Breeding and Biotechnology of Plant and Flower
Reza Shafiee; Mohammad Reza Abdollahi; Asghar Mirzaie Asl; Seyyed Saeid Moosavi; Hassan Sarikhani
Abstract
Introduction
Given the economic importance of growing flowers and plants in the world, the use of new technologies and methods in the improvement of ornamental plants in order to market them can play a significant role in marketing of these products and their trade in the international markets. Marigold ...
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Introduction
Given the economic importance of growing flowers and plants in the world, the use of new technologies and methods in the improvement of ornamental plants in order to market them can play a significant role in marketing of these products and their trade in the international markets. Marigold (Tagetes spp.), is an annual and essential plant that is cultivated as an ornamental plant in many parts of the world (Neher, 1968). But in recent years, marigolds have been used as a commercial source of essential oils, biological compounds, pigments and cosmetics (Anonymous, 1976), control of malaria mosquitoes (Wanzala and Ogoma, 2013), antihypertensive, anti-inflammatory, antidepressant, antimicrobial, antimicrobial (Senatore and Feo, 1999), and control of nematode (Prasad et al., 1992). Anther or microspore culture is known to be an effective method for producing haploid plants (Henry and De Baizer, 1980). Hybrid seed production requires pure line (as a parent), and the double haploid method can reduce the production period of pure lines to 5-6 years. Production of hybrid seeds in this valuable plant is of great importance, and the double haploid method can be important in this regard.
Materials and Methods
In the anther culture of marigold, a culture medium containing 0.2 mg/l of Naphthalene acetic acid and 1 mg /l of 6-Benzylaminopurine was used. In this study, the effect of genotype, bud size and mannitol on androgenesis induction of marigold anthers was evaluated during two separate experiments. A factorial experiment was conducted in a completely randomized design with two factors including genotype at 5 levels (T1, T2, T3, T4, T5) and bud size at three levels (3-5, 5-10 and 10-15 mm) for first experiment. The second experiment was also performed as a factorial in a completely randomized design with 3 replications. In the recent experiment, the first factor included the method of mannitol application and the second factor included the different concentrations of mannitol (0.1, 0.2, 0.3, 0.4 and 0.5 M). The factor of application method was in 2 levels: 1: adding different concentrations of mannitol to the solid culture medium and 2: pre-treating the anthers with the liquid culture medium containing different concentrations of mannitol for 24 hours. The T4 genotype was used in the second experiment.
Results
In the first experiment, the effect of different bud sizes and 5 different genotypes on callus formation, mean number of shoot per anther, mean number of shoot per callus and percentage of complete plant regeneration in anther culture of marigold were studied. The results of this experiment showed that buds with the length of 5-10 mm have anthers which their microspores are at the proper growth and development stage for callogenesis and shoot production. The T3 and T4 genotypes, (both of them belonging to French species, Tagetes patula), produced the highest percentage of plant regeneration among the various cultivars. In the second experiment, we explored the impact of mannitol treatment on androgenic traits in marigold anther culture. Specifically, we examined two concentrations: 0.1 M and 0.2 M mannitol, both applied in the form of solid culture medium. Additionally, we investigated two concentrations, 0.0 M and 0.2 M mannitol, when applied as a pre-treatment in a liquid medium containing mannitol. These treatments yielded the highest percentage of callus formation. While the pre-treatment of anthers with a liquid culture medium containing 0.5 M mannitol led to the highest mean number of shoot per anther and the mean number of shoots per callus. Also, the pre-treatment with liquid medium containing 0.2 M mannitol showed the highest percentage of complete plant regeneration.
Conclusion
Results showed that in marigold, buds with the size of 5-10 mm contained microspores with mid-uninucleate stage to early bi-nucleate stage showed the highest response to the induction of androgenesis. Also, T3 and T4 genotypes belong to the French species showed the highest response to the regeneration. In another experiment, the pre-treatment of anthers with 0.2 and 0.5 M mannitol by using mannitol in a liquid culture medium for 24 hours, respectively showed the highest percentage of complete plant regeneration and the highest mean number of shoot per callus and anther. Chromosome counting results showed that 3 out of 5 examined plants were dihaploid and had 24 chromosomes in their root tip cells, while examined mother plants were tetraploid and showed 48 chromosomes in their root tip cells.
Azam Rahimian; Mahmood Esna-Ashari; Hassan Sarikhani
Abstract
Introduction: Nowadays, crop production methods are moving towards organic farming through reducing the use of chemicals in agriculture. The new generation of seaweed extracts, like Actiwave, could be a promising approach to achieve a part of this goal. Actiwave is a metabolic enhancer derived from a ...
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Introduction: Nowadays, crop production methods are moving towards organic farming through reducing the use of chemicals in agriculture. The new generation of seaweed extracts, like Actiwave, could be a promising approach to achieve a part of this goal. Actiwave is a metabolic enhancer derived from a type of brown algae, called Ascophyllum nodosum, and contains compounds that play an important role in plant metabolism. The application of this product on various plants has often improved their vegetative and reproductive characteristics. One of the important problems in our agricultural lands is the presence of bicarbonate ion in soil (calcareous soils) or irrigation water, which increases pH around plant’s root followed by chlorosis between the veins in the young leaves resulting in a reduction or halt in plant growth. The induction of chlorosis in calcareous soils is attributed to iron deficiency due to reduction of iron absorption or availability. The use of biostimulants may help to improve plant growth under such conditions. The aim of this study was to investigate the possibility of replacing iron chelates in nutrient solution with Actiwave in two optimal and alkaline pH and its effect on some of the vegetative and biochemical properties of two strawberry cultivars in a soil-less system.
Materials and Methods: Strawberry seedlings of Camarosa and Salva cultivars were cultivated in pots containing coco-peat and perlite (1:1), followed by plant’s irrigation and nourishment through a plant nutritional program. The project was conducted in a factorial experiment (with two factors) based on a completely randomized design with three replications. The first factor was strawberry cultivar in two levels including Camarosa and Salva, and the second factor was iron treatment in four levels consisting of Hoagland nutrient solution containing iron chelate (pH=6), iron-free nutrient solution containing 0.25 ml/l Actiwave (pH=6), Hoagland nutrient solution containing iron chelate (pH=8), and iron-free nutrient solution containing 0.25 ml/l Actiwave (pH=8). Plants were fed for two months, at the end of which, the roots and shoot’s fresh and dry weight, chlorophylls a, b and total as well as carotenoids contents, catalase and ascorbate peroxidase activities in fresh leaf samples and also total Fe and active Fe in dried leaf and root samples were measured.
Results and Discussion: The results showed that the algae extract had a significant effect on all the measured parameters, so that fresh and dry weights of the aerial parts, fresh weight of the roots, as well as chlorophyll and active iron content of leaves in the treatment containing Actiwave with pH=6 was similar to the control plants. Evaluation of the activity of catalase and peroxidase enzymes in the leaf also showed that Actiwave application reduced iron deficiency stress in plants increasing the activity of these enzymes under such conditions. The reason behind these effects can be due to the Actiwave ingredient content, which includes betaine, alginates and kahydrin (derived from vitamin K). Since betaines have cytokine activity, they can increase root growth by increasing the amount of cell division. Increasing the chlorophyll content, which can be the result of reduced chlorophyll degradation, is probably due to the presence of betaines in the seaweed extract. In addition, the presence of kahydrin and alginic acid in Actiwave, with the acidification of the rhizosphere, stimulated the release of more ions around the roots, resulting in more ions received by the plant and increased their assimilation. As observed in this study, It has been proven that iron concentration in the leaf is not always an appropriate indicator for checking the state of iron in plants. Some studies attribute this to limiting the growth of young leaves and the accumulation of iron in tissues or inactivation of iron in tissues occurred through the process of alkalization in leaf apoplast. It is found that activity of catalase and ascorbate peroxidase containing iron and the iron extracted with hydrochloric acid are the most effective indices for checking the iron status in plants, and the concentration of the total iron is less important in this regard. According to the results of this study, algae extract was able to replace iron chelate in nutrient solution for growing strawberry, and so made it possible to produce this fruit by reducing the use of synthetic iron compounds. In other words, application of Actiwave instead of iron chelate is recommended in nutrient solution with pH=6.
Amir Hossein Keshtkar; Noshin Fallahi; Mohammad Reza Abdollahi; Hassan Sarikhani; Hooshmand Safari; Zhaleh Mohseni Araghi
Abstract
Introduction: Polyploidy plays an important role in creation of genetic variability. Polyploidy induction by mutagenic chemicals such as colchicine is considered to enhance the potential of secondary metabolites production in herbs breeding. Colchicine is the most effective chemicals used in the polyploidy ...
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Introduction: Polyploidy plays an important role in creation of genetic variability. Polyploidy induction by mutagenic chemicals such as colchicine is considered to enhance the potential of secondary metabolites production in herbs breeding. Colchicine is the most effective chemicals used in the polyploidy induction studies. The effect of colchicine is to form cells with two or multiply number of chromosomes resulting in a lack of germination and death of a large number of plant samples. Flow cytometry analysis and cytogenetic studies were effectively used to assess the ploidy levels for fenugreek (Trigonella foenum-graecum) plants. In the beginning of 90 decade, a new type of adsorption technique called solid-phase micro extraction (SPME) has been developed by Pawliszyn and co-workers. This method compared to the traditional techniques, offers many advantages such as the high sensitivity and reproducibility, does not require solvent, and combines extraction and pre-concentration in a single step without pre-treatment of samples. Moreover, it is a fast and inexpensive method, requires low sample volume and it can be easily automated. Solid-phase micro extraction (SPME) uses a fine rod (fused silica or metal) with a polymeric coating to extract organic compounds from their matrix and it directly transfer them into the gas chromatograph injector for thermal desorption and analysis (Kaykhaii, 2008). The aim of this study was to investigate the effect of colchicine treatment on ploidy levels and compare some of the morphological, physiological, cytogenetical, flow cytometric analysis and biochemical characteristics in diploid and tetraploid fenugreek plants.
Materials and Methods: In order to investigate the effect of polyploidy induction by colchicine on Trigonella foenum-graecum medicinal species, an experiment was planned as a factorial completely randomized design with five concentrations of colchicine (0, 0.05, 0.1, 0.2, and 0.5%) for 12, 24, 48 and 72 hours. In this experiment the effect of colchicine was examined on the percentage of survival and tetraploidy of seed, root and terminal bud samples. Level of ploidy was identified in survival explants through root tip chromosome counting and flow cytometry of leaf samples. In addition to distinguish tetraploid from the diploids plants, morphological, physiological and biochemical characteristics were considered in treated plants. SAS and SPSS software programs were used to analysis of variance and comparison of means by Duncan's multiple test. Graphs were also drawn by EXCEL software.
Results and Discussion: The analysis of variance showed that all characteristic factors for survival percent and mixoploidy percentage were statistically significant. Survival percentage was decreased with increasing of colchicine concentration and increased exposure time of colchicine-treated seed. After the observation of morphological changes, the samples were considered to assess the ploidy levels by flow cytometry system. Results showed that 0.5% colchicine concentration had the highest survival rate after control treatment for the terminal bud. The highest percentage of mixoploidy was also observed in treated terminal buds with 0.1 and 0.2% of colchicine concentrations. Morphological, physiological, cytogenetic, flow cytometric analysis and biochemical studies confirmed that terminal bud treatment with 0.2% colchicine for 72 hours is the most effective treatment to induce tetraploidy in fenugreek plant. The results of GC/MS also indicated an increase in secondary metabolites content, but traits including growth rate and plant height of tetraploid reduced compared to the diploid plants. Result of this study showed a significant increase in chlorophyll a, b and total chlorophyll contents of tetraploid plants, which were higher than the levels of diploid plants.
Conclusion: Polyploidy induction using mutagenic chemicals is one of the methods to enhance the production of plant secondary metabolites. Colchicine is the most effective mutagenic chemical in inducing plant polyploidy. Although, flow cytometry is an expensive method, it is increasingly used for ploidy screening by analyzing of nuclear DNA content. In this study, both flow cytometry and chromosome microscopic examinations were used to test ploidy. The two methods were compared, and it was found that flow cytometry testing was fast and labor saving, especially in case of a large number of samples. Tetraploidy induction significantly affected different morphological, physiological, and biochemical characteristics of Trigonella foenum-graecum. These changes suggested that ploidy manipulation as a rapid and effective method for enhancing genetic diversity and metabolite production for this plant. SMPE method offers a number of practical advantages: smaller sample volume, simplicity of extraction and low cost, when compared to the other methods that are currently being used.
Roohallah Montaghemi Rad; Ebrahim Ahmadi; Hassan Sarikhani
Abstract
Introduction: The maintenance of the quality of fresh produce is still a major challenge for the consumers. The most important qualitative attributes contributing to the marketability of fresh fruit a reappearance, color, texture, flavor, nutritional value and microbial safety. Storage conditions of ...
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Introduction: The maintenance of the quality of fresh produce is still a major challenge for the consumers. The most important qualitative attributes contributing to the marketability of fresh fruit a reappearance, color, texture, flavor, nutritional value and microbial safety. Storage conditions of product after harvest and storage duration are important factors with specific effect on products qualitativeproperties such as acidity, soluble solids materials, phenolic compounds and density. The amount of soluble solids materials is an important qualitative factor that is directly related to fruit quality and ripening.The existence of phenolic compounds, phenol absorption and changes phenol in product has caused changes in color. Shape, size, arithmetic mean diameter, geometric mean diameter, mass, volume, density, and sphericity coefficient are some of the physical properties of products that are crucial to analyze the behavior of matter in the handling.Furthermore, adequate knowledge about the physical properties of the products is of high importance in machines design, transportation and processing of product.
Materials and Methods: In this study, two types of olives, which were bitter and sweeten with 1.5%NaOHbefore the storage, were collected and their physical properties (length, width, thickness, arithmetic and geometric mean diameter, sphericity coefficient, mass, volume, density and primary moisture) were evaluated. Then, tostudy the effect of storage duration,the sweetened olives were stored at two temperatures (25℃ and 4℃temperature of the refrigerator) for 3 months,and during this period, sampling was performed at 10-dayintervals and the amount of total acidity (TA was determined by diluting each 1 ml aliquot of strawberry juice in 10 ml distilled water and then titrated to pH 8.1 using 0.1 N NaOH), total soluble solids materials (TSS was determined by a handheld refractometer device and expressed as ° Brix), phenolic compounds and the density weremeasured. The study was carried out as factorial based oncompletely randomized designwith twoindependent factors of temperature and storage time.
Results and Discussion: Results obtained for physical properties showed that the means of length, width andthickness insweet olive were 9.72, 1.34 and 1.11%respectivelyhigherthan bitter type. Penetration and absorption of NaOH and processing methods of the product can be considered among the possible causes of this increase. Linear regression relations between physical properties for sweet and bitter olives showed that mass and volume of fruit areestimated accurately according to the main dimensions. The results of data variance analysis also showed thatdensity and soluble solids materialswere significantly affected by themain and interaction effects of the twofactors at 1% probability level. Moreover, the main effect of storage time on total acidity and phenolic compoundswassignificant at 5% and 1% levels, respectively. With increasing storage time, density of sweet olive increased and soluble solids materials and phenolic compounds decreased. Acidity changes of olive during storage fluctuated, and an increase was found in the values of density and soluble solids materials along with theincrease of storage temperature from 4℃ to 25℃.With respect to density, the third month of storage showedhigherincrease compared toother months, and maximum value was reported for sweet olive in temperature of 25℃ and 19thday. Increase ofstorage duration resulted in the reduction of moisture content,whichin turns leads to the reductionof product volume and increase of density. The reductionfound in acidity in the second and third months of storage resultedfrom maturation and ripening processes of the fruit. Organic acids are a source of energy,which are consumed in the time ofmetabolism increaseand fruit ripening.Decreasein phenolic compounds also occurs as the result ofphenol penetration from olive flesh into the water - salt solution.
Conclusion: Results of this study showed that length, width, thickness, mass and volume of sweet olive weregreaterthan bitter type, but,by contrast,sphericity coefficient was higher in bitter olive. Moreover, storage time, increase caused an increase in thedensity of sweet olive and decrease insoluble solids materials and phenolic compounds.There was fluctuation inacidity during different storage times. An increase was found in the values of density and soluble solids materials along with the increase in storage temperature from 4℃ to 25℃.
Azam Seddighi; M. Gholami; Hassan Sarikhani; Ahmad Ershadi
Abstract
The use of compounds inhibiting ethylene production is very important in post-harvest technology of fruits. Sweet cherry is a non-climacteric fruit with capability of ethylene production during storage period. Factors that decrease ethylene production would increase fruits storage life. In this research ...
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The use of compounds inhibiting ethylene production is very important in post-harvest technology of fruits. Sweet cherry is a non-climacteric fruit with capability of ethylene production during storage period. Factors that decrease ethylene production would increase fruits storage life. In this research sweet cherry fruits of Mashhad cv. were sprayed with different concentrations of Salicylic Acid (0.5, 1, 2, and 3 mmol/l) and Gibberelic Acid (0, 10, 20 and 30 mg/l). Fruits were sprayed about three weeks before harvest, when their color started to change from green to yellow. The experiment was performed in a completely randomized design with three replicates. The results showed that GA had less effect on decreasing ethylene production compared to SA, but did better on anthocyanin accumulation, fruit size, soluble solids content and fruit weight. GA treatment delayed fruit ripening and ethylene production but had little or no effect on the fruit color as an important ripening index. During storage period also the total amount of anthocyanin in fruits increased due to hormone treatments whereas the ethylene production decreased compared to control fruits.
