First Report of Fire Blight of Apricot (Prunus armeniaca) Caused by Erwinia amylovora in Korea.

During a survey in May 2020, symptoms of blight were observed on apricot (Prunus armeniaca cv. undetermined) in an orchard (37°06´01.5″N 127°57´44.9″E) in Chungju, South Korea, where fire blight of apple occurred. Three apricot trees in the apple orchard were heavily diseased and showed severe shoot blight and stem canker symptoms. Bacterial isolates were recovered on King's medium B from leaves and twigs that were surface-sterilized with 70% alcohol. Colonies with uniform mucoid, smooth surfaces were collected. DNA from nine isolates did not yield an amplicon in a PCR assay for detection of Erwinia pyrifoliae using primer set CPS1/CPS2c (Kim et al. 2001). Each isolate was positive in PCR assays for E. amylovora using primer sets A/B (Bereswill et al. 1992) and AJ75/76 (Llop et al. 2000) that target pEA29. Sequencing of the PCR products resulted in 99.9% (929 bp out of 930 bp) and 100% (747 bp out of 747 bp) identity with sequences of E. amylovora FB20 (GeneBank: CP050240), respectively. Amplifications of the partial 16S rRNA (GeneBank: LC557153) and hrpN (GeneBank: LC575997) genes were performed, and the products were sequenced. The primers used to amplify 16S rRNA were 518F: 5'-CCAGCAGCCGCGGTAATACG-3' and 800R: 5'-TACCAGGGTATCTAATCC-3', and those for the hrpN genes were HRPN1: 5'-ATGAGTCTGAATACAAG-3' and HRPN3c: 5'-GCTTGCCAAGTGCCATA-3'. BLAST analyses showed 99.8% (1439 bp out of 1442 bp) and 100% (1136 bp out of 1136 bp) identities, respectively, to the sequences of E. amylovora FB20. The ability of the isolates to induce a hypersensitive reaction on tobacco (Nicotiana tabacum cv. Xanthi) leaves was also evaluated. Bacterial suspensions (1.5 ⅹ 108 CFU) of 2 isolates were injected into tobacco leaves, and after 48 h, both isolates caused a hypersensitive response. To confirm pathogenicity of isolates, 3-mm-deep holes in five immature apricot (cv. Goldcot) and five immature apple (cv. Fuji) fruits were inoculated with 10 µl bacterial suspension (1.5 ⅹ 108 CFU/ml). The inoculated fruits were placed in a humid plastic box. After 7 days at 27℃, severe necrosis and bacterial ooze were present at the inoculated sites in three repeated tests. No symptoms were observed on fruits inoculated with sterile water. To complete Koch's postulates, bacteria were reisolated from the inoculated apricot and apple fruits. PCR using the specific primer sets stated above confirmed the identity as E. amylovora. Thus, based on disease symptoms, sequences, and pathogenicity, the bacterium causing blight of apricot was identified as E. amylovora. Natural infections of E. amylovora on apricot trees have been reported in the Czech Republic and Hungary (Korba and Sillerova 2011; Vegh and Palkovics 2013). Fire blight was observed in the Czech Republic on apricot trees near pear seedlings, which are highly susceptible to E. amylovora (Korba and Sillerova 2011). Natural infections of E. amylovora on Japanese plum planted adjacent to an apple orchard with severe fire blight has been reported in the United States (Mohan and Thomson 1996). Moreover, susceptibility to fire blight has been reported for apricot and Japanese plum cultivars (Mohan and Bijman 1999). To our knowledge, this the first report of fire blight of apricot caused by E. amylovora in Korea. This report is important because it provides evidence that apricot may be an overlooked reservoir for E. amylovora, in addition to apple, pear, and other rosaceous plants, in Korea. An intensive survey for additional host plants for the fire blight pathogen will be continued in Korea. This work was supported by a grant from the Agenda program (PJ01530202) of Rural Development Administration, Republic of Korea.

Enhancement of vitality and activity of a plant growth-promoting bacteria (PGPB) by atmospheric pressure non-thermal plasma.

The inconsistent vitality and efficiency of plant growth promoting bacteria (PGPB) are technical limitations in the application of PGPB as biofertilizer. To improve these disadvantages, we examined the potential of micro Dielectric Barrier Discharge (DBD) plasma to enhance the vitality and functional activity of a PGPB, Bacillus subtilis CB-R05. Bacterial multiplication and motility were increased after plasma treatment, and the level of a protein involved in cell division was elevated in plasma treated bacteria. Rice seeds inoculated with plasma treated bacteria showed no significant change in germination, but growth and grain yield of rice plants were significantly enhanced. Rice seedlings infected with plasma treated bacteria showed elevated tolerance to fungal infection. SEM analysis demonstrated that plasma treated bacteria colonized more densely in the broader area of rice plant roots than untreated bacteria. The level of IAA (Indole-3-Acetic Acid) and SA (Salicylic Acid) hormone was higher in rice plants infected with plasma treated than with untreated bacteria. Our results suggest that plasma can accelerate bacterial growth and motility, possibly by increasing the related gene expression, and the increased bacterial vitality improves colonization within plant roots and elevates the level of phytohormones, leading to the enhancement of plant growth, yield, and tolerance to disease.

Dynamics of nitric oxide level in liquids treated with microwave plasma-generated gas and their effects on spinach development.

In this study, we generated water and phosphate buffer treated with microwave plasma-generated gas in which the major component was nitric oxide (PGNO), and investigated the efficiency of the treated water and buffer in fertilization and sanitation. Real time NO level monitored by an electrode sensor was linearly increased over PGNO injection time, and removal of O2 from liquid before PGNO injection accelerated NO assimilation into liquids. Residual NO was still present 16 h after PGNO injection was stopped. H2O2, NO2-, and NO3- were also detected in PGNO-treated liquids. Spinach plants applied with 10 and 30 times diluted PGNO-treated water and 0.5 mM phosphate buffer showed slightly higher height and dry weight than control after 5 weeks. Plants grown with 10 and 30 times diluted PGNO-treated water exhibited the increased tolerance to water deficiency. Significant anti-microbial activity within 1 h was observed in un-diluted and in half-diluted PGNO-treated water and 0.5 mM phosphate buffer. Our results suggest that water or phosphate buffer containing NO, H2O2, NO2-, and NO3- can be produced by PGNO treatment, and that PGNO-treated water or buffer can be used as a potential fertilizer enhancing plant vitality with sanitation effect.

Effects of high voltage nanosecond pulsed plasma and micro DBD plasma on seed germination, growth development and physiological activities in spinach.

In this study, we analyzed seed germination, seedling growth, and physiological aspects after treatment with high voltage nanosecond pulsed plasma and micro DBD plasma in spinach (Spinacia oleracea L.), a green leafy vegetable known to have low germination rate. Both germination and dry weight of seedlings increased after high voltage pulse shots were applied to spinach seeds. However seeds treated with many shots (10 shots) showed a decrease in germination rate and seedling growth. Seeds treated with air DBD plasma exhibited slightly higher germination and subsequent seedling growth than those treated with N2 plasma. Seed surface was degenerated after treated with high voltage pulsed plasma and micro DBD plasma but no significant difference in the degree of degeneration was observed among micro DBD plasma treatment time. Level of GA3 hormone and mRNA expression of an amylolytic enzyme-related gene in seeds were elevated 1 day after treatment with high voltage pulsed plasma. The relative amount of chlorophyll and total polyphenols in spinach seedlings grown from seeds treated with air DBD plasma was increased in 30 s, 1 min, and 3 min treatments. Taken together, our results suggest a possibility that plasma can enhance seed germination by triggering biochemical processes in seeds.