Validation of reference genes for real-time quantitative PCR in Brachiaria grass under salt stress

Silva, Fláive Loyze Baldassarini; Santos, Tiago Benedito dos; Figueiredo, Mayara de Oliveira Vidotto; Cacefo, Viviane; Vieira, Luiz Gonzaga Esteves; Ribas, Alessandra Ferreira

doi:10.1016/j.plgene.2021.100319

Cited by 4 publications

(3 citation statements)

References 58 publications

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“…According to previous studies, genes belonging to the same family also exhibit diverse expression patterns across various developmental stages, tissues, or under different stress conditions. For example, in the root of Brachiaria grass (Brachiaria mutica) under salt stress, the ACT12 gene ranks second in terms of stability [6]. However, under salt stress in Suaeda glauca, ACT7 shows lower stability [55].…”

Section: Discussionmentioning

confidence: 99%

“…Samples were taken at 0, 0.5, 1. 5,3,6,12,24,48, and 72 h after stress treatment. After sampling, it is necessary to quickly freeze the samples in liquid nitrogen and store them at −80 • C in an ultra-low temperature environment for future use.…”

Section: Materials Culture and Stress Treatmentmentioning

confidence: 99%

“…This technique plays a crucial role in investigating plant gene expression patterns under various conditions. Traditionally, certain housekeeping genes like ACT, TUB, UBC, etc., are commonly employed as reference genes [5][6][7]. However, it is important to note that the expression levels of these genes may vary under various experimental settings.…”

Section: Introductionmentioning

confidence: 99%

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Selection of Suitable Reference Genes for RT-qPCR Gene Expression Analysis in Centipedegrass under Different Abiotic Stress

Wang,

Shu,

et al. 2023

Genes

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As a C4 warm-season turfgrass, centipedegrass (Eremochloa ophiuroides (Munro) Hack.) is known for its exceptional resilience to intensive maintenance practices. In this research, the most stably expressed reference genes in the leaves of centipedegrass under different stress treatments, including salt, cold, drought, aluminum (Al), and herbicide, were screened by the quantitative real-time PCR (RT-qPCR) technique. The stability of 13 candidate reference genes was evaluated by software GeNorm V3.4, NormFinder V20, BestKeeper V1.0, and ReFinder V1.0. The results of this experiment demonstrated that the expression of the UBC (ubiquitin-conjugating enzyme) remained the most stable under cold and Al stress conditions. On the other hand, the MD (malate dehydrogenase) gene exhibited the best performance in leaf tissues subjected to salt and drought stresses. Under herbicide stress, the expression level of the RIP (60S ribosomal protein L2) gene ranked the highest. The expression levels of abiotic stress-associated genes such as PIP1, PAL, COR413, ALMT9, and BAR were assessed to validate the reliability of the selected reference genes. This study provides valuable information and reference points for gene expression under abiotic stress conditions in centipedegrass.

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Section: Discussionmentioning

confidence: 99%

Section: Materials Culture and Stress Treatmentmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Selection of Suitable Reference Genes for RT-qPCR Gene Expression Analysis in Centipedegrass under Different Abiotic Stress

Wang,

Shu,

et al. 2023

Genes

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Relative Expression of a Salinity Stress‐Responsive Na⁺/H⁺ Exchanger (NHX) in Root and Leaf Tissues of the African Leafy Vegetable, Amaranthus dubius

Haripershad,

Nakhooda,

Shaik

2024

Food and Energy Security

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Amaranthus dubius, an African leafy vegetable (ALV), is an easy‐to‐grow, annual shrub and a highly nutritious food source, containing elevated levels of essential nutrients in the leaves. Many ALVs, including A. dubius, can tolerate salinity stress, enabling their cultivation on marginal land. However, the widespread propagation of A. dubius as a stable food source has thus far not been realised due partially to the high frequency at which hybridisation occurs, resulting in high genotypic and phenotypic variability. Therefore, to increase the agricultural output capacity of this species on salt‐affected marginal lands, it is important to screen, select and then clonally propagate the identified salinity‐tolerant genotypes to ensure true‐to‐type fidelity in the regenerated population. It is also important, thereafter, to elucidate their underlying gene expression of the stress response. The present study exposed 4‐week‐old A. dubius seedlings to 100, 200 and 400 mM NaCl to determine their degree of salt tolerance. Genotypes were then screened, selected and clonally propagated through cuttings, based on high growth rates and biomass, and salt tolerance. Generally, growth and physiological parameters decreased as substrate salinity increased. However, individual salt‐stressed genotypes demonstrated similar vigour to nonstressed plants and were able to maintain total protein and chlorophyll concentrations despite increasing salinity. The relative expression of an NHX1‐like transcript was quantified in 15 genotypes using degenerately primed real‐time qPCR. The relative expression of the putative NHX1 gene was 6.7 times greater in root tissues of seedlings treated with 400 mM NaCl (10.7 ± 1.8) compared to the roots of untreated seedlings (1.6 ± 1.3), and 2.8‐fold more than leaf tissues harvested from seedlings treated with 400 mM NaCl. Furthermore, the relative electrical conductivity (EC) of root tissues was 10 times greater than the EC of leaf tissues from the same 400 mM NaCl treatment. Numerous genotypes yielded similar chlorophyll content between 200 and 400 mM NaCl treatments, with genotypes salinity‐1 (S1) (3.5 ± 0.2 μg/cm2) and S34 (4.0 ± 0.4 μg/cm2) having the highest concentrations of chlorophyll in the 400 mM group, which was positively correlated with total protein content. Following micropropagation through direct organogenesis, selected clones maintained true‐to‐type traits such as similar chlorophyll, protein and NHX1‐like expression as their parent plants when exposed to 400 mM NaCl. This study revealed that some genotypes demonstrated salt stress tolerance capabilities rivalling established halophytes by regulating the constitutive or inducible expression of an NHX1‐like protein in roots and leaves. The correlation between protein content and NHX1‐like expression was nonlinear and nonproportional, demonstrating the complexity of this response and necessitating further exploration of specific protein families or functional groups conferring salinity tolerance in this species.

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Urea transporter DUR3 gene in grasses: In silico characterization and relative expression in Megathyrsus maximus under different nitrogen sources

Ferreira,

Bavaresco,

Vidotto Figueiredo

et al. 2024

Plant Gene

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Validation of reference genes for real-time quantitative PCR in Brachiaria grass under salt stress

Cited by 4 publications

References 58 publications

Selection of Suitable Reference Genes for RT-qPCR Gene Expression Analysis in Centipedegrass under Different Abiotic Stress

Selection of Suitable Reference Genes for RT-qPCR Gene Expression Analysis in Centipedegrass under Different Abiotic Stress

Relative Expression of a Salinity Stress‐Responsive Na⁺/H⁺ Exchanger (NHX) in Root and Leaf Tissues of the African Leafy Vegetable, Amaranthus dubius

Urea transporter DUR3 gene in grasses: In silico characterization and relative expression in Megathyrsus maximus under different nitrogen sources

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Validation of reference genes for real-time quantitative PCR in Brachiaria grass under salt stress

Cited by 4 publications

References 58 publications

Selection of Suitable Reference Genes for RT-qPCR Gene Expression Analysis in Centipedegrass under Different Abiotic Stress

Selection of Suitable Reference Genes for RT-qPCR Gene Expression Analysis in Centipedegrass under Different Abiotic Stress

Relative Expression of a Salinity Stress‐Responsive Na+/H+ Exchanger (NHX) in Root and Leaf Tissues of the African Leafy Vegetable, Amaranthus dubius

Urea transporter DUR3 gene in grasses: In silico characterization and relative expression in Megathyrsus maximus under different nitrogen sources

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Relative Expression of a Salinity Stress‐Responsive Na⁺/H⁺ Exchanger (NHX) in Root and Leaf Tissues of the African Leafy Vegetable, Amaranthus dubius