amir sahraroo

 Adventitious root cultures of medicinal plants represent a bountiful reservoir of valuable secondary metabolites.  The esteemed medicinal plant Echinacea</em> purpurea</em> (L.) Monech is highly sought after for its phytochemical properties, with a traditional use of its roots in herbal medicines and dietary supplements as an immune system stimulant for treating inflammatory, viral, and respiratory diseases. However, the heterogeneous synthesis of bioactive compounds in natural environments and the limitations of conventional plant production methods necessitate exploring alternative approaches for obtaining active plant substances and organs. Among various in</em> vitro</em> cultivation techniques, the cultivation of differentiated tissues, specifically roots, consistently demonstrates the ability to yield higher quantities of metabolites than undifferentiated tissues such as callus and suspension cells. 

 The primary aim of this study was to accurately determine the ideal combination of plant explants and plant growth regulators that would facilitate the successful establishment of adventitious root suspension cultures. Three distinct plant explants, namely leaf, petiole, and root, were carefully evaluated in this study. These explants were subjected to varying concentrations (0, 0.5, 1, 1.5, and 2 mg.L-1) of two types of plant growth regulators: indole-3-butyric acid (IBA) and 1-naphthalene acetic acid (NAA). After six weeks, the number of adventitious roots developed by the explants under different concentrations of plant growth regulators was measured. The impact of different sucrose concentrations (10, 20, 30, 40, 50, and 60 grams per liter) in the culture medium was investigated over a four-week cultivation period. Various dynamic characteristics of root growth, including fresh weight, dry weight, volume of the remaining medium, electrical conductivity, and pH, were closely examined. Furthermore, phytochemical properties such as total phenol, total flavonoid, and antioxidant activity were assessed. To administer the sucrose treatments, 0.35 grams of adventitious roots were placed inside 250 ml Erlenmeyer flasks containing 50 ml of half MS culture medium, 1 mg.L-1IBA, and varying concentrations of sucrose, maintaining a pH of 5.8. Measurements were taken from three flasks of each treatment weekly. The experiments followed a Completely Randomized Factorial Design, with statistical analysis conducted using SPSS software.

 Leaf explants exhibited superior root formation to root and petiole explants. The average number of adventitious roots in leaf explants reached 3.37, while petiole and root explants produced only 0.23 and no adventitious roots, respectively. The most effective root formation occurred in MS medium with 1 mg.L-1 IBA, resulting in an average of 4 roots per explant. Hormone concentrations of 0 and 2 mg.L-1 did not induce root formation. In the fourth week of cultivation, the highest fresh weight recorded was 29.8 grams per liter, marking a remarkable 3.4-fold increase from the initial inoculation weight. The culture medium's electrical conductivity and pH decreased as the cultivation progressed. Cultivation weeks did not significantly impact total flavonoid production and related antioxidant activity in adventitious roots. However, the cultivation period significantly affected the total phenol content at 57.56 mg of gallic acid per gram of dry weight. Altogether, lower sucrose concentrations proved more efficient for biomass production, bioactive compound content, and antioxidant activity. The highest biomass accumulation and total phenol content occurred at 3% sucrose. In comparison, in the fourth week of cultivation, the highest total flavonoid content and the highest free radical inhibition percentage were observed at 2% and 1% sucrose, respectively. Notably, a negative correlation was observed between the electrical conductivity of the remaining culture medium and root biomass in lower sucrose concentrations as the culture duration increased.

In light of the immense potential harbored by adventitious root suspension cultures of E. purpurea</em> as an alternative source for the production of secondary metabolites, particularly caffeic acid derivatives, the discoveries made in this research have the potential to advance the field of mass production for active root organs, thereby facilitating the synthesis of these precious compounds through the implementation of appropriate laboratory bioreactor systems.

Diversifying products and increasing added value by producing new medicinal and health products from tea is a strategic method to improve the production situation and economize tea. In recent years, tea genotypes resulting from the natural selection of the purple-leafed variety have been introduced in some tea-growing countries. Considering that there is a possibility of the presence of such genotypes in the tea gardens of Gilan, the present research was carried out to identify and select the constant purple-colored tea plants that contain higher amounts of anthocyanins. For this purpose, through field monitoring, purple tea plants with stable color were identified and labeled from the cities of Gilan province, and then the leaves (one bud and two leaves) were examined to analyze the amount of pigments. The results showed that the environmental factors, soil nutrition and pruning operations in different places had a positive effect on the amount of pigments during the growth and development of purple tea plants. So that sample numbers 10 (12.5983 μg·g–1, DW), 19 (50.4865 μg·g–1, DW)) and 16 (10.4451 μg·g–1, DW) respectively in Lahijan and Astana Ashrafieh had the highest anthocyanin activity and the lowest anthocyanin activity in sample number 3 (1390/93 μg·g–1, DW) in Langrod city. The highest amount of chlorophyll a, total and carotenoid in sample number 35 (1.07, 1.53 and 2.49 mg·g −1 DW) respectively and the lowest amount of chlorophyll a, total and carotenoid in sample number 5, respectively (0.46, 0.71 and 1.26 mg·g −1 DW) were obtained in Astana Ashrafieh, and factors such as (radiation intensity, soil fertility, altitude above sea level and pruning operation) were involved in this phenomenon. Therefore, extractions with a higher amount and less temporal changes in the amount of anthocyanin can be monitored and studied for future research to produce tea seedlings rich in anthocyanin.

Gotu kola (Centella asiatica L.)  is one of the valuable medicinal plants, whose biodiversity is facing the risk of extinction in the world due to the limitation of distribution areas and the reduction of surface waters. In order to investigate somatic embryogenesis in gotu kola, an experiment was carried out in 2018 in the tissue culture laboratory of the Faculty of Agriculture of Gilan University. Leaves were used as explants and MS medium as basal medium. The effect of different concentrations of plant growth regulators (2,4- dichlorophenoxyacetic acid (2,4-D) in five concentrations of 0, 0.25, 0.5, 1, 1.5 and 2 mg/L in combination with benzyl adenine (BAP) in four concentrations of 0, 0.25, 0.5 and 1 mg/L) were tested to determine the appropriate culture medium for inducing gotu kola embryogenic callus. For the induction of somatic embryos the good status obtained calli were transferred to different concentration of 2,4-D including 0, 0.1, 0.2 and 0.3 mg/L. In the next step the derived somatic embryos were subjected to  0, 0.25, 0.5 and 1 mg/L of BAP for germination. The results showed that the treatment with 2 mg/L of 2,4-D had the highest percentage of callus formation (96.61%), fresh weight (3.96 g/cm2 of leaf) and average callus diameter (3.56 cm) in dark conditions. Also, the treatment of 0.1 mg/L of 2,4-D had the highest average number of embryos formed in different stages of spherical, heart, and torpedo shaped, and the treatment of 0.1 mg/L of 2,4-D had the highest percentage of embryogenesis (55.27%), which was not significantly different from the treatment of 0.2 mg/L of 2,4-D. The results showed that the treatment of 1 mg/liter of BAP had the highest percentage (56.8%) of embryo germination and also the highest average number (12.4) of produced seedlings. In general, the use of 2,4-D to create embryogenic calli and BAP to germinate the formed embryos was satisfactory.

Estimates of combining ability help determine the value of hull-less pumpkin (Cucurbita pepo L. var. styriaca) lines in producing new hybrids with high yield and better quality. This research was conducted to estimate the general and specific combining abilities and heterosis in six inbred lines proposed for hull-less pumpkins at the University of Guilan. Analysis of variance indicated that responses for all traits differed significantly. The highest fruit yield and seed length were related to the hybrid 14×16, the highest number of seeds was related to the hybrid 16×23, the highest percentage of oil was related to the hybrid 11×14 and the highest yield of oil was related to 11×23. Considering the high value of combining ability and the heterosis compared to the average of the parent and high parent, it is possible to increase the oil yield and the seed yield by using the 11×23 hybrid. The 16×23 hybrid proposes to increase the number of seeds. The best specific combining ability (SCA) for seed number was associated with the 16×23. The 11 × 14 had the best SCA for the oil yield per hectare. The 11× 23 had the best SCA for the seed yield per hectare. Significant specific combining ability and high heterosis for traits indicated low additive and high dominant gene action on hull-less pumpkin traits, therefore crossing between lines to release new hybrid cultivars is necessary for Iran's pumpkin breeding program.

Perilla frutescens var. crispa, a medicinal and aromatic plant belonging to the Lamiaceae family is widely used for the treatment of constipation, asthma, and skin disorders, and also as spice. As climatic, environmental, and edaphological conditions of the plant habitats affect active substances of medicinal plants, quantitative and qualitative changes in the essential oil content and composition of the plants cultivated in the Research Farms of four regions of Iran (Rasht, Noor, Khorramabad, and Neishabour) were evaluated in the present study in 2020. Aerial organs of the plants were collected at the vegetative stage and their essential oils were isolated by hydro-distillation. The essential oils were quantitatively and qualitatively analyzed by gas chromatography (GC) and GC-mass spectrometry (GC-MS). Among the studied areas, the highest and lowest altitudes were Neishabour and Rasht, respectively. The climatic classification of Rasht and Noor was also more similar. The essential oil contents (%w/w) of the plant samples in Rasht, Noor, Neishabour, and Khorramabad were 1.45, 1.33, 1.26, and 1.16, respectively. In total, 28, 27, 25, and 27 compounds were identified in Noor, Rasht, Khorramabad, and Neishabour regions, respectively. (E)-Caryophyllene (42.4-58.7%), α-cadinol (8.2-16.5%), (E)-nerolidol (3.8-9.5%), α-dehydro-elsholtzia ketone (2.1-7.5%), cis-dihydro-α-terpinyl acetate (1.0-7.5%), and egoma ketone (0.2-5.5%) were the major essential oil compounds at different regions. Sesquiterpene hydrocarbons (44.9-61.2%) were the major class of the compounds of essential oils. Cluster analysis based on the essential oil compositions classified the studied areas into two distinct groups. Noor and Rasht were clustered in one group and Khorramabad and Neishabour were clustered in the other group. Results showed that temperate regions are suitable to increase the essential oil contents of the plants, while arid and semi-arid regions can improve the quality of essential oils.

Medicinal plants are used in all civilizations and cultures are one of the most essential defensive lines to protect human health and fight human diseases in the world. From the distant past to now, the plants have long been considered as a source of renewable medicines.Perilla (Perilla frutescens var. crispa)is an annual plant native to East Asian, which used as a medicinal plant due to the presence of active metabolite, especially rosmarinic acid. The medicinal organs used in this plant are its leaves and flowering branches. Growth, drug yield, as well as quantity and quality of secondary metabolites of medicinal plants are affected by ecological factors. The present study was conducted to evaluate the agro-morphological and phytochemical traits of Purple Perilla grown in three different locations of Sari, Rasht, and north of Tehran.

 The experiment was conducted as a completely randomized design (CRD) with three replications. Purple Perilla  were planted as seedlings in early spring in the trial field with the density of 25×50 cm (inter-plant space × inter-row space) in three locations including Sari, Rasht, and north of Tehran in 2018. Plants were harvested at the flowering stage in late September. Argo-morphological and phytochemical traits including plant height, inflorescence length, number of branches, stem diameter, internode length, leaf length, leaf width, plant fresh weight, stem fresh weight, leaf fresh weight, flowers fresh weight, plant dry weight, leaf dry weight, flower dry weight, total phenolics and flavonoids as well as rosmarinic acid content were measured. The total phenolic contents of extracts of Perilla plants were determined according to the method of Ciocalteus Reagent (FCR) Folinthe aluminum chloride method was used for the determination of the total flavonoid content of the extracts. The rosmarinic acid content was estimated with method HPLC. Data set were subjected to variance analysis using SPSS statistical software (version 19). Also, Duncan’s multiple range test was used to compare treatment means at P<0.05.

 Results of variance analysis showed that almost all morphological traits (except stem fresh weight, plant dry weight, and plant fresh weight) and all phytochemical traits were significantly (P<0.05) influenced by locations. Mean comparisons of the effect of different locations for morphological traits showed that the maximum plant height (137.67 cm), leaf length (15.28 cm), fresh weight of leaf (108.3 g), dry weight of leaf (41.63 g) were observed in Sari, while the minimum plant height (87.34 cm), leaf length (10.8 cm), fresh weight of leaf (69.74 g), dry weight of leaf (26.13 g) were detected in north of Tehran. The Rasht had the highest flower fresh weight, flower dry weight, and number of branches with an average of 108.67 g, 25.99 g, and 33, respectively. The lowest flower fresh weight, flower dry weight, and number of branches were observed in the north of Tehran with an average of 47.07, 12.71, and 22 g, respectively. The results revealed that the highest internode length, inflorescence length, and leaf width belonged to the Sari with an average of 9.12, 13.17, and 98.8 mm, respectively, while the Rasht had the lowest internode length, inflorescence length, and leaf width with an average of 6.28, 9.8, and 6.97 mm, respectively. Also, the maximum and minimum stem diameter were observed in the north of Tehran and Sari (8.95 and 7.65 mm), respectively. Mean comparisons of the effect of different locations for phytochemical traits showed that the maximum total phenolic contents and rosmarinic acid produced in the Rasht with an average of 49.03 and 52.20 (mg/ g dry matter), respectively, while the Sari had the maximum flavonoids content (14.8 mg/ g dry matter).

 Drug yield (leaf and flower fraction) of the plants grown in Sari, Rasht, and north of Tehran was 41.63, 32.05, 26.13 and 12.71, 25.99, 18.47, respectively The highest total phenolics (49.027 mg/ g dry matter) and rosmarinic acid content (46.99 mg/ g dry matter) were observed in Rasht region, while the highest total flavonoids was observed in Sari (14.8 mg/ g dry matter).  In terms of argo-morphological and phytochemical traits, Sari and Rasht were suitable places for the growth of purple perilla.

Asparagus (Asparagus officinalis L.) is an export product and one of the most lucrative products compared to other vegetables; and it is a valuable economic, pharmaceutical, food and industrial crop. The most important agriculture aspect of asparagus is the quality of the seedlings; they depend on the percentage of rooting after planting and are very important for future crop quality. This study was conducted to determine the best level of municipal waste compost and fertigation on asparagus seedling growth with thick buds and large and dense crown, strong roots, and a high number of summer branches.

The experiment was conducted in a completely randomized design with three replications and with two factors of substrate composition (at four levels) and fertigation (at two levels) with three replications and at least 20 seedlings per replication. Asparagus seeds were planted in four different litter compositions that of 50% by volume of garden soil and sand used as a base bed and mixed with different amounts of municipal waste compost (0, 10, 20, 30% by volume). After germination, half of the seedlings in each bed were fed with fertigation containing nitrogen, potassium, and pure phosphorus at the rate of 150, 100, 50 grams per cubic meter of irrigation water. Finally, after a three-month feeding period, the important traits of seedlings including root length, number of buds, number of summer shoots, summer stem height, SPAD index, number of roots, root volume, root diameter, percentage of ash, percentage of dry matter, fresh weight, potassium, phosphorus, phenol, flavonoids and antioxidants were measured.

The results of data analysis of variance showed that the effect of fertigation and the effect of compost levels of composition on plant height, number of top summer branch, fresh weight of the plant, plant dry weight, crown thickness, number of buds, root length, number of roots, root volume, chlorophyll index (SPAD), total phenol, total flavonoids, and potassium were significant. The results showed that the highest average of these traits was related to fertigation treatment and the lowest average of these traits was obtained without fertigation treatment. The results of comparing the average simple effects of compost levels, the highest average of these traits from culture medium containing 30% municipal waste compost and the lowest average of these traits was obtained from culture medium containing no municipal waste compost. Also, the interaction effect of compost levels in fertigation on plant dry weight and canopy thickness was significant. The highest average of these traits was related to treatment with fertigation and culture medium containing 30% municipal waste compost.

It seems that due to the long production period of asparagus seedlings, paying attention to feeding the seedlings through fertigation and enriching the culture medium can help improve the quality of seedlings. As the results of our experiment showed, the use of municipal solid waste compost has an effect on germination, growth and number of summer shoots, number of buds, increase in volume, length, diameter and number of roots, crown thickness and chlorophyll index, fresh weight and plant dryness, percentage of dry matter, amount of phenol and absorption of potassium could have a great effect on the growth and quality of the seedlings of this plant. On the other hand, with the increase in plant growth and as a result, more yield, the plant needs irrigation to expand the roots and increase the number of summer stems because it needs a lot of nutrients. Therefore, it is better to use mineral fertilizer in growing asparagus. In this condition, the plant produces more roots, and the underground stems that lead to produce more and stronger seedlings. Stronger asparagus seedling growth faster than the weeds and overcomes and reaches economic performance sooner. Therefore, carefulness in the use of municipal solid waste compost and the amount of animal and mineral fertilizers, by affecting the yield and as a result the profitability of this plant, will encourage farmers to cultivate this crop. On the other hand, it is possible to use municipal solid waste compost in other ways. There is no need to use a large number of chemical fertilizers and the safe product is healthier than other plant products that use a large number of chemical fertilizers for their production.

Gotu kola (Centella asiatica L.) is one of the most valuable medicinal plants. Due to the reduction of surface water and the limitation of dispersion, the natural populations of this plant species are in extinction risk. The purpose of this experiment was to introduce a tissue culture protocol for rapid micropropagation of the Gotu kola plant. For callus induction, explants were established on Murashige and Skoog (MS) medium supplemented with different concentration of 6-benzylamino purine (BA) at six levels (0, 1, 2, 3, 4, and 5 mgL-1) and α-naphthaleneacetic acid (NAA) at four levels (0, 0.25, 0.5, and 1 mgL-1). The best callogenesis was obtained with maximum callus induction (83.33%), fresh weight (4.26 g), and daily growth rate (0.14 g/day) by combined application of 4 mg L-1 BA and 1 mg L-1 NAA. Different concentrations of BA (0, 1, 2, 3, 4 and 5 mgL-1) and NAA (0.1 mg L-1) were used for shoot regeneration. Combination of BA (4.0 mgL-1) with NAA (1.0 mgL-1) showed the highest shoot proliferation (74.99%), maximum number of shoots (7.66), petiole (6.66) and the longest shoots (1.56 cm). The effects of, IBA (0, 0.5, 1, 1.5, 2, 2.5 mgL-1) were examined on adventitious root formation. MS medium supplemented with IBA (1 mgL-1) showed the best percentage of rooting (84.66%), maximum number of roots Per plantlet (65), and the longest roots (3.33 cm) along with the highest percentage of survival (74.99%).

Leucojum aestivum L. is a perennial and bulbous plant. Its bulbs are used in the medical industry because of having alkaloids. This study was conducted as split plot in a completely randomized design with 3 replications in 2018 at the University of Gilan in field conditions. The main factor included bulbs collected from three regions of Lahijan, Tonekabon and Langrood. The sub-factor included 10 levels of organic fertilizers, biochar (3, 6 and 12 tons per hectare), inoculation of Glomus intraradices, Glomus mosseae and Glomus intraradices + Glomus mosseae fungi, inoculation of Azospirillum, Azotobacter and Pseudomonas bacteria and the control. Data collected and analysed for morphological traits and absorbed macro and micro as elements by plants. There was significant difference between genotypes in terms of the plant height, leaf area per plant, leaf weight per plant, bulb weight, flowering stem weight, root weight and total plant weight (p<0.01). A significant difference was observed in the types of fertilizers used for all morphological traits and yield (p<0.01) and for phosphorus, potassium and iron content (p<0.05). The genotype by treatments interaction effect was significant for leaf number, leaf area per plant, leaf weight per plant, bulb weight, flowering stem weight, root weight and total weight showed a difference (p<0.01). Lahijan genotype plants had the highest plant height (57.6 cm), leaf length (52.5 cm) and plant leaf weight (27.2 g/per plant). The highest root weight (9.5 g) and total plant weight (74.1 g/per plant) was for biochar treatment of 12 ton/h. The study results indicated that Leucojum aestivum L. had good adaptation to crop conditions, and using biochar fertilizer and some biofertilizers improved various crop traits of this plant.

Pennyroyal (Mentha pulegium L.), Labiatae family, is widely used as a vegetable and medicinal herb. This plant commonly grows in the wet soils such as ditches, riversides and ponds. It is indigenous to the Central, Southern and Western Asia, North Africa, Ethiopia and the Canary Islands. It can also be found in Alborz, North and Northeast regions of Iran. The aerial parts of the plant are used as pharmaceutical and flavoring substances. The aim of this research was to find the best level of nitrogen fertilizer regime, harvesting time and plant density for pennyroyal plant in Rasht area.

This study was conducted as a split-plot experiment based on randomized complete block design with three replications in Faculty of Agriculture, University of Guilan. The main plots included two harvesting times, the subplots were four levels of Nitrogen fertilizer including control (without fertilizer), 50, 100 and 150 kg ha-1 Nitrogen and the sub-subplots were plant density at 3 levels; 10, 14 and 18 plants m-2.

The results showed that triple interaction of nitrogen, plant density and harvest time had significant effect on dry weight. The highest dry weight (1909 kg ha-1) were obtained by 150 kg ha-1 net nitrogen with the density of 18 plants per m2 for the first harvest. Moreover, 50 kg ha-1 nitrogen fertilizer treatment with 10 plants per m2 showed lowest amount of dry weight (396.66 kg ha-1) in the second harvest time. However, the interaction of nitrogen and planting density revealed significant effect on fresh yield and leaf area. The highest yield (8071 kg ha-1) was related to 150 kg ha-1 nitrogen treatment with the density of 18 plants m-2. About physiological traits, the results claimed that treatments significantly affected the essential oil percentage, total chlorophyll, total phenol, nitrogen and calcium elements. The highest essential oil content (2.66%) was exhibited by the treatment of 150 kg ha-1 nitrogen with plant density of 10 plants m-2. In addition, the essential oil yield was also influenced by the interaction effect of harvest time and planting density. The highest yield (25665/075 milliliters ha-1) was found by the plant density of 18 plants m-2 from the first harvesting time.

Finally, according to our results, the best treatment was first harvesting time, 150 kg ha-1 nitrogen with the density of 18 plants m-2, which can be recommended for the planting program of aromatic pennyroyal.

An experiment was conducted to study the micropropagation of Satureja khuzistanica. The seeds were planted in 1/2MS medium. After germination and growth of plantlets, they were subcultured in fresh medium. Then, they cultured into medium supplemented by different concentrations of BA and Kinetin. Results showed that, the highest proliferation (2.4 shoots) was observed at concentrations of 0.5 mg/l IBA and 3 mg/l BA while the lowest proliferation (1.12 and 1.04 shoots) was produced at concentration of 0.25 mg/l IBA, 1.5 mg/l BA and also 0.5 mg/l Kinetin. The tallest shoots (5.76 cm) were obtained at concentration of 0.5 mg/l IBA and 1.5 mg/l BA whereas the smallest ones (2.44 cm) were observed at concentration of 1 mg/l Kinetin. Lowest percentage of survived plantlets (46.6 %) were observed in the media with IBA at concentration of 0.25 mg/l and BA at concentration of 1.5 mg/l. Quarter salt strength MS basal medium induced rich rooting response (85.2%), when compared with ½ salt strength MS (46.66%) and MS (17.6%), media. The highest rooting percentage (64.30%) and number of roots (7.7 and 8. 4) were obtained at 0.25 and 0.50 mg/l IBA, respectively. Overall, media containing 0.25 mg/l IBA plus 3 mg/l BA can be recommended for micropropagation of S. khuzistanica.

In the present study, an efficient protocol has been developed for callus induction and production of RA in callus culture of Satureja khuzistanica for the first time. In vitro callus induction was achieved from young shoot tip explants cultured on MS and B5 media supplemented with different concentrations of IBA (0.1, 1.0, 2.0 and 5.0 mgl-1) solely or in combination with cytokinins BAP and KIN (1.0, 2.0 and 5.0 mg l-1). B5 medium supplemented with 1.0 mgl-1 IBA plus 5.0 mgl-1 BAP and MS medium fortified with 2.0 mgl-1 IBA and 2.0 mgl-1 BAP were the most favorable media for callus formation with the highest induction rate (96%). Maximum growth index (2.89 and 2.63) and maximum callus biomass (2.34 and 2.33 g fresh weight) were obtained from the callus cultured on B5 medium supplemented with 1.0 mgl-1 IBA plus 5.0 mgl-1 BAP and MS medium fortified with 1.0 mgl-1 IBA plus 1.0 mgl-1 KIN, respectively. Determination and quantification of RA in cultured calli were performed by HPLC UV/MS analysis. Calli induced from the plant and maintained on supplements of IBA and BAP in the absence of light produced RA 7.5% based on dry weight (DW). No differentiation was observed in any callus during the course of this study.