been used centuries in ... Read More Abstract Background and ObjectivesNettle (Urtica dioica L.) is considered a valuable plant with high level of various beneficial phytochemical, which has been used centuries in traditional medicine. Since it contains various secondary metabolites and shows noticeable activity against different Gram-positive and Gram-negative bacteria, nettle has been identified as an appropriate choice for wider applications in the food and pharmaceutical industries in the recent years. This plant is native to Asia, Europe, Africa and North America and grows naturally in different regions of Iran. Although nettle is an important medicinal plant worldwide, there is no information about optimization of its tissue culture conditions. This study was carried out to investigate the effects of different plant growth regulators, explant types and light condition on the callogenesis properties of nettle plant under in vitro condition. Materials and MethodsA factorial experiment was carried out on the base of completely randomized design with three replicates. Different explants including young leaves, old leaves, nodes and seeds of nettle plant were cultured in the solid MS medium supplemented with 30 g l-1 sucrose and different concentrations of plant growth regulators of 2,4-D (1, 2 and 3 mg l-1) and BA (0.1, 0.5 and 1 mg l-1). Callus-related characteristics, including time of callus induction, callogenesis percentage, and their color and texture were analyzed. Analysis of variance was carried out by SAS software ver. 9.3.1 and mean comparison was conducted using Duncan multiple range test. ResultsAccording to the results, callus induction was significantly influenced by explant type. The highest callus induction was attained from young leaves, nodes and old leaves, respectively. The results also showed that callus induction and its properties were significantly influenced by light condition. Preservation of cultures in dark condition resulted in higher callus induction than light condition. In addition, the effect of plant growth regulators was significant on callus induction and its properties. The highest percentage of callus induction was obtained from young leaf explants in the medium containing 1 mg l-1 2,4-D and 1 mg l-1 BA in dark condition followed by node explants in the same plant growth regulator treatment. However, medium supplemented with 1 mg l-1 2,4-D and 0.1 mg l-1 BA showed to be more effective for old leaf explants. Based on the results of this research, young leaf explants in the MS medium containing 1 mg l-1 2,4-D and 1 mg l-1 BA in dark condition are suggested for callus induction from nettle plant. DiscussionType, color and amount of produced callus in different plant species are highly dependent on the genotypes, compositions of tissue culture media as well as the levels of both endogenous and exogenous plant growth regulators. Auxin and cytokinin are two main plant growth regulators playing important roles in callogenesis of different plant species. There is an interaction between the level of plant growth regulators and color of produced callus. Based on the results of present study, it seems that in the nettle plant, node explants endure a higher level of plant growth regulators and hence produce a brighter callus than the leaf segment explants when they are treated with high concentrations of these chemicals.

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.

Optimization of in vitro Callus Induction and Somatic Embryogenesis of Horseradish (Armoracia rusticana) Supplemented with 2,4-D and IAA

Sara Ahmadi; Kambiz Mashayekhi; Seyyed Javad Mousavizadeh

Volume 43, Issue 1 , June 2020, , Pages 25-38

https://doi.org/10.22055/ppd.2019.26639.1635

Abstract

Abstract Background and Objectives Horseradish has a male sterility that produces a small amount of seed with a low germination. It then propagates with root cuttings, which ... Read More Abstract Background and Objectives Horseradish has a male sterility that produces a small amount of seed with a low germination. It then propagates with root cuttings, which is also subject to a limited number of mother rootstocks. Therefore, the mass proliferation of this plant is important through tissue culture, especially somatic embryogenesis protocols. Somatic embryogenesis is a method to develop embryo via plant somatic cells during in vitro culture. The purpose of the present experiment is to find the best method for callus and somatic embryogenesis inducing horseradish, which is carried out in two B5 and NL media in both solid and liquid phases. Materials and Methods This research was carry out in the tissue culture lab of the department of horticultural sciences at gorgan university agricultural sciences and natural resources from 2016 to 2019. Calli are developed on callus induction media (B5 + 1 µM 2,4-D + 2 µM kin, NL + 1 µM IAA + 2 µM kin), and then transferred to somatic embryo induction phase (B5 + 3 µM 2,4-D + 4 µM kin, NL + 3 µM IAA + 4 µM kin) for 8 weeks. Elimination of IAA, 2,4-D and kin from induced calli done onto realization phase, and globular, heart and torpedo embryos observed 4 weeks later. Results Based on the results, the best surface sterilization was achieved by 70% ethanol for 30 seconds and 70% aqueous sodium hypochlorite (v/v) for 60 minutes. The callus and somatic embryo were observed in both solid and liquid B5 and NL media. In the study of callus induction, according to the measurement of appearance quality and the process of callus formation, the growth and increase of callus in solid phase are higher than that of liquid phase. The highest callus induction record is seen in solid B5 medium containing 2 µM Kin and 1 µM 2,4-D. Generally, calli on embryo induction phase are bright green to yellowish and compact consisting of separate clusters of somatic embryonic cells. The results of somatic embryogenesis revealed that globular, heart, torpedo and total embryos in B5 containing 1 µM 2,4-D are more than NL containing 1 µM IAA. Discussion Optimizing the growth of somatic embryos is important for mass propagation of plants that face problems in seed germination or have a long growth period. Due to the success of the production of horseradish somatic embryos, the results of this experiment can provide the possibility of production and propagation of horseradish plants in a short time. Further studies are required to determine the somaclonal variation of horseradish mature somatic embryos and plantlets.

Effect of Pomegranate Peel Extract, Rosmarinus and Artemisia Essential Oils on Vase Life of Cut Chrysanthemum (Chrysanthemum morifolium R.)

Masoumeh Ebrahimipour Bafghi; Maryam Dehestani-Ardakani; Jalal Gholamnezhad

Volume 43, Issue 1 , June 2020, , Pages 129-143

https://doi.org/10.22055/ppd.2019.25748.1596

Abstract

Abstract Background and Objectives Chrysanthemum (Chrysanthemum morifolium R.) is one of the most popular cut flowers in the world. Stem end blockage and water stress are two ... Read More Abstract Background and Objectives Chrysanthemum (Chrysanthemum morifolium R.) is one of the most popular cut flowers in the world. Stem end blockage and water stress are two problems in shortening the vase life of cut Chrysanthemum. Essential oils are noble alternative substitutes for silver and chemical compounds because of their antimicrobial activities and environmentally friendly nature of the extracts. A pomegranate peel extract (PGE) was evaluated as a natural antifungal preparation for the control of postharvest rots. Materials and Methods This study was conducted to investigate the effect of pomegranatepeelextract of three cultivars (‘Shahvar dane sefid’, ‘Rabab dane ghermez’ and ‘Malas Poost Siah Yazdi’) in four concentration of 0, 15, 25 and 50 mg/l and Rosmarinous and Artemisia essential oils in four concentrations of 0, 25, 50 and 75 mg/l on vase life of cut chrysanthemum in a completely randomize design (CRD). Evaluated characteristics were consisted of vase life, stem end bacteria, petals water content, fresh weight loss, electrolyte leakage, protein content and peroxidase enzyme activity. Results According to the results, flowers treated by 15 mg/l pomogrante peel extract of ‘Shahvar dane sefid’ showed the highest effect on vase life of chrysanthemum with 34.10% higher than control. The lowest population of bacterial in stem end of flowers was obtqined by 25 mg/l Rosmarinus essential oil with 70.14% lower than control. By increasing the extract and essential oils concentrations, bacterial population in stem end of cut flower significantly increased, while peroxidase activity significantly decreased. The lowest fresh weight loss was observed in stems treated with 75 mg/l pomegranate peel extract of ‘Shahvar dane sefid’, 15 mg/l Pomogrante Peel Extract of ‘Rabab Daneh Ghermez’, 25 mg/l Artemissia essential oil and 25 and 50 mg/l Rosmarinous essential oil. The highest petals water content (75.66%) was obtained in 75 mg/l Rosmarinus essential oil. The lowest petals water content (44.66%) was obtained in 75 mg/l Artemisia essential oil. The lowest electrolyte leakage was observed in cut stem floweres that were treated by 15 and 25 mg/l pomegranate peel extract cv.’Malas Poost Siah Yazdi’. Discussion Generally, compared to control, all treatments were able to extend vase life and decrease fresh weight and microbial stem end of cut flowers. Also, among three pomegranate peel extracts, extracts of cv. ‘Shahvar dane sefid’ showed the highest effect on extending vase life, decreasing the fresh weight, electrolyte leakage and stem end bacteria, and thus, recommended for extending the vase life of cut flowers.

In Vitro Regeneration of Ornamental Crocus (Crocus vernus L.) by Using Plant Growth Regulators

Sara Parvin; Mohammad Hosein Daneshvar; amin lotfi jalal- abadi

Volume 42, Issue 4 , December 2019, , Pages 429-440

https://doi.org/10.22055/ppd.2019.25898.1603

Abstract

Abstract Background and Objectives Crocus flower, which belongs to Iridaceae family, is one of the most important ornamental bulbous plants. The propagation of Crocus vernus ... Read More Abstract Background and Objectives Crocus flower, which belongs to Iridaceae family, is one of the most important ornamental bulbous plants. The propagation of Crocus vernus L. by corm is not commercially affordable due to transmission of fungal and bacterial diseases as well as the long dormancy period which takes 4 to 5 months. Thus, in vitro culture of Crocus vernus is a suitable solution for commercial reproduction. Plant tissue culture technique would be a better alternative for improving quality and production. Materials and Methods In order to micropropagate ornamental saffron, the effect of different culture media on direct organogenesis experiment was conducted in factorial arrangement in a completely randomized design with 3 replications in the tissue culture lab of the Department of Horticulture at Agricultural Sciences and Natural Resources University of Khuzestan from 2015 to 2016. The explants used in this experiment were latheral and terminal buds which were evaluated in 10 hormons treatments and control in the Murashige and Skoog medium (MS). To eliminate surface contamination, the explants were first immersed under water for 30 minutes after placing the corms. Next, they were placed in high temperature Benlate fungicide solution (55° C) for 30 minutes. Then, they were put in 70% ethanol (ethyl alcohol) for 30 seconds and 15% Sodium hypochlorite solution for 6 minutes. The explants were finally washed three times with sterile distilled water under the air flow bench. The effects of plant growth regulators including BAP (0.5, 1.0, 2.0 mg/l), TDZ (0.5, 0.75, 1.0 mg/l), Kin (0.5, 1.0, 2.0 mg/l) and 0.1mg/l IBA in direct regeneration were investigated. Results The results showed that the kind of plant growth regulators (PGR) and the explant type are very important in regeneration of Crocus vernus so that we can not secure any shoots without PGR. In direct organogenesis experiment, the maximum number of multiple shoots (15.84) was obtained from apical bud explant in MS medium supplemented with 5/0 mg/l TDZ along with 5/0 mg/l BAP, 5/0 mg/l KIN and 1/0 mg/l IBA, after 10 weeks, that had a significant difference with other treatments at 1% Duncan test. We observed 2.o mg/l BAP along with 1/0 mg/l IBA had the greatest effect on the number of corms. Discussion The results showed the shoot regeneration is controlled by the ratio of cytokinin with auxin. It should be mentioned that cytokinin plays a role in the synthesis of RNA, stimulation of the production of protein, and the activity of some enzymes. The results also indicated that the use of cytokinin along with auxin has a beneficial effect on shoot regeneration.

Micro Propagation of Penta Rootstock (Prunus domestica) in the Two Culture Media (MS and B5)

Ziba Balapoor; Hossein Hosseini Moghaddam; Mehdi zarei; mehdi Mollashahi

Volume 42, Issue 4 , December 2019, , Pages 441-454

https://doi.org/10.22055/ppd.2019.24937.1570

Abstract

Abstract Background and Objectives Culture medium and growth regulators type can influence the growth traits and for determining the most effective medium and hormone compositions ... Read More Abstract Background and Objectives Culture medium and growth regulators type can influence the growth traits and for determining the most effective medium and hormone compositions an extensive studies are needed; otherwise the culture medium may have a negative effect on the growth traits. In this research the effects of two culture media (B5 and MS) supplemented with two growth regulators (IBA and BAP) with different concentrations (0.5, 0.9, and 1.5 for IBA and 0.4, 0.8, and 1.2 mg/l for BAP) alone or in combination was evaluated. Studied traits were number of leaf per shoot, shoot length, number of regenerated shoots based on completely random plan with 5 replications. Materials and Methods Explants provided in the beginning of spring from mother plants growing in open environment. Shoot tips and axillary buds (0.3-0.4 cm long) used as the explants. After washing with tap water, explants were immersed in ethylic alcohol for few seconds (3-4 seconds) then immersed in mercuric chloride (0.005 W/V) for 5 minutes which supplemented with few drops of tween 20 and finally washed three times with sterile distilled water. Disinfected explants cultured on medium under air laminar flow bench. Growth chamber was adjusted to 16 hours light and 25 to 27ᵒC. Results The results of variance analysis demonstrated significant differences between media, hormone level and interaction between hormone concentrations for traits studied. The results showed the longest shoot (6.39 cm) and the highest shoot number (13.65) obtained in MS medium supplemented with 0.8 and 0.4 mg/l BAP respectively. In this experiment the highest root number (8.41) and root length (14.75 cm) was observed in B5 and MS medium containing 0.9 and 1.5 mg/l, IBA. Discussion Overall the most shoot proliferation of this rootstock occurred in MS medium. New shoots in MS medium showed normal and better growth comparing to shoots obtained in B5 medium, that can be one of the results of higher amount of nitrogen (ammonium nitrite) in MS medium. In B5 medium with all hormone treatments weak growth observed for this plant. In fact this medium was made for herbaceous plants and this is one of the main reasons why penta rootstock did not showed good growth symptoms on this medium.

The Effect of Different Concentrations of Cadmium Chloride on Oxidative Stress in Shoot Cultures of Lemon Balm

Sayyedeh Razieh Nourbakhsh Rezaei; Leila Shabani; Majid Rostami; Mohammad Abdoli

Volume 42, Issue 4 , December 2019, , Pages 509-522

https://doi.org/10.22055/ppd.2019.24806.1567

Abstract

Abstract Background and Objectives Melissa officinalis is a medicinal plant belonging to the Lamiaceae family. The essential oil of this plant is used in many fields. Although ... Read More Abstract Background and Objectives Melissa officinalis is a medicinal plant belonging to the Lamiaceae family. The essential oil of this plant is used in many fields. Although more than 100 types of chemical substances have been identified in this plant, the most important compositions of its essential oil include citral, citronella, geraniol and linalool. Heavy metals are one of the main causes of non-biotic stress for living organisms due to increased use in the field of industrial and agricultural development and its high accumulation and toxicity. Cadmium is an unnecessary heavy metal, which, due to its high mobility and low concentration, easily enters the food chain from the soil. Cadmium induces oxidative stress by stimulating the synthesis of free oxygen radicals in the plant. In this study, the effect of cadmium chloride on oxidative stress induction in lemon balm was investigated. This experiment was carried out in 2016-2017 at Shahrekord University. Materials and Methods This research was conducted in a completely randomized design with three replications in vitro condition. We investigated the effects of different concentrations of cadmium chloride (0, 10, 20 and 40μm) on the biochemical parameters of the lemon balm. In this study, sterile stems of lemon balm propagated on the medium (1/2 MS) were used. After 60 days, seedlings were removed from solid MS medium and the roots were cut and cultured in liquid medium of 1/2 MS with different concentrations of cadmium. The cultivation was carried out in Erlenmeyer flask (250 cc) and was kept in an incubator shaker device. Sampling for the experiment was conducted one week after the growth of the stems in the medium. Results Based on the results obtained in this study, fresh weight of shoots grown in 10 and 20μM of cadmium chloride increased compared to the control. This heavy metal significantly reduced the amount of chlorophyll a, b and carotenoids, while the total chlorophyll content decreased only at 40μm concentration relative to control. Cadmium significantly increased hydrogen peroxide levels in all treatments. Also, it increased the activity of catalase and superoxide dismutase enzymes. Malondialdehyde increased in all treatment concentrations, but this increase was not significant at 40μm of cadmium chloride. Chloride cadmium reduced the absorption of molybdenum and iron, yet increased the adsorption of manganese and zinc. Discussion The results of this study revealed that the cadmium chloride in lemon balm shoots may have the potential to accumulate compounds such as hydrogen peroxide which, as a messenger molecule, produces antioxidants and can help the plant tolerate stressed conditions. It seems that two concentrations of 10 and 20 μm of cadmium chloride have no toxic effects for the stems and the plant has managed to withstand this tension with defense mechanisms.

Determination of the Optimal Dosage of Physical and Chemical Mutagenesis Agents in Callus Tissue of Kiwifruit Leaf

Hassan Saeiahagh; Mousa Mousavi; Ranjith Pathirana

Volume 42, Issue 4 , December 2019, , Pages 523-534

https://doi.org/10.22055/ppd.2019.26394.1623

Abstract

Abstract Background and Objectives Induced mutations have been used in a wide range of horticultural crops for more than half a century. This approach has opened up great opportunities ... Read More Abstract Background and Objectives Induced mutations have been used in a wide range of horticultural crops for more than half a century. This approach has opened up great opportunities for biotechnologists and plant breeders to produce new varieties with several new traits including resistance to disease, and tolerance to adverse environmental conditions or even produce new horticultural characters in some crops. The optimization of dosage of the physical and chemical mutagenic agents for inducing mutations is the first step in using this technique in plant breeding. This study was thus conducted to determine the optimum dosage of two mutagenic agents including gamma radiation as a physical agent and ethyl methanesulphonate (EMS) as a chemical agent in kiwifruit callus in in-vitro conditions. To that end, simple sequence repeat (SSR) markers were used to detect the mutations in the regenerated plantlets. Materials and Methods Having induced the callus in tissue culture media containing naphthaleneacetic acid, 6-benzylamino purine and meta-Topolin, the produced calli were exposed to different dosage of gamma radiation (0, 10, 20, 30, 40, 50 Gy) or different EMS concentrations (0, 0.4, 0.6, 0.8, 1, 1.2 % for 60 min). Then, the callis were transferred to the shoot regeneration media. The number of dead calli, and the size of live calli as well as the number of shoots generated in live calli were then recorded. Afterwards, the regenerated shoots were transferred to elongation media, and the DNA was extracted. The SSR markers analysis was followed on the randomly selected plantlets to detect the SSR markers using a Genetic Analyzer. Results As expected, with increasing the dosage of gamma radiation, the number of dead calli increased and the callus growth was thus retarded. The lowest number of dead calli was, indeed, detected in the control and the 50 Gy dose resulted in the highest number of dead calli. The morphology of shoots generated in the treated calli and the SSR marker analysis showed that 50 Gy dose could be used as the optimum radiation dose to produce an acceptable mutation rate in kiwifruit callus. In turn, EMS as a chemical mutagenic agent showed a different pattern in increasing the number of shoots regenerated with increasing dosage, however the same trend happened with gamma radiation in the number of dead calli and live callus size. Considering the effect of EMS on callus and the number of shoots regenerated per callus, a treatment with 1% EMS for 60 min can be used to produce a high density mutant population in kiwifruit. The highest mutation rate (12.5 %) was observed in the 50 Gy gamma ray treatment with changes in 12 pairs of SSR markers among 25 loci. For EMS, this value was 2.6 % in 1% concentration treatment. Discussion To detect the induced mutations using Gamma ray and EMS, the SSR markers can be applied. However, for the EMS mutation detection, more studies are needed to evaluate the efficiency of other techniques such as single nucleotide polymorphism (SNPs).

The Effect of Lead on the Regeneration and the Metal Accumulation in Brassica Oleracea Var. Acephala by In Vitro Culture

Omid Sadeghi; Mina Taghizadeh; Mousa Solgi

Volume 42, Issue 2 , June 2019, , Pages 265-278

https://doi.org/10.22055/ppd.2019.24671.1560

Abstract

Abstract Background and Objectives Among the major environmental pollutants, lead is the most substantial contaminant due to causing toxicity in plants and organisms. Recently, ... Read More Abstract Background and Objectives Among the major environmental pollutants, lead is the most substantial contaminant due to causing toxicity in plants and organisms. Recently, the remediation of these metals has been considered in plants. Plant in vitro culture is a key tool in phytoremediation research. In general, Plant tissue culture and in vitro selection techniques are used to increase the tolerance and the accumulation of heavy metals which have been reported in numerous plant species and populations. The application of ornamental plants for phytoremediation in metal-contaminated soil is a new alternative. Ornamental kale is an important bedding plant in many landscapes of cold temperate regions. The present study was aimed to evaluate the Pb resistance and remediation of Brassica OleraceaVar. Acephala affected by in vitro culture as a bedding plant. This experiment was carried out in 2016 at Arak University. Materials and Methods In this study, the stalk explants of Brassica OleraceaVar. Acephala was used for callus induction. Callus and plantlet were exposed to different concentrations of lead in the media. The Callus induction medium was MS (Murashige and Skoog) supplemented with 0.1 mgL-1 2,4-D and BA. The regeneration medium was MS medium supplemented with 0.5 mgL-1 NAA in combination with 1 mgL-1 BA. The rooting medium was half-strength MS medium without plant growth regulators and sucrose. All of the media containing 30 gL-1 sucrose, 7 gL-1 agar, and pH were adjusted to 5.8. All of the regeneration stages from callus induction to rooting and the acclimatization stages were done by different concentration of Pb(NO3)2 (0, 10, 25, and 50 mgL-1). The traits, such as callus browning, fresh and dry weight, callus formation, survival rate, root and shoot length, leaf number, and the lead accumulation, were evaluated during various stages of callus regeneration and rooting in plantlets. A Completely Randomized Design (CRD) arrangement with three replications was used in this experiment. Data were analyzed using the ANOVA procedure of SAS statistical software (version 9.2). Results Based on the results, not only did different concentrations of lead not have the toxic effects but also they had a stimulation effect in some concentrations on callus growth and regeneration. Lead concentrations increased leaf number and length of shoots in the regenerated ornamental kale. Lead accumulation in cultures was increased by increasing the concentration of lead in the medium at all stages of the regeneration. The highest accumulation of Pb was obtained 2395.7 ppb in callus treated by 25 mg L-1 Pb in media. Discussion The results of the present study showed the occurrence probability of the somaclonal variation due to 2,4-D supplemented in the media. In the regenerated plant, results revealed that the growth traits and Pb uptake were affected by Pb concentrations. Some in vitro derived plantlets showed an increase in uptake potential of lead in their organs (6.1 to 2.7 times compared to the control) and that it is remarkable and suitable for phytoremediation studies.

Production of Cucumber Doubled Haploid Plants via Ovule Culture

Abouzar Asadi; Alireza Zebarjadi; Mohammad Reza Abdollahi

Volume 42, Issue 1 , April 2019, , Pages 77-88

https://doi.org/10.22055/ppd.2019.23322.1514

Abstract

Abstract Background and Objectives Production of pure lines is one of the most important tools due to production of high-yield seeds. Pure lines can be produced in a short time ... Read More Abstract Background and Objectives Production of pure lines is one of the most important tools due to production of high-yield seeds. Pure lines can be produced in a short time using in vitro techniques and therefore, reduce several years in required time for conventional plant breeding programs. The purpose of this study was investigation of different concentrations of TDZ on production of haploid plants in cucumber ovule culture. Materials and Methods The experimental layout was conducted in factorial arrangement in randomized complete design with three replications in greenhouse and laboratory of Razi university during 2015-2017. Factors were two cucumber genotypes (Esfahani and Beit Alfa) and different concentrations of TDZ (0, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08 mg/l). Unfertilized ovaries were harvested 1 day before an thesis and they were sliced into 1 mm cross sections under sterile conditions and placed on solid MS medium. Immediately after placing the unfertilized ovary slices of each genotype on induction medium, they were exposed to a thermal shock pretreatment at 35±1°C for 3 days. The first visual structures formed after 3 days in culture. After two weeks of culture, the frequency of embryo formation was recorded. After embryogenesis, the embryos were sub-cultured in medium containing 1.5 mg/L-1 GA3 and finally cultured in medium containing 0.05 mg/L-1 NAA and 1.5 mg/L-1 BAP for organogenesis. Results Results indicated a statistically significant effect at 1% among TDZ concentrations. Genotypes and interaction between genotype and TDZ concentration did not have a significant effect on embryo induction. According to the results of means comparison, M8 medium with the average of 23.33 for Esfahani genotype and 20.66 for Beta Alfa genotype had the highest embryo induction and M1 showed the lowest embryo induction for both genotypes. Ploidy level of calli and embryos were identified by Flucytometry assay. Haploid embryos were transferred to the regenerative medium. The chromosome content of haploid plants was doubled spontaneously in all regenerated plants. Double haploid plants originated from megaspore and they were equivalent to haploid, which relates to homozygosity and can be used directly in breeding program. Discussion The high number of embryos was obtained in high concentration of TDZ (0.08 mg/l). Flucytometry assay is a way to recognizing ploidy level in plant tissues; thus it could be useful in choosing haploid tissue. Spontaneous chromosome doubling may occur via somatic cell fusion, endoreduplication, endomitosis and possibly many other mechanisms. In conclusion, the higher rates of embryo formation frequency achieved and doubled haploids obtained in the present study are promising for future work.

The Influence of Ultrasound and Growth Regulators on in vitro Micropropagation of Lilium Ledebourii Boiss

Z. Azimzadeh; M. Mohebodini; E. CHamani; M. Erfani

Volume 40, Issue 4 , March 2018, , Pages 11-20

https://doi.org/10.22055/ppd.2018.13439

Abstract

Abstract Background and Objectives Lilium ledebourii is one of the wild species of Lilium genus, growing in northern Iran. This species has high potential for export. Plant tissue culture ... Read More Abstract Background and Objectives Lilium ledebourii is one of the wild species of Lilium genus, growing in northern Iran. This species has high potential for export. Plant tissue culture techniques are widely used in plant propagation, and using these methods can effectively provide micro-propagation of this plant in the large scale. High percentage of regeneration is necessary for plant protection, used in the breeding programs. Materials and Methods The experiment was factorial based on completely randomized design with four replications and was carried out in tissue culture Lab of Mohaghegh Ardabili University in 2015. For this purpose, segments of explant that were treated with ultrasound were cultured on MS medium supplemented with different concentrations of NAA and BA alone and/or in combination with each other. This experiment conducted Concentrations of NAA at four level Zero, 0.01, 0.1 and 1 mgl-1 and Concentrations of BAP at four levels Zero, 0.01, 0.1 and 1 mgl-1 and Ultrasound at four levels zero, 5, 10, 20 and 30 seconds. In order to remove possible contamination from the mediums, all media were autoclaved for 30 minutes at 121 °C. At the end of the experiment, the number of bulblet produced, root length, fresh weight of bulblet and etc were recorded. The cultures were kept at 20°C or 25°C under 16 h photoperiod or in darkness. Results The results showed the highest number of bulblets produced in 0, 0.01, 0.1 mg L-1 NAA and 5secend ultrasound, 0.1 mg L-1 NAA and 0, 5, 20 and 30 second ultrasound compared with control. The highest frequency of bulblet regeneration was produced in MS medium supplemented with 0, 0.01, 0.1 mg L-1 NAA and 5 second ultrasound. On the other hand, 0.1 mg L-1 NAA and 5 second ultrasound increased number and weight of bulblets. Different concentrations of NAA had also significant effects. Discussion On the other hand, ultrasound increased the number and weight of bulblets. Mechanical stress and micro streaming by acoustic cavitation might be considered as the most possible cause of the various physiological effects of ultrasound on cells. Low-energy ultrasound can modify cellular metabolisms or facilitate the uptake of nutrient, and make them easily through the cellular walls and membranes. Finally, it seems that ultrasound in combination with growth regulators have the potential to produce the highest average number of bulblets in the scale explant, using plant growth regulators in combination with an abiotic stress in in vitro.

Effect of hormonal compositions on micropropagation of fifteen cultivars of Gerbera (Gerbera jamesonii Bolus ex Hooker f.)

M. Kharrazi; A. Sharifi; F. Keykha Akhar; A. Bagheri; Y. M. Moradian

Volume 40, Issue 4 , March 2018, , Pages 91-102

https://doi.org/10.22055/ppd.2018.13443

Abstract

Background and objectives Gerbera is one of the most important ornamental plants that used as cut flower and potted plant. The common methods of propagation does not have performance ... Read More Background and objectives Gerbera is one of the most important ornamental plants that used as cut flower and potted plant. The common methods of propagation does not have performance for supplying the global demand of this ornamental plant. So the most common method for commercial propagation of gerbera is micropropagation. Material and methods In this study, effect of different hormonal treatments and cultivars on micropropagation and rooting of gerbera shoot tip explant was evaluated. Shoot tip explants were first washed with running tap water for 30 min then they were surface sterilized by dipping in 1.5% sodium hypochlorite solution for 5 min and rinsed with sterile distilled water, followed by immersing in 0.1 % mercuric chloride solution for 5 min. After Sterilization with mercuric chloride solution, capitulum was washed with sterile distilled water. Subsequent washing was done with sterile distilled water three times. Sterilization with sodium hypochlorite solution and mercuric chloride and final rinse with sterile distilled water were done under laminar air flow hood. In the proliferation phase, MS medium containing 1 mg/l BA or KIN in combination with 0.1 mg/l IAA or hormone-free medium, 3% sucrose, 8 g/l agar was used. For rooting of propagated plantlets, ½ MS medium containing 1 mg/l IBA, IAA, 2IP or ½ MS hormone-free medium, 3% sucrose, 8 g/l agar was used. After 4 weeks different parameters such as number of plantlet, height of plantlet, number of main root, root length, number of secondary root were measured. Cultivars that was used in this experiment are as follows: 1: ‘Aventura’, 2: ‘Barones’, 3: ‘Mayfair’, 4: ‘Cacharelle’, 5: ‘Real’, 6: ‘Sorbet’, 7: ‘Dalma’, 8: ‘Goldy’, 9: ‘Panama’, 10: ‘Lancaster’, 11: ‘Dune’, 12: ‘Bellavue’, 13: ‘Blinddate’, 14: ‘Applause’, 15: ‘Sunway’. Results and discussion The results showed that in all studied cultivars, the medium containing BA lead to producing the highest number of plantlet. So using the medium containing BA in propagation phase is recommended for obtaining the highest number of plantlet. Also the results showed that the rooting of gerbera plantlets was affected by cultivar and suitable hormonal composition for rooting of each cultivar was different. However, the appropriate acclimation of produced plantlets is so important. Although the length and number of roots in ½ MS medium was at the average of other treatments, but propagated plantlets from this medium, had a suitable acclimation capacity. Moreover, additional cost for utilization of rooting hormones was reduced and therefore commercial production cost decreased. So application of this medium for rooting of different gerbera cultivars is recommended. Finally acclimation of rooted plantlet was done in cocopeat and perlite medium, with 95% success.

optimizing in vitro regeneration of Hypericum (Hypericum perforatum L.)

Z. Parsamanesh; F. Bayat; M. Hedayat

Volume 40, Issue 4 , March 2018, , Pages 103-113

https://doi.org/10.22055/ppd.2018.13440

Abstract

Background and objectives: Hypericum perforatum L. (St. John’s wort) has been received considerable interest worldwide due to its biochemical characteristics and unique secondary ... Read More Background and objectives: Hypericum perforatum L. (St. John’s wort) has been received considerable interest worldwide due to its biochemical characteristics and unique secondary metabolites. In particular, aromatic polycyclic diones, such as hypericin and pseudohypericin have an interest as their antiviral, anticancer and antidepressant activities. To date, field grown plant material has generally been used for commercial St. John’s Wort production but the quality of these products may be affected by different environmental conditions, pollutants, fungi, bacteria, viruses, and insects which can alter the concentration of medicinal metabolite. In vitro systems have been reported as an effective tool for the development of genetically uniform plants. In order to approach optimal micropropagation of Hypericum perforatum, it will be necessary to optimize shoot proliferation stage in in vitro culture. Material and methods: In the present study, in vitro regeneration of Hypericum perforatum L. using different plant growth regulators on MS media has been developed. Callus was induced from leaves and shoots explants of St. Johns Wort on MS medium containing different levels of growth regulator (0, 2 and 4 mg/l BA; 1 and 2 mg/l Kin plus 0.1, 0.2 and 0.3 mg/l 2,4-D) in factorial experiment with completely randomized design in three replication. To shoot induction equal size of formed callus were separated and evaluated on MS media containing BA (0, 0.5 and 1 mg/l) and Kin (0, 0.5 and 1 mg/l). The shoot with terminal node were used on MS media containing IBA (0, 0.2, 0.4, 0.6, 0.8 and 1 mg/l ) to root induction. Results: After 4 weeks callus and shoot/root induction of explants were measured. Data analysis indicated that leaf explants produced higher callus mass than single node explant, also in leaf explants 2,4-D increment had no significant effect on callus mass induction while in single node, callus induction increased as 2,4-D increased. In presence of BA, callus induction from both explants increased with 2,4-D concentration raising. Single node explants produced highest mass of callus in media containing 2 mg/l BA plus 0.1 mg/l of 2,4-D. The highest number of shoots and the longest shoot achieved in presence of 1 mg/l BA with 0.5 mg/l Kin and 0.5 mg/l BA plus 1 mg/l Kin. Discussion: This experiment results revealed that different explants can be used to achieve high regeneration efficiency using different level of growth regulators. Also it seems that external growth regulators isn't essential lin Hypericum branch for rooting.

Study on the effects of medium, sterilization and hormonal treatment on micropropagation of some apple (Mallus domestica Borkh.) rootstocks

N. Mohamadzadeh Moghadam; H. Hamidi

Volume 40, Issue 1 , June 2017, , Pages 41-54

https://doi.org/10.22055/ppd.2017.14039.1189

Abstract

AbstractBackground and ObjectivesIn order to increase yield in apple orchards, multiplication of rootstocks is essential, and it depends on the availability of a simple and easy proliferation ... Read More AbstractBackground and ObjectivesIn order to increase yield in apple orchards, multiplication of rootstocks is essential, and it depends on the availability of a simple and easy proliferation method of rootstocks. The study investigated the effect of disinfection, culture condition and plant growth regulators on the rate of contamination, number of lateral branches, stem length, number of leaves, callus creation and the rate of branch creation of rootstocks MM106, MM111 and B9.Materials and MethodsA factorial experiment was carried out as a completely randomized design with three replications. Surface sterilization of explants with different concentrations of mercuric chloride and sodium hypochlorite (mercuric chloride 0.1% for 3 minutes and ethanol 70% for 30 seconds; mercuric chloride 0.1% for 5 minutes and ethanol 70% for 30 seconds; NaOCl 0.75% for 15 minutes and ethanol 70% for 30 seconds) was done and then explants were cultivated in modified MS, WPM and DKW mediums. The evaluated characters were infection percent of explant, number of lateral branches, stem length, percentage of callus induction and number of leaves. Data were analyzed using SAS software. The analysis of variance on the test data was performed at 5% level and comparison to the middle of the Duncan test.ResultsResults showed that using a mercuric chloride (0.1%) and ethanol (70%) respectively for 3 Minutes and 30 Seconds achieved the least contamination. Results showed significant differences between plant hormone and rootstocks for traits. In this test, the modified MS medium with 0.1 mg/L BA hormones with the largest percentage growth had more successful establishment. Average comparisons showed that rootstock MM106 in terms of all traits had a significant difference with other rootstocks. The highest stem length, number of leaves and callus creation were shown in rootstock MM106. In order to investigate the effect of hormone levels on branch creation, different levels of BAP (6-Benzylaminopurine) and IBA (Indole-3-butyric acid) were used. The best medium was the culture containing 0.1 milligrams per liter IBA plus 2 milligrams per liter BAP.DiscussionsIn general, the present results showed that genotypes respond differently to in vitro conditions. The rootstock B9 by the above method showed better response to branch creation trait than other genotypes.

Effect of Silver Nitrate on In vitro Propagation of Pomegranate (Punica granatum L.) cvs. ‘Malas Yazdi’ and ‘Robab’

babak valizadeh kaji; Ahmad Reza Abbasifar

Volume 39, Issue 4 , March 2017, , Pages 53-64

https://doi.org/10.22055/ppd.2016.12430

Abstract

Background and Objectives Traditional propagation method of pomegranate by hard wood and soft wood cuttings does not ensure disease-free and healthy plants and it depends on the season. ... Read More

Effect of Ovary Conditioned Medium on Callogenesis and Gametic Embryogenesis in another Culture of Different Cultivars of Cucumber (Cucumis Sativus L.)

S. Najafi; M.R. Abdollahi; H. Sari Khani; S.S. Moosavi

Volume 38, Issue 4 , March 2016, , Pages 105-116

https://doi.org/10.22055/ppd.2016.11846

Abstract

In this research, the effect of ovary co-culture on androgenesis efficiency in anther cultures of four cultivars of cucumber was studied. A factorial experiment based on Completely ... Read More

Evaluation response of three apple rootstocks to drought stress in vitro

S. Jalali Samghabadi; R. Jalili Marandi

Volume 38, Issue 3 , December 2015, , Pages 65-78

https://doi.org/10.22055/ppd.2015.11454

Abstract

Water deficit is one of the most important environmental factors that limit crop productivity. Therefore, the need for evaluation of crop tolerance to drought has become urgent. It ... Read More

Volume 47 (2024)

Volume 46 (2023)

Volume 45 (2022)

Volume 44 (2021)

Volume 43 (2020)

Volume 42 (2019)

Volume 41 (2018)

Volume 40 (2017)

Volume 39 (2016)

Volume 38 (2015)

Volume 37 (2014)

Volume 36 (2013)

Volume 35 (2012)

Volume 34 (2011)

Volume 33 (2010)

Volume 32 (2009)

Main Subjects = H: Tissue Culture

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