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Remusatia vivipara lectin and Sclerotium rolfsii lectin interfere with the development and gall formation activity of Meloidogyne incognita in transgenic tomato

  • Yogesh S. BhagatEmail author
  • Ramesh S. Bhat
  • Rohini M. Kolekar
  • Ashlesha C. Patil
  • S. Lingaraju
  • R. V. Patil
  • S. S. Udikeri
Original Paper
  • 29 Downloads

Abstract

Root knot nematodes are serious threats to growth and yield of solaneous crops including tomato. In this study, a binary vector carrying Remusatia vivipara (rvl1) and Sclerotium rolfsii (srl1) lectin genes were introduced independently into Lycopersicon esculentum cv. Pusa Ruby via Agrobacterium tumefaciens for resistance against root knot nematode, Meloidogyne incognita. In total, one hundred and one rvl1 and srl1-transformed plants exhibiting kanamycin resistance were confirmed to carry transgenes as detected by polymerase chain reaction (PCR) with 4.59% transformation efficiency. Genetic analysis of T1 progeny confirmed Mendelian segregation of the introduced genes. Three events each of rvl1 and srl1 transgenic tomato were randomly selected for further confirmation by Southern and TAIL-PCR analyses. All three events of srl1 transgenics showed single copy transgene, whereas two rvl1 transgenic events showed single copy of transgene, while remaining event showed two copies of transgenes. Site of integration obtained for rvl1 and srl1 transgenic events by TAIL-PCR revealed that all the three events of rvl1 and srl1 transgenics differed for their site of integration and insertion sites did not contain any predicted gene. Moreover, expression of the rvl1 and srl1 transgenes was detected by haemagglutination assay in all three events of rvl1 and srl1, but not in non-transgenic tomato plant. Homozygous progenies of these events were grown and inoculated with M. incognita. Development and reproduction of M. incognita was severely affected in transgenic tomato plants expressing RVL1 and SRL1 exhibiting the high levels of resistance compared to non-transgenic plants. Therefore, these transgenic lines demonstrate a promising potential for variety development of tomato lines with enhanced resistance against M. incognita.

Keywords

Remusatia vivipara lectin Sclerotium rolfsii lectin Transgenic tomato Resistance to root knot nematode 

Notes

Acknowledgements

The authors thank Dr. Ramesh Aggarwal (CCMB, Hyderabad, India) and Dr. Thumballi Ganapathi (BARC, Mumbai, India) for useful discussions and for providing facility to carry out southern hybridization. We thank DBT, New Delhi (India) and UAS, Dharwad (India) for financial support to carry out this research work.

Supplementary material

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Supplementary material 1 (DOC 3071 kb)
11248_2019_121_MOESM2_ESM.doc (82 kb)
Supplementary material 2 (DOC 81 kb)

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© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.Department of Biotechnology, College of AgricultureUniversity of Agricultural Sciences, DharwadDharwadIndia
  2. 2.Department of Biotechnology, College of AgricultureUniversity of Agricultural Sciences, BangaloreBengaluruIndia
  3. 3.Insititute of Organic FarmingUniversity of Agricultural Sciences, DharwadDharwadIndia
  4. 4.Department of Horticulture, College of Agriculture, BijapurUniversity of Agricultural SciencesDharwadIndia
  5. 5.Agriculture Research Station, Dharwad FarmUniversity of Agricultural Sciences, DharwadDharwadIndia

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