Overexpression of a New Chitinase Gene EuCHIT2 Enhances Resistance to Erysiphe cichoracearum DC in Tobacco Plants

Int J Mol Sci. 2017 Nov 7;18(11):2361. doi: 10.3390/ijms18112361.

Abstract

In this study, we cloned a new chitinase gene, EuCHIT2, from Eucommia ulmoides Oliver (E. ulmoides) using rapid amplification of cDNA ends (RACE) technology and constructed an overexpression vector, pSH-35S-EuCHIT2, to introduce it into tobacco (Nicotiana tabacum cv. Xanthi). Resistance to Erysiphe cichoracearum de Candolle (E.cichoracearum DC) and molecular mechanisms in the transgenic tobacco were determined by drop inoculation, spore counting, determination of physicochemical indicators, and analysis of gene expression. The chitinase activity and resistance to E. cichoracearum DC were significantly higher in the transgenic tobacco than in wild-type tobacco (p < 0.05). The activities of peroxidase (POD) and catalase (CAT), after inoculation with E. cichoracearum DC, were higher in the transgenic tobacco than in the wild-type. Conversely, the malondialdehyde (MDA) content was significantly lower in the transgenic tobacco than the wild-type before and after inoculation. In addition, our study also indicated that the resistance to E. cichoracearum DC might involve the salicylic acid (SA) and jasmonic acid (JA) pathways, because the expression levels of pathogenesis-related gene 1 (PR-1a) and coronatine-insensitive 1 (COI1) were significantly increased and decreased, respectively, after inoculation with E. cichoracearum DC. The present study supports the notion that PR-1a and POD participate in resistance to E. cichoracearum DC in the transgenic tobacco plants.

Keywords: Erysiphe cichoracearum DC; Eucommia ulmoides; chitinase; transgenic tobacco.

MeSH terms

  • Chitinases / genetics
  • Chitinases / metabolism*
  • Cyclopentanes / metabolism
  • Nicotiana / genetics
  • Nicotiana / metabolism*
  • Oxylipins / metabolism
  • Plant Diseases / genetics
  • Plant Proteins / genetics
  • Plant Proteins / metabolism
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / metabolism
  • Salicylic Acid / metabolism

Substances

  • Cyclopentanes
  • Oxylipins
  • Plant Proteins
  • jasmonic acid
  • Chitinases
  • Salicylic Acid