Plant Productions

Effect of Biological and Chemical Fertilizers and Salicylic Acid Spraying on Yield and Some Physiological Traits of Wheat Under Drought Stress

Fatemeh Chaharlang Badil; khosrow Azizi; Hamid Reza Eisvand; Ali heidar Nasrollahi; Ahmad Ismaili

Volume 45, Issue 3 , December 2022, , Pages 397-408

https://doi.org/10.22055/ppd.2022.40567.2026

Abstract

IntroductionWheat (Triticum aestivum L.) is one of the most important and strategic crops worldwide. Drought stress is one of the important and abiotic stresses that reduces the growth ... Read More IntroductionWheat (Triticum aestivum L.) is one of the most important and strategic crops worldwide. Drought stress is one of the important and abiotic stresses that reduces the growth and yield of plants.in arid and semi-arid regions. Today, the economic damage and destructive effects of the environment due to the excessive use of chemical fertilizers in agriculture are known worldwide, and it is obvious that a suitable alternative must be found for these fertilizers. Currently, biofertilizers are used as an alternative to chemical fertilizers based on the principles of sustainable agriculture and stress tolerance of plants. Salicylic acid is a natural phenolic compound that is involved in reducing the harmful effects of abiotic environmental stresses and regulating vital physiological processes in plants. Materials and MethodsThis study was carried out to assess the effect of fertilizer nutrition and foliar application of salicylic acid on wheat tolerance of Chamran 2 cultivar to drought stress experiment as a factorial split plot in the form of a randomized complete block design in the 2019-2020 crop year in the research farm of the Faculty of Agriculture of Lorestan University in three replications was performed The experimental site was located at a latitude 33°26′ N, a longitude 45°17′ E, and an altitude of 1210 m with a semi-arid climate, mean annual precipitation of 462.8 mm, mean annual evaporation of 1842.52 mm, and mean annual temperature of 16.6 °C .Main plot includes irrigation levels in two levels of without stress (100% of field capacity) and drought stress (50% of field capacity) and sub-plots including fertilizer in five levels including no use of biofertilizer + 50% chemical fertilizer, application of 100% chemical fertilizer, seed inoculation of nitroxin biofertilizer + 50% chemical fertilizer, seed inoculation of mycorrhizal + 50% chemical fertilizer, (simultaneous use of biofertilizers (nitroxin + mycorrhiza) + 50% chemical fertilizer, and foliar application treatment at two levels (foliar application with water and foliar application with a concentration of 1 mM salicylic acid). Results and DiscussionThe results showed that drought stress increased grain protein and malondialdehyde and decreased other evaluated traits. Fertilizer nutrition and salicylic acid foliar application treatment alone increased the evaluated traits and decreased malondialdehyde content compared to the no use of biofertilizer + 50% chemical fertilizer and no foliar application of salicylic acid. Combined treatment of fertilizer (nitroxin + mycorrhiza) + 50% chemical fertilizer and salicylic acid could reduce the effect of drought stress on the traits evaluated in this study on wheat of Chamran 2 cultivar. As a result can this treatment suggested to the farmer in order to reduce the consumption of chemical fertilizers in the cultivation of this wheat variety under drought stress conditions. ConclusionIn general, it can be concluded that the application of biofertilizers (nitroxin + mycorrhiza) + 50% chemical fertilizer and salicylic acid foliar application can be used as an effective and alternative fertilizer to reduce the use of chemical fertilizers and water shortage in Khorramabad for sustainable agriculture. As a result, this treatment can be recommended to the farmer to reduce the use of chemical fertilizers in the cultivation of this wheat cultivar under drought stress conditions.

Isolation, Cloning, Bioinformatics Analysis of Protein Properties and Gene Expression Analysis of Dihydrosanguinarine Oxidase (DBOX) from Opium Poppy (Papaveer somniferum L.)

Kamran Samiey; Ahmad Ismaili; Farhad Nazarian Firoz-abadi; Seyed Mohsen Sohrabi

Volume 43, Issue 1 , June 2020, , Pages 67-80

https://doi.org/10.22055/ppd.2020.21559.1463

Abstract

Abstract Background and Objectives Opium poppy (Paoaver somniferum L.) is one of the important medicinal plants considered the only source for several high-value pharmaceutically ... Read More Abstract Background and Objectives Opium poppy (Paoaver somniferum L.) is one of the important medicinal plants considered the only source for several high-value pharmaceutically BIAs. Benzylisoquinoline alkaloieds (BIAs) are a very large and complex group of plant alkaloids. Today’s various methods of genetic engineering are applied to improve the biosynthetic pathways in medicinal plants. For successful implementation of techniques such as gene silencing and over-expression, it is important to get accurate and complete information about genetic characteristics of related genes in the biosynthesis of secondary metabolites. DBOX is one of the important genes in biosynthesis of sanguinarin and papaverin alkaloids of poppy. According to the cause, the present study was performed on DBOX to explain genetic characteristic and gene expression pattern. Materials and Methods Specific primers were designed based on databases, and complete sequences of DBOX gene were amplified using DNA and cDNA as a template, and then were cloned in pTZ57R/T plasmid. After sequencing of cloned fragments, CDD, CELLO and ProtParam tools were used to determine ORFs, domains and the location of protein accumulation. Phylogenetic relationship between the gene in this plant with other plants was determined. Finally, the pattern of gene expression in different tissues was assessed using real time RT-PCR technique and RNA-seq data derived from SRA data bank. Results Sequencing results showed the 1614-bp fragment that has 100% identity with the DBOX gene of P. somniferum. The protein encoded by this gene was analyzed by bioinformatics methods. Three domains including FAD binding, Berberine and FAD/FMN-containing dehydrogenase in the protein sequence were identified and the existence of an N-terminal signal peptide was also determined in this protein. The results also showed that the highest concentration of protein was present in the plasma membrane and highest function of this protein was in the extracellular. In order to analyze and determine the expression pattern of DBOX gene in different tissues, 5-gene libraries of RNA-seq data of opium poppy (with ERX651037, ERX651023, ERX651056, ERX651082 and ERX651062 access numbers) derived from NCBI-SRA were analyzed. The numbers of sequences from the alignment between different tissues were compared using the Fisher exact test and Chi-squared 2X2 tests. Results of the gene expression analysis showed that the highest level of gene expression of this gene performed in roots and the lowest level of gene expression occurred in the stems. In addition, results of the real-time RT-PCR experiment were similar to in silico tests and the present results demonstrated that DBOX gene transcript levels in root were significantly more than other plant tissues. Discussion Results showed that DBOX gene is an intron less gene. According to existing domains, this gene is a member of a FADOX gene family. Due to the presence of FAD and berberin like domains in protein sequence of this gene, its key function in biosynthesis of BIA alkaloids was confirmed. Different expression patterns of DBOX gene in root confirmed accumulation of high value sanguinarin alkaloid in this tissue.

The Study on Gene Expression of 3-Hydroxy-3-Methylglutaryl-Coa Reductase (HMGR) and γ-Terpinene Synthesis in Thymus Vulgaris L. Under Water Deficit Stress

Fatemeh Mirzaei; Ahmad Ismaili; Foad Fatehi; Ardeshir Ghaderi; Soheila Afkar

Volume 42, Issue 2 , June 2019, , Pages 165-180

https://doi.org/10.22055/ppd.2019.20928.1436

Abstract

Abstract Background and Objectives Drought is a major abiotic stress that limits agricultural crop production. Thyme (Thymus vulgaris L.) is an aromatic and medicinal plant which ... Read More Abstract Background and Objectives Drought is a major abiotic stress that limits agricultural crop production. Thyme (Thymus vulgaris L.) is an aromatic and medicinal plant which is very important for the herbal industry. This study was aimed to investigate the variation some of the secondary metabolites and morphological traits as well as changes in expression of genes involved in the biosynthesis of thymol (including HMGR and TPS2) by real-time PCR. One of the main goals of the present study was to find out the relationship between the transcript of these genes and related metabolites in thyme. Materials and Methods This study was conducted in the greenhouse of Iranian Institute of Medicinal Plants of Jahad Daneshgahi (Karaj, Iran) in 2016. The experimental design was a randomized complete block design with 5 replication and 4 water deficit stress treatments, including 100% (T1, control), 70% (T2, mild stress), 40% (T3, moderate stress), and 20% (T4, severe stress) of field capacity (FC). Total RNA was extracted from leaves using TRIzol reagent following the manufacturer’s instructions. First strand synthesis of cDNA was performed immediately using 1 µg of total RNA with simultaneous use of oligo-dT (50 µM) primers and random hexamer and reverse transcriptase enzyme following the manufacturer protocol. In order to examine gene expression by real-time PCR, the method of fluorescence dye SYBR green (Fermentase, USA) was used. The amount of thymol and carvacrol were measured by HPLC. Results Results of morphological traits showed that increasing the water deficit stress caused a decrease in root biomass, shoot fresh weight, plant height, root length, and the number of side branches. Results of gene expression profiling showed that the highest expression of TPS2 and HMGR genes and the highest amount of thymol and carvacrol was obtained in treatment T2 (70% FC). Discussion TPS2 and HMGR gene expression changes were similar to the changes of monoterpenes thymol and carvacrol in all treatments and the highest number of monoterpenes and gene expression were obtained in mild water deficit stress treatment in comparison with control, moderate, and severe stresses. Altogether, according to the results of this study, mild stress had a significant effect on increasing the expression of HMGR and TPS2 and led to a higher production of monoterpenes.

Effect of Different Elicitor Treatments on Hairy Root of Medicinal Plant Poppies (Papaver somniferum L.)

Sh. Siahmansour; A. Ismaili; F. Nazarian Firouzabadi

Volume 41, Issue 1 , June 2018, , Pages 29-42

https://doi.org/10.22055/ppd.2018.13548

Abstract

Abstract Background and Objectives As a source of alkaloids, Poppy (Papaver somniferum L.), is one of the most important plants in the field of pharmaceutical industry. Various methods ... Read More Abstract Background and Objectives As a source of alkaloids, Poppy (Papaver somniferum L.), is one of the most important plants in the field of pharmaceutical industry. Various methods including use of elicitors, adding precursor, optimizing culture, culture of hairy roots and its metabolites engineering are used to increase the production of secondary metabolites in plant cell cultures. In recent years, cultivation of hairy roots, to the production of valuable metabolites in a number of species of medicinal plants, has taken on commercial scale; but in most cases, this technique produces low amount of alkaloid in commercial scale. Hence, it is needed to optimize the production rate using biological and non-biological elicitors. Materials and Methods This experiment was conducted in 2015 in biotechnology laboratory of Lorestan University, Khorramabad, Iran. Sterile explants were prepared from seedling and were then inoculated with Agrobacterium. Inoculated explants were kept in a dark condition for 2 days and then washed and transferred to glasses containing½ MS medium complemented with cefotaxime. Finally, the glasses were kept in tissue culture room until production of hairy roots. The produced hairy roots were used as explants for elicitors' experiments in flask containers. In this study, in order to increase biomass of hairy roots, 4 types of elicitor treatments (salicylic acid, silver nitrate, copper sulphate and yeast extract) were evaluated in separate experiments based on a completely randomized design with 3 replicates. Results Confirmation of transgenic hairy roots was tested by PCR using the rolC gene primers to ensure the removal of bacteria from the produced roots. Results of PCR products were separated by electrophoresis and expected fragments were observed in gel. The results showed that the concentration of 5.0 mg/mL of yeast extract elicitor had the most impact on morphological traits. Totally, results showed that biotic elicitors had better effects in comparison to abiotic elicitors. Also, high concentrations of elicitors in all biotic and abiotic cases had a negative impact on biomass of hairy roots. Discussion It was shown that biotic elicitors had better effects than abiotic elicitors. The remarkable result of this study was that the high concentration of elicitors had negative effects on biomass hairy root production in poppies plants. It seems that bio-active compounds elicitors induce the plant responses but high concentration of biotic and abiotic elicitors may cause cell death and browning of hairy root tissues.

Induction of Transgenic Hairy Roots in Medicinal Plant Poppy (Papaver Somniferum L.) by Agrobacterium Rhizogenes-Mediated Transformation

A Jalilian; A A; F N; S.Z H

Volume 39, Issue 4 , March 2017, , Pages 1-14

https://doi.org/10.22055/ppd.2017.12057

Abstract

Background and Objectives Opium poppy plant (Papaver somniferum L.) is one of the oldest known medicinal plants. Since the potential of medicinal components production in medicinal ... Read More Background and Objectives Opium poppy plant (Papaver somniferum L.) is one of the oldest known medicinal plants. Since the potential of medicinal components production in medicinal plants is very limited in natural conditions, plant tissue culture, as a new technology, has an important role in industrial production of secondary metabolites in medicinal plants. Among different tissue culture systems, hairy root is very stable in hormone-free culture conditions and these roots stably produce secondary metabolites over a long period due to their inherent genetic and biochemical stability. Hence, the objective of this study was to evaluate the effect of some different parameters on optimization of hairy production in P. somniferum. Materials and methods In the present research, for the optimization of hairy root in papaver, a factorial experiment was conducted based on a completely randomized design with 4 replications. The factors included different Agrobacterium rhizogenes strains (ATCC15834, A4 and GMI9534), medium (MS and ½ MS medium), two temperatures (17 and 25 °C) at co-cultivation period, and two different explants (excised shoot and hypocotyl). Different traits were evaluated including percentage of induced hairy roots, number of lateral branches/1cm, number of hairy root per explants and days after inoculation (DAI). Results Hairy roots were induced at the wound site of explants after weeks of culture. Analysis of the results showed that strain ATCC15834, ½ MS medium, temperature 25 °C and excised shoot explant were the best treatment combination for hairy roots induction in the poppy plant. Transformed hairy roots were confirmed by polymerase chain reaction (PCR) using rolB gene specific primers. Discussion All of the A. rhizogenes strains led to hairy root induction, but there were no hairy roots formed from control explants. Differences in virulence, morphology and growth rate can be partially explained by the variety of plasmids harbored by the A. rhizogenesis strains. In the present study, excised shoot explants were highly and significantly susceptible to infection by each strain of A. rhizogenes and hypocotyl explants showed low rate of hairy root induction.

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Author = A. Ismaili

Effect of Biological and Chemical Fertilizers and Salicylic Acid Spraying on Yield and Some Physiological Traits of Wheat Under Drought Stress

Abstract

Isolation, Cloning, Bioinformatics Analysis of Protein Properties and Gene Expression Analysis of Dihydrosanguinarine Oxidase (DBOX) from Opium Poppy (Papaveer somniferum L.)

Abstract

The Study on Gene Expression of 3-Hydroxy-3-Methylglutaryl-Coa Reductase (HMGR) and γ-Terpinene Synthesis in Thymus Vulgaris L. Under Water Deficit Stress

Abstract

Effect of Different Elicitor Treatments on Hairy Root of Medicinal Plant Poppies (Papaver somniferum L.)

Abstract

Induction of Transgenic Hairy Roots in Medicinal Plant Poppy (Papaver Somniferum L.) by Agrobacterium Rhizogenes-Mediated Transformation

Abstract