[논문]Defense-Related Responses in Fruit of the Nonhost Chili Pepper against Xanthomonas axonopodis pv. glycines Infection

Chang, Sung Pae; Jeon, Yong Ho; Kim, Young Ho

doi:10.5423/ppj.oa.12.2015.0256

Defense-Related Responses in Fruit of the Nonhost Chili Pepper against Xanthomonas axonopodis pv. glycines Infection 원문보기

The plant pathology journal, v.32 no.4, 2016년, pp.311 - 320

Chang, Sung Pae (Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University) , Jeon, Yong Ho (Department of Bioresource Sciences, Andong National University) , Kim, Young Ho (Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University)

Abstract ▼ AI-Helper

Xanthomonas axonopodis pv. glycines (Xag) is a necrotrophic bacterial pathogen of the soybean that causes bacterial pustules and is a nonhost pathogen of the chili pepper. In the current study, chili pepper fruit wound inoculated in planta with Xag 8ra formed necrotic lesions on the fruit surface and induced several structural and chemical barriers systemically in the fruit tissue. The initial defense response included programmed cell death of necrotizing and necrotized cells, which was characterized by nuclear DNA cleavage, as detected by TUNEL-confocal laser scanning microscopy (CLSM), and phosphatidylserine exposure on cell walls distal to the infection site, as detected by Annexin V FLUOS-CLSM. These two responses may facilitate cell killing and enhance transportation of cell wall materials used for cell wall thickening, respectively. The cells beneath the necrotic tissue were enlarged and divided to form periclinal cell walls, resulting in extensive formation of several parallel boundary layers at the later stages of infection, accompanying the deposition of wall fortification materials for strengthening structural defenses. These results suggest that nonhost resistance of chili pepper fruit against the nonhost necrotrophic pathogen Xag 8ra is activated systematically from the initial infection until termination of the infection cycle, resulting in complete inhibition of bacterial pathogenesis by utilizing organspecific in situ physiological events governed by the expression of genes in the plant fruit organ.

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문제 정의

However, resistance responses other than HR have not been documented in the nonhost defense response of other organs of the chili pepper against necrotrophic pathogen infection. Therefore, the aim of this study was to investigate the initial and latestage histopathological responses of the chili pepper fruit to Xag infection to better understand the overall defense mechanisms of a nonhost plant in response to a necrotrophic bacterial pathogen.

제안 방법

1% toluidine blue O and observed under a compound light microscope (Axiophot). The embedded samples were sectioned at 80 to 90 nm thickness with a diamond knife on an MT-X ultra-microtome to make ultrathin sections, which were stained with 2% uranyl acetate and lead citrate for 7 minutes each and examined under a JEM-1010 electron microscope (JEOL Ltd., Tokyo, Japan) at an acceleration voltage of 80 kV. At least three samples were observed per treatment each time.
5% uranyl acetate overnight. They were then dehydrated in an ethanol series (30%, 50%, 80%, 95%, and three times of 100%) each for 10 minutes and further dehydrated in propylene oxide for 15 minutes. The dehydrated tissues were embedded in Spurr’s epoxy resin (Spurr, 1969) and allowed to polymerize at 70^oC for 8 hours in an oven.

성능/효과

In conclusion, nonhost resistance of the chili pepper fruit against Xag 8ra infection is activated systematically from the initial infection until termination of the infection cycle, initially inducing PCD and PCD-related defense structures leading to HR and cell wall thickening due to nuclear aberration and PS exposure, respectively, and later in the infection process inducing boundary layer formation beneath the infection site as a structural barrier against further pathogen invasion. Defense-related materials such as lignin, suberin and cutin were deposited onto the thickened and necrotized cell walls, thereby strengthening the cell wall structures.
Our results demonstrate that PS exposure occurred mainly at the cell walls distal to the infection site, bounding occurred with thickened necrotized cell walls, and the middle lamella adjacent to modified cells with an intact cytoplasm contained numerous small vacuoles and thickened cell walls to which vesicles were incorporated at subsequent infection stages (Fig. 2E, F, 4A–D).

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