Introgression of bacterial leaf blight (BLB) resistant gene, Xa7 into MARDI elite variety, MR219 by marker assisted backcrossing (MABC) approach / Introdução do gene resistente à ferragem da folha bacteriana (BLB), xa7 na variedade MARDI elite, MR219 por abordagem de backcrossing assistida por marcador (MABC)

Abstract Bacterial leaf blight (BLB) is one of the major rice diseases in Malaysia. This disease causes substantial yield loss as high as 70%. Development of rice varieties which inherited BLB resistant traits is a crucial approach to promote and sustain rice industry in Malaysia. Hence, this study aims were to enhance BLB disease resistant characters of high yielding commercial variety MR219 through backcross breeding approach with supporting tool of marker-assisted selection (MAS). Broad spectrum BLB resistance gene, Xa7 from donor parent IRBB7 were introgressed into the susceptible MR219 (recurrent parent) using two flanking markers ID7 and ID15. At BC3F4, we managed to generate 19 introgressed lines with homozygous Xa7 gene and showed resistant characteristics as donor parent when it was challenged with Xanthomonas oryzae pv. oryzae through artificial inoculation. Recurrent parent MR219 and control variety, MR263 were found to be severely infected by the disease. The improved lines exhibited similar morphological and yield performance characters as to the elite variety, MR219. Two lines, PB-2-107 and PB-2-34 were chosen to be potential lines because of their outstanding performances compared to parent, MR219. This study demonstrates a success story of MAS application in development of improved disease resistance lines of rice against BLB disease.

Resumo A mancha bacteriana das folhas (BLB) é uma das principais doenças do arroz na Malásia. Essa doença causa perdas substanciais de rendimento de até 70%. O desenvolvimento de variedades de arroz que herdaram características de resistência ao BLB é uma abordagem crucial para promover e sustentar a indústria do arroz na Malásia. Portanto, o objetivo deste estudo foi aumentar os caracteres BLB resistentes a doenças da variedade comercial MR219 de alto rendimento por meio de uma abordagem de cruzamento retrocruzamento com ferramenta de apoio de seleção assistida por marcador (MAS). O gene de resistência a BLB de amplo espectro, Xa7 do pai doador IRBB7, foi introgressado no MR219 suscetível (pai recorrente) usando dois marcadores flanqueadores ID7 e ID15. No BC3F4, conseguimos gerar 19 linhagens introgressadas com o gene Xa7 homozigoto e apresentamos características de resistência como genitor doador quando desafiado com Xanthomonas oryzae pv. oryzae por inoculação artificial. O pai recorrente MR219 e a variedade controle, MR263, estavam gravemente infectados pela doença. As linhas melhoradas exibiram características morfológicas e de desempenho de rendimento semelhantes às da variedade elite, MR219. Duas linhas, PB-2-107 e PB-2-34, foram escolhidas como linhas potenciais por causa de seus desempenhos excelentes em comparação com a mãe, MR219. Este estudo demonstra uma história de sucesso de aplicação de MAS no desenvolvimento de linhas de arroz melhoradas com resistência a doenças contra a doença BLB.

A disease predictive model based on epidemiological factors for the management of bacterial leaf blight of rice / Um modelo preditivo de doenças baseado em fatores epidemiológicos para o manejo da queima bacteriana das folhas do arroz

Rice is a widely consumed staple food for a large part of the world's human population. Approximately 90% of the world's rice is grown in Asian continent and constitutes a staple food for 2.7 billion people worldwide. Bacterial leaf blight (BLB) caused by Xanthomonas oryzae pv. oryzae is one of the devastating diseases of rice. A field experiment was conducted during the year 2016 and 2017 to investigate the influence of different meteorological parameters on BLB development as well as the computation of a predictive model to forecast the disease well ahead of its appearance in the field. The seasonal dataset of disease incidence and environmental factors was used to assess five rice varieties/ cultivars (Basmati-2000, KSK-434, KSK-133, Super Basmati, and IRRI-9). The accumulated effect of two year environmental data; maximum and minimum temperature, relative humidity, wind speed, and rainfall, was studied and correlated with disease incidence. Average temperature (maximum & minimum) showed a negative significant correlation with BLB disease and all other variables; relative humidity, rainfall, and wind speed had a positive correlation with BLB disease development on individual varieties. Stepwise regression analysis was performed to indicate potentially useful predictor variables and to rule out incompetent parameters. Environmental data from the growing seasons of July to October 2016 and 2017 revealed that, with the exception of the lowest temperature, all environmental factors contributed to disease development throughout the cropping season. A disease prediction multiple regression model was developed based on two-year data (Y = 214.3-3.691 Max T-0.508 Min T + 0.767 RH + 2.521 RF + 5.740 WS), which explained 95% variability. This disease prediction model will not only help farmers in early detection and timely management of bacterial leaf blight disease of rice but may also help reduce input costs and improve product quality and quantity. The model will be both farmer and environmentally friendly.

O arroz é um alimento básico amplamente consumido por grande parte da população humana mundial. Aproximadamente 90% do arroz do mundo é cultivado no continente asiático e constitui um alimento básico para 2,7 bilhões de pessoas em todo o mundo. O crestamento bacteriano das folhas (BLB) causado por Xanthomonas oryzae pv. oryzae é uma das doenças devastadoras do arroz. Um experimento de campo foi realizado durante os anos de 2016 e 2017 para investigar a influência de diferentes parâmetros meteorológicos no desenvolvimento do BLB, bem como o cálculo de um modelo preditivo para prever a doença bem antes de seu aparecimento em campo. O conjunto de dados sazonais de incidência de doenças e fatores ambientais foi usado para avaliar cinco variedades/cultivares de arroz (Basmati-2000, KSK-434, KSK-133, Super Basmati e IRRI-9). O efeito acumulado de dados ambientais de dois anos; temperatura máxima e mínima, umidade relativa do ar, velocidade do vento e precipitação pluviométrica foram estudados e correlacionados com a incidência da doença. A temperatura média (máxima e mínima) apresentou correlação significativa negativa com a doença BLB e todas as outras variáveis; umidade relativa, precipitação e velocidade do vento tiveram uma correlação positiva com o desenvolvimento da doença BLB em variedades individuais. A análise de regressão stepwise foi realizada para indicar variáveis preditoras potencialmente úteis e para descartar parâmetros incompetentes. Os dados ambientais das safras de julho a outubro de 2016 e 2017 revelaram que, com exceção da temperatura mais baixa, todos os fatores ambientais contribuíram para o desenvolvimento da doença ao longo da safra. Um modelo de regressão múltipla de previsão de doença foi desenvolvido com base em dados de dois anos (Y = 214,3-3,691 Max T-0,508 Min T + 0,767 RH + 2,521 RF + 5,740 WS), que explicou 95% de variabilidade. Este modelo de previsão de doenças não só ajudará os agricultores na detecção precoce e gestão atempada da doença bacteriana das folhas do arroz, mas também pode ajudar a reduzir os custos de insumos e melhorar a qualidade e a quantidade do produto. O modelo será agricultor e ambientalmente amigável.

Flavonoids­targeted metabolomic analysis following rice yellowing.

Flavonoids are the main metabolites responsible for yellowing of rice. However, the accumulation pattern of flavonoids and the metabolic basis of flavonoid biosynthesis during rice yellowing remain unclear. Thus, flavonoid-targeted metabolomics was used to investigate the composition and concentration of flavonoids in rice during yellowing. The results indicated the differential flavonoids at Month 3 and Month 5 of storage were more in composition and concentration with higher antioxidant capacity. Accumulated flavonoids were mainly flavones, flavonols, isoflavones, and anthocyanidins, of which rutin, farrerol, naringenin, cyanidin 3-rutinoside, and diosmetin were the indicators of rice yellowing. Metabolic association among flavonoids demonstrated the formation of yellow pigments was jointly induced by flavones, flavonols, isoflavones, and anthocyanidins metabolism. Examination of flavonoid metabolism presented in this study enhanced current understanding of the relationship between flavonoid metabolites and development of rice yellowing. It also offers a theoretical basis for targeted prediction of rice yellowing in the future.

Formation of rice protein fibrils is highly sensitive to the different types of metal ions: Aggregation behavior and possible mechanisms.

The effects of Ca2+, Cu2+, and Fe3+ on rice protein (RP) fibril formation were investigated in this study. Low Ca2+ concentration (≤150 mM) moderately unfolded the conformation of RP, promoting the exposure of hydrophobic sites and RP fibril assembly. Fibril formation was especially promoted with earlier addition of Ca2+. Cu2+ and Fe3+ inhibited RP fibril formation in a dose-dependent manner, and the inhibitory effect of Fe3+ was stronger due to higher affinity with RP. Additionally, the addition of Cu2+ and Fe3+ reduced α-helix and ß-sheet contents of RP, respectively, hindering the formation of stacked ß-sheet, the main internal structure of fibrils. These two ions also resulted in the formation of random aggregates within 15-50 nm, which further inhibited the conversion of proteins to fibrils. Moreover, Cu2+ and Fe3+ prevented the recruitment of nucleus into fibril-growth-sites, and formed fibrils were disrupted into fragments when these ions were added.

Acceleration mechanism of the rehydration process of dried rice noodles by the porous structure.

Rehydration of dried rice noodles (DRNs) is a time-consuming process, which is dominated by the compactness of noodle structure. Therefore, DRNs with differentiated porous structures were prepared, and their effect on the rehydration process was investigated. Porous structure can shorten rehydration time by reducing the time needed for water to migrate into the noodle core, or the water amount required for rehydration. Magnetic resonance imaging showed that although larger pores facilitate absorbing more water, the time for water to migrate into the noodle center is longer than that of medium size pores, as water needs to fill the periphery large hole before inward migration. SAXS analysis demonstrated that the presence of flexible starch molecular chains reduce the water required to achieve the maximum tensile strain of samples, thus shortening the rehydration time. Understanding the acceleration mechanism of porous structure on rehydration contributes to designing improved process of instant noodle products.

In vitro gastrointestinal digestion and colonic fermentation of media-milled black rice particle-stabilized Pickering emulsion: Phenolic release, bioactivity and prebiotic potential.

This is a pioneer study that investigated the digestive characteristics of Pickering emulsions stabilized by media-milled black rice particles during in vitro digestion and colonic fermentation. Free fatty acid release of the emulsions improved from 28.42 ± 3.13% to 33.68 ± 4.05% after media milling. The phenolics released from media-milled sample were close to those from unground sample. Media-milled sample exhibited higher DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging ability and α-glucosidase inhibition rate than unground sample. Media milling increased the generation of short-chain fatty acids (SCFAs) during colonic fermentation, especially acetic acid (23% improvement in media-milled sample over unground sample). It also inhibited the growth of harmful bacteria namely Escherichia Shigella and Streptococcus, and promoted the growth of beneficial bacteria including Bifidobacterium and Blautia. These findings revealed that media-milled black rice particle-stabilized Pickering emulsions possessed intrinsic bioactivity and prebiotic potentials in the gastrointestinal tract for the first time.

Relationship Between Rumen Microbial Composition and Fibrolytic Isozyme Activity During the Biodegradation of Rice Straw Powder Using Rumen Fluid.

Rumen fibrolytic microorganisms have been used to increase the rate of lignocellulosic biomass biodegradation; however, the microbial and isozymatic characteristics of biodegradation remain unclear. Therefore, the present study investigated the relationship between rumen microorganisms and fibrolytic isozymes associated with lignocellulosic biomass hydrolysis. Rice straw, a widely available agricultural byproduct, was ground and used as a substrate. The biodegradation of rice straw powder was performed anaerobically in rumen fluid for 48 h. The results obtained revealed that 31.6 and 23.3% of cellulose and hemicellulose, respectively, were degraded. The total concentration of volatile fatty acids showed a 1.8-fold increase (from 85.4 to 151.6| |mM) in 48 h, and 1,230.1| |mL L-1 of CO2 and 523.5| |mL L-1 of CH4 were produced. The major isozymes identified by zymograms during the first 12| |h were 51- and 140-kDa carboxymethyl cellulases (CMCases) and 23- and 57-kDa xylanases. The band densities of 37-, 53-, and 58-kDa CMCases and 38-, 44-, and 130-kDa xylanases increased from 24 to 36 h. A microbial ana-lysis indicated that the relative abundances of Prevotella, Fibrobacter, and Bacteroidales RF16 bacteria, Neocallimastix and Cyllamyces fungi, and Dasytricha and Polyplastron protozoa were related to fibrolytic isozyme activity. The present results provide novel insights into the relationships between fibrolytic isozymes and rumen microorganisms during lignocellulose biodegradation.

Bioavailability evaluation of epoxiconazole and difenoconazole in rice and the influence of dissolved organic matter in reducing uptake and translocation.

The aim of this study was to assess the bioavailability of epoxiconazole (EPO) and difenoconazole (DIF) in rice plants by evaluating their uptake, translocation, and accumulation. The results showed that the concentration of DIF in the roots was approximately three times higher than EPO, and both accumulated mainly in the roots. In addition, EPO continued to be transported from stems to leaves, causing a rise in its concentration in leaves. Contrastingly, only a minimal amount of DIF was transported to the leaves. This phenomenon is mainly governed by their differing octanol-water partition coefficient. The effects of dissolved organic carbon (DOC) on the accumulation of EPO and DIF in the roots were similar to those of the freely dissolved concentration measured by OECAMs. The concentrations of EPO and DIF in the roots and OECAMs consistently decreased with increasing DOC levels. Furthermore, a significant linear relationship was observed between the EPO and DIF concentrations in root and OECAMs. We also confirmed the accuracy and usefulness of the OECAMs method in predicting the bioavailability of EPO and DIF in rice roots. Therefore, OECAMs show good potential for use as a passive sampler to evaluate the bioavailability of EPO and DIF.

Enhanced As extraction from paddy soils with high As contamination risk by rice plant upon Si fertilization.

Owing to flooded growing conditions and specific physiological characteristics, rice plant is more efficient in As uptake and accumulation, which provides a cost-effective and time-efficient pathway to deplete bioavailable As from paddy soils. In the present study, the enhancing effect of silicon (Si) fertilization on As extraction from heavily contaminated paddy soils by rice was explored Upon incorporation of one weak acid Si fertilizer (AcSF), soil As solubility was significantly promoted by 1.3-1.4-fold, while a slightly increase in porewater As was observed with alkaline soluble Si fertilizer Na2SiO3 (AlSF). With both Si fertilizers applied before transplanting, a relatively low Si/As molar ratio (<100) in soil porewater was obtained, As a result, soil As uptake by rice plant with Si fertilizers was enhanced by 37.2%-171.7% compared to control (CK). Notably, up to 91.6% of the total As in rice plant retained in root with Si fertilization, suggesting the importance of root removal. By harvesting the whole rice plant including roots, soil bioavailable As measured by diffusive gradients in thin films (DGT) declined by 26.9%-31.3% in AlSF treatments relative to CK. Total soil As depletion by the whole rice plant was significantly enhanced from 2.8% in CK to 7.0%-11.2% in Si fertilizer treatments. In this way, 197.5 mg As m-2-232.5 mg As m-2 could be eliminated from soil following one rice-growth season, which was 2.3-2.7-fold higher compared to CK. These results identified the effectiveness of soluble Si fertilizer in enhancing soil As depletion by rice from paddy soils with high As contamination risk, which could serve as a cost-effective strategy with little technical-restriction.

Landscape structure influences natural pest suppression in a rice agroecosystem.

Agricultural landscapes are constantly changing as farmers adopt new production practices and respond to changing environmental conditions. Some of these changes alter landscape structure with impacts on natural pest control, pesticide use, and conservation of biodiversity. In rice agroecosystems the effect of landscape structure on natural enemies and pest suppression is often poorly understood. Here we investigate the effect of landscape composition and configuration on a key pest of rice, the brown planthopper (Nilaparvata lugens). Using N. lugens as sentinel prey coupled with predator exclusions, we investigated landscape effects on herbivore suppression and rice grain yield at multiple spatial scales in two regions of Bangladesh. Ladybird beetles and spiders were the most abundant natural enemies of N. lugens with landscape effects observed at all scales on ladybird beetles. Specifically, ladybird beetles were positively influenced by road edges, and fallow land, while spiders were strongly influenced only by rice phenology. Predator exclusion cages showed that N. lugens abundance significantly increased in caged plots, reducing rice gain yield. We also used an estimated biocontrol service index that showed a significant positive relationship with landscape diversity and a significant negative impact on pest density and yield loss. These results suggest that promoting fallow lands and fragmented patches between rice fields could lead to more sustainable insect pest management in rice agroecosystems, potentially reducing the practice of prophylactic insecticide use.

A de novo evolved gene contributes to rice grain shape difference between indica and japonica.

The role of de novo evolved genes from non-coding sequences in regulating morphological differentiation between species/subspecies remains largely unknown. Here, we show that a rice de novo gene GSE9 contributes to grain shape difference between indica/xian and japonica/geng varieties. GSE9 evolves from a previous non-coding region of wild rice Oryza rufipogon through the acquisition of start codon. This gene is inherited by most japonica varieties, while the original sequence (absence of start codon, gse9) is present in majority of indica varieties. Knockout of GSE9 in japonica varieties leads to slender grains, whereas introgression to indica background results in round grains. Population evolutionary analyses reveal that gse9 and GSE9 are derived from wild rice Or-I and Or-III groups, respectively. Our findings uncover that the de novo GSE9 gene contributes to the genetic and morphological divergence between indica and japonica subspecies, and provide a target for precise manipulation of rice grain shape.

Resource utilization of rice straw to prepare biochar as peroxymonosulfate activator for naphthalene removal: Performances, mechanisms, environmental impact and applicability in groundwater.

Herein, biochar was prepared using rice straw, and it served as the peroxymonosulfate (PMS) activator to degrade naphthalene (NAP). The results showed that pyrolysis temperature has played an important role in regulating biochar structure and properties. The biochar prepared at 900°C (BC900) had the best activation capacity and could remove NAP in a wide range of initial pH (5-11). In the system of BC900/PMS, multi-reactive species were produced, in which 1O2 and electron transfer mainly contributed to NAP degradation. In addition, the interference of complex groundwater components on the NAP removal rate must get attention. Cl- had a significant promotional effect but risked the formation of chlorinated disinfection by-products. HCO3-, CO32-, and humic acid (HA) had an inhibitory effect; surfactants had compatibility problems with the BC900/PMS system, which could lead to unproductive consumption of PMS. Significantly, the BC900/PMS system showed satisfactory remediation performance in spiked natural groundwater and soil, and it could solve the problem of persistent groundwater contamination caused by NAP desorption from the soil. Besides, the degradation pathway of NAP was proposed, and the BC900/PMS system could degrade NAP into low or nontoxic products. These suggest that the BC900/PMS system has promising applications in in-situ groundwater remediation.

Torrefaction interpretation through morphological and chemical transformations of agro-waste to porous carbon-based biofuel.

In the current study, two agro-waste lignocellulosic corncob (CC) and rice husk (RH) were thermally torrefied at 200-300 °C into a porous carbon-enriched biofuel. The scanning electron microscopy (SEM) of produced biofuel confirmed the rounded, homogenous, and spherical structure of the produced biofuels with higher porosity at a temperature between 250 and 300 °C with 60 min retention time. Brunauer-Emmett-Teller (BET) analysis indicated the high surface area (CC: 1.19-2.87 m2 g-1 and RH: 1.22-2.67 m2 g-1) and pore volume (CC: 1.23-2.81 ×10-3 m3 g-1 and RH: 1.46-2.58 ×10-3 m3 g-1). Crystallinity index decline percent (CC= 62.87% and RH=57.10%) estimated thermal stability and rise in amorphous cellulose reformation during (250-300 °C)/60 min that would efficiently hydrolyze during oxidative pyrolysis carbon reactive sites the rise in surface area and total pore's volume, having higher conversion rate as compared to raw materials. Carbon content was upgraded to 94% by eliminating hydrogen and oxygen from lignocellulosic agro-waste to produce energy-dense CC and RH. The lignin macromolecule transformation extent was estimated by O/C trend, which was equal to 63% and 47% for CC and RH, respectively, at 300 °C for 60 min. Due to low bulk density and pre-grinding energy requirements, torrefied biofuel with decomposed fibrous structure have lower transportation costs.

Effects of high manganese-cultivated seedlings on cadmium uptake by various rice (Oryza sativa L.) genotypes.

Cadmium (Cd) contamination in paddy soil threatens rice growth and food safety, enriching manganese (Mn) in rice seedlings is expected to reduce Cd uptake by rice. The effects of 250 µM Mn-treated seedlings on reducing Cd uptake of four rice genotypes (WYJ21, ZJY1578, HHZ, and HLYSM) planted in 0.61 mg kg-1 Cd-contaminated soil, were studied through the hydroponic and pot experiments. The results showed that the ZJY1578 seedling had the highest Mn level (459 µg plant-1), followed by WYJ21 (309 µg plant-1), and less Mn accumulated in the other genotypes. The relative expression of OsNramp5 (natural resistance-associated macrophage protein) was reduced by 42.7 % in ZJY1578 but increased by 23.3 % in HLYSM. The expressions of OsIRT1 (iron-regulated transporter-like protein) were reduced by 24.0-56.0 % in the four genotypes, with the highest reduction in ZJY1578. Consequently, a greater reduction of Cd occurred in ZJY1578 than that in the other genotypes, i.e., the root and shoot Cd at the tillering were reduced by 27.8 % and 48.5 %, respectively. At the mature stage, total Cd amount and distribution in the shoot and brown rice were also greatly reduced in ZJY1578, but the inhibitory effects were weakened compared to the tillering stage. This study found various responses of Cd uptake and transporters to Mn-treated seedlings among rice genotypes, thus resulting in various Cd reductions. In the future, the microscopic transport processes of Cd within rice should be explored to deeply explain the genotypic variation.

Regulation of straw decomposition and its effect on soil function by the amount of returned straw in a cool zone rice crop system.

The degradation process of returned straw in rice fields can improve soil organic matter and promote sustainable agriculture. The degradation process of returned straw is a humification process as well as a mineralization process involving microorganisms and enzymes. However, the degradation process of returned straw, the effect on straw decomposing microorganisms and the regulatory mechanism on potential functionality under cool climate flooding conditions are currently unknown.For this purpose, we investigated the biodegradation of straw from a biodegradation point of view at 20, 40, 71, 104, and 137 d after return under conventional (130 kg hm-2), 1/3 straw return (2933 kg hm-2), 2/3 straw return (5866 kg hm-2), and full straw return (8800 kg hm-2) applications in cool climate rice fields.. The test found Paludibacteraceae and Archaeaceae were the dominant bacteria for straw degradation, and their relative abundance was highest when 2/3 of straw was returned to the field. The straw degradation extracellular enzyme activity was higher in the late return period (104 d). At this time, the potential functionality of the soil differed significantly among the different return amounts, with the best extracellular enzyme activity and potential functionality at the 2/3 straw return amount. Therefore, the optimal amount of rice straw returned to the field is 5866 kg hm-2 at the current conventional N application rate (130 kg hm-2) in the cold zone.

Effects of the salinity-temperature interaction on seed germination and early seedling development: a comparative study of crop and weed species.

BACKGROUND: Weeds represent a great constraint for agricultural production due to their remarkable adaptability and their ability to compete with crops. Climate change exacerbates the abiotic stresses that plants encounter. Therefore, studying plant responses to adverse conditions is extremely important. Here, the response to saline stress at different temperatures of three weed species (Chenopodium album, Echinochloa crus-galli and Portulaca oleracea) and three crops (maize, soybean and rice) was investigated. RESULTS: The germination percentage of soybean notably decreased as salinity and low temperatures increased. In contrast, maize and rice consistently maintained a high germination percentage, particularly when subjected to low salinity levels. Regarding weed species, the germination percentage of C. album was not significantly affected by salinity, but it decreased in E. crus-galli and P. oleracea with increasing salinity. The mean germination time for all species increased with salinity, especially at lower temperatures. This effect was most pronounced for soybean and E. crus-galli. C. album exhibited significant reduction in stem growth with high salinity and high temperatures, while in E. crus-galli stem growth was less reduced under similar conditions. CONCLUSION: This study showed that successful germination under saline stress did not ensure successful early development and emphasizes the species-specific nature of the temperature-salinity interaction, perhaps influenced by intraspecific variability. Increasing salinity levels negatively impacted germination and seedling growth in most species, yet higher temperatures partially alleviated these effects.

Oryzibacter oryziterrae gen. nov., sp. nov., isolated from rice paddy soil.

A novel Gram-stain-negative, aerobic, motile and pleomorphic rod-shaped bacterial strain, designated COJ-58T, was isolated from rice paddy soil. Strain COJ-58T grew optimally at 20-30 °C, at pH 5.0-8.0 and with 0-1.0 % (w/v) NaCl. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain COJ-58T forms a distinct lineage within the family Pleomorphomonadaceae, with highest similarity to Pleomorphomonas carboxyditropha SVCO-16T (95.9 %), Pleomorphomonas koreensis Y9T (95.8 %), Pleomorphomonas oryzae F-7T (95.7 %) and Pleomorphomonas diazotrophica R5-392T (95.6 %), respectively. The average nucleotide identity, digital DNA-DNA hybridization, average amino acid identity and percentage of conserved proteins values between the genomes of strain COJ-58T and its closely related taxa are ≤77.2 %, ≤21.6 %, ≤68.3 % and ≤61.3 %, respectively. The genome size of strain COJ-58T is 4.9 Mb and the genomic DNA G + C content is 63.7 mol%. The major fatty acids are C18 : 1 ω7c, C16 : 0 and summed feature 2 (C14 : 0 3-OH and/or iso-C16 : 1 I). The differential phenotypic and genotypic characteristics of strain COJ-58T indicate that it represents a novel genus and species, for which the name Oryzibacter oryziterrae gen. nov., sp. nov. is proposed, with strain COJ-58T (=KACC 22108T=JCM 34744T) as the type strain.

Integrated genome-wide differentiation and association analyses identify causal genes underlying breeding-selected grain quality traits in japonica rice.

Improving grain quality is a primary objective in contemporary rice breeding. Japanese modern rice breeding has developed two different types of rice, eating and sake-brewing rice, with different grain characteristics, indicating the selection of variant gene alleles during the breeding process. Given the critical importance of promptly and efficiently identifying genes selected in past breeding for future molecular breeding, we conducted genome scans for divergence, genome-wide association studies, and map-based cloning. Consequently, we successfully identified two genes, OsMnS and OsWOX9D, both contributing to rice grain traits. OsMnS encodes a mannan synthase that increases the white core frequency in the endosperm, a desirable trait for sake brewing but decreases the grain appearance quality. OsWOX9D encodes a grass-specific homeobox-containing transcription factor, which enhances grain width for better sake brewing. Furthermore, haplotype analysis revealed that their defective alleles were selected in East Asia, but not Europe, during modern improvement. In addition, our analyses indicate that a reduction in grain mannan content during African rice domestication may also be caused a defective OsMnS allele due to breeding selection. This study not only reveals the delicate balance between grain appearance quality and nutrition in rice but also provides a new strategy for isolating causal genes underlying complex traits, based on the concept of "breeding-assisted genomics" in plants.

Association Mapping of Candidate Genes Associated with Iron and Zinc Content in Rice (Oryza sativa L.) Grains.

Micronutrient deficiencies, particularly of iron (Fe) and zinc (Zn), in the diet contribute to health issues and hidden hunger. Enhancing the Fe and Zn content in globally staple food crops like rice is necessary to address food malnutrition. A Genome-Wide Association Study (GWAS) was conducted using 85 diverse rice accessions from the Democratic Republic of Congo (DRC) to identify genomic regions associated with grain Fe and Zn content. The Fe content ranged from 0.95 to 8.68 mg/100 g on a dry weight basis (dwb) while Zn content ranged from 0.87 to 3.8 mg/100 g (dwb). Using MLM and FarmCPU models, we found 10 significant SNPs out of which one SNP on chromosome 11 was associated with the variation in Fe content and one SNP on chromosome 4 was associated with the Zn content, and both were commonly detected by the two models. Candidate genes belonging to transcription regulator activities, including the bZIP family genes and MYB family genes, as well as transporter activities involved in Fe and Zn homeostasis were identified in the vicinity of the SNP markers and selected. The identified SNP markers hold promise for marker-assisted selection in rice breeding programs aimed at enhancing Fe and Zn content in rice. This study provides valuable insights into the genetic factors controlling Fe and Zn uptake and their transport and accumulation in rice, offering opportunities for developing biofortified rice varieties to combat malnutrition among rice consumers.