Identification and validation of reference genes for qRT-PCR based studies in horse gram (Macrotyloma uniflorum)

2021 ◽  
Author(s):  
Ragini Sinha ◽  
Meenu Bala ◽  
Pragya Prabha ◽  
Alok Ranjan ◽  
Rakesh K. Chahota ◽  
...  
Keyword(s):  
Reference Genes ◽  
Horse Gram ◽  
Qrt Pcr ◽  
Macrotyloma Uniflorum

Preparation and Food Product Development From Macrotyloma Uniflorum (Horse Gram) Bovine Milk Blends

2011 ◽  
Vol 4 (5) ◽  
pp. 275-277
Author(s):  
Preeti Verma ◽  
◽  
Sheel Sharma ◽  
Vibha Sharma ◽  
Shilpi Singh ◽  
...  
Keyword(s):  
Product Development ◽  
Bovine Milk ◽  
Food Product ◽  
Horse Gram ◽  
Macrotyloma Uniflorum ◽  
Food Product Development

Public-transcriptome-database-assisted selection and validation of reliable reference genes for qRT-PCR in rice

2019 ◽  
Vol 63 (1) ◽  
pp. 92-101 ◽  
Author(s):  
Zhe Zhao ◽  
Zixu Zhang ◽  
Zhi Ding ◽  
Hengjun Meng ◽  
Rongxin Shen ◽  
...  
Keyword(s):  
Reference Genes ◽  
Qrt Pcr ◽  
Transcriptome Database

scholarly journals Choice of a Stable Set of Reference Genes for qRT-PCR Analysis in Amblyomma maculatum (Acari: Ixodidae)

2012 ◽  
Vol 49 (6) ◽  
pp. 1339-1346 ◽  
Author(s):  
Rebecca Browning ◽  
Steven Adamson ◽  
Shahid Karim
Keyword(s):  
Reference Genes ◽  
Stable Set ◽  
Pcr Analysis ◽  
Amblyomma Maculatum ◽  
Qrt Pcr

scholarly journals Identification of the most suitable reference gene for gene expression studies with development and abiotic stress response in Bromus sterilis

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Madhab Kumar Sen ◽  
Kateřina Hamouzová ◽  
Pavlina Košnarová ◽  
Amit Roy ◽  
Josef Soukup
Keyword(s):  
Reference Gene ◽  
Herbicide Resistance ◽  
Reference Genes ◽  
Gene Selection ◽  
High Yield ◽  
Suitable Reference Gene ◽  
Grass Species ◽  
Experimental Conditions ◽  
Qrt Pcr ◽  
Suitable Reference

AbstractBromus sterilis is an annual weedy grass, causing high yield losses in winter cereals. Frequent use of herbicides had led to the evolution of herbicide resistance in this species. Mechanisms underlying herbicide resistance in B. sterilis must be uncovered because this problem is becoming a global threat. qRT-PCR and the next-generation sequencing technologies can elucidate the resistance mechanisms. Although qRT-PCR can calculate precise fold changes, its preciseness depends on the expression of reference genes. Regardless of stable expression in any given condition, no gene can act as a universal reference gene. Hence, it is necessary to identify the suitable reference gene for each species. To our knowledge, there are no reports on the suitable reference gene in any brome species so far. Thus, in this paper, the stability of eight genes was evaluated using qRT-PCR experiments followed by expression stability ranking via five most commonly used software for reference gene selection. Our findings suggest using a combination of 18S rRNA and ACCase to normalise the qRT-PCR data in B. sterilis. Besides, reference genes are also recommended for different experimental conditions. The present study outcomes will facilitate future molecular work in B. sterilis and other related grass species.

Genetics of growth habit in horse gram (Macrotyloma uniflorum (Lam.)Verdc.)

2021 ◽  
Author(s):  
Basalapura Rangegowda Chandana ◽  
Sampangi Ramesh ◽  
Gonal Basanagouda ◽  
Rotti Kirankumar ◽  
Kyasampalli Venkatesh Reddy Ashwini
Keyword(s):  
Goodness Of Fit ◽  
Growth Habit ◽  
Production Systems ◽  
Grain Legumes ◽  
Selection Strategy ◽  
Horse Gram ◽  
Macrotyloma Uniflorum ◽  
Determinate Growth ◽  
Determinate Growth Habit ◽  
Growth Habits

Abstract Growth habit is a plant architectural trait in grain legumes with no exception of horse gram. Determinacy and indeterminacy are the two types of growth habits reported in horse gram. Relative advantages of the two types of growth habit depend on the production systems to which cultivars are targeted. Dependable information on genetics of growth habit provide clues for adopting the most appropriate selection strategy to breed high yielding horse gram varieties with desired growth habit. Taking cues from the past studies, we hypothesize that growth habit in horse gram is controlled by two genes displaying inhibitory epistasis and indeterminacy is dominant over determinacy. To test this hypothesis, we monitored the inheritance of growth habit in F1, F2 and F3 generations derived from two crosses involving parents differing for growth habit. Contrary to our hypothesis, determinate growth habit of F1s of both the crosses suggested dominance of determinacy over indeterminacy. A good fit of observed segregation of F2 plants to that of the hypothesized segregation in the ratio of 13 determinate: 3 indeterminate plants, besides confirming dominance of determinacy, suggested classical digenic inhibitory epistatic control of growth habit. These results were further confirmed in F3 generation based on goodness of fit between observed numbers of plants segregating for determinacy and indeterminacy and those expected in the ratio of 49 determinate: 15 indeterminate plants. To the best of our knowledge, this is the first report on the inheritance of growth habit in horse gram.

scholarly journals A probe-based qRT-PCR method to profile immunological gene expression in blood of captive beluga whales (Delphinapterus leucas)

PeerJ ◽  
2017 ◽  
Vol 5 ◽  
pp. e3840 ◽  
Author(s):  
Ming-An Tsai ◽  
I-Hua Chen ◽  
Jiann-Hsiung Wang ◽  
Shih-Jen Chou ◽  
Tsung-Hsien Li ◽  
...  
Keyword(s):  
Gene Expression ◽  
Immune System ◽  
Reference Genes ◽  
Animal Health ◽  
Rna Extraction ◽  
Disease Onset ◽  
Delphinapterus Leucas ◽  
Amplification Efficiency ◽  
Beluga Whales ◽  
Qrt Pcr

Cytokines are fundamental for a functioning immune system, and thus potentially serve as important indicators of animal health. Quantitation of mRNA using quantitative reverse transcription polymerase chain reaction (qRT-PCR) is an established immunological technique. It is particularly suitable for detecting the expression of proteins against which monoclonal antibodies are not available. In this study, we developed a probe-based quantitative gene expression assay for immunological assessment of captive beluga whales (Delphinapterus leucas) that is one of the most common cetacean species on display in aquariums worldwide. Six immunologically relevant genes (IL-2Rα, -4, -10, -12, TNFα, and IFNγ) were selected for analysis, and two validated housekeeping genes (PGK1 and RPL4) with stable expression were used as reference genes. Sixteen blood samples were obtained from four animals with different health conditions and stored in RNAlater™ solution. These samples were used for RNA extraction followed by qRT-PCR analysis. Analysis of gene transcripts was performed by relative quantitation using the comparative Cq method with the integration of amplification efficiency and two reference genes. The expression levels of each gene in the samples from clinically healthy animals were normally distributed. Transcript outliers for IL-2Rα, IL-4, IL-12, TNFα, and IFNγ were noticed in four samples collected from two clinically unhealthy animals. This assay has the potential to identify immune system deviation from normal state, which is caused by health problems. Furthermore, knowing the immune status of captive cetaceans could help both trainers and veterinarians in implementing preventive approaches prior to disease onset.

scholarly journals Identification and validation of miRNA reference genes in poplar under pathogen stress

2021 ◽  
Author(s):  
Lichun Zhang ◽  
Xiaoqian Yang ◽  
Yiyi Yin ◽  
Jinxing Wang ◽  
Yanwei Wang
Keyword(s):  
Gene Expression ◽  
Reference Gene ◽  
Small Rna ◽  
Reference Genes ◽  
Expression Profiles ◽  
Small Rna Sequencing ◽  
Rna Seq ◽  
Internal Standards ◽  
Qrt Pcr ◽  
Canker Pathogen

Abstract Quantitative real time polymerase chain reaction (qRT-PCR) is a common method to analyze gene expression. Due to differences in RNA quantity, quality, and reverse transcription efficiency between qRT-PCR samples, reference genes are used as internal standards to normalize gene expression. However, few universal genes especially miRNAs have been identified as reference so far. Therefore, it is essential to identify reference genes that can be used across various experimental conditions, stress treatments, or tissues. In this study, 14 microRNAs (miRNAs) and 5.8S rRNA were assessed for expression stability in poplar trees infected with canker pathogen. Using three reference gene analysis programs, we found that miR156g and miR156a exhibited stable expression throughout the infection process. miR156g and miR156a were then tested as internal standards to measure the expression of miR1447 and miR171c, and the results were compared to small RNA sequencing (RNA-seq) data. We found that when miR156a was used as the reference gene, the expression of miR1447 and miR171c were consistent with the small RNA-seq expression profiles. Therefore, miR156a was the most stable miRNAs examined in this study, and could be used as a reference gene in poplar under canker pathogen stress, which should enable comprehensive comparisons of miRNAs expression and avoid the bias caused by different lenth between detected miRNAs and traditional referece genes. The present study has expanded the miRNA reference genes available for gene expression studies in trees under biotic stress.

Validation of reference genes for qRT-PCR analysis in peel and flesh of six apple cultivars (Malus domestica) at diverse stages of fruit development

2019 ◽  
Vol 244 ◽  
pp. 165-171 ◽  
Author(s):  
Lifang Zhu ◽  
Chengquan Yang ◽  
Yaohua You ◽  
Wei Liang ◽  
Nannan Wang ◽  
...  
Keyword(s):  
Malus Domestica ◽  
Fruit Development ◽  
Reference Genes ◽  
Pcr Analysis ◽  
Qrt Pcr ◽  
Apple Cultivars
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