کوروش فلکرو

In this study, tea shoots of Kashef and Lahij cultivars were plucked from tea shrubs of Lahijan Fajr research station in three seasons (spring, summer, and autumn), and tea was made in four methods on the laboratory scale (green Orthodox, green Crush Tear Curl (CTC), black Orthodox, and black CTC). The chemical properties of tea samples, including the amounts of total polyphenols, antioxidant activity, theaflavin, thearubigins, and organoleptic score, were determined. The experiment was conducted in a factorial split plot based on a completely randomized design with three replications. The main factor was harvest time and sub-factors were the cultivar (in two levels) and tea-making method (in four levels). The results showed that the highest amounts of qualitative characteristics such as sensory score, theaflavin, and ratio of theaflavin to thearubigin, antioxidant activity, and polyphenol for all tea making methods were observed in the Kashef cultivar. The black CTC tea obtained the higest scores for quality characteristics, especially for the ratio of theaflavin to thearubigin compared to Orthodox tea in both cultivars. The antioxidant activity of green tea was not significantly different between cultivars (p≤0.05). In both cultivars, Orthodox green tea had a higher score than CTC green tea due to differences in the color of brewed beverage.

Tea (Camellia sinensis L. O. Kuntze) belongs to the Theaceae family and as one of the most important economic plants in the world, it has many medicinal properties and is consumed as a fun drink. Induction of polyploidy using mutagenic chemicals is one of the methods of breeding medicinal plants in order to increase the ability to produce secondary metabolites. In order to investigate the effect of colchicine treatment on the induction of polyploidy and to compare the changes of different traits in tetraploid and diploid plants (control) of Kashef cultivar, an experiment was carried out as a factorial in the form of a completely randomized design with two factors contains different concentrations of colchicine (0, 0.2, 0.5 and 1%) and duration times (3 and 6 consecutive days) with three replications. Morphological, microscopic and chromosomal counts were used to determine the ploidy level. The number of chromosomes in diploid and tetraploid plants was recorded as 30 (2n=2x=30) and 60 (2n=4x=60), respectively. The results showed that the concentration of 0.5 % colchicine in a period of 6 consecutive days with an efficiency of 12.82 % is the most suitable treatment for the production of tetraploid plants. Induction of polyploidy caused significant changes in the leaf size, chlorophyll and carotenoid contents and increased phytochemical compounds such as polyphenol, caffeine and antioxidant activity in tetraploid shoots compared to diploid. Based on the results of this experiment, increasing the ploidy level caused an increase in antioxidant compounds and some phytochemical compounds in tea medical plant.

One of the important crop in the north of Iran that now many of them are at risk of disappearing, is tea plant (Camellia sinensis). In this study 60 accessions using 24 RAPD markers were investigated. Used RAPD markers amplified 219 bands that 81.28% of them showed polymorphic pattern. Polymorphic Information Content (PIC) were calculated for all markers (0.29-0.50). Total calculated PIC for all markers was 0.49 that evidenced the usefulness and high level of efficiency of used markers for investigating the diversity of available accessions. The pairwise similarity coefficient between the accessions varied from 0.43 to 0.94 whit average 0.65 by using Jaccard’s coefficient. According to cluster analysis samples were divided into three main groups at 0.61 similarity and second group also created three sub-group (at 0.65 similarity). The notable point in this cluster analysis is that the samples did not follow the geographical pattern. Similar result was observed in bi-plot analysis. In comparing the similarity and genetic distance of the studied populations, it was found that there is a high genetic similarity between the tea populations in Iran (0.919-0.950). In general, the results of this study showed that when the samples are compared individually, they show high diversity, but when we compared them demographically, it is clear that the diversity between the populations are low due to the way propagation, cross-pollination of tea plants and the number of imported primary plants are restored.

Tea is an evergreen shrub. Oldness of the most cultivated tea shrubs as well as release of new tea cultivars and clones in the country has implemented a need for replacement of old tea shrubs with new tea cultivars. The easiest and cheapest method for propagation of tea is through cutting process but cuttings of tea cannot easily produce root. therefore, they need treatment with hormonal growth regulators. In this study, effect of plant growth regulators IBA and NAA alone and in mixture in three levels, 1000, 2000 and 3000 mgl-1 and ZnSO4 in three levels, 2000, 4000 and 6000 mgl-1 on rooting of semi-hardwood cutting of clone 100 was compared with control (no treatment) in two different planting dates (August and October). This study was conducted based on a factorial experiment using a completely randomized design with three replications. Results showed that effects of plant growth regulators on rooting and loss of cuttings was significant compared to control. The best and the most effective plant growth regulator in usual culture medium was combination of IBA+NNA in level of 2000 mgl-1 (1000 mgl-1 each). The best timing for cutting in tea shrubs was in August.

The herbal tea [Camellia sinensis (L.) O.Kuntze)] is a non-alcoholic beverage with many medicinal properties that has entered Iran in the last century and cultivated in the northern region. Understanding the genetic diversity of plants for breeding programs and the protection of germplasm is very important so that it can be stated that the collection and evaluation of domestic and foreign germplasm reserves is the most important stage in plant breeding programs. Many studies have been done on the nucleus genome of this plant, but the investigation of the organelles genome relationships in this plant is very limited. For the first time in this research, we have tried to investigate the diversity of this germplasm in Iran by using the genome of chloroplasts.

In this research, 35 tea-plant samples from six populations (east tea cultivation district (Nashtarood), central tea cultivation district (Lahijan), west tea cultivation district (Fashalem), imported genotypes from Georgia, imported clones from Japan, and imported clones from Sri Lanka) in three collections of the tea institute were studied. At first sampling of young and fully developed leaves was performed and the DNA genomes were extracted. By using five pairs of specific primers for the chloroplast genome (DT, LF, HK, SC and rbcl) and four restriction enzymes (BglII, HinfI, AluI and PstI) with Polymerase Chain Reaction- Restriction Fragment Length Polymorphism (PCR- RFLP) method, the variety in the chloroplast genome of these plants was investigated. NTSYS and POPGENE were used for cluster and population analysis.

In this study, about 6980bp of the tea chloroplast genome was amplified in polymerase chain reaction and examined by restriction enzymes. Of the 20 primer/ enzyme combinations, four combinations (DT/ HinfI, DT/ AluI, LF/ PstI and HK/ HinfI) were shown polymorphic pattern and these four compounds put samples into seven haplotypic groups (H1, H2, H3, H4, H5, H6 and H7). All these grouping were created due to the occurrence of insertion-deletion mutations in the range of 10-40bp. Mean of genetic variation within (HS), Total (HT) and degree of genetic differentiations (GST) were 0.25, 0.46 and 0.45, respectively.

The results of investigation of 35 tea samples showed that there was no cognitive genetic structure between the different sample regions. These results also confirmed the possibility of utilization Polymerase Chain Reaction- Restriction Fragment Length Polymorphism (PCR- RFLP) technique to investigation of genetic diversity and identify genotypes and varieties of tea.

In order to evaluate water deficient tolerance of nine selected tea clones, an experiment was performed in tea collections, Shahid Eftekhari Fashalam Tea Research Station in Shaft. The experiment was conducted as split plot based on randomized complete block design with three replications. Clones were considered as main plot in nine levels and irrigation treatments were as sub plot in two levels (irrigation and non-irrigation). Sixty days after the onset of water stress, water stress index, relative water content and quantitative traits included number of shoots, fresh and dry weight of shoot, length of shoot, length and width of 5th leaf and green leaf yield were evaluated in two stress and non-stress conditions for screening of tea clones. The results of variance analysis showed significant differences between clones, irrigation treatments and clone×irrigation interaction for all measured traits. Under water stress, relative water content, yield, yield components and other growth characteristics of clones decreased, so that the highest decrease belongs to green leaf yield. The results of water stress index and changing percent of measured traits showed that the clones of 276 and 100 promising clone with the highest water stress and the lowest changing percent of quantitative traits, have the higher potential to tolerate water deficit stress conditions compared with other clones and useful for tea breeding programs as tolerate clones, also because of the low water stress index and high changing percent of quantitative traits of clones 399 and 278, can be introduced them as susceptible to water deficit stress.

Tea (Camellia sinensis (L.) O. Kuntze) is one of the most important crops in nourth of Iran. Todays, many tea plants are being destroyed for various reasons, so having information about their genetics is helpful in designing breeding programs to reach appropriate plants for specific purposes. Therefore, in this research, the genetic diversity of some tea genotypes were identified in major cultivation regions of this crop in nourth of Iran with comparison by eight imported clones.

In this study genetic diversity of 42 tea plants were investigated by useing of two morphological and ISSR markers. Morphological study was performed by descriptor that registered form for tea and 31 traits were checked out. After selecting young and well expanded leaves, their genomic DNA were extracted and 10 ISSR primer were used for investigation of genetic relationships between 42 tea genotypes. Collected data was analysed by Euclidean distances for Morphological markers and SM similarity coefficient for ISSR and clusters were designed based on UPGMA algorithm. PCA anayesd were done by SPSS.

Comparative analysis on 31 morphological characteristics in tea genotypes and clone showed moderate variations and showed a narrow range. In cluster analysis at a difference of 6.6, the samples were divided into six groups; the main group consisted of the sixth group, which contained 88% of the samples. The results of PCA on morphological characters showed that the first, five principal components accounted for 54.21% of the total variance. Used of 10 ISSR primers produced 92 scorable bands that 72 of them were showed polymorphism pattern (78.26%). The PIC analysis showed ranging from 0.43 to 0.50. cophenetic test showed that SM similarity coefficient and UPGMA algorithm was the best for cluster analyses. According to ISSR data calculated similarity were range between 0.28- 0.93. In cluster analyses, samples at similarity 0.55 divided to four group that fourth group was the main created group and coverd 66.66% of samples. The results of PCA on ISSR data showed that the first, five principal components accounted for 72.98% of the total variance.

A significant variability was observed in the selected tea genotypes at morphological and molecular levels. Morphological study showed that distribution of tea plant in the past, although based on desirable characteristics, but due to the limited initial source of this plant, there is little variation among the genotypes of the regions. In relation to the ISSR marker, the polymorphism percentage and t polymorphic information content from the primers used in this study indicate the ability of these markers to differentiate the tea genotypes. From these results it could be understood that these series of characters and primers can distinguish genetic differences very well. Using these markers, genetic variation was observed between the genotypes of tea, but this diversity was not such as to be able to disrupt the genotypes of different regions. It seems that this separation could be achieved by increasing the number of primer combinations and using other markers such as SSR and SNP. Also, the results of this study showed that Iranian tea genotypes have high genetic variation because they are mostly reproduced sexually.

The diversity of 18 tea (Camellia sinensis L.) clones was evaluated in a randomized complete block design with two replications at Fashalam tea station (Rasht) in 2017. Physiological, biochemical and crop traits such as chlorophyll a, chlorophyll b, total chlorophyll, carotenoid, reducing sugar, total protein content, proline, tea leaf yield and number of shoots were measured. The results of variance analysis showed a significant difference in the level of one percent between tea clones for all studied traits. Mean comparison of clones showed that 100 and 399 clones had the highest amount of green leaf yield and number of shoots and in terms of proline and chlorophyll a were in second rank. The lowest coefficient phenotypic and genotypic variation related to total protein trait (2.97 and 2.58) and the most related to proline (85.88 and 85.84), and other traits were in this range. The cluster analysis by Ward method was able to place 18 clones of the study in three groups and the percentage grouping accuracy of clones by discriminant analysis was 100%. Average of green leaves yield and different types of chlorophyll traits in clones of the first cluster and proline and protein traits in clones of third cluster showed a high value. The clones of these two groups could be used in planning breeding projects and deciding to select suitable parents in crossing programs to increase the leaf yield and proline (which has a role in stress tolerance) in tea. Also, clones 100 and 399 showed a high average for green leaf yield and second rank for proline traits and therefore were introduced as promising clones.

The improving of black tea quality has always been in the goals of the tea industry in Iran. In this study, two tea processing techniques (Orthodox and CTC) were used to make black tea with passing Orthodox tea through CTC roller. The treatments consisted of the Orthodox rolling time (one and two times) and the time of passes CTC (two and four times). The first and second on sieve tea were used as the first and second controls after drying. Chemical quality of made tea from combining methods consisted of theaflavin, thearubigin, theaflavin to thearubigin ratio and aqueous extract were improved (p<0.05). It was no significant in time of passes through CTC. The sensory evaluation indicated the use of combination method improved sensory characteristics in made tea, including appearance, color, taste, aroma and total score, while tea appearance was undesirable (p<0.05). Made tea from first on sieve tea passed CTC has significantly better than other treatments in sensory quality. The value of made tea from combining methods were higher than two controls.

In order to determine the best resting period (without-plucking) and collar pruning time in tea gardens, a factorial experiment with two factors using a randomized complete block design with four replications was done during four years at the Fouman (Fashaml) Shahid Eftekhari Research Station. The first factor was resting period in three levels including without-plucking in autumn, plucking in autumn and without-plucking in the last picking turn of autumn, and the second factor was collar pruning time at four levels including 6th of December, 6th of January, 6th of February and 6th of March. The results of analysis of variance and mean comparisons showed that the without-plucking treatment in autumn had the greatest effect on starch storage in tea plants. Also, the effect of plant resting time on number of shoots, yield of green leaf and tannin percentage were significant, so that the highest mean shoot number, yield of green leaf and tannin content was observed in without planking treatment during autumn. The effect of pruning time on starch storage, yield of green leaf and tannin content was significant and the highest starch content and yield of green leaf was achieved in pruning treatment of 6th of January.

To investigate karyotipic variation in tea genotypes, karyotype of 16 tea genotypes were studied. Chromosomal parameters including total length of the chromosome, long arm, short arm, arm ratio and centromeric index were measured and analysed. Analysis of variance showed significant variation among genotypes in total length of chromosomes, length of long and short arms. The results of karyotypic formula showed that the chromosomes were metacentric and sub-metacentric. The satellites and nucleotide organizer regions were observed in genotypes 277 and 581. Except for the genotype 2021, which was triploid (2n = 3x = 45) and is reported for the first time, the remaining 15 genotypes were diploid (2n = 2x = 30). Based on karyotypic analysis and classification of karyotypes, genotypes 277 and 444 were classified 1B and 2B, and 14 genotypes were classified in classes 1A and 2A. Thus genotypes 277 and 444 were asymmetric. Principal component analysis based on karyotypic parameters showed that total length of chromosomes and length of long arm played the most important role in the first component. The first component explained 54.83% of total karyotypic variation. Cluster analysis also classified the genotypes into five groups.

Increasing the activity of antioxidant defense system against harmful agents such as reactive oxygen species، resulting from drought، is a common response in plants. In order to study these responses، the effects of two withholding irrigation in treatments (10 and 20 days) on the progress of the phenolic and flavonoid compounds activity malondialdehyde rate and also the amounts of chlorophyll a، total and carotenoids in three clone of tea (DN، 100 and 258) were investigated. Results showed that the amounts of phenol had the most increase in clones DN and 100 in 20 and 10 day-treatments respectively. However، changes in neither of the treatments had no significant effect in clone 258. The amounts of flavonoid and antioxidant capacity increased in clone DN with 20 day-treatment however; it decreased in clone 258 and stayed constant in clone 100. The most values of proline appeared in 20 day-treatments in all clones. Results of malondialdehyde contents were increased in clones 258 and 100، but it did not change in clone DN. Decreasing in amount of chlorophyll a and total chlorophyll were observed in clones DN and 100 with 20 day-treatments and remained unchanged in clone 258. Carotenoids remained constant in clones and treatments. On the basis of obtained data it seemed that clone DN had more ability against drought and consequently its defense mechanism was excited more than others.

Tea (Camellia sinensis L.) is an important beverage crops containing caffeine worldwide. Releasing cultivars with high yield and quality demands knowledge about tea genetics and cytogenetics. Karyotypical study of nine tea genotypes was carried out using squash method on root tips. The best microscopic slides was first obtained using root tip of tea cuttings pretreated by α-bromo Naphthaline for 8 h، fixed by farmer solution and then stained with hematoxilin. Eight morphological traits were taken into consideration، each within three meiotic chromosomes using Micromeasure software. Analysis of variance showed significant differences (1%) among genotypes، chromosomes and the interactions، for all of the measured parameters. All studied teas were diploid and had 2n=2X=30 metacenteric and submetacenteric chromosomes. Total length of chromosomes ranged from 1. 10 to 4. 42 μm. Also، in terms of total length of chromosomes، genotypes 4 and 8 had the highest correlation and genotypes 2 and 9 had the lowest. Moreover some cytogenetic statistics was used in order estimate the karyotypic symmetry of genotypes. The results obtained showed that the genotypes 1 and 2 had the most asymmetric and genotypes 3 and 8 had the most symmetric، based on the TF%. Ordination based on principal components analysis (PCA) revealed presented that more than 97% of total diversity with two components was described. Centromeric index and long arm length had the most roles in the components، respectively. Cluster analysis using UPGMA method، also grouped the genotypes in four clusters and 100 promising clones were separated from the others، similar to result of PCA Bi-plot، relatively.