Molecular biology and (trans) gene expression technology in plant breeding
Fatemeh Keykha akhar; Masoud Fakhrfeshani; Hadi Alipour; Maryam Ameri
Abstract
Algae are an enormous biological group, forming 50% of photosynthetic organisms. In addition to having chlorophyll for the absorption of light photons, algae are rich in red, orange, and yellow carotenoids, which mainly protect cells against harmful radiation and free radicals. Moreover, these organisms ...
Read More
Algae are an enormous biological group, forming 50% of photosynthetic organisms. In addition to having chlorophyll for the absorption of light photons, algae are rich in red, orange, and yellow carotenoids, which mainly protect cells against harmful radiation and free radicals. Moreover, these organisms have phycobiliproteins (red and blue pigments), which are involved in capturing and passing light energy to chlorophylls during photosynthesis and have a wide range of antioxidant properties. Algae also play a key role in substituting artificial colorants with natural colorants due to the adverse side-effects of chemical colorants, especially since natural colors are commonly used by individuals and various industries. Recently, algal pigments have been widely used in medical, nutraceutical, cosmeceutical, and pharmaceutical industries owing to their antioxidant, antidiabetic, anti-obesity, anti-inflammatory, antiaging, antimalarial, and neuroprotective properties. The growing demand for algal bioproducts highlights the importance of evaluating the trends influential factors in their production. The current review study provided an introduction to algal pigment classification, distribution, function, application, and biological production. In addition, we have discussed crucial biochemical pathways, enzymes, and gene/biotechnological modifications, such as transformation and expression regulation, which noticeably affect the metabolism of their sink and source.
Molecular biology and (trans) gene expression technology in plant breeding
Dariush Taghavi; Mohammad Majidi; Saeed Mollaei; Bahman Panahi
Abstract
Shikonin is a naphthoquinone with some important medicinal properties and is found in gromwell plant. There are so many biotechnological approaches proposed for the productivity enhancement, and elicitation is recognized as one of the most effective strategies for increasing the production of secondary ...
Read More
Shikonin is a naphthoquinone with some important medicinal properties and is found in gromwell plant. There are so many biotechnological approaches proposed for the productivity enhancement, and elicitation is recognized as one of the most effective strategies for increasing the production of secondary metabolites in plant in vitro cultures. Moreover, a deeper understanding of the mechanisms and factors affecting shikonin biosynthesis can led to the design of more intelligent and efficient biological production systems. To this aim, in the present study, the expression of some genes related to the shikonin biosynthetic pathway including PAL, 4CL, HMGR, GPPS and PGT in in vitro cultures of Lithospermum officinale in response to methyl jasmonate (MJ) at different times, were investigated by real-time PCR. The results showed that MJ had a significant effect on increasing gene expression levels in elicited samples compared to control samples. Additionally, we found that the studied genes respond to MJ with different pattern, in which the highest increase in gene expression level was observed for PGT while the lowest increase was observed for GPPS. Maximum and minimum transcript levels were obtained in most genes at 4 and 96 h post-elicitation, respectively. It was also found that phenylpropanoid pathway genes respond better to MJ than terpenoid pathway genes. The results of the present study would increase our knowledge about elicitor signal transduction pathways, and may be particularly useful for enhancement of shikonin production in plant cell cultures of L. officinale.
Molecular biology and (trans) gene expression technology in plant breeding
Abolfazl Mazandarani; Saeid Navabpour; Ahad Yamchi
Abstract
The main source of protein and micronutrients in wheat grains is the flag leaf and to a lesser extent the lower leaves. As healthy leaves reach the final stage of growth, senescence, they remobilize the nutrients necessary before tissue destruction and death. This experiment was carried out in Golestan ...
Read More
The main source of protein and micronutrients in wheat grains is the flag leaf and to a lesser extent the lower leaves. As healthy leaves reach the final stage of growth, senescence, they remobilize the nutrients necessary before tissue destruction and death. This experiment was carried out in Golestan province, and the Wheat cultivars studied were included Euclide and Antonius. Sampling was carried out from flag leaf, other leaves, stem, and grain at 7 stages, Anthesis, 7, 11, 15, 19, 23, and 27 Day After Anthesis (DAA). The total chlorophyll content in the Antonius cultivar was higher in both flag leaf and other leaves than Euclide cultivar. The expression of TaNAM-B1 and TaSAG12 genes, which have been identified as signaling genes for senescence in wheat, showed results consistent with the results of chlorophyll content in leaves. Increased expression of both genes after anthesis was observed earlier in Euclide cultivar than the Antonius cultivar and had higher expression in most stages. In light of the results, the change in concentrations of Cu, Zn, and Fe in the Euclid cultivar was more in all organs than in Antonius one. Also, given the importance of minerals in the food basket, it can be noted that Euclid cultivar, in which leaf senescence begins earlier and more minerals are stored, can produce grains with higher nutritional value than Antonius cultivar.
Molecular biology and (trans) gene expression technology in plant breeding
Arezoo Pourfarid; Ali Pakdin-Parizi; Reza Ghorbani-Nasrabad; Heshmatollah Rahimian
Abstract
Arbuscular mycorrhizal fungi (AMF) symbiosis could mitigate the adverse effects of abiotic stresses in various plants. The aim of this study was to investigate the effect of AMF-inoculation on expression of several stress-responsive genes in two rice cultivars under different water conditions. The seedlings ...
Read More
Arbuscular mycorrhizal fungi (AMF) symbiosis could mitigate the adverse effects of abiotic stresses in various plants. The aim of this study was to investigate the effect of AMF-inoculation on expression of several stress-responsive genes in two rice cultivars under different water conditions. The seedlings of Tarom-Hashemi and Nemat rice cultivars were transplanted in soil with or without G. mosseae spores. At the tilling stage, the AMF-inoculated (+AMF) and AMF-uninoculated (−AMF) plants were subjected to flooded and water deficit conditions (70% field capacity (FC) and 50%FC). The genes expression was evaluated by qRT-PCR and reported relative to control (flooded, -AMF) plants. The results showed lower expression of osDREB2A in +AMF plants in comparison with –AMF plants under water deficient conditions. The expression of OsPIP1;2 was significantly increased in roots of +AMF to –AMF plants. But, the expression of this gene was decreased in shoots of +AMF and –AMF plants in comparison with control plants. The stress-responsive gene transcripts, OsPIP2;3, OsGH3-8, OsLTP, OsAOS2 and OsADC1 in +AMF rice cultivars was higher than -AMF plants at both water deficit conditions. Expression of OsP5CS in +AMF and –AMF plants was increased in comparison with control plants, though, their differences was not significant. In 70%FC, OsEXP15 gene expression of +AMF and –AMF root plants was increased in comparison with control plants. However, under 50%FC the gene expression was decreased and not changed in -AMF and +AMF plants, respectively. It seems AMF induced changes in rice genes expression may enhance tolerance to water deficit conditions.
Molecular biology and (trans) gene expression technology in plant breeding
Hossein Moradi Beidokhti; Valiollah Babaeizad; Heshmatollah Rahimian; Mohammad Ali Tajick Ghanbary; Ali Pakdin-Parizi
Abstract
Citrus bacterial blast disease is one of the prevalent diseases in most citrus-growing regions in the world. Plants use a wide range of mechanisms to defend against pathogens, and the plant-pathogen interaction induces the expression of genes involved in the plant resistance. Furthermore, symbiotic association ...
Read More
Citrus bacterial blast disease is one of the prevalent diseases in most citrus-growing regions in the world. Plants use a wide range of mechanisms to defend against pathogens, and the plant-pathogen interaction induces the expression of genes involved in the plant resistance. Furthermore, symbiotic association between plant and mycorrhizal fungi could effectively promote growth and protect the plant against adverse environmental conditions. In the present study, Serendipita indica-root colonized sour orange seedlings were infected by Pseudomonas viridiflava and the expression patterns of PR1, PR2, PR3, PR4, PR5, PAL, POX, and LOX genes in the early stages of citrus blast disease were investigated using the qRT-PCR at different sampling times. According to the results, the response of defense genes to bacterial infection was time dependent. In the S. indica-colonized sour orange plants, the highest level of PR1, PR4, PAL, POX, PR3, and PR5 genes expression was observed at 48 h after infection, but the expression of PR2 and LOX genes was increased at 72 h after infection compared to the control plants. It seems that S. indica can induce systemic effects and prepare the host plant to increase the expression of defense genes more rapidly once it receives a signal for the presence of the pathogen.
Molecular biology and (trans) gene expression technology in plant breeding
Meysam Bastami; Hosseini Ramin
Abstract
The regulatory sequence of rice alpha amylase 3D gene (Ramy3D) is amongst the most successful expression systems used for recombinant protein expression in plants. In the current study a 995 bp fragment consisting of Ramy3D promoter and its 5′ untranslated region was amplified from the genomic ...
Read More
The regulatory sequence of rice alpha amylase 3D gene (Ramy3D) is amongst the most successful expression systems used for recombinant protein expression in plants. In the current study a 995 bp fragment consisting of Ramy3D promoter and its 5′ untranslated region was amplified from the genomic DNA of an Iranian rice cultivar ″Nemat″, using polymerase chain reaction. The amplified fragment was ligated into the pTG19-T vector and the cloned fragment was sequenced. For in silico characterization, the rice specific consensus sequences of TATA-box and YR Rule motifs were scanned against the cloned fragment sequence using FIMO program and the cis acting elements existing in the promoter region were investigated using PlantCare database. A TATA-box motif with the rice specific pattern was identified at upstream position of the transcription start site. The identification of TATA-box in Ramy3D promoter is consistent with its metabolic and tissue specific regulation manner. Several cis regulatory motifs responsible for the metabolic and hormonal regulation of Ramy3D gene were identified including ABRE, G-Box, GC-box, GATA motif and TATCCA T/C motif. In addition, several motifs involved in response to various stimuli such as plant hormones, light and biotic and abiotic stresses were identified which include circadian motif, as-2-box, WUN-motif, TGACG-motif, Skn-1 motif, O2-site, MBS, LAMP-element, I-box, HSE, GCC Box, GATT motif, CGTCA-motif and GAG-motif.
Molecular biology and (trans) gene expression technology in plant breeding
Nahid Sadeghi Ghahderijani; Ali Niazi; Esmaeil Ebrahimie; Ali Moghadam; Mohammad Sadegh Taghizadeh
Abstract
Agrobacterium-mediated gene transfer method is one of the used methods for genetic transformation in the plant regeneration program. Transformation efficiency can be optimized depending on the strain of bacteria, the genotype of plant and conditions of growth. In this study, the gfp gene was transferred ...
Read More
Agrobacterium-mediated gene transfer method is one of the used methods for genetic transformation in the plant regeneration program. Transformation efficiency can be optimized depending on the strain of bacteria, the genotype of plant and conditions of growth. In this study, the gfp gene was transferred into sugar beet, tobacco, and soybean by Agrobacterium rhizogenes strain AR15834. The effects of bacterial concentrations, antibiotic concentrations and the types of explants and genotypes on the gene transfer efficiency and transgenic hairy roots production were investigated. The explants were inoculated with the bacteria at the adjusted concentrations and two days after the transformation, the explants were transferred to a solid MS medium containing different concentrations of kanamycin antibiotic. According to the results and the examined factors, the optimal conditions to achieving of the maximum production of transgenic hairy roots included bacterial concentration with OD600 = 0.2, cotyledon explant, 50 mg/L kanamycin concentration and Djakel genotype for soybean; bacterial concentration with OD600 = 0.2, leaf with petiole explant and SBSI004 genotype for sugar beet, and bacterial concentration with OD600nm = 0.8 and 100 mg/L kanamycin concentration for tobacco.
Plant breeding for abiotic & biotic stresses
Khalil Malekzadeh; Ali Niazi; Farajollah Shahriari-Ahmadi; Amin Mirshamsi-Kakhaki; Mohammad Zare-Mehrjerdi
Abstract
Salinity is one of the most important abiotic stresses that limit crop growth and production. Salt stress influences plants in two ways: by affecting ion toxicity and increasing osmotic stress. Ion homeostasis, the excretion of Na+ and using antioxidant systems are the major strategies of salt tolerance ...
Read More
Salinity is one of the most important abiotic stresses that limit crop growth and production. Salt stress influences plants in two ways: by affecting ion toxicity and increasing osmotic stress. Ion homeostasis, the excretion of Na+ and using antioxidant systems are the major strategies of salt tolerance in plants. Na+ and K+ transporters with enzymes that are involved in detoxification of reactive oxygen species play key roles in salt tolerance in plants. The aim of this study was to investigate the responses of high affinity K+ transporter2;1 gene (HKT2;1) which is involved in regulation of ion homeostasis and L-gulonolactone oxidase (GLOase) which is involved in the ascorbic acid biosynthesis pathway, under different concentrations of NaCl over different time points in Aeluropus littoralis shoots. Results from Real Time PCR data showed that expressions of both genes were influenced by external and internal concentrations of Na+ and the internal K+ content. AlHKT2;1 was significantly upregulated by increasing Na+ concentration at all time points. Furthermore, its highest expression level in shoots occurred after 6 days in 300mM NaCl in shoots which was 25folds more than untreated shoots. AlGLOase expression levels increased 54 h after initiation of salt stress. These results indicate that AlHKT2;1 and AlGLOase respond to different salinity conditions and probably are part of the mechanisms involved in tolerance to high salt concentrations in A. littoralis.
Molecular biology and (trans) gene expression technology in plant breeding
Sahand Amini; Cyrus Ghobadi; Ahad Yamchi
Abstract
Under osmotic stresses, proline accumulation is an important response of plants to these conditions. Proline is a compatible osmolyte which affects many cellular and molecular aspects of plant in both normal and stressful situations. Proline is shown to be involved in plant development in normal condition ...
Read More
Under osmotic stresses, proline accumulation is an important response of plants to these conditions. Proline is a compatible osmolyte which affects many cellular and molecular aspects of plant in both normal and stressful situations. Proline is shown to be involved in plant development in normal condition and in conferring resistance to plant under biotic and abiotic stresses. Therefore, many surveys have already been designed to unveil its mechanisms and signaling pathway, so that it might be an insight into resolving growing challenge of agriculture, drought and soil salinity. Δ1-pyrroline-5-carboxylate synthetase (P5CS), one of two main enzymes in proline biosynthesis pathway from glutamate precursor, has been demonstrated to play significant role in proline accumulation in plants under water stresses. Regarding the role of P5CS under osmotic stress, there are controversial observations in various plants, hence making it still unknown, whether P5CS is rate-limiting enzyme in the pathway or not. Obviously, transgene P5CS is proved to give higher resistance to transgenic plants under drought and salinity, by elevating proline content. In this literature, proline and its identified various functions in plants, characteristics of P5CS enzyme, signals, inducers and inhibitors of P5CS gene, expression pattern of P5CS under differential conditions in studied plant species are discussed. Finally, we have reviewed generated transgenic plants overexpressing P5CS and consequences of these transformations.
Molecular biology and (trans) gene expression technology in plant breeding
Maziar Habibi; Saeid Malekzadeh-Shafaroudi; Hassan Marashi; Nasrin Moshtaghi; Mohammadreza Nasiri; Saeed Zibaee
Abstract
Foot and Mouth Disease (FMD) is a very dangerous livestock disease which causes a serious loss in the production of milk and meat. Therefore, producing an effective recombinant subunit vaccine virus this disease is of great importance. Transient gene expression is a valuable tool to reach rapid and acceptable ...
Read More
Foot and Mouth Disease (FMD) is a very dangerous livestock disease which causes a serious loss in the production of milk and meat. Therefore, producing an effective recombinant subunit vaccine virus this disease is of great importance. Transient gene expression is a valuable tool to reach rapid and acceptable recombinant vaccine. An Agrobacterium-mediated transient gene expression assay was carried out in spinach (Spinacia oleracea) leaves for expression of a chimeric gene encoding a part of capsid protein of Foot and Mouth Disease virus called VP1. The plant leaves were transformed via agroinfiltration procedure. The presence of foreign gene and its expression in transformed plants were confirmed through polymerase chain reaction (PCR), real time PCR, protein Dot blot and ELISA. The results obtained in this examination showed quite a high level of gene expression in spinach leaves, showing that transient gene expression can be applied as an effective and time-saving procedure for the production of recombinant proteins. The procedures for transformation, detection of recombinant protein and its application for molecular experiments are described in the study.
Molecular biology and (trans) gene expression technology in plant breeding
Hamid Najafi zarrini; Mahsa Mohammad Jani Asrami
Abstract
There is no functional annotation for the majority of the several hundreds of receptor-like kinases in plants. A direct way of inferring the function of these proteins is to study the phenotype that results from loss of function mutants such as T-DNA mutant lines. In this research a function (phenotype) ...
Read More
There is no functional annotation for the majority of the several hundreds of receptor-like kinases in plants. A direct way of inferring the function of these proteins is to study the phenotype that results from loss of function mutants such as T-DNA mutant lines. In this research a function (phenotype) to At2g37050 gene that encodes a receptor like kinase in Arabidopsis T-DNA line was assigned. This phenotype has a shorter primary root length at later stages of development. Transcription study of the gene showed some tissue specificity with more expression level in the root in comparison with other tissues. To study genes co-expressed with At2g37050, ATTED-II web tool was used. It was found that the CLASP gene is co-expressed with At2g37050with aPearson correlation > 0.6. In kinematic analysis of the difference in root growth, the length between the root tip and the first epidermal cell with a visible root hair bulge for 8 day-old seedlings of wild type plants was 1327± 76.50 µm (n=6) and for the mutant plants, was 1109± 72.28. This parameter of the wild type and the mutant plants shows that loose of function of At2g37050 gene, reduce cell elongation in the elongation zone of root.
Molecular biology and (trans) gene expression technology in plant breeding
Behnaz Dolatabadi; Gholamali Ranjbar; Masoud Tohidfar; Ali Dehestani
Abstract
Fusarium wilt caused by Fusarium oxysporum f.sp. Lycopersici is one of the major obstacles to the production of tomato which causes huge losses in tomato products worldwide. In order to increase the tolerance to this disease, a triple structure containing PR1, chitinase and glucanase genes controlled ...
Read More
Fusarium wilt caused by Fusarium oxysporum f.sp. Lycopersici is one of the major obstacles to the production of tomato which causes huge losses in tomato products worldwide. In order to increase the tolerance to this disease, a triple structure containing PR1, chitinase and glucanase genes controlled by 35S promoter was transferred to tomato. Eight days after planting on pre-culture medium, explants were inoculated by Agrobacterium tumefaciens strain LBA4404 containing the aforementioned plasmid. When the regenerated shoots grew to 2-3 cm, they were cut and transferred to rooting medium.The plantlets were then transferred to pots filled with a soil mixture of peat moss and perlite for further acclimatization. The putative transgenic plant lines were analyzed by multiplex PCR and the transcription of the transgenes was confirmed by RT-PCR method using the specific primers. The estimated value for the frequency of the simultaneous transfer of chitinase, glucanase and PR1 genes to tomato was 2.7%. Protein extracts of transgenic plants expressing chitinase, glucanase and PR1 genes inhibited in vitro hyphal growth of F. oxysporum f.sp. lycopersici. Compared with non-transgenic control plants, despite some alterations in chlorophyll content no other morphological changes were observed in transgenic plants. The total content of chlorophyll “a” and “b” in transgenic plants were 31.8 and 36.2 % higher than that of control plants, respectively, which may be attributed to metabolic changes due to simultaneous expression of three transgenes.