مجله بیوتکنولوژی کشاورزی
سال شانزدهم شماره 1 (پیاپی 54، بهار 1403)

‎‏ ‏The purpose of this research is to investigate the efficiency of the ISSR molecular marker in ‎distinguishing the studied watermelon populations, to determine the degree of similarity and genetic ‎distance of the populations in order to use the phenomenon of heterosis and to select suitable parents in ‎watermelon hybrid seed production programs.‎

In this research, the genetic diversity of 24 different watermelon populations ‎from all over Iran was investigated using 10 ISSR primers. DNA extraction was done using the CTAB ‎method. After PCR, molecular data were scored as zero and one. Indexes related to markers include ‎total number of bands, number of polymorphic bands, polymorphic percentage, marker index (MI), ‎resolution strength index (RP) and polymorphic information content (PIC). It was measured. In order to ‎perform cluster analysis, NTSYS and Excel software were used, and GenALEx software was used to ‎analyze the molecular variance and to analyze the main coordinates.‎

In this research, a total of 202 bands were produced, of which 184 were polymorphic bands. ‎The average number of total bands and polymorphic bands in this study was 20.2 and 18.4, respectively. ‎The highest percentage of polymorphism was related to the ISSR6 primer with 100% and the lowest ‎polymorphism was related to the ISSR5 primer with 80%. The average percentage of polymorphism in ‎this study was also 0.90. The value of PIC index in this research varied between 0.26 and 0.44 and the ‎average PIC was 0.35. After cluster analysis, the studied populations were divided into five groups at a ‎similarity level of 55%. The results of molecular variance analysis and principal coordinate analysis in ‎the studied populations showed that 14% of the variation is between the studied populations and 86% ‎of the variation is within the studied populations. The results of analysis to main coordinates also ‎confirmed the results of cluster analysis‏.‏

According to the obtained results, the ISSR molecular marker has the necessary efficiency ‎to distinguish the studied populations, and on the other hand, considering the considerable genetic ‎diversity between and within the studied populations, the genotypes used in this research can be ‎considered as the population The first one was used in watermelon hybrid seed production programs ‎and also Faryab and Minab populations can be used as cross parents considering that they had the least ‎genetic similarity in order to use heterosis in watermelon hybrid seed production projects.‎‏ ‏

Late spring cold stress in the early reproductive growth of wheat causes damage due to a sudden drop in temperature. The genetic diversity of plants determines their potential to increase efficiency and, as a result, their use in breeding programs. This study was conducted with the aim of evaluating population structure, diversity of wheat genotypes, and selection of superior genotypes under cold and normal stress conditions.

Sixty-seven bread wheat genotypes were evaluated through pot cultivation in the field under two normal conditions and late spring cold stress. The experiment followed a randomized complete block design with two replications. Stress was applied by transferring the pots to a growth room with a temperature of -3ºC at the beginning of the reproductive stage. Phenotypic traits, including performance and performance components, were measured. Genotyping was done using a single nucleotide polymorphism (SNP) array with 9K density. A total of 17,093 SNP markers were used in this study.

The polymorphic information content for SNP markers ranged from 0.03 to 0.38, with an average of 0.26. Population structure analysis grouped the genotypes into five subpopulations. Molecular variance analysis showed that 75% of the observed variance in the population was related to intra-population differences. The studied genotypes were divided into four groups based on SNP markers and five and six groups based on phenotypic traits under normal and stress conditions, respectively. The genotypes in the first group, which exhibited higher average traits than other groups under stress conditions, were identified as tolerant to late spring cold.

The results indicated that main spike weight, seed yield per plant, seed weight in main spike, and weight of 100 seeds were the most important factors for grouping under late spring cold stress conditions. The calculated molecular indices, including marker indices and diversity indices, demonstrated that the SNP markers used performed relatively well in calculating genetic diversity within the studied population. In cluster analysis based on the genotypic matrix, cold-tolerant genotypes formed separate groups, indicating different tolerance mechanisms among these genotypes. These findings provide useful information for wheat breeders regarding the selection of diverse genetic resources to improve cold tolerance.

Bevacizumab (trade name Avastin®) is a humanized monoclonal antibody (mAb) that is widely used in the treatment of various cancers. It is one of the most expensive and best-selling cancer drugs in the world. Bevacizumab is produced by recombinant DNA technology in a mammalian expression system, the Chinese hamster ovary. In recent years, plant expression systems have attracted global attention due to their various benefits for producing recombinant pharmaceutical proteins. The successful expression of the mAbs is dependent on the proper folding and assembly of the light chains (LC) and heavy chains (HC). The efficient and controlled co-expression of the peptide chains is an essential consideration in designing antibody expression vectors. This study aimed to construct a plant expression vector for the simultaneous expression of HC and LC peptide chains of bevacizumab mAb using the IntF2A fusion domain.

In order to construct the recombinant vector, the sequences of genes encoding bevacizumab and IntF2A were codon-optimized and artificially synthesized for expression in tobacco plants. The bicistronic recombinant vector was constructed by the routine restriction digestion-ligation method. Finally, the recombinant construct was used for the transient expression of bevacizumab in tobacco plants through agroinfiltration approach. RT PCR analysis was used to determine the presence of transgene transcripts. The production and accumulation level of the recombinant antibody was evaluated through western blotting analysis.

Recombinant plasmid construction is verified by colony PCR, restriction digestion, and sequencing analyses. Expression of the transgenes as a single-transcript unit was confirmed by RT-PCR analysis. Western blot analysis showed the production of the full-length heterotetrameric antibody in agroinfiltrated leaves. The level of antibody accumulation was estimated at an average of 67.20 mg of bevacizumab per kg of fresh weight (3.36% TSP).

The results showed that the IntF2A-based bicistronic vector provides an efficient simultaneous expression of genes encoding the polypeptide chains of antibody and high accumulation of the full-length mAb in plant cells. The constructed vector can be used for the possibility of development and manufacturing of bevacizumab antibody in plant expression system.

When continuous selection is carried out over years, it creates effects on the genome level, which can be detected by using some strategies. This study was carried out with the aim of scanning the whole genome to identify regions of the genome in Thoroughbred and Turkoman horses that have been targeted by natural or artificial selection.

DNA was extracted from blood samples using the optimal salt method. The quality and quantity of extracted DNA of all samples was determined by nanodrop device with absorption ratio on DNA solution. For this purpose, 44 Thoroughbred horses and 67 Turkoman horses were genotyped by genomic arrays of 60k SNP chips. By two general methods of population differentiation and linkage disequilibrium methods, the selection signatures at the genome level were looked into. In order to identify the population genetic structure of the studied animals, principal component analysis was done in R program.

The study of population differentiation using the fixation index method (Fst) corrected for the sample size (θ) showed that there are evidences of selection in several loci in these two breeds. A number of five genomic regions were identified in which there were signatures of selection. These areas are located on chromosomes 4, 5, 10, 13 and 15. In order to evaluate the signatures of selection based on linkage disequilibrium methods, the extended haplotype homozygosity test (EHH) was used. The results confirmed the existence of population segregation in these genomic regions. Finally, the investigation of QTLs in the bovine orthologous regions showed that these regions are related to the traits of body length, body weight, chest depth and other important economic traits in horses.

The present research was effective in identifying regions of the genome of the two breeds of Turkoman and Thoroughbred horses being divergently selected, and identifying the genes that exist in these regions. Also, useful information was obtained on the existence of genetic diversity and signatures of selection between these two horse breeds.

Exendin-4 (Ex4) is a 39-amino acid peptide isolated from the salivary secretions of the lizard Heloderma susceptum and it has similar biological function as glucan-like peptide (GLP-1) receptor. The most important physiological role of GLP-1 is to regulate the amount of glucose metabolism by reducing the entry of glucose into the cells. Transgenic plants can be used as factories for the production of recombinant proteins. In order to maximize the efficiency of transgenic plants, the expression of foreign genes can be regulated in terms of time and place under the regulation of different promoters. Considering the nutritional value of carrot tuber and its fresh consumption, in this study, CTBEx4 gene was expressed in carrot plant using a root-specific promoter. In this study, in order to specifically express the CTBEx4 gene in carrot root, the Major Latex-Like promoter (MLL) related to the specific expression in sugar beet storage root tissue was replaced by the CaMV35S promoter in the pBI121 construct and were used for the transformation of carrot explants.

Using the CTAB method, DNA was extracted from the leaves of the sugar beet plant, then the PCR reaction was performed using specific MLL primers. The obtained fragment was replaced by the CaMV35S promoter in the pBI121 construct using restriction enzymes. Both pBI121-Mll-CTBEx4 and pBI121-CamV35S-CTBEx4 constructs were used to transform carrot leaf explants using Agrobacterium tumefaciens bacteria. Transgenic explants were indirectly regenerated into plants in MS culture media in vitro and then transferred to pot. Finally, the CTBEx4 gene expression of the transgenic plants was analyzed at mRNA and protein levels using RT-PCR and ELISA.

The results of the expression analysis using RT-PCR and ELISA methods indicated that the expression of CTBEx4 gene in carrot root tissue was regulated by MLL promoter, while no expression was observed in leaf tissue. Also, the results showed that the CTBEx4 gene was expressed under CaMV35S promoter regulation in both root and leaf tissues, but its expression in roots was lower (30%) than that under MLL promoter regulation. The obtained results indicated the ability of the MLL promoter to specific expression the CTBEx4 gene in the roots of carrot plants.

The MLL promoter was able to express the CTBEx4 gene in the root tissue of carrot plants at the mRNA and protein level, Also, CTBEx4 protein under MLL promoter had higher expression level compared to CaMV35S promoter.

Coronatine insensitive 1 (COI1) gene, insensitive to bacterial toxin, the main member of jasmonates (JAs), is closely related to biotic and abiotic resistance of plants. COI1 gene acts as the main controller of plant responses regulated by JA. To better understand the molecular basis of JA function, mutant and wild type in Arabidopsis were investigated under biotic and abiotic stresses.

In this research, coi1 gene mutation and wild type were evaluated under Pseudomonas aeruginosa, IAA, ABA, salinity and drought stresses. Also, the physiological traits of mutant and wild type were investigated under the mentioned stress. In addition, the expression profile of JA-related genes in Arabidopsis leaves under five stresses was analyzed.

The expression level of five genes in the leaves decreased in the mutant and increased in the wild type. The transcript levels of five genes related to (JA fatty acid desaturase, defective anther opening 1, allene oxide synthase, PDA reductase and 13-lipoxygenase) were well correlated with JA accumulation under five stresses. Compared to the wild type, the activities of catalase (CAT), peroxidase (PRO) and ascorbate peroxidase (APX) were high in coi1 mutant leaves under five stresses. Under these stresses, Arabidopsis plants produce much higher levels of anthocyanins in the wild type compared to the coi1 mutant. The expression of other anthocyanin biosynthetic genes was also induced by JA and dependent on COI1. These results showed that jasmonates may regulate the growth of Arabidopsis. For the first time in this study, the effect of Pseudomonas aeruginosa in coi1 mutant and wild type has been compared. The findings of the present study showed that Pseudomonas aeruginosa had the effect of cronotine by strengthening a COI-dependent pathway for pathogen proliferation, and the wild type had a high gene expression of jasmonic acid compared to the mutant. These results confirmed that the COI pathway was caused the proliferation of pathogens and the mutant type is more resistant to Pseudomonas aeruginosa than the wild type.

The results of the present study provide new insight into JA accumulation and its potential roles during development. These results indicated that jasmonate biosynthesis and signaling could be regulated by a complex regulatory network

Phytases are classified as 3-phytases (E.C.3.1.3.8), 5-phytases (E.C. 3.1.3.72) and 6-phytases (E.C. 3.1.3.26) based on the position of the first phosphate residue removed from the myo-inositol ring of phytate. Yeasts are particularly suited to expression of foreign proteins for numerous features mean while some of them shows probiotic properties. Pichia pastoris, has been developed for the production of various recombinant proteins and growth into high cell densities in an inexpensive medium. Accordingly, Pichia pastoris is an appropriate secretory system for obtaining larger quantities of correct products, in compared to other host cells. The coexpression of digestive enzymes in a single recombinant cell system would thus be advantageous. The objective of this study is to determine whether combining fungal phytases (3-phytase) with bacterial phytase (6-phytase) was more effective than each phytase alone in degrading of plant phytate. we tested the new vector, aimed to clone and coexpress the phytase genes isolated from E. coli and aspergillus niger and transformed into Pichia pastoris. Evaluations were made for the biochemical properties of the active expressed phytase.

The nucleotide sequence of phyA and appA2, 2a peptide and alpha factor were obtained from NCBI database. A coexpression system for the extracellular production of phytase of Aspergillus Niger (3-phytase) and phytase of E. coli (6-phytase) will be established in Pichia pastoris yeast. Plasmid that used in this study was pPIC9k. The genes for each enzyme are fused in-frame with the “a-factor” secretion signal and linked by the 2A-peptide-encoding sequence. After receiving the synthetic plasmid with fragments, plasmid that contained phyA and appA2 was cloned in DH5@ and after that digested by SacI and electrotransfered into Pichia pastoris. Positive cells are resuspended at BMMY containing methanol as an inducer. To follow the induction, methanol was injected into the culture every 24 hours in order to reach a definitive concentration of 0.5 percent. The recombinant protein was analyzed using sds-page.

The results showed that the recombinant phytase gene was transferred to Pichia pastoris and the results of PCR confirmed that. The molecular weight of the produced recombinant phytases was estimated to be around 45 and 80 kDa for appA and phyA, respectively. The recombinant protein has phytase activity equal to 160.97 U/mlConclusions;The designed phytase genes containing phyA and appA were successfully cloned in DH5a and the recombinant plasmid was transferred to Pichia pastoris for high expression. Recombinant yeast will be used for further experiments.

Most of the garlic clones are non-bolting and sterile and propagate asexually; however, some of the clones are bolting, and there are some fertile clones among the bolting ones. A correct understanding of the MADS-BOX genes family expression pattern and structure, including AGL6, is a valuable project and will improve garlic breeding programs. So far there are no reports relating AGL6 identification as a flowering and fertility integrator gene in garlic. So the aim of this study is to investigate the relative expression of AGL6 in Iranian bolting, semi- and non-bolting garlic clones and to identify its structure and nucleotide sequence.

At first the relative expression of the AGL6 was investigated in the different organs of non-bolting, semi-bolting and bolting Iranian garlic clones using quantitative Real time- PCR. Then the isolation and cloning of the AGL6 partial sequences from the garlic floret was carried out using RT-PCR. The sequence of the recombinant plasmid was evaluated using BLAST, Vector NTI, ORF finder and MEGA6 software.

According to the obtained results, AGL6 was expressed in the meristem, inflorescence, tepal and carpel of green and purple flowers of the bolting garlic. But there was no sign of its expression in the stamen. AGL6 expression level in the meristem of bolting garlic was higher than the meristem of the semi-bolting and non-bolting garlic in all sampling stages, especially in the flowering induction stage. AGL6 was also expressed in the inflorescence of the semi-bolting and bolting garlic, but its expression in the inflorescence of the semi- bolting garlic was 4 times less than the inflorescence of the bolting garlic. Sequence analysis of the colonized fragment showed the coding region with a 728 bp length, which was registered in the NCBI database with the accession number of OK086759.1. This nucleotide sequence encodes a protein with 243 amino acids and has MADS-box and K-box domains. Results showed the high similarity of the sequence obtained in this research with other AGL6 gene homologues in other plant species, especially petaloid monocots such as onion, lily, narcissus and saffron in NCBI. Hence the resulting sequence was named AsAGL6.In this research, for the first time, the expression pattern of a key gene of the flowering and fertility pathway (AsAGL6) and its nucleotide structure were determined in garlic. The results of this research can be effectively used in the design of classical and molecular garlic breeding programs.

This study was conducted to investigate the genetic diversity of tomato’s cultivars in Iran. Moreover, the efficiency and the application of ISSR molecular markers in evaluating tomato diversity were assessed.

Ninety-nine tomato cultivars, including previously popular, prevalent and cultivars under study for cultivation in Iran were evaluated. Accordingly, 20 ISSR primers were practiced. DNA isolation were carried out using the fresh leaf samples and then determined the quantity and quality of extracted DNA. After PCR, the amplicons were separated by horizontal electrophoresis. The gels were stained with ethidium bromide and visualized in a UV transilluminator for subsequent analysis in digitalized images. The obtained information was analyzed via statistical software.

Amongst 20 primers were applied, the 17 primers were polymorphic. They have made 275 polymorphic amplicons which size's varied from 250 to 3000 base pair (bp). The average polymorphism rate was 97% and nine primers were 100% polymorphic. Having high polymorphic information content, marker index, effective multiplex ratio, and resolution power, UBC 879 primer was properly effective in differentiating the cultivars. The mean Jaccard similarity coefficient was 0.41. The highest genetic similarity was observed between cv. H1423 and cv. Kimia while the lowest genetic similarity was between cv. Lina and cv. Pil ZTP1; 0.96 and 0.13 respectively. The UPGMA algorithm was applied in cluster analysis based on Jaccard similarity coefficient. This analysis grouped cultivars into five clusters. According to the analysis of molecular variance (AMOVA), the distinction between the formed groups was significant. In this study about 35% of the data variation was explained by the first and second components. The overall grouping pattern of clustering corresponds with principal component analysis. The cultivars were divided into five groups. By providing spatial representation of relative genetic distances among individuals, PCA analysis determined the consistency of differentiation among cultivars. cv. Lina and cv. ZTP8 were placed in one group alone and have the greatest genetic distance from other varieties.

This research claimed that Iranian tomato cultivars to some extent have high genetic diversity. In addition, it has been shown that ISSR molecular markers are appropriate for investigating the genetic diversity amongst tomato genotypes due to generation of high level of polymorphism. Thus, these markers have substantial efficiency in distinguishing tomato genotypes.

First lambing age and lambing interval are among the most important reproductive traits affecting profitability of sheep breeding systems. The present study aimed to conduct a genome wide association studies (GWAS) for identifying the loci associated with first lambing age and lambing interval in Zandi native sheep using the 50K arrays.

For this purpose, phenotypes records and genotypic data related to first lambing age and lambing interval were obtained from 96 Zandi sheep. Different steps of quality control and identification of independent SNP markers were performed by PLINK software. After quality control, 94 animals and 36070 markers were identified for further analysis. Genome wide association study was performed with reproductive traits using GEMMA software. Using the Genome Data Viewer software from NCBI database based on last assembly (ARS-UI_Ramb_v2.0) the SNP were assigned to genes if they were within the genomic sequence of the gene or within a flanking region of 300 kb up- and downstream of the gene. Finally, from online bioinformatics databases for the assignment of the genes to functional categories, DAVID and PANTHER databases were used.

In this research, seven SNP markers on chromosomes 1, 2, 3, 6, 11 and 23 were identified. Also, we identified different sets of candidate genes related to reproductive traits: UMPS, ITGB, SMARCAL1, APAF1, UGT8 and ST8SIA5 in Zandi sheep. Some of the found genes, are consistent with some of the previous studies related to reproductive traits. Some of the genes were found are consistent with some previous studies and to be involved biological pathways related to fertilization, ovulation rate, estrogen biosynthesis, conception rate, skeletal muscle development, early development of the fetus and puberty.

The results of our research can be used to understand the genetic mechanism controlling reproductive traits and considering, this study supported previous results from GWAS of reproductive traits, also revealed additional regions, using these findings could potentially be useful for genetic selection in sheep for age at first lambing.

The objectives of genetic and metabolic engineering can be greatly advanced through comprehending the molecular processes of the stress response. Researchers may access an abundance of data through transcriptome studies. Using advanced statistical methods such as meta-analysis to integrate data from many sources offers a new way to find differentially expressed genes (DEGs), get beyond biological complexity, and provide more accurate results. The current work used transcriptome data from RNA-seq meta-analysis to uncover Catharanthus roseus DEGs in response to stressors and the production of important secondary metabolites.

The EMBL-EBI database provided the RNA-seq data, which was then pre-processed and mapped quality reads using STAR software to the reference genome of C. roseus. Each data set's changes in gene expression were examined independently using the edgeR package, and the output from these analyses was then utilized for a meta-analysis with the use of the metaRNAseq program. Key genes in reaction to stress and pathways were finally examined, along with the biological function and biological pathways involved in the different and significant expression of these genes in response to stress. Co-expression analysis was carried out using the WGCNA software and hub genes associated with stress response and the production of important plant-related metabolites were found.

The findings of the meta-analysis show that 772 genes with average expression log2FC≥|1 and FDR≤0.05 were found, and that 305 and 467 of these genes, respectively, had up- and downregulated expression under stress. Of these, only a meta-analysis enables the identification of 27 genes that were not identified as DEGs in individual analyses. The functional enrichment results of DEGs demonstrate their crucial importance in metabolic processes, response to stimuli, and cellular processes. They also lead multiple pathways, such as the MAPK signaling pathway, the biosynthesis of secondary metabolites, the transduction of plant hormones, and metabolic pathways under stress. Through co-expression network analysis, stress-responsive hubs genes and associated with the TIA biosynthesis pathways were found that may be potential genes involved in the synthesis of therapeutic compounds exclusive to plants.

In this study, we identified key and novel genes by investigating the molecular responses of C. roseus plant under various stresses using meta-analysis and systems biology approaches. These genes have the potential to be employed as candidate genes in genetic and metabolic engineering projects aimed at enhancing the plant's capability to produce therapeutic metabolites in the future.

Breast cancer is still the most common cancer among women all over the world. Using surgery, chemotherapy, and radiation therapy is known as the best traditional method to treat this disease. In recent years, with the advancement of nanotechnology in medical science and treatment, new methods of cancer treatment have been investigated. Considering the medicinal and therapeutic properties of F.vulgare due to the presence of valuable compounds such as flavonoids for the treatment of cancer, in this research, first, the morphological and structural characteristics of metal nanoparticles biosynthesized from the aqueous extract of F.vulgare were discussed, then the antioxidant and anticancer properties of essential oil, aqueous extract of this plant and the biosynthesized nanoparticles were investigated.

In this research, after the biosynthesis of iron, copper, zinc, and silver nanoparticles from the aqueous extract of fennel plant (F. vulgare) and their properties in terms of size with XRD and confirmation of the structure by FTIR, the amount of secondary metabolites, the amount of antioxidant and anticancer activity of the essential oil And the aqueous extract of fennel plant (F. vulgare) and iron, copper, zinc and silver nanoparticles biosynthesized from it were evaluated on the Sum-159 and the Hs-578T cell lines.

According to the results obtained from DLS of biosynthesized metal nanoparticles, the sizes of iron, copper, zinc, and silver nanoparticles were 35, 32, 33, and 34 nm, respectively. The results of FTIR also confirmed the existence of the synthesized metal nanoparticles. Also, according to the results of HPLC and spectrophotometry, the presence of biochemical compounds such as flavonoid and anthocyanin and metabolites such as rutin, quercetin, and kaempferol in the aqueous extract of fennel plant was confirmed. In terms of antioxidant activity, a significant difference was observed between different treatments, it was also observed that with the increase in the concentration of the treatments, the amount of their antioxidant activity also increased, and the highest amount of antioxidant activity of each treatment was related to the highest concentration of each treatment. The highest amount of antioxidant activity (87.41%) was related to copper nanoparticles at a concentration of 500 mg/ml. In terms of the IC50 value of the mentioned treatments, it was also found that the Sum-159 cell line had higher resistance compared to the Hs-578T cell line against equal concentrations of different treatments.

Over the course of evolution, the immune system has finely tuned itself into a formidable defense mechanism against infectious agents. A fundamental aspect of its functionality lies in the ability to discern between self and nonself, crucial for triggering immune responses. However, cancer disrupts this equilibrium, characterized by aberrations in the DNA code that empower cells to proliferate uncontrollably while evading immune surveillance. Immunotherapy emerges as a promising avenue, seeking to train the immune system to identify and eradicate these rogue cancer cells, thereby presenting a novel strategy in the battle against cancer. Despite the effectiveness of immune checkpoint inhibitors across various advanced cancer types, the overall response rate in individuals undergoing such treatments hovers around thirty percent. Recognizing the need for advancements, recent research delves into nanoparticle-based approaches aimed at enhancing cancer therapies and vaccinations. The focus centers on developing biotechnology that employs durable artificial nanoparticles to transform cancerous cells into tumor-based Antigen-Presenting Cells (APCs). This transformative process involves stimulating coexpression of costimulatory molecules and immunostimulatory cytokines. What distinguishes this nanomedicine is its capacity to induce a generalized tumor-specific and cell-mediated immune response without assuming specific antigens expressed by tumors. This innovation holds tremendous potential for advancement in translational medicine, offering a versatile and adaptive solution to the challenges posed by cancer immunotherapies. By leveraging the capabilities of artificial nanoparticles, researchers aspire to elevate the efficacy of cancer treatments and propel the field towards a new era of more targeted and potent interventions. In conclusion, the intricate interplay between the immune system and cancer underscores the necessity for innovative therapeutic approaches. The exploration of nanoparticle-based strategies represents a frontier in cancer research, holding the promise of refining immunotherapies and ushering in a new era of precision medicine for the benefit of patients grappling with this complex and formidable disease.

In the realm of biotechnological enhancement of common beans, an imperative challenge lies in devising a reliable and effective in vitro regeneration strategy, given the inherent difficulty of regenerating this crop in laboratory settings. This research, aiming to address this challenge, leverages the power of Machine Learning (ML) models, specifically employing algorithms for Artificial Neural Networks (ANN). The primary objective is to establish an efficient and repeatable in vitro regeneration process while simultaneously optimizing and predicting future outcomes. The study incorporates various variables such as bean genotype, explants, and different doses of 6-benzylaminopurine (BAP) and CuSO4. A Recurrent Regression Neural Network (RRNN) is employed to model and anticipate the results of in vitro crop regeneration, specifically focusing on common beans. The experimental setup involves preconditioning common bean embryos with 10, 15, and 20 mg/L BAP for 25 days, followed by growth in a post-treatment environment comprising 0.3, 0.6, 0.9, and 1.2 mg/L BAP for 7 weeks. Subsequently, the plumular apice is isolated for in vitro regeneration. Notably, the RRNN model is also integrated with a Genetic Algorithm (GA) to optimize the regeneration process further. The results are compelling, with RRNN exhibiting the lowest Mean Squared Error (MSE) of 0.061, signifying superior predictive accuracy in total regeneration. In comparison, Support Vector Regression (SVR), Random Forest (RF), and Extreme Gradient Boosting (XGB) models exhibit higher MSE values of 0.081, 0.081, and 0.097, respectively. These findings underscore the efficacy of the RRNN algorithm, outperforming other models across all parameters. The superior performance of RRNN suggests its potential application in making precise predictions regarding common bean regeneration. In the context of a common bean breeding program, these outcomes can be harnessed to optimize and predict plant tissue culture methods, thereby enhancing biotechnological techniques employed in the cultivation of common beans. The integration of ML models, particularly RRNN, stands as a promising avenue for advancing crop regeneration strategies and contributing to the efficiency of biotechnological interventions in agriculture.

Nanotechnology, the manipulation of matter at the anatomical and molecular levels, has paved the way for the creation of materials with unique and enhanced properties. One of its significant applications is in the realm of nano-biotechnology, where various subfields, such as nanostructures, play a crucial role in fields like biotechnology and structural and cellular genomics. Nano-biotechnology offers a groundbreaking approach to research by allowing scientists to introduce chemicals and components into cells and fabricate novel materials through innovative techniques like assembling. Particularly noteworthy is its contribution to the in vivo administration of Nucleic Acid (NA) therapeutics. Advanced nanoscale biotechnology strategies have played a pivotal role in this domain by providing precise control over crucial parameters such as dimension, charge, drug loading, and response to external signals of delivery transporters. The integration of cutting-edge nanoscale biotechnology in the field of cancer immunotherapy holds significant promise for overcoming obstacles in NA treatment. This involves addressing challenges related to the clinical and preclinical research of different nanocarriers. This article serves as a concise overview, delving into the characteristics and encountered difficulties associated with various nanocarriers during clinical and preclinical investigations. The paper discusses a range of NA methods and elucidates how state-of-the-art nanoscale biotechnology actively supports and enhances NA treatment. By manipulating the physical and chemical attributes of delivery transporters, nanotechnology allows for the optimization of therapeutic outcomes, making it an invaluable tool in the advancement of medical research. In conclusion, the marriage of nanotechnology and biotechnology, specifically in the field of nano-biotechnology, opens up new avenues for scientific exploration and medical advancements. The precision and versatility offered by nanoscale techniques hold immense potential in revolutionizing the landscape of NA therapeutics, paving the way for more effective and targeted treatments. The ongoing progress in cutting-edge Nanobiotechnology (N-Bio-Tech) exemplifies the relentless pursuit of solutions to some of the most significant challenges in medical science, particularly in the realm of cancer immunotherapy.

The dairy industry, a prolific producer of waste and by-products, boasts a diverse array of goods that contribute significantly to our daily diet. However, the copious waste generated in the process, characterized by high Chemical Oxygen Demand (COD) and a rich nutrient profile comprising lactose, proteins, and fats, necessitates exploration of alternative biotechnological applications. This exploration is particularly crucial when considering both aerobic and anaerobic processing methods. Probiotic food items, having garnered widespread consumer acceptance, are experiencing substantial growth in the nutritional food industry. Recognizing this trend, the food industry is actively engaged in expanding the scope of probiotic meals beyond traditional dairy products. This expansion holds promise for delivering numerous health benefits to consumers. Consequently, this research delves into the extensive range of dairy products available, elucidating ways to harness by-product waste from the dairy industry. However, this pursuit is not without its challenges. Microbiologists, engineers, and technologists face hurdles in preventing the infiltration of unwanted microbes during processing. The study not only examines the methods to eliminate these intruding microbes and their digestive enzymes but also addresses the intricacies involved in thwarting the development and operations of any surviving microorganisms following treatment. Probiotics, known for their positive impact on gut health, have found application in various food matrices, extending beyond dairy to encompass non-dairy products. The research underscores the versatility of probiotics and their adaptability to different food categories. Furthermore, it briefly touches upon the innovative utilization of byproduct waste, converting what would otherwise be considered trash into valuable resources. In conclusion, the dairy industry's waste management presents a dual opportunity: to mitigate environmental impact through responsible waste disposal and to extract valuable nutrients for application in diverse biotechnological realms. This comprehensive exploration contributes to our understanding of the potential of dairy waste by-products, shedding light on sustainable practices and innovative applications that align with the evolving landscape of the nutritional food industry.

Bioinformatics has evolved into an indispensable tool across various realms of biology, playing a pivotal role in advancing scientific pursuits. Despite its ubiquitous presence in traditional life science courses, integrating hands-on training with wet-lab investigations becomes imperative to foster student engagement and comprehension. This holistic approach not only bridges theoretical knowledge with practical applications but also cultivates a deeper appreciation for the interdisciplinary nature of bioinformatics. The ever-accelerating progress in omics studies has unveiled unprecedented avenues for comprehending biological structures. This surge has sparked a revolutionary transformation in the field, propelling marine studies beyond the realms of hypothetical organisms to encompass an expanding array of marine life. At the forefront of this marine exploration is the research's central focus on the practical task of unearthing novel enzymes in marine environments. The inquiry delves into the cutting-edge concept of metagenomics, a contemporary methodology that broadens the horizons of biotechnology by incorporating non-culturable microorganisms into its purview. A crucial facet of the research involves the introduction of a viral cosmid library screening probe, responding to the challenges posed by the study of marine microbiology and bioengineering. This innovative tool becomes instrumental in deciphering the genetic makeup of elusive marine microorganisms, further enriching our understanding of their biochemical processes. The paper meticulously delineates the objectives of marine microbiology and bioengineering projects, underscoring the need for a collaborative effort to achieve them successfully. In this context, the research not only outlines the goals but also provides a strategic roadmap for their implementation through a synergistic collaborative approach. This approach harnesses the collective expertise of scientists and researchers, fostering an environment conducive to breakthrough discoveries. The fusion of experimental research techniques enhances students' proficiency in fundamental bioinformatics, instilling in them a problem-solving mindset crucial for navigating the complexities of contemporary biological research. In conclusion, the integration of hands-on training, coupled with the exploration of cutting-edge methodologies like metagenomics, marks a paradigm shift in the study of marine microbiology and bioengineering. This research not only contributes to the growing body of knowledge in these domains but also underscores the significance of collaborative efforts in pushing the boundaries of scientific exploration.