نشریه زیست فناوری گیاهان دارویی
سال پنجم شماره 2 (پاییز و زمستان 1398)

Chavil is one of the most important medicinal plants and although there are different accessions of it in Iran, there is no report on the study of its genetic diversity. In this study, molecular markers were used to evaluate the genetic diversity of eight Chavil ecotypes. After preparing the ecotypes from habitats and Isfahan Pakan Bazr Company, DNA extraction was performed and then the polymerase chain reaction was performed using 12 ISSR primers and the obtained molecular data were analyzed. The polymerase chain reaction produced bands using used primers its polymorphism content were varied from 23 to 100 percent. The F6, F8, F9 and F11 primers showed the highest and the F10 primer showed the lowest polymorphism percentage. Also, the highest PIC values were obtained for F6 and F11 primers. The average Shannon index in this study was 0.38, with the lowest data for the F10 primer and the highest for the F11 primer. The cluster analysis of molecular data divided the eight ecotypes into four different groups. The highest genetic similarity between Zardkooh and Gayoneh ecotypes and the lowest genetic similarity between Wezg and Abnahr ecotypes was observed. Ecotypes migration between four neighboring provinces of Chaharmahal and Bakhtiari, Kohgiluyeh and Boyer-Ahmad, Fars and Isfahan appears to have taken place. The present study showed that the ISSR markers were appropriate for the study of the genetic diversity of Chavil.

Thymol is one of the most widely used secondary metabolites in Thymus vulgaris, as a drug. One of the key genes which is involved in its synthesis is the cytochrome p450 gene. In order to isolate and cloning p450 gene, the extraction of RNA was carried out subsequently the cDNA synthesized. Amplification of cDNA was carried by using designed primers from conserve region in the presence of the HF enzyme that has the property of 3′ to 5′ exonuclease. In first stage cloning was done by T/A method in plasmid pTG19. Selection of recombinant colony was performed by PCR system and digestion with BamHI. The cloned fragments were sequenced for the final confirmation. In order to cloning in the plant expression vector pBI121GUS-9 , the fragments that were cut with the BamHI enzyme were cloned in homologous site of the pBI121GUS-9 vector under the promoter 35s and the Nos terminator. Confirmation of recombinant colonies was performed with colony PCR and digestion. This designed recombinant vector can be used for further research and over- expression of thymol.

The common hollyhock (Althea rosea) is a wild, annual plant. Its flowers, fruits and roots have medicinal properties. The cultivation of this medicinal plant is mostly challenged by a lack of proper seed germination and, as a result, an unfavorable establishment of seedlings in primary conditions of cultivation. Breaking the seed dormancy and stimulating seed germination in this species are known to have several ways. In three replications, this experiment was performed in a completely randomized design on three hollyhock ecotypes from Sepidan Shiraz, Kordan Karaj and Isfahan. In the first experiment, germination was evaluated after the seeds were exposed to 4 °C in different time periods (0, 10 hours, 7, 14, 21 and 28 days), while another treatment group involved the exposure of seeds to 35 and 46 °C in a steam bath for 6 hours. In the second experiment, the proliferation of hollyhock was optimized in vitro using a combination of different plant growth regulators, including 2 mg/L benzyl adenine (M4), 1 mg/L benzyl adenine in combination with 0.10 mg/L naphthalene acetic acid (M13), 1 mg/L thidiazuron (M6), 1.5 mg/L thidiazuron in combination with 0.10 mg/L naphthalene acetic acid (M17), 0.5 mg/L kinetin (M8), and 2 mg/L kinetin in combination with 0.10 mg/L naphthalene acetic acid (M20). The analysis of variance for the first experiment showed that the effect of ecotype (Sepidan Shiraz, Kordan Karaj and Isfahan) was significant (P≤0.05) on germination. This included the percentage and rate of germination, root volume and height of seedlings. The effect of temperature on seed germination was significant (P≤0.05), while also affecting other traits (P≤0.01). The highest values of proliferation, shoot length and rooting percentage were observed in the Shiraz ecotype when treated with M4 in vitro. The lowest values of these parameters were observed in the Karaj ecotype when treated with M20. Root length grew significantly in the Isfahan ecotype under M4 and M17 treatments, whereas the shortest roots were observed in the Karaj ecotype under the M20 treatment. The highest number of leaves was observed in the Shiraz ecotype by the M4 treatment, whereas the lowest number of leaves was recorded in the Karaj ecotype as a result of the M8 treatment.

Plants extracts can be a new sources for pest and disease controlling. In this research, the antibacterial effects of essential oil from 9 plants including, Juncus, Euonymus japonicas, Taxus baccata, Dana racemose, Datura stramonium, Lythrum Salicaria, Cupressus sempervirens, Ilex aquifolium and Hyssopus officinalis with different concentrations, means 12.5, 25, 50 and 100 % used for three bacteria such as E.coli, Pseudomonas syringae. pv syringae, Rathayibacter tritici through in the form of disc diffusion. The experimental was as completely randomized factorial design with three replications. At the end, qualitative and quantitative analysis of the components of the essential oil were analyzed by GC-MS. The results of variance analysis of the inhibitory effects in different concentrations showed that, plant species, the different concentrations of essential oil, the type of bacteria and mutual interaction have shown the significantly meaning, in terms of inhibitory effects (P <0.01). The results indicated, the most antimicrobial activities related to the Juncus essential oil with the zone of 13.3 mm for E.coli. The part of Juncus essential oil so examined by GC-MS system and the result has shown 14 different compounds. The maximum and minimum substances were Menthol (41%), Menthone (12%) and Pulegone (8.62%), respectively. From this research, we conducted that the juncus.s essential oil putatively can be as a source of disease control agent.

The aim of this reserch was determining of the Achillea wilhelmsii C. Koch essential oils composiation that was growned in natural counditions.Collected plants were harvested at full bloom stage in july 2019 and dried in the shade of the room condition then pulverized in the form of a homogeneous mixture and its essential oil was extracted by distillation with water. The components of the essential oil were identified and measured using the gas chromatography device connected to the mass spectrometer. The results showed that the essential oil obtained from the dried vegetative body of the plant had a yellow color with a yield of 0.89%. The results of GC-MS showed that the essential oil of this plant in the desired area consisted of a combination of 106 substances. Percentage of total essential oil, the most important main compounds identified in essential oil were: α-Thujene (2.21%) Camphene (2.38%), Sabinene (0.53%), Eucalyptol(1,8-cineole) (5.71%), Terpinen-4-ol (2/55%), Nonanal (4/25%), Camphor (9/87%), (+) 2 Bornanone (5/59%), Endo-borneol (4/05%), α-Terpineo (2/46%), Rosefuran (3/52%) Caryophyllene (0.64%), α-Farnesen (1.83), Terpinolene (3.44%) and Caryophyllene oxide (5.62) -Based on our knowledge this is the first report of Zanjan Achillea wilhelmsii C. Koch.

This research was conducted to evaluate the diversity and molecular confirmation of oat gaze by DNA barcoding method. Moringa seeds were cultivated in Baluchestan province of Baluchestan province including Bennett, Kishigh, Desh, Madhadi village, Nesfaran, Condar, Tang Fanvjh entrance, mouth and seven girls. After three months, the DNA was isolated from the leaf samples and the ITS2 region was amplified and sequenced. The ClustalW sequences were combined with Mega7 software, clustering and genetic similarity and spacing. The intrinsic similarities of the Moringa Baluchistan specimens were detected in the range of 99.9%. In the present study, ITS2 did not distinguish between Balochistan's ten variants in terms of genetic variation within the group. Therefore, in order to evaluate the ability to evaluate the genetic diversity of the species, another test was performed, so that the sequences of other species of Moringa family were used in NCBI, and the Moringa sequence of the present study was compared. The Moringa-Baluchestan sample sequence with Ethiopic peregrine sequences showed homology of about 92%. The emergence in this method showed that the studied populations were divided into five groups and it was determined that the similar species were placed in the same branch. The Moringa-Baluchestan sample sequence was grouped with sequences of peregrine species in one group.Since the Moringa species in Balochistan have been introduced based on morphological keys in the Iranian flora, peregrine has been introduced. Based on the results of this study, the same was also recognized and confirmed through molecular.