Proceedings of the Korean Society of Plant Pathology Conference
/
2003.10a
/
pp.89.1-89
/
2003
Disease suppression by ectomycorrhizal(ECM) fungi has been demonstrated on red pine seedlings. Culturing of pathogenic fungi on petri plates containing culture filtrates of ECM fungi showed that culture filtrates of the ECM fungus Hebeloma cylindrosporum may inhibit the mycelial growth of all tested soil-borne plant pathogenic(SBPP) fungi upto 60%, In order to examine the effects of ECM fungi on SBPP fungi and on red pine seedlings, both symbiotic and pathogenic fungi were inoculated into the soil with red pine seedlings by three inoculation methods; pre-inoculation of SBPP fungi 10 days before inoculation of ECM fungi, simultaneous inoculation of both fungi, post-inoculation of SBPP fungi 60 days after inoculation of ECM fungi. Seedling mortality, seedling growth, and ectomycorrhizal formation by the combined treatments were examined and compared. Pine seedlings were dead by the pre-inoculation of pathogenic fungi, except Rhizina undulate which required 9-12 days, within 6 days after inoculation. Among pathogenic fungi tested, Fusarium oxysporum was the most pathogenic with the mortality of 44%. However, no dead seedlings were shown by simultaneous inoculation of both fungi or pre-inoculation of ECM fungi. In addition, pine seedlings treated by simultaneous or post-inoculation of SBPP fungi were relatively higher than those treated by pre-inoculation in diameter at root crown and the number of ectomycorrhizal roots. There were no significant differences among inoculation methods in root length and dry weight of treated seedlings. It means that ECM fungi somehow play a role in protecting primary roots of red pine seedlings against invasion by the SBPP fungi.
Infection structures were observed using a fluorescence microscope at the penetration sites on the leaves of potato plants pre-inoculated with the bacterial strains Pseudomonas putida TRL2-3, Micrococcus luteus TRK2-2, and Flexibacteraceae bacterium MRL412, which mediated an induced systemic resistance on potato plants against late blight disease caused by Phytophthora infestans. In order to compare the infection structures on the leaves expressing systemic acquired resistance, the leaves of potato plants pre-treated with DL-3-amino butyric acid (BABA) were also observed after challenge inoculation with the same pathogen. The infection structures were investigated. The total number of germination and appressorium formation of P. infestans were counted. Furthermore, the frequencies of fluorescent epidermal cells at the penetration sites, which indicate a defense response of plant cell, were estimated. There were no differences on the germination rates of the fungal cysts among the untreated control, BABA pre-treated, and bacterial strains pre-inoculated plants. However, appressorium formation was slightly decreased on the leaves of BABA pre-treated plants compared to those of untreated as well as bacterial strains pre-inoculated plants. Furthermore, the frequencies of fluorescent cells of BABA pre-treated and bacterial strains pre-inoculated were higher than that of untreated plants, indicating an active defense reaction of the host cells against the fungal attack. On the other hand, the pre-treatment with BABA caused a stronger fluorescent of epidermal cells at the penetration sites compared to the pre-inoculation with the bacterial strains. Interestingly, the frequency of fluorescent cells by BABA, however, was lower than that by the bacterial strains. Based on the results it is suggested that the infection structures showing resistance reaction on the leaves of potato plants were different between by pre-inoculation with bacterial strains and by pre-treatment with BABA against the late blight pathogen.
Suralta, Roel R.;Cruz, Jayvee A.;Cabral, Maria Corazon J.;Niones, Jonathan M.;Yamauchi, Akira
Proceedings of the Korean Society of Crop Science Conference
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2017.06a
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pp.231-231
/
2017
Drought limits rice production under upland condition. This study quantified the effect of rhizobacteria inoculation on rice root system developmental response to drought and its role in maintaining high soil water use, and dry matter production under drought using NSIC Rc192 (rainfed lowland rice variety). The source of inoculant was Streptomyces mutabilis, a recently isolated rhizobacteria containing plant growth promoting compounds such as ACC deaminase, indole-3-acetic acid and phosphatase (Cruz et al., 2014, 2015). In the first experiment, pre-germination inoculation of seeds with S. mutabilis significantly increased the shoot and root (radicle) length as well as root hair lengths, relative to the non-inoculated control. In the second experiment, rice plants inoculated with S. mutabilis and grown in rootbox with soil generally had greater total root length under drought regardless of the timing of inoculations, relative to the non-inoculated control. Consequently, improved root system development contributed to the increase in soil water uptake under drought and thus, dry matter production. Among inoculation treatments, one-time inoculation of S. mutabilis either at pre-germination or pre-drought stress at 14 days after sowing (DAS), had significantly greater shoot dry matter production than three-time inoculation at pre-germination, at thinning (3 DAS) and at pre-drought (14 DAS). This study demonstrated the effectiveness of rhizobacteria (S. mutabilis) containing growth promoting compounds for enhancing drought dehydration avoidance root traits and improving the growth of rice plants under drought condition.
Kim, Ki-Woo;Lee, In-Jung;Kim, Chang-Soo;Eom, In-Yong;Choi, Joon-Weon;Lee, Don-Koo;Park, Eun-Woo
The Plant Pathology Journal
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v.26
no.4
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pp.394-401
/
2010
Resin flow, symptom development, and lignin biosynthesis in response to wounding and fungal inoculation were investigated in Pinus rigida and Pinus densiflora. The two-year-old seedling stems were subjected to three types of treatments: (i) wounding without inoculation, (ii) wound-inoculation with a conidial suspension of Fusarium circinatum, and (iii) pre-wounding woundinoculation with the fungus 20 days after the initial wounding. Resin flow from wounding sites was more evident in P. rigida than P. densiflora in all treatments. The wound-inoculation with the fungus induced almost two-fold higher levels of resin flow than the other treatments in both species. The pre-wounding woundinoculation appeared to result in a decrease in pitch canker development in the two pine species. Some reductions in disease severity were observed in the prewounding wound-inoculated P. rigida, showing a mean disease severity of less than 85%, compared with approximately 100% in the wound-inoculated stems. Disease severity was approximately 50% in the woundinoculated P. densiflora, whereas 10% in the pre-wounding wound-inoculated stems. Higher amounts of lignin were found from bark (ca. 40%) than from xylem (ca. 30%). The wound-inoculated bark and the pre-wounding wound-inoculated bark exhibited higher amounts of lignin among the other treatments. These results suggest that the wound-inoculation apparently prompt the increase in resin flow and lignin biosynthesis from the two pine species, and the prior wounding may be involved in decreased disease severity against the further invasion of F. circinatum.
Bioassay techniques using young leaves and roots were developed to screen resistance of Atractylis spp. against Phytophthora drechsleri. Among 638 plants collected from various regions of Korea from 1994 to 1996, 67 were pre-screened in fields naturally infested with P. drechsleri, which is the causal pathogen of rhizome rot of Atractylis. Among the pre-screened sources, 18 (ca. 26.8%) were highly resistant to the pathogen in leaf inoculation. In the root inoculation test, abundant sporangia were formed in susceptible plant roots, while only a few or no sporangia were produced on the roots which were found resistant in the leaf inoculation test. Among the selected resistant plants, A. japonica 96066 and 96104 were used to cross with another species, A. macrocephala 96362 that showed high yield with good quality of rhizome but susceptible to the pathogen. The F$_1$hybrids designated as HA03 turned out to be resistant to the pathogen, indicating that resistant gene(s) was inherited. Among intra-species hybrids of A. japonica, HA07 and HA09 were resistant to the pathogen in leaf inoculation and moderate in root inoculation. However, HA08 was susceptible in both inoculation tests. This result suggests that the parent material might be genetically heterogeneous. Further genetic study should be carried out to verify this phenomenon.
Infection structures were observed at the penetration sites on the leaves of cucumber plants inoculated with Colletotrichum orbiculare using a fluorescence microscope. The cucumber plants were previously drenched with suspension of bacterial strains Pseudomonas putida or Micrococcus luteus. The plants pre-inoculated with both bacterial strains were resistant against anthracnose after inoculation with C. orbiculare. To investigate the resistance mechanism by both bacterial strains, the surface of infected leaves was observed at the different time after challenge inoculation. At 3 days after inoculation there were no differences in the germination and appressorium formation of conidia of C. orbiculare as well as in the callose formation of the plants between both bacteria pre-inoculated and non-treated. At 5 days, the germination and appressorium formation of the fungal conidia were, however, significantly decreased on the leaves of plants pre-inoculated with M. luteus at the concentration with $1.0{\times}10^7\;cfu/ml$. Furthermore, callose formation of plants cells at the penetration sites was apparently increased. In contrast, there were no defense reactions of the plants at the concentration with $1.0{\times}10^6\;cfu/ml$ of M. luteus. Similarly, inoculation P. putida caused no plant resistance at the low concentration, whereas increase of callose formation was observed at the higher concentration. The results of this study suggest that the resistant mechanisms might be differently expressed by the concentration of pre-treatment with bacterial suspension.
Defense mechanisms against anthracnose disease caused by Colletotrichum orbiculare on the leaf surface of cucumber plants after pre-treatment with plant growth promoting rhizobacteria(PGPR), amino salicylic acid(ASA) or C. orbiculare were compared using a fluorescence microscope. Induced systemic resistance was mediated by the pre-inoculation in the root system with PGPR strain Bacillus amylolquefaciens EXTN-1 that showed direct antifungal activity to C. gloeosporioides and C. orbiculare. Also, systemic acquired resistance was triggered by the pre-treatments on the bottom leaves with amino salicylic acid or conidial suspension of C. orbiculare. The protection values on the leaves expressing SAR were higher compared to those expressing ISR. After pre-inoculation with PGPR strains no change of the plants was found in phenotype, while necrosis or hypersensitive reaction(HR) was observed on the leaves of plants pre-treated with ASA or the pathogen. After challenge inoculation, inhibition of fungal growth was observed on the leaves expressing both ISR and SAR. HR was frequently observed at the penetration sites of both resistance-expressing leaves. Appressorium formation was dramatically reduced on the leaves of plants pre-treated with ASA, whereas EXTN-1 did not suppress the appressorium formation. ASA also more strongly inhibited the conidial germination than EXTN-1. Conversely, EXTN-1 significantly increased the frequency of callose formation at the penetration sites, but ASA did not. The defense mechanisms induced by C. orbiculare were similar to those by ASA. Based on these results it is suggested that resistance mechanisms on the leaf surface was different between on the cucumber leaves expressing ISR and SAR, resulting in the different protection values.
Induction of systemic acquired resistance (SAR) against phytophthora blight and pathogenesis-related (PR) protein accumulation by TMV infection in pepper plant (Capsicum annuum cv. Nockwang) were examined to understand the mechanism of the systemic acquired resistance in pepper plant. The zoospore suspension of Phytophthora capsici was inoculated on stem of pepper plant in which TMV-pepper strain had been inoculated on fully expanded upper leaves, and thephytopha blight incidence was examined. Both disease severity and lesion length of phytophthora blight were much smaller in TMV pre-inoculated pepper plant than in uninoculated control plants. The phytophthora blight incidence was decreased about 50% in the TMV pre-inoculated pepper, compared to the uninoculated control plant at 10 days after P. capsici inoculation. Accumulation of PR1 and PR5 proteins in intercellular fluid of TMV-inoculated and uninoculated upper leaves were monitored by immuno-blot with tobacco P1b and PR5a, antibody during induction of SAR. PR1 and PR5 were detected from 24 hours after TMV inoculation in both TMV-inoculated and uninouclated upper leaves, and increased rapidly in TMV-inoculation in uninoculated upper leaves were defoliated. PR5 could be detected upto 20 days after TMV inoculation in uninoculated upper leaves. These results suggest that TMV infection induces SAR against phytophthora blight in pepper plant, and that PR proteins are accumulated very rapidly during induction of SAR and maintained for quite long time in pepper plant.
Pinewood nematode (PWN: Bursaphelenchus xylophilus) is known to kill pine tree species that are indigenous to countries where the pest was inadvertently imported, but some cultures from the extraction of dead pines do not damage trees. Experiments were conducted to examine the effect of pre-inoculation of these low-pathogenic pinewood nematode on resistance of pine trees against the pest species. The pre-inoculated pine saplings showed induced resistance which lasted for a year, and repeated inoculation of these low-pathogenic nematodes enhanced tree resistance. All nematode samples extracted from dying or dead pines that had been killed not more than three months before the extraction were pathogenic, and most of those extracted from pines that had been killed 2-3 years before were low-pathogenic. When inoculated in pine saplings, number of low-pathogenic nematodes settled, as studied two days after inoculation, was not different from that of pathogenic ones. However, as studied after 30 days of inoculation, rate of reproduction in low-pathogenic nematodes was far lower than that of pathogenic nematodes. The rate of reproduction of several nematode isolates growing on fungal mat media of Botrytis cinerea varied, but three of four low-pathogenic isolates showed same level of reproduction rates as pathogenic ones.
Pre-treatment with DL-3-aminobutyric acid (BABA) in the cucumber plants caused the decrease of disease severity after inoculation with anthracnose pathogen Colletotrichum orbiculare. In this study, ultrastructures of the vascular bundle and the infection structures in the leaves of BABA-treated as well as untreated cucumber plants were observed after inoculation with the anthracnose pathogen by electron microscopy. The ultrastructures of vascular bundle in the leaves of BABA-treated plants were similar to those of the untreated plants except plasmodesmata. In the BABA-treated plants, the plasmodesmata were more numerous than in the untreated plants, suggesting that the BABA treatment may cause the active transfer of metabolites through the vascular bundle. In the leaves of untreated plants, the fungal hyphae were spread widely in the plant tissues at 5 days after pathogen inoculation. Most cellular organelles in the hyphae were intact, indicating a compatible interaction between the plant and the parasite. In contrast, in the leaves of BABA pre-treated plants the growth of most hyphae was restricted to the epidermal cell layer at 5 days after inoculation. Most hyphae cytoplasm and nucleoplasm was electron dense or the intracellular organelles were degenerated. The cell walls of some plant cells became thick at the site adjacent to the intercellular hyphae, indicating a mechanical defense reaction of the plant cells against the fungal attack. Furthermore, hypersensitive reaction (HR) of the epidermal cells was often observed, in which the intracellular hyphae were degenerated. Based on these results it is suggested that BABA causes the enhancement of defense mechanisms in the cucumber plants such as cell wall apposition or HR against the invasion of C. orbiculare.
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