scholarly journals RNAi-Mediated Management of Whitefly Bemisia tabaci by Oral Delivery of Double-stranded RNAs

Proceedings ◽  
2019 ◽  
Vol 36 (1) ◽  
pp. 11
Author(s):  
Jain ◽  
Robinson ◽  
Mitter
Keyword(s):  
Rna Interference ◽  
Gene Silencing ◽  
Bemisia Tabaci ◽  
Oral Delivery ◽  
Target Genes ◽  
Insect Pests ◽  
Control Strategies ◽  
Plant Viruses ◽  
Potential Game ◽  
Viable Approach

The whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae) is a significant global pest of economically important vegetable, fibre, and ornamental crops. Whiteflies directly damage the plants by piercing and sucking essential nutrients, indirectly through honeydew secretion and by transmitting more than 200 plant viruses that cause millions of dollars in produce losses per year. Whitefly management is mostly reliant on the heavy use of chemical insecticides. However, this ultimately leads to increasing resistance development, detrimental effects on beneficial insects and biomagnification of ecologically harmful chemicals in the environment. Responding to consumer demands for more selective, less toxic, non-GM insect control strategies, RNA interference (RNAi) has emerged as a potential game-changing solution. The RNA interference (RNAi) is a homology-dependent mechanism of gene silencing that represents a feasible and sustainable technology for the management of insect pests. In the present study, twenty-two whitefly genes were selected based on their essential function in the insect and tested in artificial diet bioassays for mortality and gene silencing efficacy. The nine most effective dsRNA constructs showed moderate-to-high whitefly mortality as compared to negative controls six days post-feeding. qPCR analysis further demonstrated significant knockdown of target gene mRNA expression. Additionally, uptake and spread of fluorescently labelled dsRNA was evident beyond the midgut of the whitefly supporting the systemic spreading of RNAi effectors. Taken together, the oral delivery of dsRNA shows effective RNAi mediated gene silencing of target genes and offers a viable approach for the development of dsRNA biopesticides against hemipteran pest.

scholarly journals RNA Interference of Genes Encoding the Vacuolar-ATPase in Liriomyza trifolii

Insects ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 41
Author(s):  
Ya-Wen Chang ◽  
Yu-Cheng Wang ◽  
Xiao-Xiang Zhang ◽  
Junaid Iqbal ◽  
Yu-Zhou Du
Keyword(s):  
Rna Interference ◽  
Expression Analysis ◽  
Target Genes ◽  
Fluorescent Protein ◽  
Control Strategies ◽  
Vacuolar Atpase ◽  
Invasive Pest ◽  
Liriomyza Trifolii ◽  
Horticultural Crops ◽  
Genes Encoding

The leafminer fly, Liriomyza trifolii, is an invasive pest of vegetable and horticultural crops in China. In this study, a microinjection method based on dsRNA was developed for RNA interference (RNAi) in L. trifolii using genes encoding vacuolar-ATPase (V-ATPase). Expression analysis indicated that V-ATPase B and V-ATPase D were more highly expressed in L. trifolii adults than in larvae or pupae. Microinjection experiments with dsV-ATPase B and dsV-ATPase D were conducted to evaluate the efficacy of RNAi in L. trifolii adults. Expression analysis indicated that microinjection with 100 ng dsV-ATPase B or dsV-ATPase led to a significant reduction in V-ATPase transcripts as compared to that of the dsGFP control (dsRNA specific to green fluorescent protein). Furthermore, lower dsRNA concentrations were also effective in reducing the expression of target genes when delivered by microinjection. Mortality was significantly higher in dsV-ATPase B- and dsV-ATPase D-treated insects than in controls injected with dsGFP. The successful deployment of RNAi in L. trifolii will facilitate functional analyses of vital genes in this economically-important pest and may ultimately result in new control strategies.

scholarly journals Transmission of the Bean-Associated Cytorhabdovirus by the Whitefly Bemisia tabaci MEAM1

Viruses ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 1028
Author(s):  
Bruna Pinheiro-Lima ◽  
Rita C. Pereira-Carvalho ◽  
Dione M. T. Alves-Freitas ◽  
Elliot W. Kitajima ◽  
Andreza H. Vidal ◽  
...  
Keyword(s):  
Bemisia Tabaci ◽  
Control Strategies ◽  
Genomic Data ◽  
Plant Viruses ◽  
Asia Minor ◽  
Plant Interactions ◽  
Experimental Conditions ◽  
Lower Efficiency ◽  
Viral Particles ◽  
Pathogenicity Tests

The knowledge of genomic data of new plant viruses is increasing exponentially; however, some aspects of their biology, such as vectors and host range, remain mostly unknown. This information is crucial for the understanding of virus–plant interactions, control strategies, and mechanisms to prevent outbreaks. Typically, rhabdoviruses infect monocot and dicot plants and are vectored in nature by hemipteran sap-sucking insects, including aphids, leafhoppers, and planthoppers. However, several strains of a potentially whitefly-transmitted virus, papaya cytorhabdovirus, were recently described: (i) bean-associated cytorhabdovirus (BaCV) in Brazil, (ii) papaya virus E (PpVE) in Ecuador, and (iii) citrus-associated rhabdovirus (CiaRV) in China. Here, we examine the potential of the Bemisia tabaci Middle East-Asia Minor 1 (MEAM1) to transmit BaCV, its morphological and cytopathological characteristics, and assess the incidence of BaCV across bean producing areas in Brazil. Our results show that BaCV is efficiently transmitted, in experimental conditions, by B. tabaci MEAM1 to bean cultivars, and with lower efficiency to cowpea and soybean. Moreover, we detected BaCV RNA in viruliferous whiteflies but we were unable to visualize viral particles or viroplasm in the whitefly tissues. BaCV could not be singly isolated for pathogenicity tests, identification of the induced symptoms, and the transmission assay. BaCV was detected in five out of the seven states in Brazil included in our study, suggesting that it is widely distributed throughout bean producing areas in the country. This is the first report of a whitefly-transmitted rhabdovirus.

scholarly journals RNA interference as a gene silencing tool to controlTuta absolutain tomato (Solanum lycopersicum)

PeerJ ◽  
2016 ◽  
Vol 4 ◽  
pp. e2673 ◽  
Author(s):  
Roberto A. Camargo ◽  
Guilherme O. Barbosa ◽  
Isabella Presotto Possignolo ◽  
Lazaro E. P. Peres ◽  
Eric Lam ◽  
...  
Keyword(s):  
Rna Interference ◽  
Gene Silencing ◽  
Target Gene ◽  
Target Genes ◽  
Larval Mortality ◽  
Tuta Absoluta ◽  
Specific Gene ◽  
Gene Transcript ◽  
Pest Species ◽  
In Planta

RNA interference (RNAi), a gene-silencing mechanism that involves providing double-stranded RNA molecules that match a specific target gene sequence, is now widely used in functional genetic studies. The potential application of RNAi-mediated control of agricultural insect pests has rapidly become evident. The production of transgenic plants expressing dsRNA molecules that target essential insect genes could provide a means of specific gene silencing in larvae that feed on these plants, resulting in larval phenotypes that range from loss of appetite to death. In this report, we show that the tomato leafminer (Tuta absoluta), a major threat to commercial tomato production, can be targeted by RNAi. We selected two target genes (Vacuolar ATPase-AandArginine kinase) based on the RNAi response reported for these genes in other pest species. In view of the lack of an artificial diet forT. absoluta, we used two approaches to deliver dsRNA into tomato leaflets. The first approach was based on the uptake of dsRNA by leaflets and the second was based on “in planta-induced transient gene silencing” (PITGS), a well-established method for silencing plant genes, used here for the first time to deliverin planta-transcribed dsRNA to target insect genes.Tuta absolutalarvae that fed on leaves containing dsRNA of the target genes showed an ∼60% reduction in target gene transcript accumulation, an increase in larval mortality and less leaf damage. We then generated transgenic ‘Micro-Tom’ tomato plants that expressed hairpin sequences for both genes and observed a reduction in foliar damage byT. absolutain these plants. Our results demonstrate the feasibility of RNAi as an alternative method for controlling this critical tomato pest.

scholarly journals RNA interference-mediated gene silencing in murine T cells: in vitro and in vivo validation of proinflammatory target genes

2008 ◽  
Vol 6 (1) ◽  
pp. 3 ◽  
Author(s):  
Tatjana C Gust ◽  
Luisa Neubrandt ◽  
Claudia Merz ◽  
Khusru Asadullah ◽  
Ulrich Zügel ◽  
...  
Keyword(s):  
T Cells ◽  
Rna Interference ◽  
Gene Silencing ◽  
Target Genes ◽  
In Vivo Validation

scholarly journals RNA interference as a gene silencing tool to control Tuta absoluta in tomato (Solanum lycopersicum)

2016 ◽  
Author(s):  
Roberto A Camargo ◽  
Guilherme O Barbosa ◽  
Isabella Presotto Possignolo ◽  
Lazaro E. P. Peres ◽  
Eric Lam ◽  
...  
Keyword(s):  
Rna Interference ◽  
Gene Silencing ◽  
Target Gene ◽  
Target Genes ◽  
Larval Mortality ◽  
Tuta Absoluta ◽  
Specific Gene ◽  
Gene Transcript ◽  
Pest Species ◽  
In Planta

RNA interference (RNAi), a gene-silencing mechanism that involves providing double-stranded RNA molecules that match a specific target gene sequence, is now widely used in functional genetic studies. The potential application of RNAi-mediated control of agricultural insect pests has rapidly become evident. The production of transgenic plants expressing dsRNA molecules that target essential insect genes could provide a means of specific gene silencing in larvae that feed on these plants, resulting in larval phenotypes that range from loss of appetite to death. In this report, we show that the tomato leafminer (Tuta absoluta), a major threat to commercial tomato production, can be targeted by RNAi. We selected two target genes [Vacuolar ATPase-A and Arginine kinase] based on the RNAi response reported for these genes in other pest species. In view of the lack of an artificial diet for T. absoluta, we used two approaches to deliver dsRNA into tomato leaflets. The first approach was based on the uptake of dsRNA by leaflets and the second was based on “in planta-induced transient gene silencing” (PITGS), a well-established method for silencing plant genes, used here for the first time to deliver in planta-transcribed dsRNA to target insect genes. Tuta absoluta larvae that fed on leaves containing dsRNA of the target genes showed an ~60% reduction in target gene transcript accumulation, an increase in larval mortality and less leaf damage. We then generated transgenic ‘Micro-Tom’ tomato plants that expressed hairpin sequences for both genes and observed a reduction in foliar damage by T. absoluta in these plants. Our results demonstrate the feasibility of RNAi as an alternative method for controlling this critical tomato pest.

scholarly journals Amenability of Maruca vitrata (Lepidoptera: Crambidae) to gene silencing through exogenous administration and host-delivered dsRNA in pigeonpea (Cajanus cajan L.)

2021 ◽  
Author(s):  
Madhurima Chatterjee ◽  
Jyoti Yadav ◽  
Maniraj Rathinam ◽  
Kesiraju Karthik ◽  
Gopal Chowdhary ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Target Genes ◽  
Insect Pests ◽  
Crop Improvement ◽  
Maruca Vitrata ◽  
Economic Losses ◽  
In Planta ◽  
Biotic Stresses ◽  
Pod Borer ◽  
Plant Level

Abstract Insect pests are one of the major biotic stresses limiting yield in commercially important food crops. The lepidopteran polyphagous spotted pod borer, Maruca vitrata causes significant economic losses in legumes including pigeonpea. RNAi-based gene silencing has emerged as one of the potential biotechnological tools for crop improvement. We report in this paper, RNAi in M. vitrata through exogenous administration of dsRNA encoding three functionally important genes, Alpha-amylase (α-amylase), Chymotrypsin-like serine protease (CTLP) and Tropomyosin (TPM) into the larval haemolymph and their host-delivered RNAi in pigeonpea. Significant decline in the expression of selected genes supported by over-expression of DICER and generation of siRNA indicated the occurrence of RNAi in the dsRNA-injected larvae. Additionally, the onset of RNAi in the herbivore was demonstrated in pigeonpea, one of the prominent hosts, by host-delivered RNAi. Transgenics in pigeonpea (cv. Pusa992), a highly recalcitrant crop, were developed through a shoot apical meristem-targeted in planta transformation strategy and evaluated. Plant level bioassays in transgenic events characterized and selected at molecular level showed mortality of M. vitrata larvae as well as reduced feeding when compared to wild type. Furthermore, molecular evidences for down regulation of target genes in the insects that fed on transgenics authenticated RNAi. Considering the variability of gene silencing in lepidopteran pests, this study provided corroborative proof for the possibility of gene silencing in M. vitrata through both the strategies.

scholarly journals Bemisia tabaci on Vegetables in the Southern United States: Incidence, Impact, and Management

Insects ◽  
2021 ◽  
Vol 12 (3) ◽  
pp. 198
Author(s):  
Yinping Li ◽  
George N. Mbata ◽  
Somashekhar Punnuri ◽  
Alvin M. Simmons ◽  
David I. Shapiro-Ilan
Keyword(s):  
United States ◽  
Bemisia Tabaci ◽  
Insect Pests ◽  
The United States ◽  
Plant Viruses ◽  
Cultural Control ◽  
Economic Losses ◽  
Southern United States ◽  
Vegetable Crops ◽  
Plant Damage

Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae) is among the most economically important insect pests of various vegetable crops in the Southern United States. This insect is considered a complex of at least 40 morphologically indistinguishable cryptic species. Bemisia tabaci Middle East-Asia Minor 1 (MEAM1) was initially introduced in the United States around 1985 and has since rapidly spread across the Southern United States to Texas, Arizona, and California, where extreme field outbreaks have occurred on vegetable and other crops. This pest creates extensive plant damage through direct feeding on vegetables, secreting honeydew, causing plant physiological disorders, and vectoring plant viruses. The direct and indirect plant damage in vegetable crops has resulted in enormous economic losses in the Southern United States, especially in Florida, Georgia, and Texas. Effective management of B. tabaci on vegetables relies mainly on the utilization of chemical insecticides, particularly neonicotinoids. However, B. tabaci has developed considerable resistance to most insecticides. Therefore, alternative integrated pest management (IPM) strategies are required, such as cultural control by manipulation of production practices, resistant vegetable varieties, and biological control using a suite of natural enemies for the management of the pest.

scholarly journals Overexpression of an Osa-miR162a Derivative in Rice Confers Cross-Kingdom RNA Interference-Mediated Brown Planthopper Resistance without Perturbing Host Development

2021 ◽  
Vol 22 (23) ◽  
pp. 12652
Author(s):  
Wenzhong Shen ◽  
Shanni Cao ◽  
Jinhui Liu ◽  
Wenqing Zhang ◽  
Jie Chen ◽  
...  
Keyword(s):  
Rna Interference ◽  
Target Genes ◽  
Insect Pests ◽  
Brown Planthopper ◽  
Nilaparvata Lugens ◽  
Environmental Concerns ◽  
Reproduction Process ◽  
Brown Planthopper Resistance ◽  
Global Population ◽  
Host Development

Rice is a main food crop for more than half of the global population. The brown planthopper (BPH, Nilaparvata lugens) is one of the most destructive insect pests of rice. Currently, repeated overuse of chemical insecticides represents a common practice in agriculture for BPH control, which can induce insect tolerance and provoke environmental concerns. This situation calls for innovative and widely applicable strategies for rice protection against BPH. Here we report that the rice osa-miR162a can mediate cross-kingdom RNA interference (RNAi) by targeting the NlTOR (Target of rapamycin) gene of BPH that regulates the reproduction process. Through artificial diet or injection, osa-miR162a mimics repressed the NlTOR expression and impaired the oviposition of BPH adults. Consistently, overproduced osa-miR162a in transgenic rice plants compromised the fecundity of BPH adults fed with these plants, but meanwhile perturbed root and grain development. To circumvent this issue, we generated osa-miR162a-m1, a sequence-optimized osa-miR162a, by decreasing base complementarity to rice endogenous target genes while increasing base complementarity to NlTOR. Transgenic overexpression of osa-miR162a-m1 conferred rice resistance to BPH without detectable developmental penalty. This work reveals the first cross-kingdom RNAi mechanism in rice-BPH interactions and inspires a potentially useful approach for improving rice resistance to BPH. We also introduce an effective strategy to uncouple unwanted host developmental perturbation from desirable cross-kingdom RNAi benefits for overexpressed plant miRNAs.

scholarly journals RNA interference as a gene silencing tool to control Tuta absoluta in tomato (Solanum lycopersicum)

2016 ◽  
Author(s):  
Roberto A Camargo ◽  
Guilherme O Barbosa ◽  
Isabella Presotto Possignolo ◽  
Lazaro E. P. Peres ◽  
Eric Lam ◽  
...  
Keyword(s):  
Rna Interference ◽  
Gene Silencing ◽  
Target Gene ◽  
Target Genes ◽  
Larval Mortality ◽  
Tuta Absoluta ◽  
Specific Gene ◽  
Gene Transcript ◽  
Pest Species ◽  
In Planta

RNA interference (RNAi), a gene-silencing mechanism that involves providing double-stranded RNA molecules that match a specific target gene sequence, is now widely used in functional genetic studies. The potential application of RNAi-mediated control of agricultural insect pests has rapidly become evident. The production of transgenic plants expressing dsRNA molecules that target essential insect genes could provide a means of specific gene silencing in larvae that feed on these plants, resulting in larval phenotypes that range from loss of appetite to death. In this report, we show that the tomato leafminer (Tuta absoluta), a major threat to commercial tomato production, can be targeted by RNAi. We selected two target genes [Vacuolar ATPase-A and Arginine kinase] based on the RNAi response reported for these genes in other pest species. In view of the lack of an artificial diet for T. absoluta, we used two approaches to deliver dsRNA into tomato leaflets. The first approach was based on the uptake of dsRNA by leaflets and the second was based on “in planta-induced transient gene silencing” (PITGS), a well-established method for silencing plant genes, used here for the first time to deliver in planta-transcribed dsRNA to target insect genes. Tuta absoluta larvae that fed on leaves containing dsRNA of the target genes showed an ~60% reduction in target gene transcript accumulation, an increase in larval mortality and less leaf damage. We then generated transgenic ‘Micro-Tom’ tomato plants that expressed hairpin sequences for both genes and observed a reduction in foliar damage by T. absoluta in these plants. Our results demonstrate the feasibility of RNAi as an alternative method for controlling this critical tomato pest.

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