Document Type : Research Paper
Authors
1 Department of Cultivation and Development Research, Medicinal Plant Research Center, Institute of Medicinal Plants, ACECR, Karaj, Iran
2 Department of Horticulture, Medicinal Plants and Organic Products Research Center, Miyaneh Branch, Islamic Azad University, Miyaneh, Iran
3 -Department of Agronomy and Plant Breeding, Faculty of Agriculture, Shahed University, Tehran, Iran
4 Deparment of Argonomy and Plant Breeding, Miyaneh Branch, Islamic Azad University, Miyaneh, Iran
5 Medicinal Plant Research Center, Institute of Medicinal Plants, ACECR, Karaj, Iran
Abstract
Background and Objectives: Taxol is one of the most valuable effective substances found in some plants, which is used in chemotherapy against breast, stomach, ovary, liver, lung, cervix and pancreas cancers. Hazel leaf (Corylus avellana L.) is one of the plants in which taxol has been proven. The natural forests of Ardabil, Arsbaran and Miane regions are important areas of hazelnut germplasm. The purpose of this research is to investigate and group the subpopulation samples of hazelnuts in these regions based on the SCoT genetic marker and compare the samples of a cluster using principal component analysis in terms of the amount of taxol present in hazelnut leaves and the phytochemical characteristics of the fruit.
Methodology: Current research was done in 2018-2019. In this research, 78 samples of native trees were selected from the natural forests of Fandalo, Ardabil, Mianeh, and Arsbaran (according to the geographic distance of the samples of each region, the population of that region was considered) and the geographic location of each region was recorded with GPS. Quantitative and qualitative characteristics of hazelnuts were collected based on the size, geographic characteristics and possible diversity among the hazelnut stands in the northwest of the country and based on the description of hazelnuts of the deputy research and identification of plant varieties, Ministry of Agricultural Jihad. In this research, based on phenotypic diversity, subpopulation samples (single tree) were selected, and DNA extraction was done from the leaf part by CTAB method. After measuring the quality of DNA in the absorption spectrum of 260 and 280 nm in the nanodrop, using SCoT specific primers, the PCR reaction was performed. Then, electrophoresis was performed for the presence or absence of markers (zero and one) and the data were evaluated using POP Gene v1.32 software. From each group, depending on the number of samples, at least two samples were selected and the amount of taxol in their leaves was extracted and measured. Selected samples of hazelnut fruits were selected and the amount of antioxidant properties, phenols, fatty acids and the amount of some mineral elements were measured.
Results: Based on the presence and absence of SCoT genetic bands in the genome of selected hazelnut samples from different regions, principal component analysis (PCOA) was performed. Correlation (Mantel's correlation coefficient) between the similarity coefficient based on the grouping of genotypes using the SCoT marker and the similarity coefficients using geographic characteristics data was calculated as 0.54, which was significant (Pvalue<.05). Examination of the amount of genetic information content (PIC) based on the used SCoT primers showed that the amount of PIC was different in the groups separated based on PCoA. between genotypic groups in terms of protein content, fatty acid content, fiber, carbohydrate, total phenol, palmitic acid. There was a significant difference in the probability levels of 0.05 and 0.01, and no significant difference was seen in terms of other traits. The comparison of the averages of hazelnut genotypic groups showed that in terms of protein content, percentage of fatty acids, amount of antioxidant property and palmitic acid, the highest average was related to the genotypes of group 1. The values of the similarity coefficients in the grouping based on the SCOT indicator and the similarity coefficients obtained from the results showed that the correlation between the calculated coefficients is 0.76. Evaluation of gene flow (Nm) and fixation index (Fst) within the populations and genotypes of hazelnut showed the highest amount of gene flow Nm and the lowest amount of Fst in all populations (Fst=0.55 and Nm=0.7) They showed that the same trend was observed in the groups as well, although the amount of Nm coefficient in the genotypes of group 1 was (Nm=81). The amount of inter-population heterozygosity (Dst) was estimated to be 0.15.
Conclusion: Since the study of phytochemical traits always requires time and conducting numerous tests, the existence of reliable and reproducible markers is necessary to find genotypes with high phytochemical and taxol characteristics. The low value of the gene flow parameter (Nm) showed that the studied populations have a high tendency to differentiate. The existence of high genetic diversity within populations makes it possible to implement breeding programs. The high genetic similarity between the populations, the study of the SCoT indicator (the highest 0.97, the lowest 0.81) showed that the populations belong to the same origin and genetic background.
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