RNA‐Seq identification of candidate defense genes by analyzing <i>Mythimna separata</i> feeding‐damage induced systemic resistance in balsas teosinte

Yang, Liyan; Gao, Jing; Zhang, Yurong; Tian, Jingyun; Sun, Yi; Wang, Chuangyun

doi:10.1002/ps.5519

Cited by 9 publications

(8 citation statements)

References 33 publications

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“…Additionally, AMP and JAC1 , two maize genes that are up‐regulated in response to S. frugiperda feeding (Figure S2 ), reduced caterpillar weight gain when overexpressed in maize. Although transcriptomic studies have identified numerous maize genes that are up‐regulated in response to arthropod feeding (Bui et al ., 2018 ; Guo et al ., 2019 ; Pan et al ., 2020 ; Song et al ., 2017 ; Tzin et al ., 2015 , 2017 ; Wang et al ., 2017 ; Yang et al ., 2020 ; Zhang et al ., 2016 ), the majority of these genes have not been investigated for their roles in plant defence against herbivory. This is at least in part due to the time and cost of creating maize lines that have individual genes overexpressed.…”

Section: Discussionmentioning

confidence: 99%

A sugarcane mosaic virus vector for rapid in planta screening of proteins that inhibit the growth of insect herbivores

Chung

Bigham

Lappe

et al. 2021

Plant Biotechnology Journal

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Summary Spodoptera frugiperda (fall armyworm) is a notorious pest that threatens maize production worldwide. Current control measures involve the use of chemical insecticides and transgenic maize expressing Bacillus thuringiensis (Bt) toxins. Although additional transgenes have confirmed insecticidal activity, limited research has been conducted in maize, at least partially due to the technical difficulty of maize transformation. Here, we describe implementation of a sugarcane mosaic virus (SCMV) vector for rapidly testing the efficacy of both endogenous maize genes and heterologous genes from other organisms for the control of S. frugiperda in maize. Four categories of proteins were tested using the SCMV vector: (i) maize defence signalling proteins: peptide elicitors (Pep1 and Pep3) and jasmonate acid conjugating enzymes (JAR1a and JAR1b); (ii) maize defensive proteins: the previously identified ribosome‐inactivating protein (RIP2) and maize proteinase inhibitor (MPI), and two proteins with predicted but unconfirmed anti‐insect activities, an antimicrobial peptide (AMP) and a lectin (JAC1); (iii) lectins from other plant species: Allium cepa agglutinin (ACA) and Galanthus nivalis agglutinin (GNA); and (iv) scorpion and spider toxins: peptides from Urodacus yaschenkoi (UyCT3 and UyCT5) and Hadronyche versuta (Hvt). In most cases, S. frugiperda larval growth was reduced by transient SCMV‐mediated overexpression of genes encoding these proteins. Additionally, experiments with a subset of the SCMV‐expressed genes showed effectiveness against two aphid species, Rhopalosiphum maidis (corn leaf aphid) and Myzus persicae (green peach aphid). Together, these results demonstrate that SCMV vectors are a rapid screening method for testing the efficacy and insecticidal activity of candidate genes in maize.

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Section: Discussionmentioning

confidence: 99%

A sugarcane mosaic virus vector for rapid in planta screening of proteins that inhibit the growth of insect herbivores

Chung

Bigham

Lappe

et al. 2021

Plant Biotechnology Journal

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“…Validation of selected reference genes. JA and SA signaling pathways play key roles in plant defense against herbivorous insects 51,52 , and JA and SA responsive genes could be expressed upon herbivore attack or hormone stimuli 51,53 . A key transcription factor of JA signaling-CsMYC2, a key enzyme in the biosynthesis of JA-CsOPR3, two enzyme involved in the biosynthesis of SA-CsPAL and CsPALc were selected as target genes to validate the rationality of diurnal expression in different tissues, JA treatment and E. onukii infestation, T. aurantii infestation or E. obliqua regurgitant treatment individually.…”

Section: Scientific Reports |mentioning

confidence: 99%

Identification and evaluation of reliable reference genes for quantitative real-time PCR analysis in tea plants under differential biotic stresses

Xü

Dong

et al. 2020

Sci Rep

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The selection of reliable reference genes (RGs) for normalization under given experimental conditions is necessary to develop an accurate qRT-PCR assay. To the best of our knowledge, only a small number of RGs have been rigorously identified and used in tea plants (Camellia sinensis (L.) O. Kuntze) under abiotic stresses, but no critical RG identification has been performed for tea plants under any biotic stresses till now. In the present study, we measured the mRNA transcriptional levels of ten candidate RGs under five experimental conditions; these genes have been identified as stable RGs in tea plants. By using the ΔCt method, geNorm, NormFinder and BestKeeper, CLATHRIN1 and UBC1, TUA1 and SAND1, or SAND1 and UBC1 were identified as the best combination for normalizing diurnal gene expression in leaves, stems and roots individually; CLATHRIN1 and GAPDH1 were identified as the best combination for jasmonic acid treatment; ACTIN1 and UBC1 were identified as the best combination for Toxoptera aurantii-infested leaves; UBC1 and GAPDH1 were identified as the best combination for Empoasca onukii-infested leaves; and SAND1 and TBP1 were identified as the best combination for Ectropis obliqua regurgitant-treated leaves. Furthermore, our results suggest that if the processing time of the treatment was long, the best RGs for normalization should be recommended according to the stability of the proposed RGs in different time intervals when intragroup differences were compared, which would strongly increase the accuracy and sensitivity of target gene expression in tea plants under biotic stresses. However, when the differences of intergroup were compared, the RGs for normalization should keep consistent across different time points. The results of this study provide a technical guidance for further study of the molecular mechanisms of tea plants under different biotic stresses.With the increasing popularity of gene expression analysis in biological research, quantitative real-time polymerase chain reaction (qRT-PCR) has become a critical and powerful tool for rapid and reliable quantification of mRNA transcriptional expression levels of target genes due to its high-throughput screening, sensitivity, simplicity, specificity and accuracy 1,2 . Relative quantification of target gene expression under certain stresses has been widely studied since the beginning of this century 3 . An accurate assay of gene expression through qRT-PCR relies on every step of sample preparation and processing, e.g., the integrity of purified RNA, the efficiency of reverse transcription, and the overall transcriptional activity of the tissues or cells analysed 4 ; each step needs to be accurately normalized by stably expressed reference genes (RGs) 5,6 . Therefore, the selection of reliable RGs for normalization under given experimental conditions is a requirement for developing an accurate qPCR assay.Housekeeping genes, such as the glyceraldehyde 3-phosphate (GAPDH), the actin gene (ACTIN), translation elongation factor EF-1 alpha (EF-1α), 18 s r...

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“…Additionally, AMP and JAC1, two maize genes that are upregulated in response to S. frugiperda feeding ( Figure S1), reduced caterpillar weight gain when overexpressed in maize. Although transcriptomic studies have identified numerous maize genes that are upregulated in response to arthropod feeding (Bui et al, 2018;Pan et al, 2020;Tzin et al, 2015Tzin et al, , 2017Yang et al, 2020;Guo et al, 2019;Song et al, 2017;Wang et al, 2017;Zhang et al, 2016), the majority of these genes have not been investigated for their role in plant defense against herbivory. This is at least in part due to the time and cost of creating maize lines that have individual genes overexpressed.…”

Section: Discussionmentioning

confidence: 99%

Rapid screening of pest resistance genes in maize using a sugarcane mosaic virus vector

Chung

Bigham

Lappe

et al. 2021

Preprint

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Spodoptera frugiperda (fall armyworm) is a notorious pest that threatens maize production world-wide. Current control measures involve the use of chemical insecticides and transgenic maize expressing Bacillus thuringiensis (Bt) toxins. Although several additional transgenes have confirmed insecticidal activity in other plants, limited research has been conducted in maize, at least partially due to the technical difficulty of maize transformation. Here, we describe implementation of a sugarcane mosaic virus (SCMV) vector for rapidly testing the efficacy of transgenes for the control of S. frugiperda in maize. Four categories of proteins were tested using the SCMV vector: (i) maize defense signaling proteins: peptide elicitors (Pep1 and Pep3) and jasmonate acid conjugating enzymes (JAR1a and JAR1b); (ii) maize defensive proteins: the previously identified ribosome-inactivating protein (RIP2) and maize proteinase inhibitor (MPI), and two proteins with predicted but unconfirmed anti-insect activities, an antimicrobial peptide (AMP) and a lectin (JAC1); (iii) lectins from other plant species: Allium cepa agglutinin (ACA) and Galanthus nivalis agglutinin (GNA); and (iv) spider and scorpion toxins: peptides from Urodacus yaschenkoi (UyCT3 and UyCT5) and Hadronyche versuta (Hvt). In most cases, S. frugiperda larval growth on maize was reduced by transient SCMV-mediated overexpression of genes encoding these proteins. Additionally, experiments with some of the SCMV-expressed genes showed effectiveness against two aphid species, Rhopalosiphum maidis (corn leaf aphid) and Myzus persicae (green peach aphid). Together, these results demonstrate that SCMV vectors can be exploited as a rapid screening method for testing the efficacy and insecticidal activity of candidate genes in maize.

show abstract

RNA‐Seq identification of candidate defense genes by analyzing Mythimna separata feeding‐damage induced systemic resistance in balsas teosinte

Cited by 9 publications

References 33 publications

A sugarcane mosaic virus vector for rapid in planta screening of proteins that inhibit the growth of insect herbivores

A sugarcane mosaic virus vector for rapid in planta screening of proteins that inhibit the growth of insect herbivores

Identification and evaluation of reliable reference genes for quantitative real-time PCR analysis in tea plants under differential biotic stresses

Rapid screening of pest resistance genes in maize using a sugarcane mosaic virus vector

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