anahita pahlavan

Honey bees, classified within the insect class and widely distributed globally, consist of 26 identified subspecies across five evolutionary lines. Among these, the Iranian honey bee ( Apis mellifera meda ) is notable for its presence in Iran and neighboring regions. This subspecies exhibits unique biological traits, including a high reproductive capacity and effective propolis collection, distinguishing it from other local bee populations. Understanding genetic diversity within these populations is crucial for conservation and breeding efforts.

The aim of this study is to determine the phylogenetic relationships and natural variations of the Iranian honey bee in comparison to other bee breeds worldwide using the cytochrome oxidase I (COI) region.

The study focuses on the identification of COI gene sequences in honeybees. The mitochondrial COI gene sequences related to the Iranian honey bee and eight populations of other honey bee breeds from around the world (24 ecotypes) were extracted and aligned from the genome database (NCBI). BLAST was used for sequence retrieval, and Clustal W was employed for alignment among species. Phylogenetic analysis was performed using MEGA software, applying the Neighbor Joining method to construct a phylogenetic tree based on the edited sequences. The percentage of nucleotide substitutions and replacements was calculated using the maximum likelihood method. Additionally, molecular diversity indices were computed using DnaSP software to assess genetic variation and conservation parameters among populations.

The bioinformatics analysis of biological data revealed that transfer substitution occurred more frequently than crossover substitution. Additionally, the percentage of substitution in pyrimidine bases was higher than in purine bases. The dN/dS substitution ratio in the gene locus sequence across the examined ecotypes was estimated at 0.08, indicating purifying selection during the evolution of this gene locus. Furthermore, examination of the COI region of the mitochondrial genome identified a total of 4 haplotypes, 20 mutations, and 195 polymorphic loci within the entire studied population. The neutrality values, based on Tajima's test, indicated effects related to genetic drift, genetic bottlenecks, or balancing selection during the evolutionary history of the studied population. Phylogenetic analysis revealed nine distinct categories of evolutionary paths for the COI gene in the studied ecotypes. In the phylogenetic tree, the Iranian honey bee was placed in a separate cluster, exhibiting the highest and lowest phylogenetic distances relative to Apis koschevnikovi and Apis mellifera , respectively.

This research indicates that the conserved region of the mitochondrial cytochrome oxidase gene among various bee species is minimal, highlighting significant polymorphism and susceptibility to mutations. A total of 20 mutations and 195 polymorphic sites were identified, suggesting purifying selection processes that contribute to the gene's biological functions. The study also demonstrates that the COI region can effectively differentiate Iranian honey bee populations from others globally. Overall, these findings enhance our understanding of genetic diversity and evolutionary relationships within bee species, underscoring the importance of continued research for conservation efforts.

Malva sylvestris, commonly known as the common mallow, is a flowering plant species in the genus Malva. It is native to Europe, North Africa, and Western Asia, and has been introduced to other parts of the world. The plant has been known since ancient times and is considered the "type-species" for the genus. The leaves are edible and the plant (and seeds) are used as herbal remedies. Malva sylvestris is either a perennial or biennial plant. It spreads readily from seed and can self-seed prolifically. The seeds resemble small wheels or discs. The plant is known to be susceptible to a virus called Malva vein clearing potyvirus, which is transmitted by aphids. It also contains compounds such as malvin, malonylmalvin, and the naphthoquinone malvone A. The aim of this article is to evaluate the various (nine) ecotypes of Iranian Malva sylvestris L. by analyzing their morphological and physiological characteristics. By employing a step-by-step regression approach, the study seeks to identify significant traits that differentiate these ecotypes. The research intends to enhance understanding of the adaptive strategies of Malva sylvestris in diverse environmental conditions. Ultimately, the findings aim to contribute valuable insights for conservation efforts and the potential use of these ecotypes in horticulture and agriculture. This comprehensive evaluation will also provide a foundation for future studies on the species' ecological adaptability. In this study, 9 ecotypes of Malva sylvestris were collected in 2017 from different habitats in Iran (Mashhad, Torbat-e Heydariyeh, Fariman, Zabol, Zarand, Jiroft, Rudbar, Bandar Abbas and Khorramdasht) (Table 1) and identified in the Herbarium of Torbat-e Heydariyeh University. They were then cultivated in a completely randomized design with three replications in late February 2018 in the greenhouse of the Agricultural Research Institute of the University of Zabol and evaluated in May 2019. The seeds of each ecotype were planted in 5-liter pots (after germination and thinning, five plants of each ecotype were kept in each pot) in a growing medium consisting of an equal mixture of agricultural soil, coco peat, perlite, and well-rotted animal manure. Irrigation was calculated based on the temperature conditions in Sistan and the greenhouse, as well as the field capacity of the pot mixture, and was carried out regularly until flowering. At full flowering, the stem diameter and length of three plants from each pot were randomly measured and their means were considered for each treatment. At this stage, the number of flowers with seeds and the number of leaves of each plant were counted. Fresh and dry weights of root, stem and whole plant were measured with a digital scale (0.01 g accuracy). For dry weight measurement, fresh samples were placed in an oven at 70°C for 48 hours. Chlorophyll a, chlorophyll b and carotenoid contents were determined. The absorbance of the samples for chlorophyll a and b and carotenoids was measured at wavelengths of 663, 645 and 470 nm, respectively, using a spectrophotometer. Proline, soluble carbohydrates, and protein were measured. Pearson correlation coefficients were used to calculate simple correlation coefficients between morphological traits. Statistical analysis of traits was performed using SAS Ver. 9 and Excel software, and means were compared by Duncan's method at 1% and 5% probability levels. The highest stem diameter (9.58 mm), root length (61.22 cm), root weight (18.86 g), root dry weight (4.84 g), and proline content (0.614) belonged to Mashhad ecotype. The number of leaves had a negative and significant correlation with the fresh weight of the plant, while it had a positive and significant correlation with plant height and shoot dry weight. Other traits did not show a significant correlation with the number of leaves per plant. The highest correlation was observed between morphological traits between fresh weight and leaf dry weight (P<0.01) and in phytochemical traits between carotenoid and chlorophyll b (P<0.05). Based on stepwise regression in the presented models, root weight and plant dry weight had the most positive effect on root length, but stem diameter and plant weight had the most negative effect. Chlorophyll b had the most negative and direct effect on proline yield, but chlorophyll a, carotenoids, carbohydrates, and total protein had the most positive effects, respectively. The variance analysis results indicated significant differences among the various ecotypes of Malva sylvestris regarding morphological and phytochemical traits (P<0.01) (Tables 2 and 3). Mean comparisons revealed that the Mashhad ecotype excelled in stem diameter, root length, and fresh and dry root weight, while the Rudbar ecotype showed the highest fresh weight in aerial parts. The Bandar Abbas ecotype had superior fresh and dry weights of aerial parts, leaf count, flower count, and seed count, and the Torbat-e Heydariyeh ecotype was notable for flower and seed counts per plant (Table 4). The tallest stem (40.55 cm) was recorded in the Jiroft ecotype, while the shortest (1.81 cm) was from Mashhad. The greatest stem diameter (9.58 mm) belonged to the Mashhad ecotype, and the smallest (3.54 mm) was found in Zabol. Root length also varied, with the Mashhad ecotype having the longest (61.22 cm) and Rudbar the shortest (9.55 cm) (Table 4). The highest fresh and dry root weights were observed in the Mashhad ecotype, while the Jiroft ecotype had the lowest. In the results of step-wise correlation and regression analysis of the medicinal plant Malva sylvestris L., the highest positive regression coefficients for yield were related to the traits of proline content, root fresh weight, and plant dry weight, which indicates their more fundamental role in increasing yield and their potential for improvement. Overall, the present study showed that root-related traits had an important effect on the final yield in the Mashhad population of Malva sylvestris L., and the Mashhad ecotype also showed the most desirable performance in terms of the evaluated traits. Due to its high performance in these traits, the Mashhad ecotype is recommended for researchers, universities, and private sectors involved in the cultivation and domestication of medicinal plants.

In recent years, environmental concerns and food safety in developing countries are the most important issues. Pesticides are vastly utilized in farming to improve crop production and quality and to reduce energy costs. These pesticides are biodegradable chemical compounds that are generally applied in farming to control pests and promote agricultural production. However, their excessive use, even in low concentrations, may cause serious health problems and environmental pollution. Therefore, a great deal of the research has focused on methods used for determining the presence of pesticides in various food matrices, according to this proposing sensitive diagnostic methods is essential for reliable quantification of pesticides availability. Meanwhile, rapid detection technologies which are among the most important tools in the analysis of food products are required to be assessed for analysis of residual persistence of pesticides in food and agricultural products to effectively control their quality and safety. In this article, it is attempted to provide a complete review of some detection methods including HPLC, HPTLC, GC/ MS/ MS, CE-DAD, LPME, SPME, LLE, DLLME, AChE, SERS, LC/ Tandem/ MS/ MS, MSPD, luminescence chemistry, safety assessment, and Meisel chromatography electrosynthesis used to identify pesticide residues in different food samples.

Since rbcl gene and protein, as well as internal transcribed spacer (ITS1,4) regions, have not been used in the evaluation of Malva sylvestris L., this study was aimed to assess different Iranian Malva sylvestris L. genotypes by evaluation of morphological and phytochemical traits, along with rbcl and ITS1,4 gene sequences. Furthermore, three-dimensional and functional structures of Malva sylvestris L. rbcl protein were examined, as well. Nine Malva sylvestris L. genotype samples were collected from different regions of Iran. After species identification, completely randomized design with three replicates was used to evaluate different genotypes, based on morphological and phytochemical traits. DNA extraction was carried out using the SDS method and then final PCR products were sent to Macrogen Company, South Korea for sequencing. Sequence quality was assessed using Chromas 2.1.1 software. Then, the sequences were aligned using the ClustalW method in MegAlign 5 software and the dendrogram of the phylogenetic relationships and similarity matrices were plotted, as well. SWISS-MODEL and QMEAN servers were used for modeling and validation of rbcl protein. Ramachandran plot analysis and Pro-SA servers were used to evaluate the structure and chemical quality of the protein. Comparison of the mean physio-morphological traits between different genotypes showed the highest stem diameter (9.58 mm), root length (61.22 cm), root fresh weight (18.86 g), root dry weight (4.84 g) as well as proline content (0.614mg/gDW) in Mashhad genotype. Based on stepwise regression results in the presented models, root fresh weight and plant dry weight had the most positive effect on root length, but stem diameter and plant fresh weight had the most negative effect. Moreover, while chlorophyll b had the most negative and direct effect on proline function, chlorophyll a, carotenoids, carbohydrates and total protein contents had the most positive effects, respectively. Assessment of proteinprotein interaction networks revealed that most proteins encoded by matK, psb-tranH genes interact with rbcl protein. The results of cluster analysis, similarity matrix as well as dN/dS ratio showed high similarity and conservation of ITS and rbcl sequences among different Malva sylvestris L. genotypes. rbcl and ITS sequences were not suitable markers to evaluate phylogenetic relationships intraspecies (at subspecies level) but are useful to evaluate interspecies relationships. Furthermore, the Mashhad genotype is suitable for dry and water deficit conditions, withstanding such conditions. Therefore, it is recommended that this genotype can be used both as a parent and or directly to breed, develop and modify Malva sylvestris L. species.