Studying Salt-Induced Shifts in Gene Expression Patterns of Glucosinolate Transporters and Glucosinolate Accumulation in Two Contrasting Brassica Species.

Brassica crops are well known for the accumulation of glucosinolates-secondary metabolites crucial for plants' adaptation to various stresses. Glucosinolates also functioning as defence compounds pose challenges to food quality due to their goitrogenic properties. Their disruption leaves plants susceptible to insect pests and diseases. Hence, a targeted reduction in seed glucosinolate content is of paramount importance to increase food acceptance. GLUCOSINOLATE TRANSPORTERS (GTRs) present a promising avenue for selectively reducing glucosinolate concentrations in seeds while preserving biosynthesis elsewhere. In this study, 54 putative GTR protein sequences found in Brassica were retrieved, employing Arabidopsis GTR1 and GTR2 templates. Comprehensive bioinformatics analyses, encompassing gene structure organization, domain analysis, motif assessments, promoter analysis, and cis-regulatory elements, affirmed the existence of transporter domains and stress-related regulatory elements. Phylogenetic analysis revealed patterns of conservation and divergence across species. Glucosinolates have been shown to increase under stress conditions, indicating a potential role in stress response. To elucidate the role of GTRs in glucosinolate transportation under NaCl stress in two distinct Brassica species, B. juncea and B. napus, plants were subjected to 0, 100, or 200 mM NaCl. Based on the literature, key GTR genes were chosen and their expression across various plant parts was assessed. Both species displayed divergent trends in their biochemical profiles as well as glucosinolate contents under elevated salt stress conditions. Statistical modelling identified significant contributors to glucosinolate variations, guiding the development of targeted breeding strategies for low-glucosinolate varieties. Notably, GTR2A2 exhibited pronounced expressions in stems, contributing approximately 52% to glucosinolate content variance, while GTR2B1/C2 displayed significant expression in flowers. Additionally, GTR2A1 and GTR1A2/B1 demonstrated noteworthy expression in roots. This study enhances our understanding of glucosinolate regulation under stress conditions, offering avenues to improve Brassica crop quality and resilience.

Agrobacterium-Mediated Transformation for the Development of Transgenic Crops; Present and Future Prospects.

Plant transformation based on Agrobacterium-mediated transformation is a technique that mimics the natural agrobacterium system for gene(s) introduction into crops. Through this technique, various crop species have been improved/modified for different trait/s, showing a successful genetic transformation so far. This technique has many advantages over other transformation methods such as stable integration of transgene, cost effective. However, there are many limitations of this technology such as mostly the crops are recalcitrant to agrobacterium, low transformation efficiency, transgene integration as well as off targets. So, it's very important to explore the major limitations and possible solutions for Agrobacterium-mediated transformation in order to increase its genetic transformation efficiency. Therefore, the present review article gives a comprehensive study how the transgenic crops are developed using Agrobacterium-mediated transformation, crops that have already been modified through this method, and risks associated with transgenic plants based on Agrobacterium-mediated transformation. Moreover, the challenges and problems associated with Agrobacterium-mediated transformation and how those problems can be solved in future for a successful genetic transformation of crops using modern biotechnology techniques such as CRISPR/Cas9 systems. The present review article will be really helpful for the audience those working on Genome editing of crops using Agrobacterium-mediated transformation and will opens many ways for future plant genetic transformation.

Salt-Induced Modulation of Ion Transport and PSII Photoprotection Determine the Salinity Tolerance of Amphidiploid Brassicas.

Brassica species show varying levels of resistance to salt stress. To understand the genetics underlying these differential stress tolerance patterns in Brassicas, we exposed two widely cultivated amphidiploid Brassica species having different genomes, Brassica juncea (AABB, n = 18) and Brassica napus (AACC, n = 19), to elevated levels of NaCl concentration (300 mM, half the salinity of seawater). B. juncea produced more biomass, an increased chlorophyll content, and fewer accumulated sodium (Na+) and chloride (Cl-) ions in its photosynthesizing tissues. Chlorophyll fluorescence assays revealed that the reaction centers of PSII of B. juncea were more photoprotected and hence more active than those of B. napus under NaCl stress, which, in turn, resulted in a better PSII quantum efficiency, better utilization of photochemical energy with significantly reduced energy loss, and higher electron transport rates, even under stressful conditions. The expression of key genes responsible for salt tolerance (NHX1 and AVP1, which are nuclear-encoded) and photosynthesis (psbA, psaA, petB, and rbcL, which are chloroplast-encoded) were monitored for their genetic differences underlying stress tolerance. Under NaCl stress, the expression of NHX1, D1, and Rubisco increased several folds in B. juncea plants compared to B. napus, highlighting differences in genetics between these two Brassicas. The higher photosynthetic potential under stress suggests that B. juncea is a promising candidate for genetic modifications and its cultivation on marginal lands.

Contrasting Responses to Stress Displayed by Tobacco Overexpressing an Algal Plastid Terminal Oxidase in the Chloroplast.

The plastid terminal oxidase (PTOX) - an interfacial diiron carboxylate protein found in the thylakoid membranes of chloroplasts - oxidizes plastoquinol and reduces molecular oxygen to water. It is believed to play a physiologically important role in the response of some plant species to light and salt (NaCl) stress by diverting excess electrons to oxygen thereby protecting photosystem II (PSII) from photodamage. PTOX is therefore a candidate for engineering stress tolerance in crop plants. Previously, we used chloroplast transformation technology to over express PTOX1 from the green alga Chlamydomonas reinhardtii in tobacco (generating line Nt-PTOX-OE). Contrary to expectation, growth of Nt-PTOX-OE plants was more sensitive to light stress. Here we have examined in detail the effects of PTOX1 on photosynthesis in Nt-PTOX-OE tobacco plants grown at two different light intensities. Under 'low light' (50 µmol photons m-2 s-1) conditions, Nt-PTOX-OE and WT plants showed similar photosynthetic activities. In contrast, under 'high light' (125 µmol photons m-2 s-1) conditions, Nt-PTOX-OE showed less PSII activity than WT while photosystem I (PSI) activity was unaffected. Nt-PTOX-OE grown under high light also failed to increase the chlorophyll a/b ratio and the maximum rate of CO2 assimilation compared to low-light grown plants, suggesting a defect in acclimation. In contrast, Nt-PTOX-OE plants showed much better germination, root length, and shoot biomass accumulation than WT when exposed to high levels of NaCl and showed better recovery and less chlorophyll bleaching after NaCl stress when grown hydroponically. Overall, our results strengthen the link between PTOX and the resistance of plants to salt stress.

Knowledge of radiation legislation and radiation exposure in common radiological investigations among final year medical students, foundation doctors, specialist radiology registrars and radiographers at a UK university teaching hospital.

OBJECTIVE: Junior doctors routinely request radiological investigations for patients. Prior studies have noted that among this group there is a lack of knowledge on radiation legislation and radiation exposure in common radiological investigations. However, no studies have compared this against radiology trainees and radiographers. We compared knowledge of radiation legislation and radiation exposure in common radiological investigations among final year medical students (FYMS), foundation year doctors (FY1, FY2) against specialist radiology trainees (SRT) and radiographers (RG). METHODS: A 12-question multiple choice questionnaire (MCQ) was distributed to FYMS, FY1, FY2, SRT and RG at a UK teaching hospital. Questions assessed knowledge of radiation legislation and radiation-dose estimates of common radiological investigations. Mean MCQ scores were compared using one-way ANOVA and Tukey post-test to determine statistical significance (p-value < 0.05). RESULTS: 127 participants were included in the study. Mean scores (%) for FYMS (49.3%), FY1 (52.6%) and FY2 (51.1%) were significantly lower compared to SRT (64.4%) and RG (66.3%) (p-value < 0.05). Mean test scores between FYMS, FY1 and FY2 did not significantly differ (p-value > 0.05). CONCLUSION: FYMS, FY1 and FY2 knowledge of radiation legislation and radiation exposure in common radiological investigations was poor compared to SRT and RG. Patients require knowledge of radiation risk to provide informed consent as per IRMER regulations, thus we propose formal teaching on the subject matter to promote radiation safety culture among medical undergraduates and postgraduates. ADVANCES IN KNOWLEDGE: First study to compare knowledge of radiation legislation and radiation exposure in common radiological investigations between medical students and junior doctors to radiology trainees and radiographers.

Chloroplasts as Cellular Factories for the Cost-effective Production of Cellulases.

Chloroplasts are vital photosynthetic organelles in plant cells that carry out several important cellular functions including synthesis of amino acids, fatty acids, and lipids and metabolism of nitrogen, starch, and Sulphur to sustain the homeostasis in plants. These organelles have got their own genome, and related genetic machinery to synthesize required proteins for various plant functions. Genetic manipulations of the chloroplast genome for different biotech applications has been of great interest due to desired features including the availability of operonal mode of gene expression, high copy number, and maternal mode of inheritance (in the most field crops). Their capacity to often express transgenes at high levels make it a cost-effective platform for the production of foreign proteins, particularly high-value targets of industrial importance, at large scale. This article reviews briefly the research work carried out to produce cellulolytic enzymes in higher plant chloroplasts. The challenges and future opportunities for the same are also discussed.