Evaluation of suitable reference genes for normalization of quantitative real-time PCR analysis in rice plants under Xanthomonas oryzae pv. oryzae--infection and melatonin supplementation

Chen, Xian; Laborda, Pedro; Dong, Yan; Liu, Fengquan

doi:10.1186/s43014-020-00035-9

Cited by 9 publications

(3 citation statements)

References 32 publications

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“…Housekeeping genes such as Tubulin , Actin , and 18S rRNA are often used as reference genes. However, these reference genes show expression instability in various species, at different developmental stages, or in different tissues 8 , 9 , 14 , 18 . Therefore, the reference genes most appropriate for specific sample types and experimental conditions need to be selected.…”

Section: Discussionmentioning

confidence: 99%

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Identification of RT-qPCR reference genes suitable for gene function studies in the pitaya canker disease pathogen Neoscytalidium dimidiatum

Wang

Sang

et al. 2022

Sci Rep

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Neoscytalidium dimidiatum is the main causal agent of pitaya canker. Most studies of virulence and pathogenicity genes have measured expression levels using real-time quantitative polymerase chain reaction (RT-qPCR). Suitable reference genes are essential for ensuring that estimates of gene expression levels by RT-qPCR are accurate. However, no reference genes can be robustly applied across all contexts and species. No studies to date have evaluated the most effective reference genes for normalizing gene expression levels estimated by RT-qPCR in N. dimidiatum. In this study, RT-qPCR data for individual candidate reference genes were analyzed using four different methods: the delta Ct method and the geNorm, NormFinder, and BestKeeper algorithms. We evaluated the utility of eight candidate reference genes (18S rRNA, Actin (1), Actin (2), Actin, GAPDH (1), GAPDH (2), UBQ, and Tubulin) for normalizing expression levels estimated by RT-qPCR in N. dimidiatum at different developmental stages, at different temperatures, and during interaction with pitaya. All candidate reference genes were suitable for gene expression analysis except for Actin (2). Tubulin and Actin (1) were the most stably expressed reference genes under different temperatures. Actin (1) and Actin were the most stably expressed reference genes in N. dimidiatum at different developmental stages. Tubulin and UBQ were the most stably expressed reference genes during interaction with pitaya. Actin and 18s rRNA were the most stably expressed across all experimental conditions. Subsequently, Tubulin and UBQ were further investigated in analyses of pectinase expression during the pitaya–N. dimidiatum interaction. Our results provide insights that will aid future RT-qPCR studies of gene expression in N. dimidiatum.

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Section: Discussionmentioning

confidence: 99%

“…mali 12 , Magnaporthe oryzae 13 , Xanthomonas oryzae pv. oryzae 14 , and Ustilaginoidea virens 15 . However, no studies to date have systematically screened for stably expressed reference genes for RT-qPCR analysis in N. dimidiatum .…”

Section: Introductionmentioning

confidence: 99%

Identification of RT-qPCR reference genes suitable for gene function studies in the pitaya canker disease pathogen Neoscytalidium dimidiatum

Wang

Sang

et al. 2022

Sci Rep

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“…MEL triggers the activation of plant defense responses, including the expression of defense-related genes and the production of hormones and secondary metabolites. These responses strengthen the plant’s defense system against viral infections [ 6 , 126 , 127 ]. The exact mechanism through which MEL confers resistance against viral diseases in plants is still under investigation.…”

Section: Role Of Mel In the Mitigation Of Biotic Stressesmentioning

confidence: 99%

Melatonin: The Multifaceted Molecule in Plant Growth and Defense

Khan,

Hussain,

Yun

et al. 2024

IJMS

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Melatonin (MEL), a hormone primarily known for its role in regulating sleep and circadian rhythms in animals, has emerged as a multifaceted molecule in plants. Recent research has shed light on its diverse functions in plant growth and defense mechanisms. This review explores the intricate roles of MEL in plant growth and defense responses. MEL is involved in plant growth owing to its influence on hormone regulation. MEL promotes root elongation and lateral root formation and enhances photosynthesis, thereby promoting overall plant growth and productivity. Additionally, MEL is implicated in regulating the circadian rhythm of plants, affecting key physiological processes that influence plant growth patterns. MEL also exhibits antioxidant properties and scavenges reactive oxygen species, thereby mitigating oxidative stress. Furthermore, it activates defense pathways against various biotic stressors. MEL also enhances the production of secondary metabolites that contribute to plant resistance against environmental changes. MEL’s ability to modulate plant response to abiotic stresses has also been extensively studied. It regulates stomatal closure, conserves water, and enhances stress tolerance by activating stress-responsive genes and modulating signaling pathways. Moreover, MEL and nitric oxide cooperate in stress responses, antioxidant defense, and plant growth. Understanding the mechanisms underlying MEL’s actions in plants will provide new insights into the development of innovative strategies for enhancing crop productivity, improving stress tolerance, and combating plant diseases. Further research in this area will deepen our knowledge of MEL’s intricate functions and its potential applications in sustainable agriculture.

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Revisiting rice transformation for a fail-safe protocol and its application for various gene functional and molecular studies

Manna,

Rengasamy,

Reddy

et al. 2024

J Plant Growth Regul

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Evaluation of suitable reference genes for normalization of quantitative real-time PCR analysis in rice plants under Xanthomonas oryzae pv. oryzae--infection and melatonin supplementation

Cited by 9 publications

References 32 publications

Identification of RT-qPCR reference genes suitable for gene function studies in the pitaya canker disease pathogen Neoscytalidium dimidiatum

Identification of RT-qPCR reference genes suitable for gene function studies in the pitaya canker disease pathogen Neoscytalidium dimidiatum

Melatonin: The Multifaceted Molecule in Plant Growth and Defense

Revisiting rice transformation for a fail-safe protocol and its application for various gene functional and molecular studies

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