Evaluation of putative reference genes for gene expression normalization in soybean by quantitative real-time RT-PCR

<p>Abstract</p> <p>Background</p> <p>Real-time quantitative reverse transcription PCR (RT-qPCR) data needs to be normalized for its proper interpretation. Housekeeping genes are routinely employed for this purpose, but their expression level cannot be assumed to remain constant under all possible experimental conditions. Thus, a systematic validation of reference genes is required to ensure proper normalization. For soybean, only a small number of validated reference genes are available to date.</p> <p>Results</p> <p>A systematic comparison of 14 potential reference genes for soybean is presented. These included seven commonly used (<it>ACT2, ACT11, TUB4, TUA5, CYP, UBQ10, EF1b</it>) and seven new candidates (<it>SKIP16, MTP, PEPKR1, HDC, TIP41, UKN1, UKN2</it>). Expression stability was examined by RT-qPCR across 116 biological samples, representing tissues at various developmental stages, varied photoperiodic treatments, and a range of soybean cultivars. Expression of all 14 genes was variable to some extent, but that of <it>SKIP16, UKN1 </it>and <it>UKN2 </it>was overall the most stable. A combination of <it>ACT11, UKN1 </it>and <it>UKN2 </it>would be appropriate as a reference panel for normalizing gene expression data among different tissues, whereas the combination SKIP16, UKN1 and MTP was most suitable for developmental stages. <it>ACT11, TUA5 </it>and <it>TIP41 </it>were the most stably expressed when the photoperiod was altered, and <it>TIP41, UKN1 </it>and <it>UKN2 </it>when the light quality was changed. For six different cultivars in long day (LD) and short day (SD), their expression stability did not vary significantly with <it>ACT11, UKN2 </it>and <it>TUB4 </it>being the most stable genes. The relative gene expression level of <it>GmFTL3</it>, an ortholog of Arabidopsis <it>FT </it>(<it>FLOWERING LOCUS T</it>) was detected to validate the reference genes selected in this study.</p> <p>Conclusion</p> <p>None of the candidate reference genes was uniformly expressed across all experimental conditions, and the most suitable reference genes are conditional-, tissue-specific-, developmental-, and cultivar-dependent. Most of the new reference genes performed better than the conventional housekeeping genes. These results should guide the selection of reference genes for gene expression studies in soybean.</p

Comprehensive Analysis of NAC Domain Transcription Factor Gene Family in Populus trichocarpa

<p>Abstract</p> <p>Background</p> <p>NAC (<b>NAM, ATAF1/2 </b>and <b>CUC2</b>) domain proteins are plant-specific transcriptional factors known to play diverse roles in various plant developmental processes. NAC transcription factors comprise of a large gene family represented by more than 100 members in <it>Arabidopsis</it>, rice and soybean etc. Recently, a preliminary phylogenetic analysis was reported for NAC gene family from 11 plant species. However, no comprehensive study incorporating phylogeny, chromosomal location, gene structure, conserved motifs, and expression profiling analysis has been presented thus far for the model tree species <it>Populus</it>.</p> <p>Results</p> <p>In the present study, a comprehensive analysis of NAC gene family in <it>Populus </it>was performed. A total of 163 full-length NAC genes were identified in <it>Populus</it>, and they were phylogeneticly clustered into 18 distinct subfamilies. The gene structure and motif compositions were considerably conserved among the subfamilies. The distributions of 120 <it>Populus </it>NAC genes were non-random across the 19 linkage groups (LGs), and 87 genes (73%) were preferentially retained duplicates that located in both duplicated regions. The majority of NACs showed specific temporal and spatial expression patterns based on EST frequency and microarray data analyses. However, the expression patterns of a majority of duplicate genes were partially redundant, suggesting the occurrence of subfunctionalization during subsequent evolutionary process. Furthermore, quantitative real-time RT-PCR (RT-qPCR) was performed to confirm the tissue-specific expression patterns of 25 NAC genes.</p> <p>Conclusion</p> <p>Based on the genomic organizations, we can conclude that segmental duplications contribute significantly to the expansion of <it>Populus </it>NAC gene family. The comprehensive expression profiles analysis provides first insights into the functional divergence among members in NAC gene family. In addition, the high divergence rate of expression patterns after segmental duplications indicates that NAC genes in <it>Populus </it>are likewise to have been retained by substantial subfunctionalization. Taken together, our results presented here would be helpful in laying the foundation for functional characterization of NAC gene family and further gaining an understanding of the structure-function relationship between these family members.</p

Genome-Wide Identification, Evolutionary Expansion, and Expression Profile of Homeodomain-Leucine Zipper Gene Family in Poplar (Populus trichocarpa)

BACKGROUND: Homeodomain-leucine zipper (HD-ZIP) proteins are plant-specific transcriptional factors known to play crucial roles in plant development. Although sequence phylogeny analysis of Populus HD-ZIPs was carried out in a previous study, no systematic analysis incorporating genome organization, gene structure, and expression compendium has been conducted in model tree species Populus thus far. PRINCIPAL FINDINGS: In this study, a comprehensive analysis of Populus HD-ZIP gene family was performed. Sixty-three full-length HD-ZIP genes were found in Populus genome. These Populus HD-ZIP genes were phylogenetically clustered into four distinct subfamilies (HD-ZIP I-IV) and predominately distributed across 17 linkage groups (LG). Fifty genes from 25 Populus paralogous pairs were located in the duplicated blocks of Populus genome and then preferentially retained during the sequential evolutionary courses. Genomic organization analyses indicated that purifying selection has played a pivotal role in the retention and maintenance of Populus HD-ZIP gene family. Microarray analysis has shown that 21 Populus paralogous pairs have been differentially expressed across different tissues and under various stresses, with five paralogous pairs showing nearly identical expression patterns, 13 paralogous pairs being partially redundant and three paralogous pairs diversifying significantly. Quantitative real-time RT-PCR (qRT-PCR) analysis performed on 16 selected Populus HD-ZIP genes in different tissues and under both drought and salinity stresses confirms their tissue-specific and stress-inducible expression patterns. CONCLUSIONS: Genomic organizations indicated that segmental duplications contributed significantly to the expansion of Populus HD-ZIP gene family. Exon/intron organization and conserved motif composition of Populus HD-ZIPs are highly conservative in the same subfamily, suggesting the members in the same subfamilies may also have conservative functionalities. Microarray and qRT-PCR analyses showed that 89% (56 out of 63) of Populus HD-ZIPs were duplicate genes that might have been retained by substantial subfunctionalization. Taken together, these observations may lay the foundation for future functional analysis of Populus HD-ZIP genes to unravel their biological roles

GPT-4 Vision on Medical Image Classification -- A Case Study on COVID-19 Dataset

This technical report delves into the application of GPT-4 Vision (GPT-4V) in the nuanced realm of COVID-19 image classification, leveraging the transformative potential of in-context learning to enhance diagnostic processes

Functional conservation and divergence of Miscanthus lutarioriparius GT43 gene family in xylan biosynthesis

Background: Xylan is the most abundant un-cellulosic polysaccharides of plant cell walls. Much progress in xylan biosynthesis has been gained in the model plant species Arabidopsis. Two homologous pairs Irregular Xylem 9 (IRX9)/9L and IRX14/14L from glycosyltransferase (GT) family 43 have been proved to play crucial roles in xylan backbone biosynthesis. However, xylan biosynthesis in grass such as Miscanthus remains poorly understood

Synthetic Datasets for Autonomous Driving: A Survey

Autonomous driving techniques have been flourishing in recent years while thirsting for huge amounts of high-quality data. However, it is difficult for real-world datasets to keep up with the pace of changing requirements due to their expensive and time-consuming experimental and labeling costs. Therefore, more and more researchers are turning to synthetic datasets to easily generate rich and changeable data as an effective complement to the real world and to improve the performance of algorithms. In this paper, we summarize the evolution of synthetic dataset generation methods and review the work to date in synthetic datasets related to single and multi-task categories for to autonomous driving study. We also discuss the role that synthetic dataset plays the evaluation, gap test, and positive effect in autonomous driving related algorithm testing, especially on trustworthiness and safety aspects. Finally, we discuss general trends and possible development directions. To the best of our knowledge, this is the first survey focusing on the application of synthetic datasets in autonomous driving. This survey also raises awareness of the problems of real-world deployment of autonomous driving technology and provides researchers with a possible solution.Comment: 19 pages, 5 figure

Genome-Wide Analysis of Sorghum GT47 Family Reveals Functional Divergences of MUR3-Like Genes

Sorghum (Sorghum bicolor) is an important bioenergy crop. Its biomass mainly consists of the cellulosic and non-cellulosic polysaccharides, both which can be converted to biofuels. The biosynthesis of non-cellulosic polysaccharides involves several glycosyltransferases (GT) families including GT47. However, there was no systemic study on GT47 family in sorghum to date. Here, we identified 39 sorghum GT47 family members and showed the functional divergences of MURUS3 (MUR3) homologs. Sorghum GT47 proteins were phylogenetically clustered into four distinct subfamilies. Within each subfamily, gene structure was relatively conserved between the members. Ten gene pairs were identified from the 39 GT47 genes, of which two pairs might be originated from tandem duplication. 25.6% (10/39) of sorghum GT47 genes were homologous to Arabidopsis MUR3, a xyloglucan biosynthesis gene in primary cell walls. SbGT47_2, SbGT47_7, and SbGT47_8, three most homologous genes of MUR3, exhibited different tissue expression patterns and were selected for complementation into Arabidopsis mur3-3. Physiological and cell wall analyses showed that SbGT47_2 and SbGT47_7 may be two functional xyloglucan galactosyltransferases in sorghum. Further studies found that MUR3-like genes are widely present in the seed plants but not in the chlorophytic alga Chlamydomonas reinhardtii. Our results provide novel information for evolutionary analysis and functional dissection of sorghum GT47 family members

Derun dil

Safveti Ziya'nın Malumat'ta tefrika edilen Derun Dil adlı roman