Host-delivered RNAi-mediated root-knot nematode resistance in Arabidopsis by targeting splicing factor and integrase genes

Kumar, Anil; Kakrana, Atul; Sirohi, Anil; Subramaniam, Kuppuswamy; Srinivasan, Raji; Abdin, M.Z.; Jain, Pradeep

doi:10.1007/s10327-017-0701-3

Cited by 24 publications

(13 citation statements)

References 31 publications

(15 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The results reported by Kumar et al . regarding RNAi targeting of splicing and integrase genes showed the same pattern of a decreased number of egg masses compared to the number of females 32 . The reduction in fecundity was further confirmed by analysis of the number of eggs per egg mass in the RNAi lines, demonstrating a strong decline in the number of eggs by up to 74%, similar to the decrease reported for 16D10 11 .…”

Section: Discussionmentioning

confidence: 79%

Development of nematode resistance in Arabidopsis by HD-RNAi-mediated silencing of the effector gene Mi-msp2

Joshi

Kumar

Singh

et al. 2019

Sci Rep

Self Cite

View full text Add to dashboard Cite

Root-knot nematodes (RKNs) are devastating parasites that infect thousands of plants. As RKN infection is facilitated by oesophageal gland effector genes, one such effector gene, Mi-msp2, was selected for a detailed characterization. Based on domain analysis, the Mi-MSP2 protein contains an ShKT domain, which is likely involved in blocking K+ channels and may help in evading the plant defence response. Expression of the Mi-msp2 gene was higher in juveniles (parasitic stage of RKNs) than in eggs and adults. Stable homozygous transgenic Arabidopsis lines expressing Mi-msp2 dsRNA were generated, and the numbers of galls, females and egg masses were reduced by 52–54%, 60–66% and 84–95%, respectively, in two independent RNAi lines compared with control plants. Furthermore, expression analysis revealed a significant reduction in Mi-msp2 mRNA abundance (up to 88%) in female nematodes feeding on transgenic plants expressing dsRNA, and northern blot analysis confirmed expression of the Mi-msp2 siRNA in the transgenic plants. Interestingly, a significant reduction in the reproduction factor was observed (nearly 40-fold). These data suggest that the Mi-msp2 gene can be used as a potential target for RKN management in crops of economic importance.

show abstract

Section: Discussionmentioning

confidence: 79%

Development of nematode resistance in Arabidopsis by HD-RNAi-mediated silencing of the effector gene Mi-msp2

Joshi

Kumar

Singh

et al. 2019

Sci Rep

Self Cite

View full text Add to dashboard Cite

show abstract

“…However, transgenic tobacco plants expressing dsRNA of splicing factor gene through CaMV35S promoter exhibited about 95% reduction in gall formation as well as in number of nematode females (Yadav et al, 2006 ). Similarly, the splicing factor gene driven by 35S promoter in Arabidopsis transgenic lines exhibited up to 71% reduction in gall number (Kumar et al, 2017 ). In the last few years, several promoters have been identified in various crops including tobacco (Opperman et al, 1994 ), LEMMI9 in tomato (Escobar et al, 1999 ), Hahsp17.7G4 in Helianthus annuus (Escobar et al, 2003 ), AtCel-1 in Arabidopsis (Sukno et al, 2006 ), AtWRKY23 (Grunewald et al, 2008 ), and ZmRCP-1 in maize, banana, and plantains (Onyango et al, 2016 ).…”

Section: Discussionmentioning

confidence: 99%

“…RKN infestation causes an estimated annual crop loss of hundreds of billions of dollars (Abad et al, 2008 ; Elling, 2013 ). The demonstration of host-delivered RNA interference (HD-RNAi) in plant-parasitic nematodes (PPNs) offers an effective strategy to control nematode infection in crop plants (Tamilarasan and Rajam, 2013 ; Dutta et al, 2015 ; Dong et al, 2016 ; Niu et al, 2016 ; Banerjee et al, 2017 ; Kumar et al, 2017 ). Two house-keeping genes ( splicing factor and integrase ) were successfully silenced in nematodes through host-mediated RNAi in tobacco plants (Yadav et al, 2006 ).…”

Section: Introductionmentioning

confidence: 99%

Identification, Validation and Utilization of Novel Nematode-Responsive Root-Specific Promoters in Arabidopsis for Inducing Host-Delivered RNAi Mediated Root-Knot Nematode Resistance

et al. 2017

Self Cite

View full text Add to dashboard Cite

The root-knot nematode (RKN), Meloidogyne incognita, is an obligate, sedentary endoparasite that infects a large number of crops and severely affects productivity. The commonly used nematode control strategies have their own limitations. Of late, RNA interference (RNAi) has become a popular approach for the development of nematode resistance in plants. Transgenic crops capable of expressing dsRNAs, specifically in roots for disrupting the parasitic process, offer an effective and efficient means of producing resistant crops. We identified nematode-responsive and root-specific (NRRS) promoters by using microarray data from the public domain and known conserved cis-elements. A set of 51 NRRS genes was identified which was narrowed down further on the basis of presence of cis-elements combined with minimal expression in the absence of nematode infection. The comparative analysis of promoters from the enriched NRRS set, along with earlier reported nematode-responsive genes, led to the identification of specific cis-elements. The promoters of two candidate genes were used to generate transgenic plants harboring promoter GUS constructs and tested in planta against nematodes. Both promoters showed preferential expression upon nematode infection, exclusively in the root in one and galls in the other. One of these NRRS promoters was used to drive the expression of splicing factor, a nematode-specific gene, for generating host-delivered RNAi-mediated nematode-resistant plants. Transgenic lines expressing dsRNA of splicing factor under the NRRS promoter exhibited upto a 32% reduction in number of galls compared to control plants.

show abstract

“…Limitations of the host-derived RNAi strategy include efficient delivery of siRNA molecules to target pathogens and the effectiveness of these molecules in triggering the RNAi pathway in those pathogens (Li et al 2011). To date, the effectiveness of nematode control using this strategy has been variable (Klink et al 2009; Kumar et al 2017; Sindhu et al 2009; Vieira et al 2015). To achieve new and stable resources for SCN resistance in soybean breeding programs and to demonstrate consistent effects against SCN, we obtained stable transgenic soybean plants expressing RNAi silencing vectors and analyzed the small interfering RNA(siRNA) expression levels in multiple lines via small RNA-seq.…”

Section: Introductionmentioning

confidence: 99%

Host-derived gene silencing of parasite fitness genes improves resistance to soybean cyst nematodes in stable transgenic soybean

Tian

Vodkin

et al. 2019

Theor Appl Genet

View full text Add to dashboard Cite

Key message Soybean expressing small interfering RNA of SCN improved plant resistance to SCN consistently, and small RNA-seq analysis revealed a threshold of siRNA expression required for resistance ability. Abstract Soybean cyst nematode (SCN), Heterodera glycines , is one of the most destructive pests limiting soybean production worldwide, with estimated losses of $1 billion dollars annually in the USA alone. RNA interference (RNAi) has become a powerful tool for silencing gene expression. We report here that the expression of hairpin RNAi constructs, derived from two SCN genes related to reproduction and fitness, HgY25 and HgPrp17, enhances resistance to SCN in stably transformed soybean plants. The analyses of T 3 to T 5 generations of stable transgenic soybeans by molecular strategies and next-generation sequencing confirmed the presence of specific short interfering RNAs complementary to the target SCN genes. Bioassays performed on transgenic soybean lines targeting SCN HgY25 and HgPrp17 fitness genes showed significant reductions (up to 73%) for eggs/g root in the T 3 and T 4 homozygous transgenic lines. Targeted mRNAs of SCN eggs collected from the transgenic soybean lines were efficiently down-regulated, as confirmed by quantitative RT-PCR. Based on the small RNA-seq data and bioassays, it is our hypothesis that a threshold of small interfering RNA molecules is required to significantly reduce SCN populations feeding on the host plants. Our results demonstrated that host-derived gene silencing of essential SCN fitness genes could be an effective strategy for enhancing resistance in crop plants. Electronic supplementary material The online version of this article (10.1007/s00122-019-03379-0) contains supplementary material, which is available to authorized users.

show abstract

Host-delivered RNAi-mediated root-knot nematode resistance in Arabidopsis by targeting splicing factor and integrase genes

Cited by 24 publications

References 31 publications

Development of nematode resistance in Arabidopsis by HD-RNAi-mediated silencing of the effector gene Mi-msp2

Development of nematode resistance in Arabidopsis by HD-RNAi-mediated silencing of the effector gene Mi-msp2

Identification, Validation and Utilization of Novel Nematode-Responsive Root-Specific Promoters in Arabidopsis for Inducing Host-Delivered RNAi Mediated Root-Knot Nematode Resistance

Host-derived gene silencing of parasite fitness genes improves resistance to soybean cyst nematodes in stable transgenic soybean

Contact Info

Product

Resources

About