Medicinal Plants
S. Sadeqifard; M. Azizi; S. Karimi Zinkanlu
Abstract
IntroductionGrowth regulators play a vital role in developmental stages of plants and their use can be improved the yield of crops. One of the most important compounds that used as growth regulators in recent years is salicylic acid. The application of salicylic acid in the activation of systemic acquired ...
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IntroductionGrowth regulators play a vital role in developmental stages of plants and their use can be improved the yield of crops. One of the most important compounds that used as growth regulators in recent years is salicylic acid. The application of salicylic acid in the activation of systemic acquired resistance, and metabolite synthesis and antioxidant enzymes has been proved. Also, salicylic acid as a natural compound has potential to prevent ethylene production and its effect. In order to evaluate the effect of salicylic acid in different concentration on yield and oils content of Ziziphora, a split plot experiment with three replications during 2014-2015 in the field of Agricultural Faculty of Shirvan and Ferdowsi University of Mashhad laboratories was conducted. Materials and MethodsThis research was conducted during 2014-2015 under field conditions in a split plot design in time, at Shirvan Agricultural Research Station (latitude 40 º / 37, longitude 93 º / 57 and with a height of 1097 m above sea level) and in laboratory Department of Horticultural Sciences, Ferdowsi University of Mashhad. Each plot had an area of 4 square meters in furrow planting way which the distance between rows were 50 cm and on rows 15 cm and were carried out on 25 March 2014. Irrigation was carried out immediately after planting, and a second irrigation was done 20 days later, once the seeds had germinated. Irrigation and weed control are also done manually once a week. Salicylic acid (Sigma Aldrich, 99.5%) in the three phases of plant growth (The onset of vegetative growth, throughout vegetative growth, and at the flowering stage), at concentrations (0, 10-2, 10-4 and 10-6 M) in three repeats were sprayed. The first spraying (The onset of vegetative growth) was done on April 3, 2014, the second spraying (throughout vegetative growth) on May 4, and the third (at the flowering stage) in early June 2014. The plants were harvested on July 20. To measure the dry weight of the plants, the samples were dried in the shade for a week. Essential oil content was determined using a 30-gram dry sample in 500 ml of water through hydro-distillation with a Clevenger apparatus for 4 hours. For biochemical characteristics first samples extract were obtained. To prepare extracts, dry samples were milled then one gram of each sample was transferred to the 50 ml Erlen and 10 ml of 80% methanol was added and shaken for 24 hours. After 24 hours the extract was filtered using filter paper then used for measuring phenols, flavonoids and antioxidant activity. Phenol measurement: phenolic content was measured using the Folin reagent. In 5.0 mL of each extract (10 milligrams per ml) 5.2 ml Folin reagent was added. After five minutes, 300 ml of a molar sodium carbonate solution is added and maintained in a 40 °C bath for 30 minutes. The sample absorbance at 760 nm was measured by a spectrophotometer against the blank. Gallic acid was used as the standard for calibration curve. Flavonoids measurement: Flavonoid content of the samples were measured using aluminum chloride reagent. In 5.0 ml of methanol extract (10 milligrams per ml), 5.1 ml of methanol, 1.0 ml of 10% aluminum chloride, 1.0 ml of a molar potassium acetate and 8.2 ml of distilled water was added. The mixture kept in dark for 30 minutes and absorbance at a wavelength of 415 nm was read against the blank. Quercetin was used as the standard for calibration curve evaluation. JMP statistical software was used to analyze the data. Results and DiscussionBased on the results, the application of salicylic acid significantly affected fresh and dry weight, plant height, and phenol and flavonoid content (P < 0.01), but had no significant effect on essential oil percentage or yield. The highest fresh weight (264.763 grams per square meter) and dry weight (93.11 grams per square meter) were obtained with the application of 10⁻² molar salicylic acid during vegetative growth, while the lowest fresh and dry herb yields were observed in the control group. Maximum height of plants (29.01 cm) was related to 10-2 M salicylic acid during vegetative growth and the lowest of plant height (23/24 cm) was related to control treatment at the time of flowering. The highest amount of phenols (0.504 mg equivalents gallic acid per gram of dry weight) was detected in plants treated with 10-2 M salicylic acid during vegetative growth and the lowest phenols (0.248 mg equivalents gallic acid per gram of dry weight) was reported in 10-2M salicylic acid and during vegetative growth. The highest amount of flavonoids (0.31 mg equivalent Quercetin per gram of dry weight) was obtained in 10-4 M salicylic acid treatments during vegetative growth and the lowest of them (0.176 mg equivalent Quercetin per gram of dry weight) was in 10-6 M salicylic acid treatment at the beginning of vegetative growth. ConclusionsIn conclusion the results showed that the highest amount of fresh and dry weight and plant height was under the conditions of 10-2 M salicylic acid concentration and during vegetative growth and the best yield of essential oils, phenols and flavonoids in during vegetative growth and 10-4M salicylic acid concentration was obtained.
Medicinal Plants
Zhaleh Zandavifard; Majid Azizi
Abstract
Introduction: St. John’s wort (Hypericum perforatum L.) is a medicinal plant which used mainly in treatment of mild depression, neurological disorders and has been recently shown to have anticancer potential. The principle medicinal components of St. John’s wort are hypericin, pseudohypericin, ...
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Introduction: St. John’s wort (Hypericum perforatum L.) is a medicinal plant which used mainly in treatment of mild depression, neurological disorders and has been recently shown to have anticancer potential. The principle medicinal components of St. John’s wort are hypericin, pseudohypericin, and hyperforin. Light is one of the most important environmental factors affecting plant growth, survival, reproduction and distribution. The light quality, light intensity, duration and photoperiod directly affect plant growth. Light quality refers to the color or wavelength reaching the plant's surface. A prism (or raindrops) can divide sunlight into respective colors of red, orange, yellow, green, blue, indigo and violet. Red and blue have the greatest impact on plant growth. Green light is least effective (the reflection of green light gives the green color to plants). Blue light is primarily responsible for vegetative leaf growth. The principle objective of the current study was to evaluate the effects of different spectral quality including red, blue, green and white on the growth factors and production of hypericin.Materials and Methods: This experiment was conducted on the basis of Completely Randomized Design with four treatments and 10 replications in the growth chamber in the Department of Horticulture, Ferdowsi University of Mashhad (FUM), Iran. In this study, seeds were obtained from the research greenhouse of FUM. Seeds after soaking in running water for 24 hours were planted in small pots (250g). After the seedlings have reached to height of 25cm, each 10 pots were put inside the boxes (20×30cm) made of colored filters. Experiment continued in a growth chamber with day and night temperature 25 and 21°C, respectively, relative humidity 45%, 16 hours of light with the intensity of 1000 lux and 8 hours of darkness for 50 days. Then morphological parameters including plant height, number of stems, number of leaves, number and length of internodes, fresh and dry weight of shoot and root were measured. To count the number of black nodules, the upper, middle and lower parts of seedling were evaluated individually. Hypericin content of the H. perforatum plantlets were measured according to the previous work of Azizi & Omidbaigi, 2002. Data were analyzed statistically by using SAS and Excel software. The significant differences between means were assessed by Tukeyʼs test at P < 0.05.Results and Discussion: The results showed that morphological parameters including plant height, leaf number, internode length, root fresh weight, dry weight of stem and root were affected significantly by light treatments. Minimum and maximum of plant height was related to white and red lights, respectively. Increasing plant height under the influence of red light was due to the variation in levels of growth regulators. Red and blue light by changing of the GA hormone level in the plant and affected elongation of plant stem. Minimum and maximum of leaf number was related to blue and white light respectively. Also, other morphological traits including internode length, fresh and dry weights of stem and root showed significant differences. Internode length for white light was less than red, green and blue light. Maximum of the fresh and dry weight of shoot and root of seedlings was observed under white light. Results suggest that photosynthetic compounds move in plant under the influence of light quality. Also the number of black nodules in three different parts of plant and hypericin content were compared in plantlets under the effect of light quality. Results analysis also confirmed that different lights had the significant impact on the number of black nodules in upper and middle leaves of H. Perforatum seedlings. Seedlings treated with the red light had the highest number of black nodules in the middle section of H. perforatum. In the top third of St. John’s wort seedlings, red, blue and green light was inducted the highest number of black nodules formation than white light. The highest level of hypericin was related to red, white, blue and green lights, respectively. The number of black nodules in the plant and the hypericin content have positive correlation. It seems that in the seedlings treated with the red light, carbohydrates made from photosynthesis most used in biosynthesis of secondary metabolites than plant growth. In fact, carbon allelochemical compounds such as terpenes and phenolics have such metabolism direction to explain the increasing in secondary metabolites.Conclusion: Different light spectra affects plant hormones levels and with alteration in the primary and secondary metabolites lead to a change in the morphological and biochemical traits of plant. In general, it can commented that the quantity and quality of light is able to affect the growth and the active metabolites of medicinal plants and using red and white mixture of light during seedling growth period have an effective role on generation of more strong seedlings with higher potential production of active ingredients.
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.
Roghayeh Fathi; Mehdi Mohebodini; Esmaeil Chamani
Abstract
Introduction: Transformed hairy roots obtained by infection of plants with Agrobacterium rhizogenes strains are used as a tissue culture tool for secondary metabolite production since hairy roots are genetically and biologically stable and they are able to produce metabolite within a short time. Chicory ...
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Introduction: Transformed hairy roots obtained by infection of plants with Agrobacterium rhizogenes strains are used as a tissue culture tool for secondary metabolite production since hairy roots are genetically and biologically stable and they are able to produce metabolite within a short time. Chicory (Cichorium intybus L.) is a medicinal plant from Asteraceae. This plant contains many important metabolites include chicoric asid, inulin, scoline, coumarin and flavonoids.
Materials and Methods: The seeds were surface-sterilized by rinsing with 2% benomil for 30 min and later sterilized with 5% sodium hypochlorite for 20 min and subsequently with 70% ethanol for 90 s. the wounded leaf of 20 and 28-day-old seedlings inoculated with Agrobacterium rhizogenes 11325. Bacterial colonies were cultured on liquid LB medium for 1 day. The explants were submerged in the solution for 15 min and were shaken gently. After that, the explants were drained on sterile filter paper and then transferred to MS solid medium. The explants were incubated at (25 ± 2) ◦C under dark condition for three different co-cultivation period (24, 48 and 72 hours). In the next step, the explants were transferred to MS solid medium supplemented with 500 mg/L cefotaxime. The explants were incubated at (25 ± 2) ◦C under a 16-h photoperiod condition for four weeks. At the end of incubation time, the percentage of hairy root induction, root number and root length were investigated. Genomic DNA of Cichorium intybus L. hairy root was extracted following the method of CTAB and subjected to PCR analysis. Non-transformed root DNA was used as a negative control and pRi11325 plasmid DNA was used as positive control. The pair of primers specific to the rolB fragment sequences was: 5-ATGGATCCCAAATTGCTATTCCCCACGA -3 and 5-TAGGCTTCTTTCATTCGGTTTACTGCAGC-3. The amplification protocol for the rolB was a 5 min melting at 94 ◦C followed by 35 cycles of a 5 min melting at 94 ◦C, a 45 s annealing at 55 ◦C and a 1 min elongation at 72 ◦C, and final elongation at 72 ◦C for 7 min. PCR products were electrophoretically separated on 0.8 % agarose gels (w/v) and stained with gel red. To determine the best media composition for growth of hairy roots, approximately 100 mg FW roots were cultured in basal liquid MS, solid MS and liquid 1/2MS medium supplemented with 500 mg/L cefotaxime on a rotary shaker (90 rpm) at 25 ◦C in dark for 5 weeks, and increase in fresh weight and dry weight was recorded at 5 week later. Three replicates were made for each experimental set. Non-transformed roots were cultured on same mediums.
Results and Discussion: Each of the two different explants – leaf and petiole showed different levels of hairy root induction in different co-culture time. Hairy root induction was observed 7 days after co-cultivation. There were low adventitious roots formed from control explants. 72 hours co-cultivation period was found to be more efficient in inducing hairy root phenotype in both explant types. The infection of explants by A. rhizogenes is a process that implies a succession of events, including the transfer and integration of T-DNA, into the host plant genome. Therefore, the time of contact between A. rhizogenes and the explants determines the transformation efficiency. A maximum of transformation frequency (53.33%) was observed in leaf explants in 72 hours co-cultivation followed by 33.33% in petiole explants in 72 hours co-cultivation. In leaf explants maximum root number (8.5 roots per explant) and maximum length of root (9.16 cm) induced from 20-day-old seedling in 72 hours co-culture. In petiole explants maximum root number (3.66 roots per explant) and maximum root length (11.46 cm) induced from 28-day-old seedling in 72 hours co-culture. The “hairy root” phenotype characterized by high branching and fast growth was previously reported in “reactive species” like Datura innoxia but not for “difficult species”. The high phenolic content of woody plant species could be responsible for the reduced hairy root phenotype as reported for gingko or pine. Three different culture media (solid MS, liquid MS and 1/2 MS media) were tested to determine the best suitable media for the maximum Biomass of hairy roots line.
Conclusions: Genetically transformed hairy roots obtained by infection of plants with Agrobacterium rhizogenes are suitable source for production of bioactive molecules due to their genetic stability and generally show fast growth in culture media free of growth hormones. This study describes the protocol for hairy root induction which could further be useful for the production of secondary metabolites and biomass.
Sadrohlah Ramezani; Alireza Abbasi; Abdolali Shojaeiyan; Noorollah Ahmadi; Rozariya Cozzolino; Soniya Piacente
Abstract
Introduction: There are 58 species belonging to sage genus as annual and perennial plant in different regions of Iran that 18 species of them are endemic to Iran and they have different medicinal properties such as antibiotic, sedative, carminative, antispasmodic and commonly used in treatment of respiratory ...
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Introduction: There are 58 species belonging to sage genus as annual and perennial plant in different regions of Iran that 18 species of them are endemic to Iran and they have different medicinal properties such as antibiotic, sedative, carminative, antispasmodic and commonly used in treatment of respiratory problems: infections, cough, cold and sore throat and cosmetics industries. The present study has aimed to evaluate the composition of essential oils achieved by Solid Phase Micro-Extraction method from aerial parts of two Salvia species native to Iran: Salvia limbata and Salvia multicaulis.
Materials and Methods: The experiments were carried out at the Research Station of Agriculture College, TarbiatModares University in Tehran, Iran during the years 2011-2013. The seeds of Salvialimbata and Salvia multicauliswere collected in Ardabil and Isfahan provinces in 2009. The seeds were sown in planting trays(filled with soil and cocopeat 1:1) under controlled greenhouse condition (temperature: 26±1°C, light: 3000 lux, relative humidity: 65%) in the last week of February 2011. The soil of experimental pots (soil and coco peat 2:1) was a clay silt loam with pH of 7.4. After two months,seedlings with uniform height and stem diameter with two true leaves were transferred to a growth chamber adjusted to 30/20 °C, 50% relative humidity, light intensity of approximately 3000 Lux and 16 h photoperiod.Aerial parts of two cultivated plantsincludingSalvia limbata and Salvia multicaulisat flowering stage were harvested in June 2012 and kept at 80°C until further experiments. Volatile compounds were extracted by solid phase micro-extraction (SPME) method for the first time in Iran for these species. Before the SPME, the leaves lyophilized and then were used. The optimization of SPME extraction and desorption conditions were performed by analyzing dried leaves of Salvia officinalis L., used as the matrix. The sample preparation procedure was as: 15 mg of dried sage leaves mixed into a 20 ml screw-on cap HS vial to 5 ml of 5% Ethanol and 0.5 mg of Na2SO4. The vials were sealed after stirringwith a Teflon (PTFE) septum and an aluminum cap (Chromacol, Hertfordshire, UK) for the production of headspace and the successive analysis. The sample vial was put in the instrument dry block-heater and held at 40°C for 20 min to come into equilibrium. The extraction and injection processes were automatically performed using an auto sampler MPS 2 (Gerstel, Mülheim, Germany). The fiber was, then, automatically inserted into the vial’s septum for 10 min, to allow the volatile compounds absorption onto the SPME fiber surface. Each SPME fiber was conditioned before its first use, as recommended by the manufacturer. In order to desorb the volatile metabolites, the SPME fiber was introduced into the injector port of the gas chromatograph device, model GC 7890A, Agilent (Agilent Technologies, Santa Clara, USA) coupled with a mass spectrometer 5975 C (Agilent) wherein the metabolites were thermally desorbed and transferred directly to a capillary column HP-Innowax (30m×0.25 mm×0.5µm Agilent J&W) and analyzed. The identification of VOMs was accomplished by comparing the retention times of the chromatographic peaks with those, when available, of authentic standards run under the same conditions. For volatiles for which reference substances were not available, the identification was performed by matching their retention indices (RI) determined relative to the retention time of a series of n-alkanes (C8–C20) with linear interpolation, with those of authentic compounds or literature data (Van Den Dool&Kratz, 1963). Confirmation of metabolites identification was also conducted by searching mass spectra in the available database (NIST, version 2005; Wiley, version 2007).
Results and Discussion: 66 volatile components were identified in Salvia limbata, which Sabinene (19.16%), β-Pinene (19%), α-Pinene (16.3%), α-Terpinolene (14.41%), 1,8-Cineole (10.86%) and Limonene (3.73%) were highest amounts. 58 volatile compounds were identified inSalvia multicaulis thatCamphene (28.85%), α-Pinene (12.33%), Camphor (10.73%), Limonene (9.01%), 1,8-Cineole (5.47%), β-Pinene (4.58%) and Bornyl Acetate (3.75%) had the maximum amounts, respectively. The main part of volatile constituents of Salvia limbata and Salvia multicaulisplantsbelonged to monoterpenes (91.57 and 84.28% respectively) and sesquiterpenes (5.12 % and 7.58 %, respectively).
Conclusion: The results obtained in Salvia limbataand Salvia multicaulisin respect to main components are similar to previous papers (3, 16). However, there is some compound such as α-Terpineol in Salvia limbatawhich has not been reported. According to the obtained results by solid phase micro-extraction is closer to plant compounds.
Nastaran Hemmati; Azim Ghasemnezhad; Javad Fattahi Moghadam; Pooneh Ebrahimi
Abstract
Introduction: all fruits that called citrus are from rutaceae family and aurantioideae subfamily. This subfamily have more than 33 different genus that only three of its genus (citrus, poncirus and fortunella) have economic aspects and in citrus producing country are important. It's reported that orange ...
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Introduction: all fruits that called citrus are from rutaceae family and aurantioideae subfamily. This subfamily have more than 33 different genus that only three of its genus (citrus, poncirus and fortunella) have economic aspects and in citrus producing country are important. It's reported that orange skin has a phenolic compounds which play a role in natural defense mechanism. Also various compounds of phenolic and antioxidant have a major role in fruit tolerance to stressful condition suh as cold and drought. Metabolites found in citrus fruits have antioxidant properties and it's very useful in pharmaceutical, food and cosmetics industries. Oranges, like other citrus fruits, are an excellent source of vitamin C; Vitamin C is a powerful natural antioxidant. Consumption of foods rich in vitamin C helps the body develop resistance against infectious agents and scavenge harmful, pro-inflammatory free radicals from the blood. Various factors such as rootstock type can effect on quality and quantity of citrus fruits. Also, the usage of rootstock causes the change in plant characteristics such as flowering time, ripening time, fruit quality and antioxidant characters of the fruits. Other factors except the rootstock such as scion, geographical and climate factors are effective on producing secondary metabolites. Also active substances or secondary metabolites are producing by the conduction of genetic processes, but their production are being effected by other factors obviously. The aim of this study is to investigating the biochemical changes grafted tree fruit that affected by rootstock with study the correlation between grafted tree and rootstock changes.
Materials and Methods: This study was done to compare the amount of total phenol, total flavonoids and antioxidant features of fruit flesh and skin with investigating the effect of cultivar and rootstock on these parameters based on completely randomized factorial design with three replications. For this purpose total phenol, total flavonoid and antioxidant activity in two citrus cultivar (morro and mars) that grafted on four rootstock (yuzu, citrumelo, sour orange and shel mahalleh) with seedling rootstocks fruit were studied in fruit skin and flesh. Fruits were harvested in the middle of December according to their total soluble solid materials (TSS) which was 10 and then transferred to the researching laboratory in Gorgan Agricultural Science and Natural Resources University. Antioxidant properties using DPPH method in 517 nm wavelength, total amount of phenol using folin siocalteu method in 765 nm wavelength and the total amount of flavonoid were done using the aluminum chloride method in 415 nm wavelength and they were measured using spectrophotometer.
Results and Discussion: the result showed that the two factors consisting rootstock and scion have significant effect on the amount of total phenol, total flavonoid and antioxidant properties of extracts of citrus skin and flesh. The greatest amount of phenolic compounds was produced in the skin of morro cultivar that grafted on shel mahalleh rootstock and the lowest amount was observed in the flesh of yuzu seedling rootstock. Total flavonoid was affected by fruit tissue, cultivar and rootstock. The maximum amount of that was seen in the skin of morro and mars cultivar that was grafted on yuzu rootstock and the minimum amount was recorded in the flesh of morro cultivar that grafted on sour orange rootstock. Also the highest antioxidant activity was produced in skin of citrumelo seedling rootstock and the lowest amount was seen in flesh of yuzu seedling rootstock. The investigation on citrus rootstock showed that, antioxidant activity, total phenol and total flavonoid had significant effect in different rootstock and cultivar fruit. These compounds were affected by climatic condition. Because the light is effective in biosynthesis of phenolic compounds, in fact, these substances have a protective role against the light, especially short wavelengths. Therefore their more accumulation is in skin. The results of this experiment and also the results of the other researcher show that the rootstock effect is related to the quality of grafted species with the species, rootstock type and their interaction.
Conclusion: Based on these experiments, there was significant difference between antioxidant compounds of grafted tree fruit with rootstock fruit but there wasn’t a clear relationship between them. It seems this difference was due to combination and physiological characteristic of each fruit. It seems that the accumulation of chemicals in citrus fruit superior than every factors depends on genetic characteristics and inherent abilities. So that some factors specially rootstock has an important and determinant role in accumulation of these substances.