Identification of Suitable Reference Genes for qRT-PCR Normalization in Tilia miqueliana Maxim

Wang, Huanli; Yan, Lingjun; Huang, Xi; Wang, Zhongwei; Yue, Yuanhao; Tang, Shijie

doi:10.32604/phyton.2022.020735

Cited by 2 publications

(1 citation statement)

References 42 publications

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“…Tilia is a unique tree species found in China, known for its strong adaptability and drought resistance [1]. It is majestic, making it suitable for planting as a street and courtyard tree, widely distributed in Jiangsu, Anhui, Zhejiang, and Jiangxi Provinces [2]. Tilia is a plant with both medicinal and edible properties.…”

Section: Introductionmentioning

confidence: 99%

The Impact of Magnetic Field and Gibberellin Treatment on the Release of Dormancy and Internal Nutrient Transformation in Tilia miqueliana Maxim. Seeds

Shi,

Cao,

Gao

et al. 2024

Forests

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The seeds of Tilia miqueliana Maxim. exhibit deep dormancy, which is categorized as combinational dormancy. This study utilized a comprehensive treatment involving magnetic fields, gibberellin (GA3), and cold stratification to promote the release of seed physiological dormancy and enhance germination rates. After being soaked in 98% H2SO4 for 15 min, mature seeds of Tilia were exposed to magnetic field treatments (150 MT, 250 MT) for different durations (25 min, 45 min, 65 min, and 85 min), as well as GA3 solution soaking (concentration: 0 μmol·L−1, 1443 μmol·L−1). Subsequently, cold stratification (0–5 °C) was applied to investigate the effects of these treatments on seed dormancy release and nutrient transformation. The results indicated that the comprehensive treatment involving magnetic field, GA3 solution soaking, and cold stratification effectively released the physiological dormancy of Tilia seeds and improved germination rates. Among the treatments, M150T85G1443 (magnetic field intensity: 150 MT, magnetic field treatment time: 85 min, GA3 soaking concentration: 1433 μmol·L−1) exhibited the most favorable outcome. After 75 days of cold stratification following the comprehensive treatments, the germination rate of M150T85G1443 seeds reached 89%. Additionally, the levels of storage substances such as starches and crude fats within the seeds decreased, while the utilization of soluble sugars and soluble proteins increased. The M150T85G1443 treatment exhibited the highest degree of variation, leading to gradual increases in metabolic activities of the seeds and a transition from dormancy to germination.

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Section: Introductionmentioning

confidence: 99%

The Impact of Magnetic Field and Gibberellin Treatment on the Release of Dormancy and Internal Nutrient Transformation in Tilia miqueliana Maxim. Seeds

Shi,

Cao,

Gao

et al. 2024

Forests

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Identification and validation of stable reference genes for RT-qPCR analyses of Kobresia littledalei seedlings

Sun,

Li,

et al. 2024

BMC Plant Biol

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Background Kobreisa littledalei, belonging to the Cyperaceae family is the first Kobresia species with a reference genome and the most dominant species in Qinghai-Tibet Plateau alpine meadows. It has several resistance genes which could be used to breed improved crop varieties. Reverse Transcription Quantitative Real-Time Polymerase Chain Reaction (RT-qPCR) is a popular and accurate gene expression analysis method. Its reliability depends on the expression levels of reference genes, which vary by species, tissues and environments. However, K.littledalei lacks a stable and normalized reference gene for RT-qPCR analysis. Results The stability of 13 potential reference genes was tested and the stable reference genes were selected for RT-qPCR normalization for the expression analysis in the different tissues of K. littledalei under two abiotic stresses (salt and drought) and two hormonal treatments (abscisic acid (ABA) and gibberellin (GA)). Five algorithms were used to assess the stability of putative reference genes. The results showed a variation amongst the methods, and the same reference genes showed tissue expression differences under the same conditions. The stability of combining two reference genes was better than a single one. The expression levels of ACTIN were stable in leaves and stems under normal conditions, in leaves under drought stress and in roots under ABA treatment. The expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression was stable in the roots under the control conditions and salt stress and in stems exposed to drought stress. Expression levels of superoxide dismutase (SOD) were stable in stems of ABA-treated plants and in the roots under drought stress. Moreover, RPL6 expression was stable in the leaves and stems under salt stress and in the stems of the GA-treated plants. EF1-alpha expression was stable in leaves under ABA and GA treatments. The expression levels of 28 S were stable in the roots under GA treatment. In general, ACTIN and GAPDH could be employed as housekeeping genes for K. littledalei under different treatments. Conclusion This study identified the best RT-qPCR reference genes for different K. littledalei tissues under five experimental conditions. ACTIN and GAPDH genes can be employed as the ideal housekeeping genes for expression analysis under different conditions. This is the first study to investigate the stable reference genes for normalized gene expression analysis of K. littledalei under different conditions. The results could aid molecular biology and gene function research on Kobresia and other related species.

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Identification of Suitable Reference Genes for qRT-PCR Normalization in Tilia miqueliana Maxim

Cited by 2 publications

References 42 publications

The Impact of Magnetic Field and Gibberellin Treatment on the Release of Dormancy and Internal Nutrient Transformation in Tilia miqueliana Maxim. Seeds

The Impact of Magnetic Field and Gibberellin Treatment on the Release of Dormancy and Internal Nutrient Transformation in Tilia miqueliana Maxim. Seeds

Identification and validation of stable reference genes for RT-qPCR analyses of Kobresia littledalei seedlings

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