Identification of suitable reference genes for normalization of RT-qPCR data in eggplant fruit and shoot borer (Leucinodes orbonalis Guenée)

Kariyanna, B; Prabhuraj, A.; Asokan, R.; Gireesh‐Babu, P.; Jalali, S. K.; Venkatesan, T.; Gracy, R. Gandhi; Mohan, M.

doi:10.2478/s11756-019-00346-4

Cited by 19 publications

(13 citation statements)

References 36 publications

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“…Gene expression analysis using RT-qPCR was carried out using three independent biological replicates for each sample 32 . Extraction of total RNA was performed by using a TIANGEN kit (Beijing, China).…”

Section: Methodsmentioning

confidence: 99%

Identification of key gene networks controlling organic acid and sugar metabolism during watermelon fruit development by integrating metabolic phenotypes and gene expression profiles

et al. 2020

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The organoleptic qualities of watermelon fruit are defined by the sugar and organic acid contents, which undergo considerable variations during development and maturation. The molecular mechanisms underlying these variations remain unclear. In this study, we used transcriptome profiles to investigate the coexpression patterns of gene networks associated with sugar and organic acid metabolism. We identified 3 gene networks/modules containing 2443 genes highly correlated with sugars and organic acids. Within these modules, based on intramodular significance and Reverse Transcription Quantitative polymerase chain reaction (RT-qPCR), we identified 7 genes involved in the metabolism of sugars and organic acids. Among these genes, Cla97C01G000640, Cla97C05G087120 and Cla97C01G018840 (r2 = 0.83 with glucose content) were identified as sugar transporters (SWEET, EDR6 and STP) and Cla97C03G064990 (r2= 0.92 with sucrose content) was identified as a sucrose synthase from information available for other crops. Similarly, Cla97C07G128420, Cla97C03G068240 and Cla97C01G008870, having strong correlations with malic (r2 = 0.75) and citric acid (r2 = 0.85), were annotated as malate and citrate transporters (ALMT7, CS, and ICDH). The expression profiles of these 7 genes in diverse watermelon genotypes revealed consistent patterns of expression variation in various types of watermelon. These findings add significantly to our existing knowledge of sugar and organic acid metabolism in watermelon.

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Section: Methodsmentioning

confidence: 99%

Identification of key gene networks controlling organic acid and sugar metabolism during watermelon fruit development by integrating metabolic phenotypes and gene expression profiles

et al. 2020

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“…The entire data were analyzed using the 2-ΔΔCt method [61]. Three independent biological replicates were used for gene expression analysis [62]. Actin "cla016178" was used as a reference gene [58].…”

Section: Rtq-pcr Expression Analysis Of Genes Involved In Compatibilimentioning

confidence: 99%

Comparative Physiological and Biochemical Mechanisms in Di, Tri, and Tetraploid Watermelon (<em>Citrullus lanatus</em> L.) Grafted by Branches

Kaseb¹,

Umer²,

Gebremeskel³

et al. 2020

Preprint

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Polyploid seeds production is laborious, complicated, and costly work. Tetraploid and triploid plants produce a fewer number of seeds/fruit, and triploid embryos are fairly weak, covered with a more hardened seed coat as compared to diploid seeds. Here we investigated the interactive effect of new grafting technique of polyploid watermelon scion onto rootstock on plants' survival rate, biochemical, and hormones contents. In this study, three different branches, apical meristem (AM), branch with 1 node (1N), and 2 nodes (2N) from di, Tri, and tetraploid watermelon plants, were used as scion and grafted onto squash rootstock. The results showed highly significant differences between polypoid watermelon when 1N using as a scion, tetraploid showed maximum survival rates, higher contents of hormones, and antioxidants (AOX) activities, compared to diploid, these may be the possible reasons for high compatibility in tetraploid and degrading the grafting zone in diploid. RTq-PCR results confirm that the expression of genes linked to compatibility is consistent with the hormonal and AOX activities. This study provides an alternative and economical approach to produce more tetraploid and triploid plants for breeding or seeds production by using branches as scions.

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“…Relative expression levels for the CE and GST genes were calculated by 2 − CT method. The 28SR3 (28S ribosomal protein S3 mitochondrial) (Kariyanna et al, 2019) gene was used as an internal control to normalize the expression of target genes.…”

Section: Genome Mining and Phylogenetic Analysismentioning

confidence: 99%

Genome Mining and Expression Analysis of Carboxylesterase and Glutathione S-Transferase Genes Involved in Insecticide Resistance in Eggplant Shoot and Fruit Borer, Leucinodes orbonalis (Lepidoptera: Crambidae)

et al. 2020

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The shoot and fruit borer, Leucinodes orbonalis (Lepidoptera: Crambidae) is the major cause of low productivity in eggplant and insecticides being the mainstay of management of L. orbonalis. However, field control failures are widespread due to the evolution of insecticide resistance. Taking advantage of the whole genome sequence information, the present study investigated the level of insecticide resistance and the expression pattern of individual carboxylesterase (CE) and glutathione S-transferases (GSTs) genes in various field collected populations of L. orbonalis. Dose-mortality bioassays revealed a very high level of resistance development against fenvalerate (48.2–160-fold), phosalone (94-534.6-fold), emamectin benzoate (7.2–55-fold), thiodicarb (9.64–22.7-fold), flubendiamide (187.4–303.0-fold), and chlorantraniliprole (1.6–8.6-fold) in field populations as compared to laboratory-reared susceptible iso-female colony (Lo-S). Over-production of detoxification enzymes viz., CE and GST were evident upon enzyme assays. Mining of the draft genome of L. orbonalis yielded large number of genes potentially belonging to the CE and GST gene families with known history of insecticide resistance in other insects. Subsequent RT-qPCR studies on relative contribution of individual genes revealed over-expression of numerous GSTs and few CEs in field populations, indicating their possible involvement of metabolic enzymes in insecticide resistance. The genomic information will facilitate the development of novel resistance management strategies against this pest.

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Identification of suitable reference genes for normalization of RT-qPCR data in eggplant fruit and shoot borer (Leucinodes orbonalis Guenée)

Cited by 19 publications

References 36 publications

Identification of key gene networks controlling organic acid and sugar metabolism during watermelon fruit development by integrating metabolic phenotypes and gene expression profiles

Identification of key gene networks controlling organic acid and sugar metabolism during watermelon fruit development by integrating metabolic phenotypes and gene expression profiles

Comparative Physiological and Biochemical Mechanisms in Di, Tri, and Tetraploid Watermelon (<em>Citrullus lanatus</em> L.) Grafted by Branches

Genome Mining and Expression Analysis of Carboxylesterase and Glutathione S-Transferase Genes Involved in Insecticide Resistance in Eggplant Shoot and Fruit Borer, Leucinodes orbonalis (Lepidoptera: Crambidae)

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