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Transcriptomic Characterization of Genes Harboring Markers Linked to Maize Yield
Background: It is currently believed that breeding priorities, including maize breeding, should focus on introducing varieties with greater utility value, specifically higher yields, into production. Global modern maize breeding relies on various molecular genetics techniques. Using the above mentioned technologies, we can identify regions of the genome that are associated with various phenotypic traits, including yield, which is of fundamental importance for understanding and manipulating these regions. Objectives: The aim of the study was to analyze the expression of candidate genes associated with maize yield. To better understand the function of the analyzed genes in increasing maize yield, their expression in different organs and tissues was also assessed using publicly available transcriptome data. Methods: RT-qPCR analyses were performed using iTaq Universal SYBR Green Supermix (Bio-Rad, Hercules, CA, USA) and CFX96 Touch Real-Time PCR Detection System (Bio-Rad, Hercules, CA, USA). Each of the performed RT-qPCR experiments consisted of three biological replicates and three technical replicates, the results of which were averaged. Results: The research results allowed us to select three out of six candidate genes (cinnamoyl-CoA reductase 1—CCR1, aspartate aminotransferase—AAT and sucrose transporter 1—SUT1), which can significantly affect grain yield in maize. Not only our studies but also literature reports clearly indicate the participation of CCR1, AAT and SUT1 in the formation of yield. Identified molecular markers located within these genes can be used in breeding programs to select high yielding maize genotypes.
Rapid Identification of Rhizobia Nodulating Soybean by a High-Resolution Melting Analysis
Soybean [Glycine max (L.) Merr.] is one of the most important and oldest crops. Due to its ability to form symbiotic interactions with nitrogen-fixing bacteria, it is a valuable source of nitrogen for agriculture and proteins for humans and livestock. In Europe, for instance, in Poland, the soybean cultivation area is still not large but is gradually increasing due to climate change. The lack of indigenous soybean microsymbionts in Polish soils forces the application of commercial strains to establish effective symbioses. Fast and reliable identification methods are needed to study the persistence, competitiveness, and dispersal of bradyrhizobia introduced as inocula. Our study aimed to apply real-time PCR coupled with high-resolution melting curve (HRM) analysis to detect and differentiate bacterial strains occupying soybean nodules. HRM-PCR was performed on crude extracts from nodules using primers specific for recA, a highly conserved nonsymbiotic gene. By comparing them with the reference strains, we were able to identify and assign Bradyrhiobium strains that had been introduced into field locations in Poland. In conclusion, HRM analysis was proven to be a fast and accurate method for identifying soybean microsymbionts and might be successfully used for identifying other legume-nodulating bacteria.
Effects of biopreparations based on Bacillus and Trichoderma, combined with mineral and organic fertilization and a Pisum sativum L. forecrop on improving the tolerance of Maize plants to drought stress
The increased frequency of extreme weather phenomena, such as heat waves and drought, adversely affects the condition of plants. The need to strive for more sustainable methods of growing plants requires undertaking researches that focus on strengthening the immunity of plants using methods that have a positive impact on both crops and the natural environment. The aim of the study was to assess the effectiveness and compare the effects of selected microbiological preparations based on Bacillus bacteria and Trichoderma symbiotic fungi, combined with mineral (NPK) and organic fertilization (manure) and a Pisum sativum L. forecrop on improving the tolerance of maize plants to drought stress. The pot experiment was carried in 2023 as a two-factor experiment in three replicates. Physiological parameters were assessed based on measurements of photosynthetic efficiency (A - CO2 assimilation rate, E - Transpiration Rate, Gs – Stomatal Conductance) and chlorophyll content (CCI) and fluorescence (F0 - initial fluorescence, Fm - maximum fluorescence, Fv/Fm - maximum photochemical efficiency of PSII, Yield - quantum yield of the photochemical reaction in PSII, ETR – electron transport rate, NPQ - Non - Photo-chemical Quenching), as well as soil respiration (NCER- Net CO2 Exchange Rate, W flux- Net H2O Exchange Rate, Ce- Soil Respiration) and biometric measurements (dry mass of shoots and roots).The measurement of photosynthesis efficiency under drought stress clearly indicated the highest, significant effect caused by Trichoderma preparation with both fertilizers. In the control, CO2 assimilation was practically inhibited due to drought (98% drop), while in the plants in which the Trichoderma preparation was used together with half dose of NPK and manure, there was only a slight decrease (1% and 13% respectively). A greatest, significant improvement in the DM of roots under drought was noted in plants in which the Pisum forecrop was applied together with NPK and manure (230% and 168% respectively). Pisum forecrop and treatments with microbiological preparation containing Trichoderma, make it possible to reduce the fertilization dose by at least half. This is particularly important in view of the global trend of increasing drought stress and efforts to improve soil quality.
Wykorzystanie systemu CRISPR-Cas9 w celu wyłączenia wybranych genów kodujących transportery MATE u Lotus japonicus
DArTseq-Based, High-Throughput Identification of Novel Molecular Markers for the Detection of Fusarium Resistance in Maize
Modern maize breeding worldwide relies on a broad range of molecular genetics research techniques. These technologies allow us to identify genomic regions associated with various phenotypic traits, including resistance to fungi of the genus Fusarium. Therefore, the aim of this publication was to identify new molecular markers linked to candidate genes that confer maize resistance to Fusarium fungi, using next-generation sequencing, association mapping, and physical mapping. In the study, a total of 5714 significant molecular markers related to maize plant resistance to Fusarium fungi were identified. Of these, 10 markers were selected that were significantly associated (with the highest LOD values) with the disease. These markers were identified on chromosomes 5, 6, 7, 8, and 9. The authors were particularly interested in two markers: SNP 4583014 and SilicoDArT 4579116. The SNP marker is located on chromosome 5, in exon 8 of the gene encoding alpha-mannosidase I MNS5. The SilicoDArT marker is located 240 bp from the gene for peroxisomal carrier protein on chromosome 8. Our own research and the presented literature review indicate that both these genes may be involved in biochemical reactions triggered by the stress caused by plant infection with Fusarium fungal spores. Molecular analyses indicated their role in resistance processes, as resistant varieties responded with an increase in the expression level of these genes at various time points after plant inoculation with Fusarium fungal spores. In the negative control, which was susceptible to Fusarium, no significant fluctuations in the expression levels of either gene were observed. Analyses concerning the identification of Fusarium fungi showed that the most abundant fungi on the infected maize kernels were Fusarium poae and Fusarium culmorum. Individual samples were very sparsely colonized by Fusarium or not at all. By using various molecular technologies, we identified genomic regions associated with maize resistance to Fusarium fungi, which is of fundamental importance for understanding these regions and potentially manipulating them.
MtABCG40 is a cytokinin transporter that limits lateral root density and nodule formation in Medicago truncatula
SUMMARY Numerous studies have already demonstrated that cytokinin (CK) distribution plays a pivotal role in shaping plant morphology in response to environmental changes. However, the contribution of short‐distance CK translocation to root mineral nutrition remains poorly understood, and the specific roles of CK transporters in root morphology are still unclear. Identifying the molecular identity of CK transporters is, therefore, essential for advancing our understanding of root plasticity, under varying soil fertility conditions and common nutritional deficiencies. In this study, we identified and characterised MtABCG40, a full‐size ATP‐binding cassette (ABC) transporter of the G subfamily in Medicago truncatula , as a CK transporter. MtABCG40 expression is root‐specific, and it is induced both by nitrogen deficiency and CK treatment. We propose that MtABCG40 mediates the lateral translocation of CKs from the xylem to surrounding cells, giving rise to lateral organs, and thereby influencing root morphology, by suppressing the lateral root and nodule formation under nitrogen‐limited conditions. Furthermore, MtABCG40 reduces apoplastic CK concentrations in the root meristematic zone, lowering the responsiveness of the root apical meristem to CKs. In summary, the activity of MtABCG40 indirectly influences CK perception and, consequently, modulates auxin‐mediated cellular responses.