• Title/Summary/Keyword: inoculation

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Observations of Infection Structures after Inoculation with Colletotrichum orbiculare on the Leaves of Cucumber Plants Pre-inoculated with Two Bacterial Strains Pseudomonas putida or Micrococcus luteus

  • Jeun, Yong-Chull;Lee, Kyung-Hoo
    • Mycobiology
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    • v.33 no.3
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    • pp.131-136
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    • 2005
  • 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.

Improvement of K+ and Na+ Ion homeostasis and salt tolerance by Co-inoculation of arbuscular mycorrhizal fungi (AMF) and spore associated bacteria (SAB)

  • Selvakumar, Gopal;Kim, Kiyoon;Roy, C. Aritra;Jeon, Sunyong;Sa, Tongmin
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.246-246
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    • 2017
  • Salinity inhibits plant growth and restricts the efficiency of arbuscular mycorrhizal fungi. The selective uptake of nutrients from the soil and their effective transport to host roots make it essential for plant growth and development under salt stress. AMF spore associated bacteria shown to improve mycorrhizal efficiency under stress. Thus, this study aimed to understand the co-inoculation efficiency of AMF and SAB on maize growth and ion homeostasis under salt stress. Two AMF strains and one SAB were inoculated with maize either alone or in combination with one another. The results of our study showed that AMF and SAB co-inoculation significantly improved dry weight and nutrient uptake of maize under salt stress. Co-inoculation significantly reduced proline accumulation in shoots and Na+ accumulation in roots. Co-inoculation treatment also exhibited the high K+/Na+ ratios in roots at 25 mM NaCl. Mycorrhizal colonization showed positive influence for regulation of ZmAKT2, ZmSOS1 and ZmSKOR gene expressions, contributing to K+ and Na+ ion homeostasis. CLSM view showed that SAB were able move and localize into inter and intra cellular spaces of maize roots. In addition, CLSM view of AMF spores showed that gfp-tagged SAB also associated on the spore outer hyaline layer.

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Biochemical Reactions of Barley Leaves at Intervals After Inoculation with Erysiphe graminis f. sp. hordei (보리 흰가루병균 접종후 보리엽내 경시적 생화학반응)

  • Song Dong Up;Cho Baik Ho;Kim Ki Chung
    • Korean Journal Plant Pathology
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    • v.2 no.1
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    • pp.37-42
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    • 1986
  • Papilla and cytoplasmic aggregates clearly formed on the epidermal cells of barley leaves in response to the primary germ-tubes of Erysiphe graminis f. sp. hordei, but their sizes were much smaller than those in response to the appressoria. Some cells of barley leaves exposed to powdery mildew for 36-48h were more deeply stained as compared to the other cells by acid fuchsin. However, the content of malondialdehydein in powdery mildewed leaves, one of the product of lipid peroxidation, did not increase by 96h after inoculation. Positive reactions for callose, protein and phenolics were recognized in the papilla and cytoplasmic aggregates at 6h after inoculation, but cutin, suberin, cellulose and lignin were not noticeable until 72h after inoculation. The total phenol content in methanol extracts increased with increasing time after inoculation. All histochemical reactions were not race-specific in barley­powdery mildew combinations tested.

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A novel method for high-frequency transgenic shoot regeneration via Agrobacterium tumefaciens in flax (Linum usitatissimum L.)

  • Beyaz, Ramazan;Darcin, E. Selcen;Aycan, Murat;Kayan, Mustafa;Yildiz, Mustafa
    • Journal of Plant Biotechnology
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    • v.43 no.2
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    • pp.240-247
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    • 2016
  • In this study, routinely used transformation method, which includes transferring explants onto co-cultivation medium after inoculating them with bacterial solution for a while, was compared with 3 different inoculation methods. In every 3 methods, hypocotyl explants excised from 7-day-old sterile flax seedlings having cotyledon leaves and no root system dried under air flow in sterile cabin for 35 min were inoculated with different volumes of bacterial solution at different inoculation periods. GV2260 line of Agrobacterium tumefaciens having 'pBIN 19' plasmid containing npt II (neomycin phosphotransferase II) gene and GUS reporter gene was used in transformation studies. After inoculation, hypocotyl segments of seedlings (0.5 cm in length) - were excised and left to co-cultivation for 2 days. Then, explants were transferred to regeneration medium supplemented with different antibiotics. The presence of npt-II and GUS genes in transformants was confirmed by PCR and GUS analysis. The highest results in all characters examined in all cultivars were obtained from the 2 inoculation method in which hypocotyls excised from seedlings inoculated with $500{\mu}l$ of bacterial solution after drying in sterile cabin for 35 min were used.

Ultrastructures of the Loaves of Cucumber Plane Treated with DL-3-Aminobutyric Acid at the Vascular Bundle and the Penetration Sites after Inoculation with Colletotrichum orbiculare

  • Jeun, Y.C.;Park, E.W.
    • The Plant Pathology Journal
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    • v.19 no.2
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    • pp.85-91
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    • 2003
  • 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.

Development of a Robot for Automation of a Callus Inoculation (식물조직배양 자동화를 위한 로봇개발 - 엔드이펙터 및 시스템의 성능시험 -)

  • Chung, Suk-Hyun;No, Dae-Hyun
    • Journal of Bio-Environment Control
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    • v.18 no.2
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    • pp.87-94
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    • 2009
  • This study was conducted to develop an automation system of inoculation processing of a lily callus. The results are summarized as followings: The end-effector was manufactured as suction and machine type. And these end-effectors can separate the callus from the mediums and divide the separated callus and then inoculate the divided callus to new mediums. Using the machine type end-effect0r, the results of the experiment showed the success rate in the division process was 100% while the separation and inoculation process was 92%. To develop the automation controller of inoculation process, the system was developed to control an external device and the manipulator. The data communication program between a robot and a personal computer was also developed using CAsyncsocket and Ethernet Interface.

Evaluation of Yeast Diversity During Wine Fermentations with Direct Inoculation and pied de cuve Method at an Industrial Scale

  • Li, Erhu;Liu, Chuanhe;Liu, Yanlin
    • Journal of Microbiology and Biotechnology
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    • v.22 no.7
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    • pp.960-966
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    • 2012
  • The diversity and composition of yeast populations may greatly impact wine quality. This study investigated the yeast microbiota in two different types of wine fermentations: direct inoculation of a commercial starter versus pied de cuve method at an industrial scale. The pied de cuve fermentation entailed growth of the commercial inoculum used in the direct inoculation fermentation for further inoculation of additional fermentations. Yeast isolates were collected from different stages of wine fermentation and identified to the species level using Wallersterin Laboratory nutrient (WLN) agar followed by analysis of the 26S rDNA D1/D2 domain. Genetic characteristics of the Saccharomyces cerevisiae strains were assessed by a rapid PCR-based method, relying on the amplification of interdelta sequences. A total of 412 yeast colonies were obtained from all fermentations and eight different WL morphotypes were observed. Non-Saccharomyces yeast mainly appeared in the grape must and at the early stages of wine fermentation. S. cerevisiae was the dominant yeast species using both fermentation techniques. Seven distinguishing interdelta sequence patterns were found among S. cerevisiae strains, and the inoculated commercial starter, AWRI 796, dominated all stages in both direct inoculation and pied de cuve fermentations. This study revealed that S. cerevisiae was the dominant species and an inoculated starter could dominate fermentations with the pied de cuve method under controlled conditions.

Changing Patterns of Acute Phase Proteins and Inflammatory Mediators in Experimental Caprine Coccidiosis

  • Hashemnia, Mohammad;Khodakaram-Tafti, Azizollah;Razavi, Seyed Mostafa;Nazifi, Saeed
    • Parasites, Hosts and Diseases
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    • v.49 no.3
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    • pp.213-219
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    • 2011
  • This experiment was conducted to assess the changing patterns and relative values of acute phase proteins and inflammatory cytokines in experimental caprine coccidiosis. Eighteen newborn kids were allocated to 3 equal groups. Two groups, A and B, were inoculated with a single dose of $1{\times}10^3$ and$1{\times}10^5$sporulated oocysts of Eimeria arloingi, respectively. The third group, C, received distilled water as the control. Blood samples were collected from the jugular vein of each kid in both groups before inoculation and at days 7, 14, 21, 28, 35, and 42 post-inoculation (PI), and the levels of haptoglobin (Hp), serum amyloid A (SAA), TNF-${\alpha}$, and IFN-${\gamma}$ were measured. For histopathological examinations, 2 kids were selected from each group, euthanized, and necropsied on day 42 PI. Mean Hp concentrations in groups A and B (0.34 and 0.68 g/L) at day 7 PI were 3.2 and 6.3 times higher than the levels before inoculation. The mean SAA concentrations in groups A and B (25.6 and 83.5 ${\mu}g$/ml) at day 7 PI were 4.2 and 13.7 times higher than the levels before inoculation. The magnitude and duration of the Hp and SAA responses correlated well with the inoculation doses and the severity of the clinical signs and diarrhea in kids. These results were consistent with the histopathological features, which showed advanced widespread lesions in group B. In both groups, significant correlations were observed for TNF-${\alpha}$ and IFN-${\gamma}$ with SAA and Hp, respectively. In conclusion, Hp and SAA can be useful non-specific diagnostic indicators in caprine coccidiosis.

Effectiveness on the Inoculation of Arbuscular Mycorrhizal Fungi in Cutting of Grapevine (포도 삽목에서 내생 균근균 접종효과)

  • Wee, Chi-Do;An, Gi-Hong;Kim, Hong-Lim;Sohn, Bo-Kyoon
    • Korean Journal of Soil Science and Fertilizer
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    • v.43 no.6
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    • pp.1002-1007
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    • 2010
  • The study was performed to investigate the influence on growth and development of grape-cuttings by arbuscular mycorrhizal (AM) fungi inoculation, AM colonization rate, and the phenomena of mycorrhizal association. Among the grape-cuttings, 'Kyoho' and 'Tamnara' cultivars inoculated with AM fungi showed significantly increase of leaf area, leaf number, total root length and root surface area than non-infected ones. But 'Cambell Early' did not showed any significant difference in total root length and root surface area even after the inoculation. The AM colonization rates in mycorrhizal inoculation treatment were 22.5-32.5% in total average after 8weeks, and were 29.6%, 28.8%, and 48.8% for 'Cambell Early', 'Tamnara', and 'Kyoho' respectively after 12weeks. The AM colonization rate marked very low level in non-colonization control plot.

Response of Barley Genotypes to Fusarium Head Blight under Natural Infection and Artificial Inoculation Conditions

  • Khanal, Raja;Choo, Thin Meiw;Xue, Allen G.;Vigier, Bernard;Savard, Marc E.;Blackwell, Barbara;Wang, Junmei;Yang, Jianming;Martin, Richard A.
    • The Plant Pathology Journal
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    • v.37 no.5
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    • pp.455-464
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    • 2021
  • Forty-eight spring barley genotypes were evaluated for deoxynivalenol (DON) concentration under natural infection across 5 years at Harrington, Prince Edward Island. These genotypes were also evaluated for Fusarium head blight (FHB) severity and DON concentration under field nurseries with artificial inoculation of Fusarium graminearum by the grain spawn method across 2 years at Ottawa, Ontario, and one year at Hangzhou, China. Additionally, these genotypes were also evaluated for FHB severity under greenhouse conditions with artificial inoculation of F. graminearum by conidial suspension spray method across 3 years at Ottawa, Ontario. The objective of the study was to investigate if reactions of barley genotypes to artificial FHB inoculation correlate with reactions to natural FHB infection. DON concentration under natural infection was positively correlated with DON concentration (r = 0.47, P < 0.01) and FHB incidence (r = 0.56, P < 0.01) in the artificially inoculated nursery with grain spawn method. Therefore, the grain spawn method can be used to effectively screen for low DON. FHB severity, generated from greenhouse spray, however, was not correlated with DON concentration (r = 0.12, P > 0.05) under natural infection and it was not correlated with DON concentration (r = -0.23, P > 0.05) and FHB incidence (r = 0.19, P > 0.05) in the artificially inoculated nursery with grain spawn method. FHB severity, DON concentration, and yield were affected by year, genotype, and the genotype × year interaction. The effectiveness of greenhouse spray inoculation for indirect selection for low DON concentration requires further studies. Nine of the 48 genotypes were found to contain low DON under natural infection. Island barley had low DON and also had high yield.