Novel multifunctional plant growth-promoting bacteria isolated from the oil palm rhizosphere under long-term organic matter application.

Most agricultural products are presently cultivated on marginal lands with poor soil properties and unfavorable environmental conditions (diseases and abiotic stresses), which can threaten plant growth and yield. Plant growth-promoting bacteria (PGPB) are beneficial bacteria that promote plant growth and biomass and act as biocontrols against diseases and stress. However, most isolated PGPBs have a single function and low survival rates owing to their limited growth behaviors. In this study, we isolated multifunctional PGPB from oil palm rhizosphere, quantitatively measured their activities, and evaluated their effectiveness in Brassica rapa (Komatsuna) cultivation. This is the first study to report the isolation of three multifunctional PGPB strains with ammonium production, phosphate-potassium-silicate solubilization, and indole-3-acetic acid (IAA) production from the oil palm rhizosphere, namely Kosakonia oryzendophytica AJLB38, Enterobacter quasimori AJTS77, and Lelliottia jeotgali AJTS83. Additionally, these strains showed antifungal activity against the oil palm pathogen Ganoderma boninense. These strains grow under high temperature, acidic and alkaline pH, and high salt concentration, which would result in their proliferation in various environmental conditions. The cultivation experiments revealed these strains improved the growth and biomass with half the dosage of chemical fertilizer application, which was not significantly different to the full dosage. Furthermore, the overall plant growth-promoting activities in quantitative assays and overall B. rapa growth in cultivation experiments were statistically correlated, which could contribute to the prediction of plant growth promotion without plant cultivation experiments. Thus, the selected PGPB could be valuable as a biofertilizer to improve soil health and quality and promote agricultural sustainability.

Whole-Genome DNA Methylation Analysis in Brassica rapa subsp. perviridis in Response to Albugo candida Infection.

DNA methylation is an epigenetic mark associated with several mechanisms in plants including immunity mechanisms. However, little is known about the regulatory role of DNA methylation in the resistance response of Brassica species against fungal diseases. White rust, caused by the fungus Albugo candida, is one of the most widespread and destructive diseases of all the cultivated Brassica species, particularly Brassica rapa L. and Brassica juncea (L.) Czern and Coss. Here, we investigate whole-genome DNA methylation modifications of B. rapa subsp. perviridis in response to white rust. As a result, 233 and 275 differentially methylated regions (DMRs) in the susceptible cultivar "Misugi" and the resistant cultivar "Nanane" were identified, respectively. In both cultivars, more than half of the DMRs were associated with genes (DMR-genes). Gene expression analysis showed that 13 of these genes were also differentially expressed between control and infected samples. Gene ontology enrichment analysis of DMR genes revealed their involvement in various biological processes including defense mechanisms. DMRs were unevenly distributed around genes in susceptible and resistant cultivars. In "Misugi," DMRs tended to be located within genes, while in "Nanane," DMRs tended to be located up and downstream of the genes. However, CG DMRs were predominantly located within genes in both cultivars. Transposable elements also showed association with all three sequence contexts of DMRs but predominantly with CHG and CHH DMRs in both cultivars. Our findings indicate the occurrence of DNA methylation modifications in B. rapa in response to white rust infection and suggest a potential regulatory role of DNA methylation modification in defense mechanisms which could be exploited to improve disease resistance.

Influence of potassium-solubilizing bacteria on the growth and radiocesium phyto-transfer of Brassica rapa L. var. perviridis grown in contaminated Fukushima soils.

The supply of K, being the chemical analog of Cs, affects the phytotransfer of radiocesium such as 137Cs from contaminated soils and its accumulation in plant tissues. Since K and Cs have high affinity to the same clay particle surfaces, the presence of potassium-solubilizing bacteria (KSB) could increase the availability of not only K+ in the rhizosphere but also of radiocesium. In this study, we obtained five KSB isolates with the highest solubilization capacities from soybean rhizosphere on modified Aleksandrov medium containing sericite as K source. Based on biochemical and 16S rRNA gene sequence analysis, we identified the bacteria as Bacillus aryabhattai MG774424, Pseudomonas umsongensis MG774425, P. frederiksbergensis MG774426, Burkholderia sabiae MG774427, and P. mandelii MG774428. We evaluated the KSB isolates based on plant growth promotion and 137Cs accumulation in komatsuna (Brassica rapa L. var. Perviridis) grown in three soils collected from Miyanoiri, Takanishi, and Ota contaminated by 137Cs from the Fukushima accident. Inoculation with KSB showed beneficial effects on plant growth and increased the overall plant biomass production (~40%). On the average, KSB inoculation resulted in the removal of 0.07 ± 0.04% of 137Cs from the soil, more than twice the control. But similar to the effect of KSB inoculation on komatsuna biomass production, different KSBs performed variably and exhibited site-specific responses independent of their K-solubilizing capacities, with higher 137Cs phyto-transfer in roots than in shoots. In terms of root transfer factor (TF), values were highest in komatsuna plants grown in Miyanoiri and Ota soils inoculated with P. frederiksbergensis and Burkholderia sabiae, while they were highest in Takanishi soils inoculated with Bacillus aryabhattai and P. umsongensis. These TF values were also much higher than previously reported values for komatsuna grown in 137Cs-contaminated Fukushima soils inoculated with other rhizobacteria. Thus, KSB inoculation significantly enhance not only the growth of komatsuna but 137Cs uptake.

Establishment of a cultivation method for leaf lettuce (Lactuca sativa var. crispa) and komatsuna (Brassica rapa var. perviridis) with high zinc content for patients with zinc deficiency and evaluation of its effectiveness.

BACKGROUND: Insufficient intake of zinc is associated with various diseases worldwide. To overcome this problem, we aimed to establish a method for cultivating leafy vegetables with high zinc content in hydroponics without inhibiting their growth. Furthermore, we evaluated the effectiveness of the cultivated leafy vegetables with high zinc content in zinc-deficient mice. RESULTS: By adjusting the zinc concentration in the hydroponic solution to 5 mg L-1 starting from 7 days before harvesting, the zinc content in leaf lettuce increased eight times of that in the control, without any inhibition of the growth. Furthermore, when oxidized glutathione (GSSG) was added simultaneously with zinc to the hydroponic solution, the zinc content further doubled (16 times of that in the control). Similar results were obtained with komatsuna and red leaf lettuce, although there was a difference in the effect of GSSG treatment. The effectiveness of leafy vegetables with high zinc content in ameliorating zinc deficiency was evaluated by feeding lettuce with high zinc content to zinc-deficient mice. High zinc content lettuce significantly increased the zinc content in the liver, kidneys, gastrocnemius, and tibia of these mice. CONCLUSION: We established a cultivation method for lettuce and komatsuna with high zinc content without inhibiting growth by adjusting the zinc concentration in the hydroponic solution at an appropriate concentration for an appropriate period. The result of feeding test indicates that the intake of leafy vegetables with high zinc content can ameliorate zinc deficiency and might be useful in protection from several diseases associated with this deficiency. © 2020 Society of Chemical Industry.

Effect of Prohydrojasmon on the Growth of Eggplant and Komatsuna.

Prohydrojasmon (PDJ) can improve the polyphenol and anthocyanin content and antioxidant activity of some crop plants, but it also shows a suppressive effect on the plant growth. This study aimed to investigate the inhibitory effect of PDJ on the growth of two crop plants: komatsuna (Brassica rapa var. periviridis) and eggplant (Solanum melongena). We applied various concentrations of PDJ drip-wise or by spraying to eggplant and komatsuna seedlings and made detailed observations of growth. In general, no significant suppressive effect of PDJ was observed in the aerial parts in both plants. However, a significant inhibitory effect was found in roots treated with PDJ at concentrations of 600 and 1000 ppm. Interestingly, komatsuna treated with PDJ at a concentration of 200 ppm in both approaches resulted in a significant increase in root weight up to 37%. At a concentration range of 200-400 ppm, PDJ showed no inhibitory effects, and in some cases slightly promoted root growth. Therefore, this could be the recommended concentration range. We conclude that application of PDJ can still be beneficial to the vegetable crops without causing serious inhibition or suppression effects on the growth, as long as it is kept at rather low concentrations.

Effects of micro wet milling on bioaccessibility of phosphatidic acid and lysophosphatidic acid in komatsuna during in vitro digestion.

Foods rich in phosphatidic acid (PA) can ameliorate stomach ulcers in mice by hydrolysis of PA to lysophosphatidic acid (LPA). In this study, PA-rich komatsuna was produced using the micro wet milling (MWM) system, which can mill food products into micrometer-scale without causing detrimental factors such as frictional heat. To evaluate the efficiency of the MWM system in increasing PA and forming LPA, the availability of PA in the MWM komatsuna to hydrolyze into LPA under in vitro simulated gastrointestinal (GI) digestion conditions were investigated. The results showed that through effective MWM milling, komatsuna was sufficiently milled into smaller particles, and PA was abundantly produced in the milled komatsuna; the increased PA promoted LPA formation during digestion, resultant a dominant molecular species of 16:0 LPA which could effectively reduce ulcer lesions. These indicated that MWM can elevate the bioaccessibility of komatsuna PA and LPA in the GI tract, which will benefit the dietary treatment of stomach ulcers.

Environmental impacts and consumer preference for sustainably cultivated Japanese mustard spinach, komatsuna.

Sustainable agriculture is spreading in Japan in response to growing concerns about the environmental burden of the agriculture sector, but less than 1% of the total crop area for each vegetable in Japan is grown sustainably. Environmentally friendly agricultural products are produced by using organic and low-input farming techniques; low-input farming aims to reduce chemical inputs, such as fertilizers, pesticides, and herbicides, by half. Here, we used komatsuna (Japanese mustard spinach, Brassica rapa var. perviridis) as a model vegetable to study the environmental impact of low-input farming and ways to promote the purchase of organically and low-input farmed vegetables. We first assessed greenhouse gas emissions resulting from organic, low-input, and conventional farming of komatsuna. We also evaluated the effectiveness of providing consumers with detailed farm management and seasonality information to market organically and low-input farmed vegetables. We estimated marginal willingness-to-pay (MWTP) using choice-based conjoint analysis, based on attributes of price, fertilizer use, pesticide use, and region of origin. For seasonality, the questionnaire incorporating these attributes was conducted twice: once assuming purchasing in season, the other out of season. The greenhouse gas emissions of organic farming per area (196.7 kg CO2-eq/10 a/year) and per yield (72.3 kg CO2-eq/t/year) were less than those of low-input (322.6 kg CO2-eq/10 a/year, 120.7 kg CO2-eq/t/year) and conventional (594.0 kg CO2-eq/10 a/year, 220.7 kg CO2-eq/t/year) farming. MWTPs were highest for pesticide-free komatsuna (76.9 yen out of season, 66.2 yen in season), followed by full organic fertilizer (66.0 yen out of season, 63.4 yen in season), half organic fertilizer (35.8 yen out of season, 19.8 yen in season), and half pesticide (29.2 yen out of season, 21.0 yen in season). Consumers showed greater preference for organically and low-input farmed komatsuna out of season than in season. Consumers were more interested in pesticide information than in fertilizer and region of origin information. Our findings suggest that providing detailed cultivation and seasonality information would be a beneficial consumer communication tool to increase the market for sustainable agricultural products.

Parasitoid wasps' exposure to host-infested plant volatiles affects their olfactory cognition of host-infested plants.

Using Cotesia vestalis, a parasitoid wasp of diamondback moth larvae and three crucifer plant species (cabbage, komatsuna, and Japanese radish), we examined the effects of exposure to host-infested plant volatiles from one plant species on a newly emerged wasp's subsequent olfactory cognition of host-infested plant volatiles from the same or different plant species. The preference of C. vestalis between infested and uninfested plant volatiles was tested in a choice chamber. Volatile-inexperienced wasps significantly preferred infested cabbage and infested radish volatiles, but not infested komatsuna volatiles. After exposure to infested cabbage volatiles, wasps showed a significant preference for infested cabbage volatiles, while the significant preference for infested radish volatiles that had been observed in inexperienced wasps was no longer observed. After exposure to infested komatsuna volatiles, wasps significantly preferred infested komatsuna volatiles, and the pre-exposure significant preferences for infested cabbage volatiles and infested radish volatiles remained. After exposure to infested radish volatiles, the significant preferences for infested cabbage and infested radish volatiles did not change. Furthermore, wasps showed a significant preference for infested komatsuna volatiles. The compound compositions of the volatile blends from the three infested plant species were grouped separately in a principal coordinates analysis. The experience-based cognition of C. vestalis for host-infested plant volatiles of three plant species is discussed.