Identification of suitable reference genes for quantitative reverse transcription PCR in Luffa (Luffa cylindrica)

Zhao, Gangjun; Wang, Meng; Gan, Yaqin; Gong, Hao; Li, Junxing; Zheng, Xianliang; Liu, Xiaoxi; Zhao, Siying; Luo, Jianxun; Wu, Haibin

doi:10.1007/s12298-022-01182-8

Cited by 4 publications

(4 citation statements)

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“…The immature luffa fruit shares similarities in taste and texture with those of the zucchini plant and is described to have a mild and slightly sweet taste, with a juicy and tender inside flesh (Rolnik & Olas, 2020). Like other plants found within this family, the immature fruit is also nutritionally dense, making it an optimal source of carbohydrates, vitamins, and minerals (Zhao et al., 2022). Luffas can be cultivated in various growing climates around the world, but they are considered a relatively important economic, cultural, and culinary staple in China, India, Southeast Asia, and Brazil.…”

Section: Introductionmentioning

confidence: 99%

An investigation into the sensory properties of luffa [Luffa cylindrica(L.)] sap

Gorman,

Baxter,

Frampton

et al. 2024

Journal of Food Science

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The seeds and sap of luffa [Luffa cylindrica (L.)] are usually discarded as waste. As such, this study aimed to identify the sensory properties of luffa sap (aqueous solution) and if it can be incorporated into a food item (orange juice) for added nutritional benefits and as an alternative sweetener. A sensory trial (n = 94) asked participants to evaluate a luffa sap sample and five different orange juice samples with increasing amounts of luffa sap (control [0%], 5%, 7.5%, 10%, 12.5%). The participants evaluated the samples using 9‐point hedonic scales, check‐all‐that‐apply, and an open‐ended comment question. The luffa sap was described as having a mild flavor as well as woody, earthy, and floral attributes and an aftertaste. The liking scores for the orange juice with the 5% luffa sap did not significantly differ from the control. However, as the amount of luffa sap increased above 5%, the liking scores decreased and were significantly different from the control. The orange juice with luffa sap samples (7.5% and above) was associated with off‐flavors, while the orange juice with 5% luffa sap and the control were associated with the attributes (sweet, fruity, orange, tropical, citrus) that increased the participants liking. Future studies should continue to investigate the sensory properties of luffa sap and its incorporation into different food products.Practical ApplicationThis is one of the first studies to investigate the sensory properties of luffa sap with participants residing in the Western world. The luffa sap was found to be woody, earthy, bitter, and floral. It was acceptable to add luffa sap to orange juice up to 5% by volume. However, it did not increase the sweetness perception of the orange juice. At a 7.5% luffa sap addition and higher levels, off‐flavors were observed in the orange juice.

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Section: Introductionmentioning

confidence: 99%

An investigation into the sensory properties of luffa [Luffa cylindrica(L.)] sap

Gorman,

Baxter,

Frampton

et al. 2024

Journal of Food Science

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“…[19,20]. However, many studies have proved that the transcription levels of the housekeeping genes may change with different species, tissues, and organs [21][22][23]. Therefore, appropriate internal reference genes should be selected according to specific experimental conditions to reduce experimental errors.…”

Section: Introductionmentioning

confidence: 99%

Screening and Validation of Appropriate Reference Genes for Real-Time Quantitative PCR under PEG, NaCl and ZnSO4 Treatments in Broussonetia papyrifera

Chen,

Wang,

Hao

et al. 2023

IJMS

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Real-time quantitative PCR (RT-qPCR) has a high sensitivity and strong specificity, and is widely used in the analysis of gene expression. Selecting appropriate internal reference genes is the key to accurately analyzing the expression changes of target genes by RT-qPCR. To find out the most suitable internal reference genes for studying the gene expression in Broussonetia papyrifera under abiotic stresses (including drought, salt, and ZnSO4 treatments), seven different tissues of B. papyrifera, as well as the roots, stems, and leaves of B. papyrifera under the abiotic stresses were used as test materials, and 15 candidate internal reference genes were screened based on the transcriptome data via RT-qPCR. Then, the expression stability of the candidate genes was comprehensively evaluated through the software geNorm (v3.5), NormFinder (v0.953), BestKeeper (v1.0), and RefFinder. The best internal reference genes and their combinations were screened out according to the analysis results. rRNA and Actin were the best reference genes under drought stress. Under salt stress, DOUB, HSP, NADH, and rRNA were the most stable reference genes. Under heavy metal stress, HSP and NADH were the most suitable reference genes. EIF3 and Actin were the most suitable internal reference genes in the different tissues of B. papyrifera. In addition, HSP, rRNA, NADH, and UBC were the most suitable internal reference genes for the abiotic stresses and the different tissues of B. papyrifera. The expression patterns of DREB and POD were analyzed by using the selected stable and unstable reference genes. This further verified the reliability of the screened internal reference genes. This study lays the foundation for the functional analysis and regulatory mechanism research of genes in B. papyrifera.

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“…The majority of genetic studies on bitter gourd have relied on traditional reference genes (TRGs), such as ACT7 (Actin-7) [21,22], GAPDH (Glyceraldehyde-3-phosphate dehydrogenase) [23], 18S rRNA (18S ribosomal RNA) [24], CYP (Peptidyl-prolyl cis-trans isomerase) [25], and EF1α (Elongation factor 1-alpha) [26]. However, accumulating evidence suggests that many of these traditional reference genes exhibit unstable expression across different plant species, tissues, experimental conditions, and developmental stages [27][28][29][30]. Furthermore, it remains unknown whether these traditional reference genes can accurately quantify the expression of fruit ripening-related genes in bitter gourd.…”

Section: Introductionmentioning

confidence: 99%

Identification and Analysis of Reference and Tissue-Specific Genes in Bitter Gourd Based on Transcriptome Data

Zheng,

Ma,

Luo

et al. 2023

Horticulturae

Self Cite

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Accurate and standardized quantification of reverse transcription PCR (qRT-PCR) results relies on the use of a dependable reference gene. The precise control of transgene expression in terms of both spatial and temporal aspects necessitates the utilization of tissue-specific gene promoters. However, the identification of stable reference genes across various tissues, particularly in fruits at different ripening stages, as well as tissue-specific genes in bitter gourds, remains largely unexplored. In this study, we employed RNA-Seq-based transcriptome datasets obtained from nine tissues to comprehensively screen for new reference genes (NRGs) and tissue-specific genes. Through the utilization of five algorithms in conjunction with qRT-PCR analysis, we successfully identified two highly stable reference genes, namely HMG1/2 and PHOS32, from a pool of 11 NRGs and five traditional reference genes (TRGs). To validate their reliability, we performed expression pattern analysis of two genes associated with fruit ripening (McACO1 and McACO2) using HMG1/2 and PHOS32, as well as an unstable reference gene, HSCP2. Furthermore, we conducted qRT-PCR validation of 12 tissue-specific genes using HMG1/2 as the reference gene. This study not only contributes to the precise normalization of target genes in bitter gourd but also provides a solid foundation for regulating transgenes through the utilization of suitable tissue-specific promoters.

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Identification of suitable reference genes for quantitative reverse transcription PCR in Luffa (Luffa cylindrica)

Cited by 4 publications

References 50 publications

An investigation into the sensory properties of luffa [Luffa cylindrica(L.)] sap

An investigation into the sensory properties of luffa [Luffa cylindrica(L.)] sap

Screening and Validation of Appropriate Reference Genes for Real-Time Quantitative PCR under PEG, NaCl and ZnSO4 Treatments in Broussonetia papyrifera

Identification and Analysis of Reference and Tissue-Specific Genes in Bitter Gourd Based on Transcriptome Data

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