As(III)-oxidizing and plant growth-promoting bacteria increase the starch biosynthesis-related enzyme activity, 2-AP levels, and grain quality of arsenic-stressed rice plants.

BACKGROUND: Grain quality is an important index of rice production, particularly when plants are grown under stress. Arsenic (As) contamination in paddy fields severely affects rice grain yield and quality. Here, the effects of As and combinations of As(III)-oxidizing bacteria (Pseudomonas stutzeri 4.25, 4.27, and 4.44) and plant growth-promoting bacteria (Delftia acidovorans KKU2500-12 and Cupriavidus taiwanensis KKU2500-3) on enzymes related to starch accumulation in grains and the grain quality of Khao Dawk Mali 105 rice cultivated in As-contaminated soil under greenhouse conditions were investigated. RESULTS: Arsenic affected the activities of starch biosynthesis-related enzymes, and decreases of up to 76.27%, 71.53%, 49.74%, 73.39%, and 47.46% in AGPase, SSS, GBSS, SBE, and SDBE activities, respectively, and 9.42-61.07% in starch accumulation in grains were detected after growth in As-contaminated soil. However, the KKU2500-3/4.25 and KKU2500-3/4.44 combinations yielded the greatest enzyme activities in grains, and compared with the results observed in uninoculated seedlings, increases in starch accumulation of up to 51.16% and 23.81% were found in the inoculated seedlings after growth in medium- and high-As-contaminated soils, at 10-17 and 10-24 days after anthesis, respectively. The bacteria increased the 2-AP content in rice under As stress, possibly via the induction of proline, a 2-AP substrate. Bacterium-inoculated rice had significantly greater 2-AP levels than uninoculated rice, and 2.16-9.93% and 26.57-42.04% increases were detected in rice plants grown in medium- and high-As-contaminated soils, respectively. CONCLUSIONS: Arsenic toxicity can be mitigated in rice growing under greenhouse conditions by maintaining starch biosynthesis, accumulating amylose, and increasing 2-AP content. The effectiveness of these bacteria should be validated in paddy fields; hence, safe rice grains with a good starch content and aroma could be produced.

Low Cd-accumulating rice grain production through inoculation of germinating rice seeds with a plant growth-promoting endophytic bacterium.

This study investigated the effects of the plant growth-promoting endophytic bacterium Cupriavidus taiwanensis KKU2500-3 on the growth of KDML105 rice plants and cadmium (Cd) accumulation in grains. The rice plants were cultivated in soils with 20 and 50 ppm Cd under greenhouse conditions for two consecutive years. At both levels, Cd reduced rice growth and development. Under Cd stress, KKU2500-3 colonized the root surface and interior of rice plants at the early growth stage, and this colonization remained until the late stage. The colonized bacteria increased the pigment contents but reduced the root-to-aboveground translocation of Cd. In soil with 20 ppm Cd, the phytochelatin content of the bacteria-inoculated rice was lower (32.3-89.3%) than that of uninoculated rice. In soil with 50 ppm Cd, the bacteria-inoculated rice exhibited higher glutathione reductase (5-63%) and proline (5-115%) levels, a higher reduced glutathione (GSH)/0.5 oxidized glutathione (GSSG) ratio (4-212%) and decreased lipid peroxidation (1-19%) compared with uninoculated rice. The root-to-grain translocation factor of inoculated rice in soil with 50 ppm Cd was significantly lower than that of inoculated rice in soil with 20 ppm Cd, and this finding was consistent with the 38.6% and 75.1% reductions in Cd accumulation observed in grains from soils with 20 and 50 ppm Cd, respectively. The Cd content of KDML105 grains grown in soil with 50 ppm Cd was 0.36 ppm, which is below the Codex standard for polished rice (0.4 ppm). The levels of available P, Zn, and SO42- also affect Cd availability in soil, and colonized KKU2500-3 showed varying responses to different Cd levels. Thus, bacterial inoculation, the Cd level and soil properties play important roles in Cd accumulation in KDML105 rice grains. The role of C. taiwanensis KKU2500-3 on the production of low-Cd-accumulating rice in paddy fields contaminated with a range of Cd levels should be further investigated.

Changes in the proteome of the cadmium-tolerant bacteria Cupriavidus taiwanensis KKU2500-3 in response to cadmium toxicity.

Cupriavidus taiwanensis KKU2500-3 is a cadmium (Cd)-tolerant bacterial strain that was previously isolated from rice fields contaminated with high levels of Cd. In 500 µmol/L CdCl2, the KKU2500-3 strain grew slower and with a more prolonged lag-phase than when grown in the absence of Cd. A proteomic approach was used to characterize the protein expression in the Cd-tolerant bacteria C. taiwanensis KKU2500-3 during growth under Cd stress. When compared with the untreated cells, a total of 982 differentially expressed protein spots were observed in the CdCl2-treated cells, and 59 and 10 spots exhibited >2- and >4-fold changes, respectively. The level of up- and downregulation varied from 2.01- to 11.26-fold and from 2.01- to 5.34-fold, respectively. Of the 33 differentially expressed protein spots analyzed by MALDI TOF MS/MS, 19 spots were successfully identified, many of which were involved in stress responses. The most highly upregulated protein (+7.95-fold) identified was the chaperone GroEL, which indicated that this factor likely contributed to the bacterial survival and growth in response to Cd toxicity. Detection of the downregulated protein flagellin (-3.52-fold) was consistent with the less effective ATP-mediated and flagella-driven motility. The flagella-losing cells were also observed in the Cd-treated bacteria when analyzed by scanning electron microscopy. Thus, the Cd-stressed cells may downregulate pathways involving ATP utilization in favor of other mechanisms in response to Cd toxicity. When the KKU2500-3 strain was grown in the presence of Cd, H2S was not detected, suggesting a possible role of the sulfur in precipitation with Cd. Apart from a general response, no specific process could be determined using the present proteomic approach. However, the potential role of protein folding-mediated GroEL, flagella-mediated motility and CdS biotransformation in Cd toxicity response observed in this study as well as the extent of Cd-tolerant mechanisms using other methods could facilitate the future application of this strain in addressing Cd environmental contamination.

Kosakonia oryziphila NP19 bacterium acts as a plant growth promoter and biopesticide to suppress blast disease in KDML105 rice.

This study demonstrates that root-associated Kosakonia oryziphila NP19, isolated from rice roots, is a promising plant growth-promoting bioagent and biopesticide for combating rice blast caused by Pyricularia oryzae. In vitro experiments were conducted on fresh leaves of Khao Dawk Mali 105 (KDML105) jasmine rice seedlings. The results showed that NP19 effectively inhibited the germination of P. oryzae fungal conidia. Fungal infection was suppressed across three different treatment conditions: rice colonized with NP19 and inoculated by fungal conidia, a mix of NP19 and fungal conidia concurrently inoculated on the leaves, and fungal conidia inoculation first followed by NP19 inoculation after 30 h. Additionally, NP19 reduced fungal mycelial growth by 9.9-53.4%. In pot experiments, NP19 enhanced the activities of peroxidase (POD) and superoxide dismutase (SOD) by 6.1-63.0% and 3.0-67.7%, respectively, indicating a boost in the plant's defense mechanisms. Compared to the uncolonized control, the NP19-colonized rice had 0.3-24.7% more pigment contents, 4.1% more filled grains per panicle, 26.3% greater filled grain yield, 34.4% higher harvest index, and 10.1% more content of the aroma compound 2-acetyl-1-pyrroline (2AP); for rice colonized with NP19 and infected with P. oryzae, these increases were 0.2-49.2%, 4.6%, 9.1%, 54.4%, and 7.5%, respectively. In field experiments, blast-infected rice that was colonized and/or inoculated with NP19 treatments had 15.1-27.2% more filled grains per panicle, 103.6-119.8% greater filled grain yield, and 18.0-35.8% higher 2AP content. A higher SOD activity (6.9-29.5%) was also observed in the above-mentioned rice than in the blast-infected rice that was not colonized and inoculated with NP19. Following blast infection, NP19 applied to leaves decreased blast lesion progression. Therefore, K. oryziphila NP19 was demonstrated to be a potential candidate for use as a plant growth-promoting bioagent and biopesticide for suppressing rice blast.

A cadmium-tolerant endophytic bacterium reduces oxidative stress and Cd uptake in KDML105 rice seedlings by inducing glutathione reductase-related activity and increasing the proline content.

The effect of the endophytic Cupriavidus taiwanensis KKU2500-3 on the Cd toxicity of KDML105 rice seedlings was investigated in a 10 µM CdCl2 hydroponic system. As demonstrated after bacterial inoculation of germinating rice seeds, KKU2500-3 colonized all rice plant parts. In RB (Rice + KKU2500-3) and RBC (Rice + KKU2500-3+Cd), KKU2500-3 effectively colonized and was detected at a markedly higher number in the root surface and interior than in shoots and leaves. The activities of antioxidant enzymes ascorbate peroxidase (APOX), glutathione reductase (GR), and superoxide dismutase (SOD) and the proline content in inoculated rice were higher in roots and aboveground tissues. RBC exhibited a higher reduced-to-oxidized glutathione ratio in roots and leaves (3-55%) but a lower malondialdehyde content (8-78%). Phytochelatins (PCs) were detected in all rice tissues, but their levels in RBC were 13-70% lower than those in RC (Rice + Cd), demonstrating that the induction of PCs in rice was unrelated to KKU2500-3. The Cd levels in roots and shoots were lower in RBC than RC, and the root-to-shoot Cd translocation factor was 0.6-62.2% lower. At 30 DAT, the Cd levels in RBC roots and shoots were 30.2% and 73.7% lower, respectively, than those in RC. Colonized KKU2500-3 activated GR and increased the proline content to overcome rice Cd toxicity. These effects may trap Cd in plant cells and reduce its translocation. Hence, KKU2500-3 synergistically interacts with rice to detoxify Cd at early growth stages, and KDML105 rice grains with low Cd accumulation could be produced if this interaction is maintained until late growth stages.