Pathogenesis-related proteins induced by agroinoculationassociated cell wall weakening can be obviated by spray-on inoculation or mannitol ex vivo culture

다국어 초록 (Multilingual Abstract)

Agroinoculation transient expression systems are commonly accompanied with elevated pathogenesisrelated (PR) protein production and leaf necrosis. We identified the major PR proteins in Nicotiana benthamiana in response to agroinoculation and determined that their occurrence was mainly due to agrobacterium infection and the method of inoculation, rather than due to viral vectors overexpressing foreign proteins. A spray-on inoculation method was optimized in this research and used to obviate the leaf necrosis and PR proteins induced by agrobacterium.
Subsequently, this method also increased the yield and purity of the protein-of-interest. A further investigation of PR protein induction by a necrosis-inducing protein, Jun a 1, suggested that the plant pathogenic response was related to biochemical integrity of plant cell wall, which was also confirmed by an osmo-stabilizing mannitol ex vivo culture experiment. These findings provide insight into the response of plants to agroinoculation and suggested a connection between cell wall weakening and PR protein elicitation.

참고문헌 (Reference)

1 변의백, "β-(1,3)-Glucan Isolated from Agrobacterium Species Induces Maturation of Bone Marrow-derived Dendritic Cells and Drives Th1 Immune Responses" 한국식품과학회 24 (24): 1533-1540, 2015

2 Bayley ST, "The influence of mannitol and auxin on growth of cell walls in avena coleoptiles" 21 : 633-641, 1957

3 J. A. L. van Kan, "Structure of tobacco genes encoding thaumatin-like proteins" Springer Nature 12 (12): 153-155, 1989

4 Zun Liu, "Plant-Expressed Recombinant Mountain Cedar Allergen Jun a 1 Is Allergenic and Has Limited Pectate Lyase Activity" S. Karger AG 153 (153): 347-358, 2010

5 Zhou Y, "Plant expression of trans-encapsidated viral nanoparticle vaccines with animal RNA replicons methods in molecular biology" 1499 : 77-86, 2017

6 Collmer A, "Pectic enzyme production and bacterial plant pathogenicity" Springer 1 : 65-72, 1990

7 Edreva A, "Pathogenesis-related proteins: research progress in the last 15 years" 31 : 105-124, 2005

8 Zhou N, "PAD4functions upstream from salicylic acid to control defense responses in Arabidopsis" 10 : 1021-1030, 1998

9 Hu H, "Localization of boron in cell walls of squash and tobacco and its association with pectin (evidence for a structural role of boron in the cell wall)" 105 : 681-689, 1994

10 Neff NT, "Inhibitory effects of a pectinenriched tomato cell wall fraction on agrobacterium tumefaciens binding and tumor formation" 83 : 525-528, 1987

11 Yiyang Zhou, "In planta Production of Flock House Virus Transencapsidated RNA and Its Potential Use as a Vaccine" Springer Nature 57 (57): 325-336, 2015

12 John A Lindbo, "High-efficiency protein expression in plants from agroinfection-compatible Tobacco mosaic virus expression vectors" Springer Nature 7 (7): 52-, 2007

13 Sabine Munch-Garthoff, "Expression of ?-1,3-glucanase and chitinase in healthy, stem-rust-affected and elicitor-treated near-isogenic wheat lines showingSr5-orSr24-specified race-specific rust resistance" Springer Nature 201 (201): 235-244, 1997

14 Wan J, "Chitin signaling and plant disease resistance" 3 : 831-833, 2008

15 Van Camp W, "Characterization of iron superoxide dismutase cDNAs from plants obtained by genetic complementation in Escherichia coli" 87 : 9903-9907, 1990

16 Ueda K, "Cell wall development of Micrasterias americana, especially in isotonic and hypertonic solutions" 21 : 617-631, 1976

17 Kauffmann S, "Biological function of ‘pathogenesis-related’ proteins: four PR proteins of tobacco have 1,3-beta-glucanase activity" 6 : 3209-3212, 1987

18 Legrand M, "Biological function of pathogenesis-related proteins: four tobacco pathogenesis-related proteins are chitinases" 84 : 6750-6754, 1987

19 Bent AF, "Arabidopsis in planta transformation. Uses, mechanisms, and prospects for transformation of other species" 124 : 1540-1547, 2000

20 Zun Liu, "An efficient Foxtail mosaic virus vector system with reduced environmental risk" Springer Nature 10 (10): 88-, 2010

21 Sukanya S. Rao, "Agrobacterium adherence involves the pectic portion of the host cell wall and is sensitive to the degree of pectin methylation" Wiley-Blackwell 56 (56): 374-380, 1982

22 Zun Liu, "A tobamovirus expression vector for agroinfection of legumes and Nicotiana" Elsevier BV 147 (147): 151-159, 2010

23 Cornelissen BJC, "A tobacco mosaic virus-induced tobacco protein is homologous to the sweet-tasting protein thaumatin" 321 : 531-532, 1986

24 Simone Hahn, "A novel and fully scalable Agrobacterium spray-based process for manufacturing cellulases and other cost-sensitive proteins in plants" Wiley-Blackwell 13 (13): 708-716, 2015

25 Taiko Tutumi, "A cytochemical study on polysaccharides in cells of micrasterias americana" International Society of Cytology 40 (40): 113-118, 1975