Simultaneous RNAi Knockdown of Three FMRFamide-Like Peptide Genes, Mi-flp1, Mi-flp12, and Mi-flp18 Provides Resistance to Root-Knot Nematode, Meloidogyne incognita

Banakar, Prakash; Hada, Alkesh; Papolu, Pradeep K.; Rao, Uma

doi:10.3389/fmicb.2020.573916

Cited by 27 publications

(15 citation statements)

References 82 publications

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“…Chemical nematicides are the mainstay due to their ability to reduce high densities of nematodes in the soil in a short period to avoid significant yield losses (Regmi and Desaeger, 2020). However, due to their high toxicity and possible environmental and health hazards, most of the chemical nematicides, fumigants, and insecticides have been withdrawn or banned from the global market (Ruiz-Suárez et al, 2015;Banakar et al, 2020;Hada et al, 2021b,c). In addition, limited label claim of some of the recently introduced synthetic chemicals such as fluensulfone, fluopyram, etc., restricts their usage in all the crops and also against different nematodes.…”

Section: Introductionmentioning

confidence: 99%

Identification, Characterization, and Evaluation of Nematophagous Fungal Species of Arthrobotrys and Tolypocladium for the Management of Meloidogyne incognita

et al. 2021

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Root-knot nematodes belonging to the genus Meloidogyne are agriculturally important pests, and biocontrol strategies offer safer alternatives for their management. In the present study, two fungal species from Indian soils were identified as Arthrobotrys thaumasia and Tolypocladium cylindrosporum based on morphological characteristics and further confirmed using molecular markers. In vitro evaluation of A. thaumasia against M. incognita and Caenorhabditis elegans showed 82 and 73% parasitism, respectively, whereas T. cylindrosporum gave 65.2 and 57.7% parasitism, respectively. Similarly, culture filtrates of A. thaumasia caused 57.7 and 53.7% mortality of M. incognita and C. elegans, respectively, whereas T. cylindrosporum caused higher mortality of 87.3 and 64%, respectively. Besides, greenhouse evaluation of both fungi against M. incognita infecting tomato significantly reduced nematode disease burden reflecting parasitic success measured as the total number of galls, egg masses, eggs per egg mass, and derived nematode multiplication factor. Application of A. thaumasia and T. cylindrosporum reduced nematode multiplication factor by 80 and 95%, respectively, compared with control. General metabolite profiling of tested fungi using gas chromatography–mass spectrometry and ultra-performance liquid chromatography–quadrupole/time of flight mass spectrometry reported for the first time here showed presence of various volatile and non-volatile compounds with nematicidal activity, viz., trimethyl-heptadiene, methyl-hexadecanol, dodecadienal, decane, terpendole E, dodecane, acetamido-6-anthraquinone, and hexadecanol. Also, other compounds such as undecane, dibutyl-disulfide, octadecenal, paganin, talathermophilin, dactylarin, tolypyridone A, tolypyridone B, pyridoxatin, and destruxin were identified, reported in the literature to possess antibacterial, antifungal, and insecticidal properties. This is the first report of the occurrence of both fungi from India and pioneer demonstration of T. cylindrosporum for root-knot nematode management.

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Section: Introductionmentioning

confidence: 99%

Identification, Characterization, and Evaluation of Nematophagous Fungal Species of Arthrobotrys and Tolypocladium for the Management of Meloidogyne incognita

et al. 2021

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“…Control of nematode parasitism by simultaneous silencing of many genes has been demonstrated in Heterodera schachtii 41 and also in M. incognita. 42 These studies established that simultaneous and/or combinatorial gene knockdown not only strengthens resistance to nematode infestation, but also extends the spectrum of resistance against diverse nematode species. 43 The findings described in this article are a proof of concept for the successful validation of the MIGS strategy for the delivery of siRNAs in plants as well as its efficacy against M. incognita.…”

Section: Discussionmentioning

confidence: 98%

Micro RNA‐induced gene silencing strategy for the delivery of siRNAs targeting Meloidogyne incognita in a model plant Nicotiana benthamiana

Hada

Patil

Bajpai

et al. 2021

Pest Management Science

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BACKGROUND: Occurrence of multiple biotic stresses on crop plants result in drastic yield losses which may have severe impact on the food security. It is a challenge to design strategies for simultaneous management of these multiple stresses. Hence, establishment of innovative approaches that aid in their management is critical. Here, we have introgressed a micro RNAinduced gene silencing (MIGS) based combinatorial gene construct containing seven target gene sequences of cotton leaf curl disease (CLCuD), cotton leaf hopper (Amrasca biguttula biguttula), cotton whitefly (Bemisia tabaci) and root-knot nematode (Meloidogyne incognita).RESULTS: Stable transgenic lines of Nicotiana benthamiana were generated with the T-DNA harboring Arabidopsis miR173 target site fused to fragments of Sec23 and ecdysone receptor (EcR) genes of cotton leaf hopper and cotton whitefly. It also contained C2/replication associated protein (C2/Rep) and C4 (movement protein) along with ⊎C1 gene of betasatellite to target CLCuD, and two FMRFamide-like peptide (FLP) genes, Mi-flp14 and Mi-flp18 of M. incognita. These transgenic plants were assessed for the amenability of MIGS approach for pest control by efficacy evaluation against M. incognita. Results showed successful production of small interfering RNA (siRNA) through the tasiRNA (trans-acting siRNA) pathway in the transgenic plants corresponding to Mi-flp18 gene. Furthermore, we observed reduced Mi-flp14 and Mi-flp18 transcripts (up to 2.37 ± 0.12-fold) in females extracted from transgenic plants. The average number of galls, total endoparasites, egg masses and number of eggs per egg mass reduced were in the range 27-62%, 39-70%, 38-65% and 34-49%, respectively. More importantly, MIGS transgenic plants showed 80% reduction in the nematode multiplication factor (MF).CONCLUSION: This study demonstrates successful validation of the MIGS approach in the model plant, N. benthamiana for efficacy against M. incognita, as a prelude to translation to cotton.

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“…Increasing evidence for FLP-mediated biological functions in host finding, invasion, and reproduction of PPN support the potential valuable repository of FLP genes as possible targets for the management of plant pathogenic nematodes ( Kimber et al, 2007 ; Dalzell et al, 2010 ; Papolu et al, 2013 ; Dong et al, 2014 ; Kumari et al, 2017 ; Banakar et al, 2020 ). Previous interrogation of H. glycines EST database by McVeigh et al (2006) identified two flp-22 EST transcripts.…”

Section: Discussionmentioning

confidence: 99%

“…Additionally, several studies have successfully applied the method of host-derived RNAi to reduce PPN infection and reproduction by targeting flp genes ( Papolu et al, 2013 ; Dutta et al, 2020 ; Hada et al, 2020 ). More recently, Banakar et al (2020) generated transgenic tobacco using hairpin fusion expressing gene cassette of FLPs containing Mi-flp-1 , Mi-flp-12 , and Mi-flp-18 , which significantly reduced the reproduction of M. incognita . Surprisingly, flp-32 of G. pallida is the first FLP-encoding gene reportedly, which negatively regulates nematode movement by activating its putative receptor (Gp-flp-32R; Atkinson et al, 2013 ).…”

Section: Introductionmentioning

confidence: 99%

FMRFamide-Like Peptide 22 Influences the Head Movement, Host Finding, and Infection of Heterodera glycines

et al. 2021

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The FMRFamide-like peptides (FLPs) represent the largest family of nematode neuropeptides and are involved in multiple parasitic activities. The immunoreactivity to FMRFamide within the nervous system of Heterodera glycines, the most economically damaging parasite of soybean [Glycine max L. (Merr)], has been reported in previous research. However, the family of genes encoding FLPs of H. glycines were not identified and functionally characterized. In this study, an FLP encoding gene Hg-flp-22 was cloned from H. glycines, and its functional characterization was uncovered by using in vitro RNA interference and application of synthetic peptides. Bioinformatics analysis showed that flp-22 is widely expressed in multiple nematode species, where they encode the highly conserved KWMRFamide motifs. Quantitative real-time (qRT)-PCR results revealed that Hg-flp-22 was highly expressed in the infective second-stage juveniles (J2s) and adult males. Silencing of Hg-flp-22 resulted in the reduced movement of J2s to the host root and reduced penetration ability, as well as a reduction in their subsequent number of females. Behavior and infection assays demonstrated that application of synthetic peptides Hg-FLP-22b (TPQGKWMRFa) and Hg-FLP-22c (KMAIEGGKWVRFa) significantly increased the head movement frequency and host invasion abilities in H. glycines but not in Meloidogyne incognita. In addition, the number of H. glycines females on the host roots was found to be significantly higher in Hg-FLP-22b treated nematodes than the ddH2O-treated control J2s. These results presented in this study elucidated that Hg-flp-22 plays a role in regulating locomotion and infection of H. glycines. This suggests the potential of FLP signaling as putative control targets for H. glycines in soybean production.

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Simultaneous RNAi Knockdown of Three FMRFamide-Like Peptide Genes, Mi-flp1, Mi-flp12, and Mi-flp18 Provides Resistance to Root-Knot Nematode, Meloidogyne incognita

Cited by 27 publications

References 82 publications

Identification, Characterization, and Evaluation of Nematophagous Fungal Species of Arthrobotrys and Tolypocladium for the Management of Meloidogyne incognita

Identification, Characterization, and Evaluation of Nematophagous Fungal Species of Arthrobotrys and Tolypocladium for the Management of Meloidogyne incognita

Micro RNA‐induced gene silencing strategy for the delivery of siRNAs targeting Meloidogyne incognita in a model plant Nicotiana benthamiana

FMRFamide-Like Peptide 22 Influences the Head Movement, Host Finding, and Infection of Heterodera glycines

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