Selection of Reliable Reference Genes for Gene Expression Normalization in Sagittaria trifolia

Tang, Jing; Li, E; Liu, Jiexia; Zhang, Zhiping; Hua, Bing; Jiang, Jinzhi; Miao, Minmin

doi:10.3390/genes14071321

Cited by 3 publications

(2 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…EXP1 (encoding one expansion) is the major factor regulating cell wall extension and plays an important role in plant organ expansion ( Wu et al., 2001 ). During the development of leaf and stem in water dropwort, the relative expression of EXP1 showed an increasing trend with the continuous development and expansion of leaf and stem in water dropwort ( Tang et al., 2023 ). To verify the stability of the two selected reference genes TBP and UBQ , the expression patterns of EXP1 at different developmental stages were analyzed.…”

Section: Resultsmentioning

confidence: 99%

Selection of suitable reference genes for qPCR normalization in different developmental stages of Oenanthe javanica

Feng,

Yang,

Yan

et al. 2023

Front. Plant Sci.

View full text Add to dashboard Cite

Gene expression analysis is widely used to unravel molecular regulatory mechanisms and identify key genes in plants. Appropriate reference gene is an important prerequisite to ensure the accuracy and reliability of qPCR analysis results. Water dropwort is a plant of the Oenanthe genus in the Apiaceae family, which has high economic benefits. However, the underlying molecular regulatory mechanisms in the growth and development of water dropwort have not been fully understood and the appropriate reference genes in different developmental stages of water dropwort not yet reported. In this study, 10 candidate reference genes (ACTIN, PP2A, SAND, EF-1α, GAPDH, UBQ, MIP, TBP, RPS-18, eIF-4α) were identified and cloned from Oenanthe javanica. The qPCR primers of candidate reference genes were designed and verified. Four statistical algorithms, geNorm, NormFinder, BestKeeper and RefFinder were used to evaluate the expression stability of 10 candidate reference genes in different developmental stages of water dropwort. The results showed that TBP and UBQ were the most stable genes in different developmental stages of water dropwort, while GAPDH was the most unstable gene. The normalization of EXP1 genes at different developmental stages further confirmed the reliability of internal reference genes. The results of this study provide a theoretical basis for selecting appropriate internal reference genes in different developmental stages of water dropwort. This study also provides technical support and reliable basis for the expression analysis of key genes in different developmental stages of water dropwort.

show abstract

Section: Resultsmentioning

confidence: 99%

Selection of suitable reference genes for qPCR normalization in different developmental stages of Oenanthe javanica

Feng,

Yang,

Yan

et al. 2023

Front. Plant Sci.

View full text Add to dashboard Cite

show abstract

“…The relative expression of each housekeeping gene was calculated using the formula Q = E −ΔΔCt , where E represents the gene amplification efficiency, typically assumed to be two (100.00% efficiency). ΔCt is calculated as Ct (min)—Ct (sample), where Ct (min) is the lowest Ct value among all samples, and Ct (sample) is the Ct value of each sample [ 54 , 55 ]. The data were subjected to two-sample anova and plotted using Origin.…”

Section: Methodsmentioning

confidence: 99%

Optimization of preparation conditions for Salsola laricifolia protoplasts using response surface methodology and artificial neural network modeling

Guo,

Xi,

Guzailinuer

et al. 2024

Plant Methods

View full text Add to dashboard Cite

Background Salsola laricifolia is a typical C3–C4 typical desert plant, belonging to the family Amaranthaceae. An efficient single-cell system is crucial to study the gene function of this plant. In this study, we optimized the experimental conditions by using Box-Behnken experimental design and Response Surface Methodology (RSM)-Artificial Neural Network (ANN) model based on the previous studies. Results Among the 17 experiment groups designed by Box-Behnken experimental design, the maximum yield (1.566 × 106/100 mg) and the maximum number of viable cells (1.367 × 106/100 mg) were obtained in group 12, and the maximum viability (90.81%) was obtained in group 5. Based on these results, both the RSM and ANN models were employed for evaluating the impact of experimental factors. By RSM model, cellulase R-10 content was the most influential factor on protoplast yield, followed by macerozyme R-10 content and mannitol concentration. For protoplast viability, the macerozyme R-10 content had the highest influence, followed by cellulase R-10 content and mannitol concentration. The RSM model performed better than the ANN model in predicting yield and viability. However, the ANN model showed significant improvement in predicting the number of viable cells. After comprehensive evaluation of the protoplast yield, the viability and number of viable cells, the optimal results was predicted by ANN yield model and tested. The amount of protoplast yield was 1.550 × 106/100 mg, with viability of 90.65% and the number of viable cells of 1.405 × 106/100 mg. The corresponding conditions were 1.98% cellulase R-10, 1.00% macerozyme R-10, and 0.50 mol L−1 mannitol. Using the obtained protoplasts, the reference genes (18SrRNA, β-actin and EF1-α) were screened for expression, and transformed with PEG-mediated pBI121-SaNADP-ME2-GFP plasmid vector. There was no significant difference in the expression of β-actin and EF1-α before and after treatment, suggesting that they can be used as internal reference genes in protoplast experiments. And SaNADP-ME2 localized in chloroplasts. Conclusion The current study validated and evaluated the effectiveness and results of RSM and ANN in optimizing the conditions for protoplast preparation using S. laricifolia as materials. These two methods can be used independently of experimental materials, making them suitable for isolating protoplasts from other plant materials. The selection of the number of viable cells as an evaluation index for protoplast experiments is based on its ability to consider both protoplast yield and viability. The findings of this study provide an efficient single-cell system for future genetic experiments in S. laricifolia and can serve as a reference method for preparing protoplasts from other materials.

show abstract

Selection of a suitable reference gene for gene-expression studies in Trichomonas gallinae under various biotic and abiotic stress conditions

Cai,

Zhu,

Liu

et al. 2024

Gene

View full text Add to dashboard Cite

Selection of Reliable Reference Genes for Gene Expression Normalization in Sagittaria trifolia

Cited by 3 publications

References 42 publications

Selection of suitable reference genes for qPCR normalization in different developmental stages of Oenanthe javanica

Selection of suitable reference genes for qPCR normalization in different developmental stages of Oenanthe javanica

Optimization of preparation conditions for Salsola laricifolia protoplasts using response surface methodology and artificial neural network modeling

Selection of a suitable reference gene for gene-expression studies in Trichomonas gallinae under various biotic and abiotic stress conditions

Contact Info

Product

Resources

About