سید الیاس مرتضوی

Gene drive technology is one of the modern technologies that have many potential uses in the agriculture, industries and medical sciences. Like other technologies, this technology has some biosafety issues related to human and animal health and the environment. These issues were discussed in this article. The most important issue is related to changing in Mendelian inheritance template in filial generations and distribution of a specific allele in a local population. Moreover, adequacy of the national and international laws and regulations related to GMOs for gene-drived organisms, and also the risk assessment methods for gene-drived organisms including identification of possible risks of the gene-drive technology and containment of the gene-drived organisms during field experiment were discussed. In the following, the possible risks in release of gene-drived organisms were considered and some related aspects including the risks related to pathogen control, the risks related to invasive species control, the risks related to control agricultural pests, and the other social and economic dimensions were investigated.

Risk assessment for a transgene is one of the key steps in the genetic transformation. In order to use cryIAc gene in production of transgenic plants, a library and in-silico research was performed to confirm the safety of the gene product for human consumption, animal feed and the environment. In the first step, the molecular mechanism of action of the CryIAc protein and its specific receptors in the midgut cells of the target organism was explored. Digestibility of the protein in the stomach and intestine liquids have been investigated based on the peer reviewed documents. It was demonstrated that the CryIAc protein could be completely digested in the stomach liquid of vertebrates, especially mammalians including the human. It was shown that addition of the pure protein into the animal diets did not result to any abnormalities in the growth pattern and also blood histochemistry of the experimental animals. Also, whole plant assays using the different diets contained the transgenic and non-transgenic plant materials in different experimental animals was discussed. These experiments did not result to any differences between the groups. Possibility of the toxic effect of the CryIAc protein was investigated via in-silico tools. The results did not show any similarity between the known toxic protein and the CryIAc protein. Allergenicity of the CryIAc protein was assessed in-silico in different ways. It was proven that there were any similarities between the protein and the other known allergens. At the end, the persistency of the CryIAc protein in the soil was explored. Since the different experiments on exposure of the pure protein or the transgenic plant tissues into the environment proved that the CryIAc protein is degraded easily in the soil, it could be concluded that the CryIAc protein is an environmental friendly protein. Eventually, the final conclusion is that the CryIAc protein is a safe protein for human consumption, animal use and the environment, and could be used safely in genetic engineering programs.

The most threatening factor in cotton production worldwide is Lepidoptera pests. The pest management using chemicals imposes huge costs and environmental side effects. Introduction of genetic engineering as a modern technology has opened a new insight for investigators to control the pests through transgenic pest resistant varieties. Cotton was the first transgenic crop which received cry genes, and introduced to farmers after numerous field trials. The field trials on evaluation of transgenic lines are a complex process in three levels comprising mini, mid and large scale trials, but the first step in this process is evaluation of transgene in the field condition along with considering other important crop properties. There are several Lepidoptera pests causing damage to cotton in nearly all cotton producing areas worldwide. However, bollworm (Helicoverpa armigera Hübner) and spiny cotton bollworms (Earias vittella Fabricius and Earias insulana Boisduval) are the most the most important pests in Iran. Also, there are many biotypes among different geographical populations. This fact implies that the transgenic lines should be tested in different locations within a country. A number of research works has been conducted to produce transgenic cotton via both modern biotechnology and traditional backcrosses in Iran. As a result, some different transgenic lines harbouring cry genes were produced. Primary evaluation of these transgenic lines was resulted to a few lines exhibited the most resistance characters against the pests. More precise evaluation of the lines for pest resistance traits and yield was done in this study.

Three transgenic lines harbouring cryIAc gene comprising A, B and CRI lines along with a non-transgenic variety as negative control (Varamin) were used in the experiment. The experiment was conducted in a Lepidoptera pest affected farm at Minab, Hormozgan from June to July of 2015. A complete randomized block design with eight replications was adopted as the statistical model. Each plot consisted of eight lines with 8 meters length in which the distance between lines were 80 cm. All agricultural operations including irrigation, fertilizer and weeding were carried out according to local recommendations. During growing season, some properties related to the pest resistance including number of bolls per plant, percentage of contaminated bolls, percentage of contaminated locule, and percentage of boll injuries were recorded, and damage index was then calculated. The varieties yield was recorded as well. All traits were recorded on two separate dates i.e. June and July, 2015. The data were analyzed using SAS software according to the adopted statistical model.

Analysis of variance showed that all recorded traits in the experiment were significant for cultivars effect, except for boll injuries percentage. Primarily, the current results demonstrate that all cultivars were exposed to the pest equally. Other traits including number of bolls per plant, percentage of contaminated bolls, percentage of contaminated locule, damage index and yield differed significantly between the cultivars. The mean comparison for number of boll per plant showed that transgenic A and B (transgenic) lines produced and kept the most boll number, and CRI transgenic line and Varamin cultivar ranked as the second. This trait is very important, because it not only acts as a yield component, but also involves in resisting against pests. The mean comparison for contaminated boll and contaminated locule traits showed that non-transgenic Varamin cultivar had the largest contamination amounts. In contrast, transgenic A and B lines exhibited the least contamination for both traits.
The most important trait expressing resistance is damage index. The lowest damage index was calculated for B line (0.69 and 0.46% in June and July, respectively) and A line (0.75 and 0.56% in June and July, respectively), and CRI line ranked as the second (2 and 5%). Meanwhile, the damage index of Varamin cultivar in this experiment was 3.8 and 11.5% in June and July, respectively. The damage index for A and B transgenic lines in the first and second recording show a downward pattern in contrast to CRI line and Varamin cultivar. Hence, Lepidoptera pests does not succeed in feeding on the A and B lines. Moreover, A and B lines exhibited the best yield (5041 and 4466 kg/ha, respectively), and the CRI line ranked as the second (3382 kg/ha). The Varamin cultivar yield was 3071 kg/ha which was the least record.

In general, two out of three transgenic lines comprising A and B lines had the best yield, the minimum contaminations and injuries by Lepidoptera pests. Varamin cultivar, as non-transgenic control line showed the least tolerance and yield and the CRI cultivar were ranked between the superior transgenic and non-transgenic lines. Both A and B transgenic lines have proper agronomic traits and the best resistance characters against Lepidotera pests. It can be concluded that these lines, as the first Iranian transgenic varieties, can be used for registration and commercialization.

In order to produce transgenic rice lines lacking selectable marker gene, biolistic co-transformation technique using gold nanoparticles was adopted. In the first step, the efficiency of different sizes of gold particles was evaluated. The results showed that the efficiency of the nanoparticles in the transformation was comparable to that of the micro particles. Subsequently, two separate plasmids including p UBC harboring cry IA (c) gene and pTRA132 carrying hph gene as selectable marker gene were mixed and coated with gold nanoparticles. Embryo genic calli originated from mature rice seeds were targeted with plasmid-coated nan projectiles. Putative transgenic cells were selected from selection media after three subcultures in MS medium were supplemented with 50 mg hygromycine B l-1 in three week's intervals. The selected calli were then grown into putative transgenic plantlets. Molecular analysis performed on the regenerated lines showed that at least one copy of both genes was inserted into the genome of some transgenic lines. RT-PCR analysis also confirmed successful transcription of the cry IA (c) gene. Accordingly, it could be concluded that the biolistic co-transformation method using gold nanoparicles could be a successful method for gene delivery into rice varieties. Nevertheless, expression of cry IA (c) gene and loci of both genes should still be investigated in future studies.

Publication of Seralini et al. 2013 about the risk of tumor induction in transgenic maize cultivar resistant to Roundup herbicides raised a wave of propaganda against transgenic crops in the world. One part of this Propaganda was a video called “Moment of Truth«in that، authors expressed their results and concluded that the transgenic maize and implicitly، all other transgenic crops have an ability to induce tumors in mice and even in human as great as a basketball ball. So، they ask EU officials to revise about production، release، importation and consumption of such products in EU countries. The video was translated and published widespread in Iran and affected on public opinion and caused public anxiety، so this article was written in response to it. The evaluation of this study indicated that the race of mice (Harlan Sprague-Dawley) which has been used in this study has a high risk of tumors incidence itself، even without feeding the mice with transgenic crop. In addition، problems such as lack of replications and statistical analysis in experiments of this study increased its uncertainties. Besides، the best conclusion based on Seralini’s resultscould be that GM maize was prevented tumors and cancer، or at least، there was no impact on incidence of tumors. However، surprisingly the authors concluded that GM maize might induce tumor in mice. Officially and scientific resources including European Food Safety Authority (EFSA) have rejected these findings، and finally in 2013، the paper was retracted by the publication Journal.

In order to produce a marker-free transgenic rice with improved tolerance to salinity and drought stresses, expression vectors pABRII-Chl and pABRII-Cyt containing "choline oxidase" gene (with or without leader sequence respectively) were constructed from pChl and pCyt and pTR 132 for co-transformation. The pChl and pCyt vectors were digested with HindIII-BamH and BamHIEcoRI enzymes. Then the resulting sequences were ligated and inserted into expression vector pTRA132, in which the HindIII-EcoRI fragment (hph gene) had been deleted. The constructs pABRII-Chl or pABRII-Cyt and pTRA132 (containing hph gene) were introduced into embryogenic calli derived from the mature seeds of a rice cv. Hashemi by biolistic transformation method. Then putative transformants were screened after 3 rounds of selection on N6 medium containing increasing concentrations of Hygromycin B from 60 to 80 mg/L. Finally, Hygromycin resistant calli were regenerated on MS medium supplemented with 50 mg/L Hygromycin B. Putative transgenic rice plants were analyzed by polymerase chain reaction PCR. Then, four of the transgenic plants were analyzed using Southern blotting. Each transgenic plant received one copy number of both choline oxidase and hph genes. Expression of the transgene was confirmed by reverse transcription PCR. The high frequency of transformation rate in this study showed that co-transformation method is a reliable method for stable transformation with the goal to make markerfree transgenic plants in subsequent steps.

One of the landmark results of the Earth Summit held in Rio de Janeiro in 1992 was the Convention on Biological Diversity. The Biodiversity Convention calls upon parties to conserve and sustainably use biological diversity while equitably sharing the benefits of the use of genetic resources. These goals are key elements of sustainable development. Biodiversity in general means the variation in all kind of life around us. The life of human beings and other creatures and even next generations somehow depends on the biodiversity conservation. Hence, the conservation and understanding the ways to suitably utilize this diversity is crucial for sustainable development and well-being of all societies. Biodiversity was considered as a valuable source of income in economy for long time. Nowadays, in addition to its traditionalusage (hunting, fisheries), biodiversity is widely being used in agriculture, industry, medicine and even for amusement and eco-tourism. Biodiversity conservation is one of the main concerns for most of the countries. It is anticipated that in future, the direct pressure (over exploitation, habitat destruction …) and indirect effects of human activities on natural ecosystems will be the main threats for biodiversity, resource renovation, and even for life on the Earth. Identification, conservation, and use of genetic resources are crucial for better utilization of biodiversity toward sustainable development. Today, public awareness about the importance of identification and conservation of biodiversity and its important role in health of biosphere is increasing. World’s population is increasing; supplying food for this increasing population is very important and this has to be done through sustainable agriculture. In this review, with emphasis on the importance of biodiversity for human health and next generations and introducing its threats, we tried to show the importance and vital role of biotechnology and genetic engineering in identification, conservation, sustainable exploitation and even increasing biodiversity.

about the socio-economic benefits of these crops. But, still some criticisms about their cultivation and usage can be heard from around the world especially in EU countries. Anti GMOs claim that genetic engineering methods are not natural and there are some concerns about gene escape, religious and ethical issues, intellectual property rights and its effects on economic independency of countries, and ultimately more poverty and social inequilibrity. Moreover, application of antibiotic resistance genes as selectable markers, and Bt genes because of some fears about their toxicity for human being and non-target organisms, and finally, application of herbicide resistance genes because of the risk of gene escape that may lead to super weed production were criticized by the anti-GMOs. Reasons claimed by these groups for their oppositions and appropriate scientific responses to these claims, as well as the main economicreasons behind these criticisms were considered.