Skip to main content
SpringerLink
Log in

A chimeric cry8Ea1 gene flanked by MARs efficiently controls Holotrichia parallela

  • Original Paper
  • Published:
Plant Cell Reports Aims and scope Submit manuscript

Abstract

Key message

Peanuts transformed with the synthetic cry8Ea1 gene flanked by MARs are a potentially effective control strategy against white grubs. Cry8Ea1 protein levels of the construct containing MARs were increased by 2.5 times.

Abstract

White grubs are now recognized as the most important pests of peanut worldwide. A synthetic cry8Ea1 gene, which was toxic to Holotrichia parallela larvae, was expressed in chimeric peanut roots using an Agrobacterium rhizogenes-mediated transformation system. The relative mRNA and protein levels of the cry8Ea1 gene were confirmed by quantitative real-time PCR and ELISA, respectively. The effects of matrix attachment regions (MARs) on the expression and activity of the cry8Ea1 gene were analyzed. The average expression level of cry8Ea1 in peanut roots was higher for the plants harboring constructs flanked by MARs from tobacco. Moreover, differing from previous studies, the synthetic cry8Ea1 gene flanked by MARs showed more variation in protein levels than mRNA levels. These composite plants containing cry8Ea1 gene flanked by MARs exhibited a high toxicity against Holotrichia parallela larvae as shown by bioassay analysis, thus offering a potential effective combination to control subterranean insects in peanuts.

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

Similar content being viewed by others

References

  • Ashok B, Swaroop S, Ahuja DB (2012) Field efficacy of neonicotinoid insecticides against white grub (Holotrichia consanguinea Blanch.) on groundnut. Indian J Entomol 74:198–200

    Google Scholar 

  • Cervera M, Navarro A, Navarro L, Peña L (2008) Production of transgenic adult plants from clementine mandarin by enhancing cell competence for transformation and regeneration. Tree Physiol 28:55–66

    Article  PubMed  CAS  Google Scholar 

  • Gatehouse J (2008) Biotechnological prospects for engineering insect-resistant plants. Plant Physiol 146:881–887

    Article  PubMed  CAS  Google Scholar 

  • Geng L, Niu L, Gresshoff P, Shu C, Song F, Huang D, Zhang J (2012) Efficient production of Agrobacterium rhizogenes-transformed roots and composite plants in peanut (Arachis hypogaea L.). Plant Cell Tiss Org Cul 109:491–500

    Article  CAS  Google Scholar 

  • George J, Redmond CT, Royalty RN, Potter DA (2007) Residual effects of imidacloprid on Japanese beetle (Coleoptera: Scarabaeidae) oviposition, egg hatch, and larval viability in turfgrass. J Econ Entomol 100:431–439

    Article  PubMed  CAS  Google Scholar 

  • Gorman C, Arope S, Grandjean M, Girod PA, Mermod N, Al-Rubeai M (2009) Use of MAR elements to increase the production of recombinant proteins. Cell Line Development 6:1–32

    Article  CAS  Google Scholar 

  • Harlow E, Lane D (1999) Using Antibodies: A laboratory manual. Cold Spring Harbor Laboratory, New York, pp 495–496

  • Hilder V, Boulter D (1999) Genetic engineering of crop plants for insect resistance—a critical review. Crop Protect 18:177–191

    Article  Google Scholar 

  • Holmes-Davis R, Comai L (1998) Nuclear matrix attachment regions and plant gene expression. Trends Plant Sci 3:91–97

    Article  Google Scholar 

  • Indrasumunar A, Searle I, Lin M-H, Kereszt A, Men A, Carroll BJ, Gresshoff PM (2011) Nodulation factor receptor kinase 1α controls nodule organ number in soybean (Glycine max L. Merr). Plant J 65:39–50

    Article  PubMed  CAS  Google Scholar 

  • Kereszt A, Li D, Indrasumunar A, Nguyen CDT, Nontachaiyapoom S, Kinkema M, Gresshoff PM (2007) Agrobacterium rhizogenes-mediated transformation of soybean to study root biology. Nat Protocols 2:948–952

    Article  CAS  Google Scholar 

  • Levin JS, Thompson WF, Csinos AS, Stephenson MG, Weissinger AK (2005) Matrix attachment regions increase the efficiency and stability of RNA-mediated resistance to tomato spotted wilt virus in transgenic tobacco. Trans Res 14:193–206

    Article  CAS  Google Scholar 

  • Li Y, Han J, Yu C, Yu W, Mu W (2012) Toxicity and control effect of seven insecticides to Holotrichia parallela. Acta Phytophylacica Sin 39:147–152

    Google Scholar 

  • Liang G, Wu K, Yu H, Li K, Feng X, Guo Y (2008) Changes of inheritance mode and fitness in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) along with its resistance evolution to Cry1Ac toxin. J Invertebr Pathol 97:142–149

    Article  PubMed  CAS  Google Scholar 

  • Liu J, Yan G, Shu C, Zhao C, Liu C, Song F, Zhou L, Ma J, Zhang J, Huang D (2010) Construction of a Bacillus thuringiensis engineered strain with high toxicity and broad pesticidal spectrum against coleopteran insects. Appl Micro Biotechnol 87:243–249

    Article  Google Scholar 

  • Liu X, Nong X, Liu C, Xi G, Zhang X, Zhang Z (2011) Biocontrol of peanut white grubs, Holotrichia parallela, using entomopathogenic fungus Metarhizium anisopliae at sowing period of peanut. Chin J Biol Control 27:485–489

    Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Nong XQ, Liu CQ, Lu X, Wang QL, Wang GJ, Zhang ZH (2011) Laboratory evaluation of entomopathogenic fungi against the white grubs, Holotrichia oblita and Anomala corpulenta (Coleoptera: Scarabaeidae) from the field of peanut, Arachis hypogaea. Biocontrol Sci Technol 21:593–603

    Article  Google Scholar 

  • Nuñez-Valdez ME, Calderón MA, Aranda E, Hernández L, Ramírez-Gama RM, Lina L, Rodríguez-Segura Z, Gutiérrez MdC, Villalobos FJ (2008) Identification of a putative Mexican strain of Serratia entomophila pathogenic against root-damaging larvae of scarabaeidae (Coleoptera). Appl Environ Microbiol 74:802–810

    Article  PubMed  Google Scholar 

  • Oliver JB, Reding ME, Youssef NN, Klein MG, Bishop BL, Lewis PA (2009) Surface-applied insecticide treatments for quarantine control of Japanese beetle, Popillia japonica Newman (Coleoptera: Scarabaeidae), larvae in field-grown nursery plants. Pest Manag Sci 65:381–390

    Article  PubMed  CAS  Google Scholar 

  • Phi-Van L, von Kries JP, Ostertag W, Strätling WH (1990) The chicken lysozyme 5′ matrix attachment region increases transcription from a heterologous promoter in heterologous cells and dampens position effects on the expression of transfected genes. Mol Cell Biol 10:2302–2307

    PubMed  CAS  Google Scholar 

  • Rogers DJ, Ward AL, Wightman JA (2005) Damage potential of two Scarab species on groundnut. Int J Pest Manage 51:305–312

    Article  Google Scholar 

  • Shu C, Yu H, Wang R, Fen S, Su X, Huang D, Zhang J, Song F (2009) Characterization of two novel cry8 genes from Bacillus thuringiensis strain BT185. Current Microbiol 58:389–392

    Article  PubMed  CAS  Google Scholar 

  • Singer S, Liu Z, Cox K (2012) Minimizing the unpredictability of transgene expression in plants: the role of genetic insulators. Plant Cell Rep 31:13–25

    Article  PubMed  CAS  Google Scholar 

  • Singsit C, Adang M, Lynch R, Anderson W, Wang A, Cardineau G, Ozias-Akins P (1997) Expression of a Bacillus thuringiensis cryIA(c) gene in transgenic peanut plants and its efficacy against lesser cornstalk borer. Transgenic Res 6:169–176

    Article  PubMed  CAS  Google Scholar 

  • Srikanth J, Santhalakshmi G, Nirmala R (2011) An improved bioassay method for entomopathogenic fungi of sugarcane pests and its evaluation in studies of virulence in subcultures. Sugar Tech 13:156–165

    Article  CAS  Google Scholar 

  • Tiwari S, Mishra D, Singh A, Singh PK, Tuli R (2008) Expression of a synthetic cry1Ec gene for resistance against Spodoptera litura in transgenic peanut (Arachis hypogaea L.). Plant Cell Rep 27:1017–1025

    Article  PubMed  CAS  Google Scholar 

  • Tiwari S, Mishra DK, Chandrasekhar K, Singh PK, Tuli R (2011) Expression of δ-endotoxin Cry1EC from an inducible promoter confers insect protection in peanut (Arachis hypogaea L.) plants. Pest Manag Sci 67:137–145

    Article  PubMed  CAS  Google Scholar 

  • USDA (2012) World Agricultural Production. WAP 10-10 edn. United States department of agriculture

  • Wang T, Xue L, Hou W, Yang B, Chai Y, Ji X, Wang Y (2007) Increased expression of transgene in stably transformed cells of Dunaliella salina by matrix attachment regions. Appl micro biotechnol 76:651–657

    Article  CAS  Google Scholar 

  • Wightman JA, Brier HB, Wright GC (1994) The effect of root damage caused by simulated white grub attack on the growth, yield and water-use of groundnut plants. Plant Soil 160:267–275

    Article  Google Scholar 

  • Ma S, Wang A (2012) Molecular farming in plants: an overview. In: Molecular farming in plants: recent advances and future prospects. Springer, Netherlands, pp 1–20

    Book  Google Scholar 

  • Zhang J, Lu L, Ji L, Yang G, Zheng C (2009) Functional characterization of a tobacco matrix attachment region-mediated enhancement of transgene expression. Trans Res 18:377–385

    Article  Google Scholar 

Download references

Acknowledgments

The Holotrichia parallela larvae were kindly provided by Cangzhou Academy of Agricultural and Forestry Sciences in China. This study was supported by 863 Projects of China (2011AA10A203), the Key Project of Chinese National Programs for Fundamental Research and Development (973 Program) (2009CB118902), and the National Transgenic Major Program (2013ZX08009003-001-004 and 2013ZX08004-004-009).

Author information

Authors and Affiliations

  1. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, 100193, China

    Lili Geng, Jing Chi, Changlong Shu, Fuping Song & Jie Zhang

  2. ARC Centre of Excellence for Integrative Legume Research, The University of Queensland, St. Lucia, Brisbane, QLD, 4072, Australia

    Peter M. Gresshoff

  3. Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

    Dafang Huang

Authors
  1. Lili Geng
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jing Chi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Changlong Shu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Peter M. Gresshoff
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Fuping Song
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Dafang Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jie Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jie Zhang.

Additional information

Communicated by P. Lakshmanan.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Geng, L., Chi, J., Shu, C. et al. A chimeric cry8Ea1 gene flanked by MARs efficiently controls Holotrichia parallela . Plant Cell Rep 32, 1211–1218 (2013). https://doi.org/10.1007/s00299-013-1417-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00299-013-1417-2

Keywords

Search

Navigation