نشریه بیوتکنولوژی و بیوشیمی غلات
سال دوم شماره 4 (پیاپی 8، زمستان 1402)

Environmental stresses such as salinity are one of the major threats to the sustainability of crop production. Climate change has increased the intensity of these environmental tensions in different regions of the world. Drought and salinity are the critical environmental stresses in arid and semi-arid regions. In recent decades, various strategies have been introduced to reduce the effects of salinity stress on crops. The use of beneficial soil microorganisms is one of these methods.

In the present study, the effect ofAzospirillum bacteria and Piriformospora indica fungus in separate and combined inoculation was investigated to identify the role of these above-mentioned plant growth-promoting microorganisms in the modulation of the molecular, biochemical, and physiological aspects in wheat under salt stress conditions. A 2×2×2 factorial experimental design was implemented in a completely randomized block design with three replications under greenhouse conditions in 2022. The experiment was conducted in the pot. The main treatments included salt stress imposition, inoculation with and without Azospirillum zaea, and endophytic fungus Piriformospora indica application. Salinity was induced using calcium chloride (65 mg) and sodium chloride (0.375 g) to achieve a salinity level of 12 dS/m. Saline water irrigation was performed 21 and 28 days after wheat germination. Normal water with a salinity of 0.3 decisiemens per meter was used for the control treatment. The wheat cultivar Pishgam was used for this experiment. Twenty-four hours after applying the second salinity treatment, leaf sampling was conducted to extract total RNA and study the expression of the delta-pyrroline-5-carboxylate synthase gene. The changes in leaf proline content and the expression of the gene responsible for proline synthesis (pyrroline-5-carboxylate synthase), malondialdehyde production, ion leakage, leaf relative water content, nitrogen content, and chlorophyll pigment levels in response to applied microorganisms under both salinity and non-salinity conditions were investigated. Additionally, succinate dehydrogenase activity and root colonization rate with fungus were measured under the influence of salinity and inoculation with Azospirillum.

The percentage of fungal colonization in wheat roots of cultivar Pishgam was 24.9%. Inoculation with Azospirillum increased the root colonization rate by 3.17%. Salinity had no significant effect on succinate dehydrogenase, but inoculation with Azospirillum significantly increased succinate dehydrogenase. The plants inoculated with P. indica and Azospirillumhad a higher relative water content in leaves under salt stress. Plants inoculated with bacteria and fungus also had higher nitrogen content. Salinity increased the amount of ion leakage, but inoculation with fungus and bacteria prevented it from increasing. Salinity did not cause a significant change in the amount of chlorophyll pigments, although inoculation with P. indicaand Azospirillum increased the amount of chlorophyll a. The effect of salinity, P. indica, Azospirillum, and their interaction on the expression level of the delta-pyrroline-5 carboxylate synthase gene was significant. Control plants showed higher levels of gene expression, while the inoculated plants with P. indica and Azospirillum had lower gene expression levels. Additionally, the interaction between Azospirillum and P. indica on the expression level of the delta-pyrroline-5 carboxylate synthase gene was significant and it caused a greater decrease compared to the plants that were only inoculated with P. indica. The effect of salinity, P. indica, and bacteria, as well as their mutual effects on leaf proline concentration, was significant. Under salt stress, the amount of proline in the plants inoculated with P. indica was lower than that of the control plants. A similar trend was observed in plants inoculated with bacteria, showing lower levels of proline under salt stress. The expression trend of the delta-pyrroline-5-carboxylate synthase gene was consistent with the trend of proline accumulation under salt stress, indicating the effect of salt induction on the expression of this gene.

The inoculated plants with P. indica and Azospirillum were less affected by induced salt stress than the control plants. The inoculated plants exhibited a lower rate of malondialdehyde and electrolyte leakage, as well as a higher level of chlorophyll pigment and leaf-relative water content. Additionally, P5CS gene expression and proline accumulation in inoculated plants were lower than in control plants.

Rice (Oryza sativa L.) feeds more than half of the world's population. Rice is considered a model plant in biological and agriculture field research due to its small genome compared with other plants. Storage proteins in the aleurone layer of rice seeds include albumin and globulin, as well as the endosperm, prolamin, and glutelin proteins. To determine the number of proteins and identify the protein spots between the tissues of two embryo samples and rice endosperm, the protein spots were matched according to their expression level in the images obtained from two-dimensional gel electrophoresis.
The profile of proteins in the endosperm and embryo of rice seeds was determined using isoelectric focusing (IEF) and polyacrylamide gel electrophoresis(SDS-PAGE) methods. The Image Master software was utilized to analyze the two-dimensional gels. Considering the importance of local rice consumption in Iran and the significant production of this economically important crop in the country, this study aimed to determine the diversity and differences in the expression of protein spots within the linear pH range of 4 to 7. The two-dimensional electrophoresis method was employed on the Hasan-Sarai variety of rice indigenous to the region of Guilan. For this research, the cultivar being examined was procured from the Guilan Rice Research Center. Subsequent stages of the experiment, including sample separation and total protein extraction, were conducted at the Payame Noor University laboratory in Asadabad. Ultimately, the protein samples obtained were sent to the Health Research Institute at the Faculty of Pharmacy, where two-dimensional electrophoresis was carried out using an isoelectric focusing device. Finally, the extracted protein samples were transferred to the Health Research Institute of the Faculty of Pharmacy to perform two-dimensional electrophoresis using an isoelectric focusing device.
Based on the results of the two-dimensional gel analysis, a large part of the protein spots are present in the pH range of 4 to 7. The analysis of protein profile based on standard proteins with specific molecular weight and isoelectric point indicates that embryo tissue and endosperm based on isoelectric point and molecular weight showed that 284 protein spots in the embryo and 123 protein spots in endosperm have been identified. Among the identified spots, 57 common protein spots showed increased and decreased expression between the two samples. The presence of remaining protein spots in the two samples showed a significant difference between protein content in the embryo and endosperm.
Image master software was used to determine the isoelectric point and molecular weight of protein spots and expression changes between common spots. Analysis was done in protein databases and possible proteins were identified in embryo tissue and endosperm. The proteins identified in rice endosperm and embryo tissue in two-dimensional electrophoresis included grain storage proteins, glutelin precursors, acidic and alkaline glutelins, globulins, prolamins, HSPs, and hypothetical proteins. Therefore, in this way, it is possible to identify the level of expression of proteins and also their biosynthetic pathways.

Corn is one of the most important crops in the cereal family, and after wheat, it has the largest cultivated area in the world. Among the types of corn, sweet corn seeds are sensitive to storage conditions due to their low starch content and poor seed vigor. Therefore, it is essential to understand their physiological and biochemical behavior during storage to prolong their longevity and prevent the loss of vigor. On the other hand, seed deterioration is a natural phenomenon, resulting in seeds losing their viability and quality even under optimal storage conditions. This undesirable phenomenon causes more damage, especially in sensitive seeds such as sweet corn. Part of this damage is related to the decrease in the speed and percentage of seed germination, which leads to a decrease in plant density and failure to achieve the desired density in the field, resulting in decreased crop yield. This experiment evaluated and tested the ability of seed priming with ascorbic acid to improve damage caused by sweet corn seed deterioration.

This experiment was carried out in the Laboratory of Seed Science and Technology, Faculty of Agriculture, Bu-Ali Sina University, as a factorial in a completely randomized design with four replications. The seed used for the experiment was Chase variety. Wheat seeds deteriorated by accelerated aging method for 72 hours at 43 °C. Then, the deteriorated seeds were primed at 25°C with 1, 1.5 ml of ascorbic acid and distilled water for 18 hours. The studied traits included germination percentage, mean germination time, germination rate, vigor index, malondialdehyde content, soluble sugars, soluble proteins, and superoxide dismutase enzyme activity.

The results showed that vitamin priming of seeds with ascorbic acid improved the germination percentage, germination rate, vigor index, soluble sugars, soluble proteins, and activity of the antioxidant enzyme superoxide dismutase in deteriorated seeds. Among the examined traits, the mean germination time and malondialdehyde content of primed seeds decreased compared to non-primed ones. The concentrations of ascorbic acid used in mean germination time, germination rate, vigor index, and soluble sugars were not significantly different. However, it had a more favorable effect than hydropriming in improving the studied parameters of sweet corn seeds. While seed priming with a concentration of 1.5 mM ascorbic acid had the most significant effect on germination percentage, soluble proteins, malondialdehyde content, superoxide dismutase activity, and its difference with other priming treatments was significant. Even though seed priming with a concentration of 1.5 mM ascorbic acid had the greatest effect on the germination percentage, soluble proteins, malondialdehyde content, superoxide dismutase activity. However, its difference was significant from other priming treatments. It increased the percentage of germination, soluble proteins, and superoxide dismutase activity by 71, 85, and 44%, respectively, and decreased the content of malondialdehyde by 52.9%.

Based on the results from this experiment, it can be concluded that applying ascorbic acid, especially the concentration of 1.5 mM, as the best priming treatment is recommended to recover the lost quality of deteriorated sweet corn seeds and improve their germination characteristics.

Drought stress is known as one of the most important factors limiting the growth and production of agricultural plants in most parts of the world and Iran. It is a multidimensional stress and affects plants at different levels. From a physiological point of view, drought stress causes several changes in the relative water content of leaves and photosynthetic pigments of plants, which in turn causes a decrease in the efficiency of photosynthesis and a significant reduction in plant growth parameters and ultimately grain yield. In Iran, drought stress usually occurs during the filling stage of wheat grains, and due to the limitation of irrigation, supplementary irrigation can play an important role in dealing with drought stress. Durum wheat (Triticum durum, Desf) is an important food product due to its heavy gluten properties and non-sticky dough, ideal for preparing pasta products such as macaroni and spaghetti.
To investigate the physiological characteristics, yield, and yield components of durum wheat cultivars under drought conditions and supplemental irrigation, 10 durum wheat genotypes were evaluated in the form of a split-plot design with the basis of randomized complete blocks in dryland and supplemental irrigation with 3 replications during Crop years 2017-2018 and 2018-2019 in Kohdasht city of Lorestan province. Grain yield and yield components (weight of 1000 seeds, number of seeds per spike, number of spikes per plant) as well as biochemical and physiological properties of photosynthetic pigments, greenness index, relative leaf water, leaf protein and light consumption efficiency were investigated.
Variance analysis showed a significant variation among the examined cultivars, moisture conditions, as well as the interaction of cultivar × humidity conditions for most of the traits except the interaction effect of cultivar × humidity conditions for chlorophyll b and leaf protein. The effect of year and interaction effects of year with other factors were not significant for most of the traits. Performing supplementary irrigation compared to rainfed conditions caused all traits to increase significantly, except greenness index and protein content. Mean comparison of interaction effect of cultivar × moisture environment with Duncan's method at 5% level showed that the genotypes reacted differently for the traits in two moisture conditions and in dry conditions, Omrabi3, Hana, Aria and Saji genotypes were superior to other genotypes in terms of agricultural and physiological traits, and with supplementary irrigation, these for genotypes maintained their superiority, and two genotypes, Shabrang and Behrang, were added to this group.
In general, it can be said that the results were almost the same in the two years studied. Supplementary irrigation compared to rainfed conditions improved the agronomic and physiological characteristics of the cultivars. Among the genotypes, there was a high diversity for all the examined traits and this diversity was different in two humidity conditions. Considering the two-year mean yield, the genotypes of Omrabi3, Saji, Arya and Hana for rainfed conditions and Omrabi3, Hana and Behrang for supplementary irrigation conditions of the studied area are recommended.

Identifying superior genotypes in breeding programs is always difficult due to environmental changes in the target areas and the interaction of these changes with the studied genotypes. Therefore, advanced agricultural lines in breeding programs should be evaluated in environments with different weather conditions in different years to ensure production and stable performance. A genotype usually has different reactions in terms of yield in various environments. The purpose of this research is to increase the area under cultivation of durum wheat and introduce new high-performing varieties to farmers.

In this regard, two new and promising lines of durum wheat obtained from the country's hot climate durum wheat breeding program, numbered D-98-8 and D-98-10, which were tested during the two crop years of 2019-2020 and 2020-2021 (EDYT-W2019) with higher yield than the available commercial cultivars, were selected. To compare the yield with Shabarang variety (as a control), they were grown in the farmers' fields in the warm climate of Khuzestan province ( Hamidiyeh and Ramhormoz cities). The seeds of the two superior durum wheat lines, D-98-8 and D-98-10, with Shebarang as the best regional and national control, were planted in November with a density of 450 seeds per square meter and using a seeder. The area of ​​each treatment in each location was 2500 square meters (without repetition) and according to the existing conditions of the area, gravity irrigation was used. During the growing season, routine operations were carried out and visits to the farm were made regularly and necessary notes were taken. To check the nutritional value, the qualitative characteristics of its bakery were analyzed.

The results of the qualitative analysis showed that the line of D-98-10 was in good condition in terms of semolina extraction percentage (74%), protein percentage (12%) and other investigated traits. Based on the field analysis, in both locations, the D-98-10 line was almost the early-mature line and showed the highest yield (by 5.6%) compared to the Shabrang control. Line D-98-8 with 130 days was a med-maturing line and Shabrang with 135 days was the late one. In both locations, the D-98-10 line showed a higher yield than the other two genotypes, while being suitable for early maturity.

In total, D-98-10 and D-98-8 lines were selected due to their superior yield, relative earliness, high 1000 seed weight and relative superiority in terms of other studied traits compared to Shabrang variety, and they could be used for further investigation and release as a new variety of durum wheat for hot regions of the country.

According to the studies conducted on different plant species, about 90% of the entire genome of a plant is transcribed as RNA. However, only 1-2% of the transcribed genome is protein-coding RNAs which their transcription would end up in, at least, a protein peptide. Non-coding RNAs are RNA molecules from which no protein is synthesized and cannot be translated. Long non-coding RNAs (LncRNAs) are also a group of non-coding RNAs having a length of about 200 nucleotides. Different studies have verified that LncRNAs are widely diverse in type and function. Accordingly, sense and antisense, intronic, intergenic, bidirectional, and enhancer LncRNAs are some known altered types of LncRNAs. Long non-coding RNAa primarily interact with messenger RNAs (mRNA), DNA, proteins, and small RNAs (e.g. microRNAs, miRNA and siRNA) making them an important agent in gene expression processes at epigenetic, transcription, post-transcription, translation and post-translation-levels. There are also some studies vertifying the involvement of such RNAs in the regulatation of the time and quantity of expression in response to various conditions. In addition, this large group of RNAs has important roles in biological processes such as chromatin rearrangement, transcription activation and promoter settings, interference with the strcuture arrangement of transcriptional productions, and the mRNA-to-protein translation process. Among the biological effects of long non-coding RNAs, their essential role in phenotyping functions such as growth and development, response to biotic and abiotic stresses (environmental stresses), regulation of crop cell differentiation and cell cycle specially cereals can be mentioned. Due to the great importance of non-coding RNAs, especially LncRNAs, in this review article we try to investigate the types and mechanisms of LncRNA functions at different epigenetic, transcriptional, post-transcriptional, translational and post-transcriptional levels. Correspondingly, the relationship between LncRNAs and the response of crops to changes in environmental conditions and their interactive role on factors and proteins involved in such biological activities in crops are investigated. It has been verified that there are fewer published articles available related to the role and function of LncRNAs in biological systems of crops, especially cereal.