Skip to main content
SpringerLink
Log in

Cloning and functional analyses of pepper CaRKNR involved in Meloidogyne incognita resistance

  • Published:
Euphytica Aims and scope Submit manuscript

Abstract

Root-knot nematodes (Meloidogyne spp.) are destructive pests of crops. Pepper (Capsicum annuum L.) contains genes that control resistance to root-knot nematodes. Using suppression subtractive hybridization and RACE strategies, a nucleotide-binding site and leucine-rich repeat (NBS-LRR) family gene, CaRKNR (FJ231739), was isolated and cloned from the nematode-resistant pepper line HDA149. CaRKNR is a novel NBS-LRR gene with an open reading frame of 3600 bp that is homologous (70.45 % identity) to the gene Mi-1.2. After Meloidogyne incognita inoculation, real-time qPCR showed that the CaRKNR expression level was increased from 0.63 to 2.16 times. Using the virus-induced gene silencing system, the CaRKNR gene’s expression level was reduced significantly than controls, and the average numbers of galls and egg masses in silenced seedlings were 44.39 and 42.01, respectively, while the controls were 0.13. This study revealed that CaRKNR was induced by M. incognita and its expression correlated with pepper resistance against root-knot nematodes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

Abbreviations

ARC:

APAF-1, R proteins, and CED-4

Blast:

Basic local alignment search tool

LRR:

Leucine-rich repeat

NB:

Nucleotide binding

NJ:

Neighbour-joining

ORF:

Open reading frame

Pfam:

Protein family

RACE:

Rapid amplification of cDNA end

RKN:

Root-knot nematode (Meloidogyne spp.)

RT-qPCR:

Reverse transcriptase quantitative polymerase chain reaction

SSH:

Suppression subtractive hybridization

VIGS:

Virus induced gene silencing

References

  • Abad P, Gouzy J, Aury JM, Castagnone-Sereno P, Danchin EG, Deleury E, Perfus-Barbeoch L, Anthouard V, Artiguenave F, Blok VC (2008) Genome sequence of the metazoan plant-parasitic nematode Meloidogyne incognita. Nat Biotechnol 26:909–915

    Article  CAS  PubMed  Google Scholar 

  • Ammiraju JS, Veremis JC, Huang X, Roberts PA, Kaloshian I (2003) The heat-stable root-knot nematode resistance geneMi-9 from Lycopersicon peruvianum is localized on the short arm of chromosome 6. Theor Appl Genet 106:478–484

    CAS  PubMed  Google Scholar 

  • Belkhadir Y, Subramaniam R, Dangl JL (2004) Plant disease resistance protein signaling: NBS-LRR proteins and their partners. Curr Opin Plant Biol 7:391–399

    Article  CAS  PubMed  Google Scholar 

  • Bleve-Zacheo T, Bongiovanni M, Melillo MT, Castagnone-Sereno P (1998) The pepper resistance genes Me1 and Me3 induce differential penetration rates and temporal sequences of root cell ultrastructural changes upon nematode infection. Plant Sci 133:79–90

    Article  CAS  Google Scholar 

  • Chen R, Li H, Zhang L, Zhang J, Xiao J, Ye Z (2007) CaMi, a root-knot nematode resistance gene from hot pepper (Capsium annuum L.) confers nematode resistance in tomato. Plant Cell Rep 26:895–905

    Article  CAS  PubMed  Google Scholar 

  • Claverie M, Dirlewanger E, Cosson P, Bosselut N, Lecouls A, Voisin R, Kleinhentz M, Lafargue B, Caboche M, Chalhoub B (2004) High-resolution mapping and chromosome landing at the root-knot nematode resistance locus Ma from Myrobalan plum using a large-insert BAC DNA library. Theor Appl Genet 109:1318–1327

    Article  CAS  PubMed  Google Scholar 

  • Djian-Caporalino C, Pijarowski L, Fazari A, Samson M, Gaveau L, O’byrne C, Lefebvre V, Caranta C, Palloix A, Abad P (2001) High-resolution genetic mapping of the pepper (Capsicum annuum L.) resistance loci Me3 and Me4 conferring heat-stable resistance to root-knot nematodes (Meloidogyne spp.). Theor Appl Genet 103:592–600

    Article  CAS  Google Scholar 

  • Djian-Caporalino C, Fazari A, Arguel M, Vernie T, VandeCasteele C, Faure I, Brunoud G, Pijarowski L, Palloix A, Lefebvre V (2007) Root-knot nematode (Meloidogyne spp.) Me resistance genes in pepper (Capsicum annuum L.) are clustered on the P9 chromosome. Theor Appl Genet 114:473–486

    Article  CAS  PubMed  Google Scholar 

  • Ernst K, Kumar A, Kriseleit D, Kloos DU, Phillips MS, Ganal MW (2002) The broad-spectrum potato cyst nematode resistance gene (Hero) from tomato is the only member of a large gene family of NBS-LRR genes with an unusual amino acid repeat in the LRR region. Plant J 31:127–136

    Article  CAS  PubMed  Google Scholar 

  • Gilbert JC, McGuire DC (1956) Inheritance of resistance to severe root knot from Meloidogyne incognitain commercial type tomatoes. Proc Am Soc Hortic Sci 68:437–442

    Google Scholar 

  • González-Candelas L, Alamar S, Sánchez-Torres P, Zacarías L, Marcos J (2010) A transcriptomic approach highlights induction of secondary metabolism in citrus fruit in response to Penicillium digitatum infection. BMC Plant Biol 10:194–211

    Article  PubMed Central  PubMed  Google Scholar 

  • Huang X, McGiffen M, Kaloshian I (2004) Reproduction of Mi-virulent Meloidogyne incognita isolates on Lycopersicon spp. J Nematol 36(1):69–75

    PubMed Central  CAS  PubMed  Google Scholar 

  • Jablonska B, Ammiraju JS, Bhattarai KK, Mantelin S, de Ilarduya OM, Roberts PA, Kaloshian I (2007) The Mi-9 gene from Solanum arcanum conferring heat-stable resistance to root-knot nematodes is a homolog of Mi-1. Plant Physiol 143:1044–1054

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Kim S, Park M, Yeom SI, Kim YM, Lee JM, Lee HA, Seo E, Choi J, Cheong K, Kim KT (2014) Genome sequence of the hot pepper provides insights into the evolution of pungency in Capsicum species. Nat Genet. doi:10.1038/ng.2877

    Google Scholar 

  • Liu Y, Schiff M, Dinesh-Kumar S (2002) Virus-induced gene silencing in tomato. Plant J 31:777–786

    Article  CAS  PubMed  Google Scholar 

  • López-pérez M, Ballester AR, González-candelas L (2014) Identification and functional analysis of Penicillium digitatum genes putatively involved in virulence towards citrus fruit. Mol Plant Pathol. doi:10.1111/mpp.1217

    PubMed  Google Scholar 

  • Meyers BC, Kozik A, Griego A, Kuang H, Michelmore RW (2003) Genome-wide analysis of NBS-LRR encoding genes in Arabidopsis. Plant Cell Online 15:809–834

    Article  CAS  Google Scholar 

  • Milligan SB, Bodeau J, Yaghoobi J, Kaloshian I, Zabel P, Williamson VM (1998) The root knot nematode resistance gene Mi from tomato is a member of the leucine zipper, nucleotide binding, leucine-rich repeat family of plant genes. Plant Cell Online 10:1307–1319

    Article  CAS  Google Scholar 

  • Nombela G, Williamson VM, Muniz M (2003) The root-knot nematode resistance gene Mi-1.2 of tomato is responsible for resistance against the whitefly Bemisia tabaci. Mol Plant Microbe Interact 16:645–649

    Article  CAS  PubMed  Google Scholar 

  • Paal J, Henselewski H, Muth J, Meksem K, Menéndez CM, Salamini F, Ballvora A, Gebhardt C (2004) Molecular cloning of the potato Gro1-4 gene conferring resistance to pathotype Ro1 of the root cyst nematode Globodera rostochiensis, based on a candidate gene approach. Plant J 38:285–297

    Article  CAS  PubMed  Google Scholar 

  • Qin C, Yu C, Shen Y, Fang X, Chen L, Min J, Cheng J, Zhao S, Xu M, Luo Y, Yang Y, Wu Z, Mao L, Wu H, Ling-Hu C, Zhou H, Lin H, González-Morales S, Trejo-Saavedra DL, Tian H, Tang X, Zhao M, Huang Z, Zhou A, Yao X, Cui J, Li W, Chen Z, Feng Y, Niu Y, Bi S, Yang X, Li W, Cai H, Luo X, Montes-Hernández S, Leyva-González MA, Xiong Z, He X, Bai L, Tan S, Tang X, Liu D, Liu J, Zhang S, Chen M, Zhang L, Zhang L, Zhang Y, Liao W, Zhang Y, Wang M, Lv X, Wen B, Liu H, Luan H, Zhang Y, Yang S, Wang X, Xu J, Li X, Li S, Wang J, Palloix A, Bosland PW, Li Y, Krogh A, Rivera-Bustamante RF, Herrera-Estrella L, Yin Y, Yu J, Hu K, Zhang Z (2014) Whole-genome sequencing of cultivated and wild peppers provides insights into Capsicum domestication and specialization. PNAS 111(14):5135–5140

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Rossi M, Goggin FL, Milligan SB, Kaloshian I, Ullman DE, Williamson VM (1998) The nematode resistance gene Miof tomato confers resistance against the potato aphid. Proc Natl Acad Sci USA 95:9750–9754

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Thies JA, Fery RL (1998) Modified expression of the N gene for southern root-knot nematode resistance in pepper at high soil temperatures. J Am Soc Hortic Sci 123:1012–1015

    CAS  Google Scholar 

  • Tytgat T, Meutter JD, Gheysen G, Coomans A (2000) Sedentary endoparasitic nematodes as a model for other plant parasitic nematodes. Nematology 2:113–121

    Article  Google Scholar 

  • Valentine T, Shaw J, Blok VC, Phillips MS, Oparka KJ, Lacomme C (2004) Efficient virus-induced gene silencing in roots using a modified tobacco rattle virus vector. Plant Physiol 136:3999–4009

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Van Der Vossen EA, Der Voort V, Rouppe JN, Kanyuka K, Bendahmane A, Sandbrink H, Baulcombe DC, Bakker J, Stiekema WJ, Klein-Lankhorst RM (2000) Homologues of a single resistance-gene cluster in potato confer resistance to distinct pathogens: a virus and a nematode. Plant J 23:567–576

    Article  PubMed  Google Scholar 

  • Zheng J, Zou X, Mao Z, Xie B (2011) A novel pepper (Capsicum annuum L.) WRKY gene, CaWRKY30, is involved in pathogen stress responses. J Plant Biol 54:329–337

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We thank INRA Versailles for providing the pepper HDA149. This work was supported by the National Science Foundation of China (30971905, 31101425), Agro-scientific Research in the Public Interest (201103018) and the China Agriculture Research System (CARS-25).

Author information

Authors and Affiliations

  1. Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Key Laboratory of Biology and Genetic Improvement of Horticultural Crops, Ministry of Agriculture, 12 Zhongguancun Southern Street, Haidian, Beijing, 100081, China

    Zhenchuan Mao, Pingping Zhu, Feng Liu, Yonghong Huang, Jian Ling, Guohua Chen, Yuhong Yang, Dongxin Feng & Bingyan Xie

Authors
  1. Zhenchuan Mao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Pingping Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Feng Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yonghong Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jian Ling
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Guohua Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yuhong Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Dongxin Feng
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Bingyan Xie
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Bingyan Xie.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 1886 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Mao, Z., Zhu, P., Liu, F. et al. Cloning and functional analyses of pepper CaRKNR involved in Meloidogyne incognita resistance. Euphytica 205, 903–913 (2015). https://doi.org/10.1007/s10681-015-1438-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10681-015-1438-8

Keywords

Search

Navigation