Virus-Induced Gene Silencing Suggests (1,3;1,4)-β-glucanase Is a Susceptibility Factor in the Compatible Russian Wheat Aphid–Wheat Interaction

The Russian wheat aphid (RWA), Diuraphis noxia (Kurdjumov), is a significant insect pest of wheat (Triticum aestivum L.) and has a major economic impact worldwide, especially on winter wheat in the western United States. The continuing emergence of new RWA biotypes virulent to existing resistance genes reinforces the need for more durable resistance. Studies have indicated that resistance in previously susceptible plants can be produced by knock-down of susceptibility genes or other genes involved in host plant susceptibility. Therefore, investigation into genes involved in compatible RWA–wheat interactions could be a feasible approach to achieving durable RWA resistance. The objective of this study was to test whether silencing (1,3;1,4)-β-glucanase, previously observed to be highly induced in susceptible compared with resistant wheat during aphid infestation, would confer resistance to a susceptible wheat genotype. Barley stripe mosaic virus–mediated virus-induced gene silencing was employed to test whether (1,3;1,4)-β-glucanase is involved in the susceptible reaction of ‘Gamtoos-S' (GS). Controlled infestation with U.S. biotype RWA2 was done to assess aphid reproduction and host symptom development. Aphids on (1,3;1,4)-β-glucanase-silenced plants reproduced less per day and had longer prenymphipositional periods than those on control GS plants. Furthermore, the (1,3;1,4)-β-glucanase-silenced plants exhibited less chlorosis and greater dry weight compared with GS. Aphid reproduction and host plant symptom development showed linear relationships with (1,3;1,4)-β-glucanase transcript levels. Our results suggest that (1,3;1,4)-β-glucanase is required for successful infestation by the RWA and may be a susceptibility factor that could be exploited as a potential target for RWA resistance breeding.

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California TRV-based VIGS vectors mediate gene silencing at elevated temperatures but with greater growth stunting

BMC Plant Biology ◽

10.1186/s12870-021-03324-8 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Jamilur Rahman ◽

Ian T. Baldwin ◽

Klaus Gase

Keyword(s):

Gene Silencing ◽

Host Plant ◽

Target Genes ◽

Elevated Temperatures ◽

Tobacco Rattle Virus ◽

Vector System ◽

Nicotiana Attenuata ◽

Cdna Clones ◽

Virus Induced Gene Silencing ◽

In Planta

Abstract Background Tobacco rattle virus (TRV) based virus-induced gene silencing (VIGS), a widely used functional genomics tool, requires growth temperatures typically lower than those of the plant’s native environment. Enabling VIGS under native conditions in the field according to applicable safety regulations could be a revolutionary advance for ecological research. Results Here, we report the development of an enhanced thermal tolerant VIGS vector system based on a TRV California isolate. cDNA clones representing the whole viral genome were sequenced and used to construct separate binary plant transformation vectors for functional elements of RNA1 (6765 nt) and RNA2 (3682 nt). VIGS of target genes was induced by transient transformation of the host plant with both vectors or by treating the host plant with sap from already VIGS induced plants. In Nicotiana attenuata the silencing efficiency of the PDS (phytoene desaturase) gene was 90% at 28 °C and 78% at 30 °C. Silencing at these temperatures was more prominent and durable than silencing induced by the widely used TRV PpK20-based pBINTRA6/pTV00 system, but was associated with a viral phenotype. Differences in the suppressor protein and RNA dependent RNA polymerase sequences between the TRV California isolate and PpK20 may be the reason for their different thermal tolerance. Conclusions The new TRV California-based VIGS vectors induce gene silencing in Nicotiana attenuata at higher temperatures than the existing pBINTRA6/pTV00 vector system, but cause greater growth defects. The new vector system opens up an avenue to study genes functions in planta under field conditions.

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A Virus-Induced Gene Silencing Screen Identifies a Role for Thylakoid Formation1 in Pseudomonas syringae pv tomato Symptom Development in Tomato and Arabidopsis

PLANT PHYSIOLOGY ◽

10.1104/pp.109.148106 ◽

2009 ◽

Vol 152 (1) ◽

pp. 281-292 ◽

Cited By ~ 22

Author(s):

Tamding Wangdi ◽

Srinivasa Rao Uppalapati ◽

Satish Nagaraj ◽

Choong-Min Ryu ◽

Carol L. Bender ◽

...

Keyword(s):

Gene Silencing ◽

Pseudomonas Syringae ◽

Symptom Development ◽

Virus Induced Gene Silencing

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Faculty Opinions recommendation of High throughput virus-induced gene silencing implicates heat shock protein 90 in plant disease resistance.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1003609.201391 ◽

2003 ◽

Author(s):

Jen Sheen

Keyword(s):

Disease Resistance ◽

Heat Shock ◽

Heat Shock Protein ◽

Gene Silencing ◽

High Throughput ◽

Plant Disease Resistance ◽

Plant Disease ◽

Heat Shock Protein 90 ◽

Virus Induced Gene Silencing

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Faculty Opinions recommendation of Deletion of fucose residues in plant N-glycans by repression of the GDP-mannose 4,6-dehydratase gene using virus-induced gene silencing and RNA interference.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.8177956.8591054 ◽

2011 ◽

Author(s):

Keith Davis ◽

Nobuyuki Matoba

Keyword(s):

Rna Interference ◽

Gene Silencing ◽

Virus Induced Gene Silencing

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Virus-induced Gene Silencing (VIGS) in Barley Seedling Leaves

BIO-PROTOCOL ◽

10.21769/bioprotoc.1506 ◽

2015 ◽

Vol 5 (12) ◽

Author(s):

Lokanadha Gunupuru ◽

Shahin Ali ◽

Fiona Doohan ◽

Steven Scofield

Keyword(s):

Gene Silencing ◽

Virus Induced Gene Silencing ◽

Barley Seedling

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Cucumber mosaic virus-induced gene silencing in banana

Scientific Reports ◽

10.1038/s41598-019-47962-3 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 2

Author(s):

Yuh Tzean ◽

Ming-Chi Lee ◽

Hsiao-Hsuan Jan ◽

Yi-Shu Chiu ◽

Tsui-Chin Tu ◽

...

Keyword(s):

Gene Silencing ◽

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Virus Induced Gene Silencing

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Morphological and Chemical Factors Related to Western Flower Thrips Resistance in the Ornamental Gladiolus

Plants ◽

10.3390/plants10071384 ◽

2021 ◽

Vol 10 (7) ◽

pp. 1384

Author(s):

Dinar S. C. Wahyuni ◽

Young Hae Choi ◽

Kirsten A. Leiss ◽

Peter G. L. Klinkhamer

Keyword(s):

Host Plant ◽

Plant Resistance ◽

Host Plant Resistance ◽

Frankliniella Occidentalis ◽

Western Flower Thrips ◽

Resistance Breeding ◽

Breeding Programs ◽

Triterpenoid Saponins ◽

Flower Thrips ◽

Highly Correlated

Understanding the mechanisms involved in host plant resistance opens the way for improved resistance breeding programs by using the traits involved as markers. Pest management is a major problem in cultivation of ornamentals. Gladiolus (Gladiolus hybridus L.) is an economically important ornamental in the Netherlands. Gladiolus is especially sensitive to attack by western flower thrips (Frankliniella occidentalis (Pergande) (Thysanoptera:Thripidae)). The objective of this study was, therefore, to investigate morphological and chemical markers for resistance breeding to western flower thrips in Gladiolus varieties. We measured thrips damage of 14 Gladiolus varieties in a whole-plant thrips bioassay and related this to morphological traits with a focus on papillae density. Moreover, we studied chemical host plant resistance to using an eco-metabolomic approach comparing the 1H NMR profiles of thrips resistant and susceptible varieties representing a broad range of papillae densities. Thrips damage varied strongly among varieties: the most susceptible variety showed 130 times more damage than the most resistant one. Varieties with low thrips damage had shorter mesophylls and epidermal cells, as well as a higher density of epicuticular papillae. All three traits related to thrips damage were highly correlated with each other. We observed a number of metabolites related to resistance against thrips: two unidentified triterpenoid saponins and the amino acids alanine and threonine. All these compounds were highly correlated amongst each other as well as to the density of papillae. These correlations suggest that papillae are involved in resistance to thrips by producing and/or storing compounds causing thrips resistance. Although it is not possible to distinguish the individual effects of morphological and chemical traits statistically, our results show that papillae density is an easy marker in Gladiolus-breeding programs targeted at increased resistance to thrips.

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A cassava common mosaic virus vector for virus-induced gene silencing in cassava

Plant Methods ◽

10.1186/s13007-021-00775-w ◽

2021 ◽

Vol 17 (1) ◽

Author(s):

Decai Tuo ◽

Peng Zhou ◽

Pu Yan ◽

Hongguang Cui ◽

Yang Liu ◽

...

Keyword(s):

Gene Silencing ◽

Mosaic Virus ◽

Gene Function ◽

Viral Vector ◽

Function Analysis ◽

Systemic Infection ◽

Cdna Clones ◽

Virus Induced Gene Silencing ◽

Efficient Gene ◽

Gene Function Analysis

Abstract Background Cassava is an important crop for food security and industry in the least-developed and developing countries. The completion of the cassava genome sequence and identification of large numbers of candidate genes by next-generation sequencing provide extensive resources for cassava molecular breeding and increase the need for rapid and efficient gene function analysis systems in cassava. Several plant virus-induced gene silencing (VIGS) systems have been developed as reverse genetic tools for rapid gene function analysis in cassava. However, these VIGS vectors could cause severe viral symptoms or inefficient gene silencing. Results In this study, we constructed agroinfection-compatible infectious cDNA clones of cassava common mosaic virus isolate CM (CsCMV-CM, genus Potexvirus, family Alphaflexiviridae) that causes systemic infection with mild symptoms in cassava. CsCMV-CM was then modified to a viral vector carrying the Nimble cloning frame, which facilitates the rapid and high-throughput cloning of silencing fragments into the viral genome. The CsCMV-based vector successfully silenced phytoene desaturase (PDS) and magnesium chelatase subunit I (ChlI) in different cassava varieties and Nicotiana benthamiana. The silencing of the ChlI gene could persist for more than two months. Conclusions This CsCMV-based VIGS system provides a new tool for rapid and efficient gene function studies in cassava.

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