مجله بیوتکنولوژی کشاورزی
سال ششم شماره 3 (پیاپی 15، پاییز 1393)

Transition from vegetative to reproductive stage in rice plays a vital role in regulation of timing of flowering and is one of its adaptation related traits to different cultivation areas or different cropping seasons. Several genes related to heading date were identified in rice, that Hd1 on the 6th rice chromosome is one of the most important ones. In this study, two rice cultivars including Sadri (as early flowering donor parent) and Neda (as late flowering recipient parent) were crossed to investigate the effect of Hd1-harboring genomic region on rice heading date, and BC2 generation was developed with two times of backcrossing. Then a single heterozygote plant in BC2 generation was self-pollinated to obtain BC2F2 population for conducting phenotypic and molecular studies. The studied population had a continuous phenotypic distribution in heading date and some individuals in the population showed transgressive segregation over late flowering parent. On the basis of an InDel region in first exon of Hd1 gene a functional specific marker was designed which produced a distinct different banding pattern among the cross parents. Hence, this marker was used toevaluate allelic pattern of Hd1 in BC2F2 population. Analysis of allelic segregation revealed that Hd1 locus in the studied population followed from the expected Mendelian segregation (with 1:2:1 ratios). Composite interval mapping (CIM) with microsatellite markers showed that genomic region at Hd1-RM527 interval had a high relationship (LOD>7.5) to heading date and explained nearly 28 percent of phenotypic variation of the trait. Additive effect of the donor parent allele on the trait was negative and was estimated nearly -3.4 days. Results of this investigation indicated that Hd1 had a significant effect on heading date in rice and it is possible to use the newly developed functional marker in this research for tracing the Hd in segregating populations and marker-assisted selection (MAS) for early maturity.

Sunflower is one of the most important oilseed crops. Evaluation of genetic diversity and grouping of genotypes and lines are computed as important agents for plant breeding programms. In this study, the genetic diversity and classification of some advanced inbred lines of sunflower developed during different breeding programs were studied using ISSR markers. Among 21 primers, 15 ISSR primers were selected for their reproducibility. A total of 70 bands were produced through ISSR primers which 27 and 43 bands out of them were monomorphic and polymorphic, respectively. Results revealed that primer UBC 807 (0.4) had the highest polymorphism information content value and primer UBC 804 (0.15) had the lowest value. Using Dice similarity coefficient the lowest amount of genetic similarity 0.65) was observed between breeding lines SF25 with SF278 and the highest ones (0.93) was observed between lines HA336 and SF315. Cluster analysis using UPGMA algorithm was divided the studied lines into 8 separate groups. Regarding to principal coordinates analysis; each component accounts a small percentage of the total variation which emphasis on good genomic distribution of selected ISSR primers.

Grapevine (Vitis sp.), belongs to Vitaceae family, is one of the most important food sources in world because of its wide consumptions. In the current investigation, 22 SSR primer pairs and 7 AFLP primer combinations were used to identify molecular makers associated with 14 flower and fruit related traits in Iranian grapevine germplasm. The traits studied, included fruit water content, percentage of pollen germination, percentage of fruit production, bunch and berry weight, fruit lobe and single seed weight, bunch and yielding system length and width, total soluble solids, acid content and fruit pH. Stepwise regression analysis showed significant relationship between some SSR alleles and all 14 traits in grapevine varieties. In general, 49 alleles produced by 18 SSR primers, showed significant association with variation of 14 traits. The highest and lowest number of associated markers achieved for berry weight and fruit lobe weight each with 7 alleles and pH, fruit acid content, fruit water content and percentage of pollen germination each with 1 alleles, respectively. Significant association between polymorphism of AFLP markers and studied traits was also detected. Forty-nine AFLP fragments showed significant association with the variations of 14 traits. Some of the associated AFLP segments were common among different traits. The fruit lobe weight, with 11 associated markers and total soluble solids, percentage of fruit production, pollen germination and bunch width each with 1 associated marker, were the traits with the highest and lowest number of associated AFLP markers, respectively.

Milk is considered as a complete food to meet parts of human daily requirements for a long time. Proteins are one of the most important ingredients of milk and play a critical role in processing technology such as cheese factory. Several factors such as genetics influence on quantity and quality of milk as well as cheese. Several genes on BTA6 are recognized to be related to milk and cheese related traits. In the current study the genetic association of three candidate genes of the region including; Osteopontin, PPARGC1α, and Kappa casein with milk and cheese related traits, were investigated in Brown Swiss cattle. Total DNA was extracted from 100 cattle by salting out procedure. Genotypes frequencies were estimated by PCR-RFLP and using specific enzymes. Four factors of efficiency fat, protein and dry matter of cheese were measured. Association between polymorphisms and breeding values and cheese quality were analyzed by GLM procedure at 5% of significant level. The results showed that the population was in Hardy-Weinberg equilibrium, except Kappa Casein locus. Significant effect was observed only in combined genotype of PPARGC1A-T19C and kappa casein with breeding values of milk production. Because of the limited samples or missing of some haplotypes, genotypes had no effect on cheese production. The increasing of the samples to demonstrate other genetic combination and using modern techniques such as dense SNPs are strongly recommended.

Sequencing and analysis of the data obtained are best and most common methods in studies of genetic diversity. PIT-1gene is known as growth hormone IGF-1 gene. this gene is a member of family of transcription factors in the poultry is located on chromosome 1, with a length of approximately 14 kb. This gene has seven exons and five introns in poultry. The purpose of this study was analysis region of intron 5 sequence of PIT-1 gene in broiler chickens Arian lines and determining its phylogenetic relationship with other chicken breeds. For this study, blood samples were collected of seven broiler chickens of both sex Arian lines. After extracting DNA from them, target gene was amplified with specific primers and after purification was sequenced. Five different haplotypes were determined based on six single nucleotide polymorphism sequence and the final sequences of each haplotype with length approximate 260 bp which includes 21/92 % adenine, 18/8 % guanine, 32/69 % cytosine and 27/31 % thymine. After ensuring the accuracy of sequencing, submitted to gene bank database (NCBI) with accession number of JQ946630- JQ946636.The phylogenic tree was drawn with consensus sequence of PIT-1 gene of Arian line chicken and other similar sequences of different chicken breeds obtained from gene bank. Results the phylogeny indicated that Arian line chicken and American Chickens are on a lineages

Dracaena (Dracaena marginata) is one of the important economic ornamental plants in Iran as well as around the world. Micropropagation and enhancement in genetic variation of various ecotypes of this ornamental plant have been considered in several research studies for several years. The main objective of this study was maintenance of the best media culture for callus induction and shoots regeneration in Dracaena "Tricolor". Therefore, small stem segments of 0.5 cm in length were cut from the maternal plants. In order to set up a proper decontamination control system for Dracaena small stem segment, various levels of sodium hypochlorite including 1%, 1.25% and 1.5% were applied in combination with different time periods of 15 and 10 minutes. Moreover, the most suitable medium cultures for callus induction, shoots regeneration and root induction were identified. A proper statistic approach has been used for data analysis in various stages of callus induction and shoots regeneration. The obtained results showed that the most appropriate decontamination treatment to obtain the highest percentage of tissue healthiness could be applied by using 1.25% sodium hypochlorite for 15 minutes. The most suitable MS medium for callus induction was contained either 0.5 mg/l 2,4-D or 1 mg/l NAA. Moreover, applying the following growth regulators including 1 mg/l Kinetin in MS based medium, and 1 mg/l NAA provided the best results for shoots regeneration and root induction.

The present a method for fast and accurate detection of abortion bacterial agents is the most priority of control and treatment for this disease. The aim of this study was the optimization of Real Time PCR for simultaneous detection of four of the most important bacteria causing abortion (Campylobacter, Brucella, Yersinia and Salmonella). 217 vaginal swab samples 132 samples from aborted and 85 samples from non-aborted ewes) were collected from some rural regions of Zanjan province. Pure samples of Brucella, Yersinia and Salmonella were also purchased from Persian Type Culture Collection (PTCC). After DNA extraction, specific primers of Brucella, Salmonella and Yersinia were examined on each purchased bacteria DNA sample. With 68 positive samples from 132 aborted ewes and 29 positive samples from 8 healthy ewes, Campylobacter was distinguished as a main factor of abortion in rural areas of Zanjan. The precision of Campylobacters primer was also assessed using Real Time PCR in different dilutions. The results showed that all bacteria in the same condition concurrently were amplified with Real Time PCR using specific primers and Campylobacters primer is able to identify and amplifying of bacteria in all dilutions even when just one copy is available.

The use of antibacterial property of nanosilver particles has significantly expanded and applied in a variety of products, consisting clothing, paints, plastics, food packaging, wound dressings, bandages, and household appliances such as refrigerators and washing machines. However, their use in animal feeding as prebiotics have been remained minimized, mostly because of the low cost antibiotics used as growth promoters. After the ban of this practice, silver compounds appear as a potential alternative to other products already in use. The results of different studies with different species revealed that Ag NPs had no negative impact on their growth and development. Our works at Shahid Bahonar University of Kerman using 0.5, 1 and 1.5 ppm Ag NPs in water improved the number of lactic acid bacteria without any negative impact on broiler performance. However, the major concerns about the safe use of additive in animal feeding are its effective role as antimicrobial, acting selectively over potential pathogens but not over symbiotic microbial communities; a low toxic effect over the animal and its human consumer; and a low risk of environmental pollution. Despite the growth of commercialization of Ag NPs, little is known about the environmental effects of the widespread use of the products containing AgNPs. As low as just a few ng L−1, can affect prokaryotes, invertebrates and fish indicating a significant potential risk to the environment. Mechanisms of toxicity are still poorly understood although it seems clear that in some cases nanoscale specific properties may cause biouptake and toxicity above that caused by the dissolved Ag ions. The present study reviews the proceeding research works on toxcicity of Ag NPs along with some recommendations for better understanding of the role of nanoscale silver particles in environmental and ecotoxicological researchs.

Plants are suitable replacements for current biodrugs and recombinant protein expression systems such as transgenic animals or microbial systems. As the plant plastid genome is highly polyploid, the transformation of chloroplast permits the introduction of thousands of copies of foreign gene per plant cell and generates extraordinarily high levels of recombinant protein. Cardiovascular diseases are the second one leading cause of human mortality after cancer. Human tissue-type plasminogen activator (tPA) is one of the most important pharmaceutical recombinant proteins involved in the breakdown of blood clots in different parts of body such as brain and heart blood vessels. The truncated form of tissue plasminogen activator (K2S) has longer plasma half life, better diffusion into the clot and higher fibrinolytic activity. In this study, in order to introduction of K2S gene in tobacco chloroplast, after construct design, the interest gene was ligated to pKCZ vector and cloned in E. coli. Following the successfully shooting of the K2S-containing vector (pKCZK2S) to leaf explants using the biolistic delivery procedure, the explants were selected on selection medium containing 500 mgL-1 spectinomycine antibiotic. After regeneration of grown shoots on selective medium, in order to achieve homoplasmy, fourth rounds of selection and regeneration in presence of spectinomycine antibiotic were performed. The presence, site-specific integration, expression of the K2S gene and homoplasmy in transplastomic plants were confirmed by PCR, RT-PCR and Southern blotting methods.

Pistachio (Pistacia vera) is one of the most important horticultural products in the world known by the name of Iran. Moreover, Iran has the richest germplasm of pistachio due to the origin of pistachio and the existence of large numbers of commercial pistachio cultivars. Nevertheless, few chromosomal studies in pistachio have been yet reported and they however showed contradicting results on the number of chromosomes. In the present study, different pistachio species including P. vera, and wild species, i.e., Pistacia khinjuk and P. atlantica subsp. mutica were studied by classical chromosome staining and GISH technique. In order to stain chromosomes, two methods of aceto-iron-haematoxylin and Feulgen were used. Results indicated that the best staining was achieved by Feulgen compared with another method. Chromosome numbers of ‘Ohadi’, ‘Akbary’, ‘Ahmad-Aghaei’, ‘Nish-Kelaghi’ and ‘Khanjary’ were 2n=30 while chromosome numbers of ‘Italiaei’, ‘Ghazvini’ and ‘Sarakhs’ wild variety were 2n=28. Chromosome numbers of both P. khinjuk and P. atlantica subsp. mutica were 2n=28. In GISH experiment, no signal of hybridization of P. khinjuk and P. atlantica subsp mutica genomic probes were detected on P. vera chromosomes. This lack of detectable hybridization sites may however indicate that these chromosomes are probably originated from different taxa.

Study on native chickens could be considered as a way to maintain population genetic variation, breeding programs development and helps to obtain information on their genetic structure. Mitochondria genome Sequencing is one of the most practical methods in order to determine phylogenetic relationships among close populations and species. The aim of this study was to investigate the nucleotide sequence of the mitochondrial DNA control region of Khorasan native chickens, its genetic and phylogenetic analysis. Blood samples were supplied from 5 Khorasan native chickens. Then 845 bp fragments of extracted DNA were amplified by using specific primers. Sequencing of amplified fragments was carried out with Sanger method. Using of similar sequence of mitochondria DNA from other chicken breeds available in the NCBI database, phylogenetic tree was drawn and matrix of genetic distances was formed between Khorasan native chickens and other breeds for mitochondrial DNA control region. According to the results no variation was observed among the nucleotide sequences. Phylogenetic test results showed the least genetic distance was recognized between Khorasan native chicken with Marandi chicken of Iran, White Leghorn, Buff Cochin and satsuma dori of Japan, Lv erwu of China, Denizli and Gerze of Turkey, Dandarawi of Egypt and India. Thereby can conclude that Khorasan native chicken of Iran has a close relationship with Asian chickens.