مجله فنآوری زیستی در کشاورزی
سال سیزدهم شماره 2 (زمستان 1393)

Total of nineteen isolates of Botrytis sp. were collected from infected strawberry plants of hydroponic cultivated greenhouse and fields surround kurdistan province. According to the morphological characteristics، isolates were identified as Botryits cinerea species. For confirming the species identification by molecular method specific primers C729 were used and PCR test repeated twice. The results showed that these isolates amplified the 700 bp band which is specific for this species. Genetic diversity of existing isolates was investigated by RAPD analysis. According to the results obtained، isolates were clustered into four groups. Considering this clustering mode، in first glance، most of the greenhouse isolates were grouped together and separated from the group of field isolates، but some of them didn’t follow the pattern and were divided differently. Likewise، study among field isolates collected surround the province and their relationships، showed that there is no correlation between molecular clustering and geographical origin of these isolatesو but also it was clear that some of the geographically further isolates were more similar to each other rather than isolates from closer geographical regions.

Pollution of natural ecosystems by petroleum compounds have been more considered because of their high persistence in environment and adverse impacts on human health. Nowadays، a wide range of physical، chemical، thermal and bioremediation methods are employed in order to remediate contaminated sites. Bioremediation technology is based on use the natural or engineered microorganisms for reclamation of contaminated sites and environmental conservation. The current study was carried out in order to isolate and characterize the oil degrading native bacteria from Tange Fani Poldokhtar region، Lorestan province، Iran. Soil and water of contaminated sites were sampled and after transfer to the laboratory، samples were cultivated on the nutrient agar، nutrient broth and 2xYP media. After purification، bacterial strains were identified on the basis of phenotypic-biochemical traits and sequence of 16S rDNA gene as Pseudomonas aeruginosa، Acinetobacter junii، Acinetobacter baumannii، Delftia tsuruhatensis، Sphingobacterium multivorum، Stenotrophomonas acidaminiphila and Comamonas koreensis. Furthermore، the bacterial strains were evaluated regarding biofilm formation، siderophore production and swarming motion، which the highest amount of biofilm formation and swarming motion was belonged to P. aeruginosa. In this study، bacterium Comamonas koreensis for the first time as oil degrading introduced.

Identification of controlling genomic loci of complex traits with providing of useful genetic information، as introduction of breeding programs of marker assisted selection are a practical approach in breeding of complex traits. In the present study، 96 recombinant inbred lines were used as mapping population to identify controlling QTLs of two important traits related to grain quality including of gelatinization temperature and protein content. Linkage map was constructed with 438 markers including 314 AFLP and 124 SSR. The results of composite interval mapping totally five QTLs detected for two traits. Between two identified QTLs for gelatinization temperature، qGT-6 was detected in vicinity of wx gene on the chromosome 6. Considering of Anbarbu allele’s role in both of QTLs in enhancing of gelatinization temperature، these QTLs can be very efficient in improving this trait. As well as for protein grain three QTLs was identified that considering of positive additive effect of qPR-1b and explain of more than 13 percent of phenotype variation it can be considered in marker assisted selection.

Fusarium graminearum is one of the most important causes of wheat scab in different part of the world. This fungus is able to produce widespread Trichothecene mycotoxin such as Nivalenol and Deoxynivalenol which are harmful for both human and animals. The wheat ear samples were collected from wheat fields in Sistan and Baluchestan province. Sampling was performed during 2011-2012. The fungal isolates were separated، purified، then identified using various keys for Fusarium spp. and 293 isolates of Fusarium species belonging to eight species were isolated and identified. Among the identified isolates، F. graminearum showed the highest frequency (68. 8%) compared to other species. When the isolates of F. graminearum were morphologically identified، 168 isolates were chosen using species-specific primer pairs (Fg16F/Fg16R). In these isolates، the presence of three genes: Tri13، Tri5 and Tri7 were detected by using PCR and specific primers for the genes. The two NIV and DON chemotype was detected among isolates of F. graminearumusing specific primers. The results of PCR reaction with specific primers showed that all tested isolates possess the genes involved in production of trichothecene. Therefore، the detection of genes involved in trichothecene production using species-specific primers to determine Fusarium isolates producer trichothecene could alternative the chemical expensive and time procedure.

Rapeseed (Brassica napus) with more than 40 percent of oil and more of 40 percent of cake protein and just 5 percent of saturated fatty acids in oil is accounted as an important oil grains. Evaluation of genetic diversity is the main and basic principles of each breeding program. In this study genetic diversity among 45 various genotypes investigated by 15 ISSR markers. For analysis of obtained data، amplified bands scored as zero (absent of band) and one (present of band) data with Darwin5. 0، MEGA3. 1 and Xlstat softwares. Cluster analysis of data obtained by use of Dice dissimilation coefficient and neighbor joining (NJ). Obtain dendrogram divided genotypes into five clusters. Number of all amplified alleles was 110 and equal to average of 7. 33 alleles for each marker. Total distance among genotypes and Information polymorphism index mean was equal to 0. 570 and 0. 350 respectively. These markers were able to demonstrate three individual bands in UBC 125، 134 and 165 markers for Savannah، Olpro and Zarfam genotypes respectively. accordance obtained results from this study، peering is showing that using of more diverse genotypes can be considered for breeding programs in order to exploit heterosis for producing of hybrid.

To investigate accuracy of linkage disequilibrium (LD) in genomic simulation studies a base population was simulated in two statuses LD with values of zero and one in first generation. Effective population size، LD and variance of markers frequency were used for determination of simulation accuracy. The results showed a high correlation between actual and expected effective population sizes. Also observed and expected LD’s were nearly similar and the given trend was improved with increasing effective population size. In addition، the variances of the observed and expected free recombination markers frequency were adapted with each other. According to the evaluated criteria the base population was accurately simulated which it could be used to generate the reference population. Based on the results، we suggest that proposed criteria should be used to verify the simulation after creating of historical population in genomic simulated studies.

Using mutation has been relevant to induce genetic variation and new genetic material، and it was used to improve different qualitative and quantitative traits in crop plants. In this research، eighteen rice mutant lines which were selected from a mutant population developed from mutagenesis of rice elite cultivar Neda by using ethyl methane sulfonate (EMS) for reduced plant height، higher tiller number، early maturation and higher yield، were evaluated at molecular level using ISSR markers. Eight out of 10 primers successfully amplified DNA from different genotypes. In total، 67 bands (in average 8. 4 bands per primer) were produced، 53. 7% of which showed polymorphism between genotypes. Average Nei`s genetic diversity in the studied population was 14. 1%. Highest similarity to original cultivar (86. 8%) belonged to high-yielding mutant group and lowest similarity (76. 7%) belonged to high-tiller mutant group. Cluster analysis using UPGMA method، placed the original cultivar alone in one cluster and 4 mutant groups in another distinct cluster. Cluster analysis was also performed separately in each sub-cluster that except for high-tiller group it could discriminate the original cultivar from any other three mutant groups. On the basis of the results of this research، it can be concluded that mutation technique was a desirable method for inducing genetic variation in rice and also it is possible to use ISSR markers for identification of mutant lines in populations developed from mutagenesis by EMS.

The objective of the present study was to design and construction of a specific chloroplast vector to produce biodegradable biopolymers and also induce abiotic tolerance in transgenic plants. To do this، three genes involved in Polyhidroxybutirate biosynthetic pathway were isolated. The genes isolation and functionality accuracy was proved using expression in recombinant E. coli. Then an inducible expression cassette was designed for this operon under control of groE promoter and Thr terminator and subsequently cloned adjacent of neo selectable marker gene integrated with large subunit of atpB under control of psbA promoter and terminator among loxP sequences. The resulting cassette was cloned in the center of a 4Kb fragment from chloroplast genome that makes possibility of transgene targeting into trnI/trnAintergenic region by homologous recombination. These recombinant plastid vectors were called pFNPi (+) and pFNPi (-). Moreover badh gene was added to the chloroplast vector. This gene coding a protein detoxifing Betaine could be used for resistance to salt، drought and cold stresses. After intron removing، codon optimization and remove of internal restriction enzyme recognition sites was placed under Prrn promoter and T7gene10 5’UTR. The final resulting vector called pFBNPi will be suitable for bacteria and plant transformation for production of biodegradable biopolymer and resistant transplastomic plant for salt، drought and cold stresses.

Faba bean is an important legume in Iran and many other countries. Bean yellow mosaic virus (BYMV) has been found in faba bean fields in a number of countries. In this survey، during 2010 growing season، a total of 240 faba bean samples with mosaic، distortion and stunting symptoms suspected for BYMV infection were collected from faba bean fields of Jiroft، Kerman province. These samples were analyzed using BYMV polyclonal antibody in serological tests (DAS-ELISA and TPIA). In faba bean fields of Jiroft، the percentage of infection with BYMV was 62. 5. Among the collected weeds، only two samples of Chenopodium quinoa were infected by BYMV. In greenhouse mechanical inoculation assays، the virus induced local lesions and systemic symptoms on a number of plant species of Chenopodiaceae، Amaranthaceae، Fabaceae، Solanaceae and Brassicaceae families. A fragment of 1700 nt length from the NIb، CP and 3´-UTR regions of a BYMV isolate (J2) was amplified in IC-RT-PCR using a Potyviridae universal primer pairs. BLAST analysis on the partial NIb region of the isolate and comparison of the nucleotide sequence showed that this isolate is similar to a Japanese isolate. This is the first report of occurrence of BYMV in faba bean fields of Kerman province.

Tomato mosaic virus is one of the most damaging viruses infecting vegetable crops. This study was conducted to estimate the distribution of ToMV in some cucurbit plants including cucumber، squash and watermelon. Toward this aim، during the years 2008 to 2010 samples were taken from numbers of cucurbit cultivations of different provinces of Iran. Leaf samples were selected from symptomatic plants and tested with DAS-ELISA method. Samples with positive reaction with antisera in ELISA were further selected for subsequent assays. Bioassay was done using with the herbaceous host plants and mechanical inoculations. RT-PCR was done with the total RNA extracted from the leaf samples. Samples were amplified by semi-nested RT-PCR using with the universal tobamovirus primer and Tomato mosaic virus (ToMV) specific primer pairs. A desired DNA fragment with a size about 675 bp was amplified for ToMV infected plants. Results of the ELISA، RT-PCR tests and bioassay confirmed the presence of ToMV in different cucurbit crops as follow: cucumber and squash in Maragheh district، watermelon in Orumia district، cucumber in Zahedan and Roodan districts and squash in Varamin district. Results of this study indicated that cultivated cucrbits in Iran are infected with ToMV. It is necessary to consider the ToMV control management strategy in general control management scheme for cucurbit plants.

Molecular farming is the use of plants to produce valuable pharmaceutical proteins in the bulky and cost-effective manner. Molecular farming and the production of recombinant proteins (such as insulin، interferon-gamma، tissue plasminogen activator، etc.) in transplastomic plants، has achieved a great success in Iran. Due to the growing demand for recombinant proteins، the selection of a proper expression system is very important. Chloroplast genome engineering، for its many advantages، is one of the suitable methods for the production of recombinant proteins. To achieve high levels of recombinant proteins in chloroplasts، plastid’s specific transcription، translation and post-translational modifications should be considered. Some of the strategies used for increasing of foreign proteins in chloroplast can be mentioned: gene insertion in the suitable site in chloroplast genome، use of strong promoters، appropriate regulatory sequences in the 5'' and 3'' UTRs، codon optimization، and utilization of factors enhancing protein stability such as chaperons and fusion proteins. In this article، besides a brief description about chloroplast and its genome، methods for enhancing recombinant protein production in transplastomic plants were discussed.

During the autumn of 1392 strawberry farms were inspected in the province of Kurdistan. In most fields، leaf spot symptoms of field bindweed were observed. Infected tissues surface sterilized with 5. 0 percent sodium hypochlorite and were cultured on the medium PDA (potato، dextrose، agar). After seven days of incubation at 25-23°C، purifiction was done from edge of colonies in the maner of hyphal tip on water-agar medium. A total of 21 isolates posses pycnidium obtained that were morphologically similar. one of the isolates as representative was choosen and its ITS1، ITS2 and 5. 8s gene was sequenced. Finally، based on the results of morphological and molecular studies the causal agent of leaf spot of field bindweed was diagnosed as Peyronellaea glomerata. This is the first report of P. glomerata leaf spot on field bindweed. This fungus can be used as biocontrol agents in the integrated control of field bindweed if does not able to disease other crop in Kurdistan area.