Alkalinity stress is a major hindrance to enhancing rice production globally due to its damaging effect on plants\' growth and development compared with salinity stress. However, understanding of the physiological and molecular mechanisms of alkalinity tolerance is limited. Therefore, a panel of indica and japonica rice genotypes was evaluated for alkalinity tolerance at the seedling stage in a genome-wide association study to identify tolerant genotypes and candidate genes. Principal component analysis revealed that traits such as alkalinity tolerance score, shoot dry weight, and shoot fresh weight had the highest contribution to variations in tolerance, while shoot Na+ concentration, shoot Na+:K+ ratio, and root-to-shoot ratio had moderate contributions. Phenotypic clustering and population structure analysis grouped the genotypes into five subgroups. Several salt-susceptible genotypes such as IR29, Cocodrie, and Cheniere placed in the highly tolerant cluster suggesting different underlying tolerance mechanisms for salinity and alkalinity tolerance. Twenty-nine significant SNPs associated with alkalinity tolerance were identified. In addition to three alkalinity tolerance QTLs, qSNK4, qSNC9, and qSKC10, which co-localized with the earlier reported QTLs, a novel QTL, qSNC7, was identified. Six candidate genes that were differentially expressed between tolerant and susceptible genotypes were selected: LOC_Os04g50090 (Helix-loop-helix DNA-binding protein), LOC_Os08g23440 (amino acid permease family protein), LOC_Os09g32972 (MYB protein), LOC_Os08g25480 (Cytochrome P450), LOC_Os08g25390 (Bifunctional homoserine dehydrogenase), and LOC_Os09g38340 (C2H2 zinc finger protein). The genomic and genetic resources such as tolerant genotypes and candidate genes would be valuable for investigating the alkalinity tolerance mechanisms and for marker-assisted pyramiding of the favorable alleles for improving alkalinity tolerance at the seedling stage in rice.

As an important driving force, introgression plays an essential role in shaping the evolution of plant species. However, knowledge concerning how introgression affects plant evolution in agroecosystems with strong human influences is still limited. To generate such knowledge, we used InDel (insertion/deletion) molecular fingerprints to determine the level of introgression from japonica rice cultivars into the indica type of weedy rice. We also analyzed the impact of crop-to-weed introgression on the genetic differentiation and diversity of weedy rice, using InDel (insertion/deletion) and SSR (simple sequence repeat) molecular fingerprints. Results based on the STRUCTURE analysis indicated an evident admixture of some weedy rice samples with indica and japonica components, suggesting different levels of introgression from japonica rice cultivars to the indica type of weedy rice. The principal coordinate analyses indicated indica-japonica genetic differentiation among weedy rice samples, which was positively correlated with the introgression of japonica-specific alleles from the rice cultivars. In addition, increased crop-to-weed introgression formed a parabola pattern of dynamic genetic diversity in weedy rice. Our findings based on this case study provide evidence that human activities, such as the frequent change in crop varieties, can strongly influence weed evolution by altering genetic differentiation and genetic diversity through crop-weed introgression in agroecosystems.

In rice (Oryza sativa L.), rice bran contains valuable nutritional constituents, such as high unsaturated fat content, tocotrienols, inositol, γ-oryzanol, and phytosterols, all of which are of nutritional and pharmaceuticals interest. There is now a rising market demand for rice bran oil, which makes research into their content and fatty acid profile an area of interest. As it is evident that lipid content has a substantial impact on the eating, cooking, and storage quality of rice, an understanding of the genetic mechanisms that determine oil content in rice is of great importance, equal to that of rice quality. Therefore, in this study, we performed a genome-wide association study on the composition and oil concentration of 161 Vietnamese rice varieties. Five categories of fatty acids in rice bran were discovered and the bran oil concentration profile in different rice accessions was identified. We also identified 229 important markers related to the fatty acid composition of bran oil, distributed mainly on chromosomes 1 and 7. Seven quantitative trait loci and five potential genes related to unsaturated fatty acid content were detected, including OsKASI, OsFAD, OsARF, OsGAPDH, and OsMADS29. These results provide insights into the genetic basis of rice bran oil composition, which is pivotal to the metabolic engineering of rice plants with desirable bran oil content through candidate genes selection.

Plant height (PH) in rice (Oryza sativa) is an important trait for its adaptation and agricultural performance. Discovery of the semi-dwarf1 (SD1) mutation initiated the Green Revolution, boosting rice yield and fitness, but the underlying genetic regulation of PH in rice remains largely unknown. Here, we performed genome-wide association study (GWAS) and identified 12 non-repetitive QTL/genes regulating PH variation in 619 Asian cultivated rice accessions. One of these was an SD1 structural variant, not normally detected in standard GWAS analyses. Given the strong effect of SD1 on PH, we also divided 619 accessions into subgroups harbouring distinct SD1 haplotypes, and found a further 85 QTL/genes for PH, revealing genetic heterogeneity that may be missed by analysing a broad, diverse population. Moreover, we uncovered two epistatic interaction networks of PH-associated QTL/genes in the japonica (Geng)-dominant SD1NIP subgroup. In one of them, the hub QTL/gene qphSN1.4/GAMYB interacted with qphSN3.1/OsINO80, qphSN3.4/HD16/EL1, qphSN6.2/LOC_Os06g11130, and qphSN10.2/MADS56. Sequence variations in GAMYB and MADS56 were associated with their expression levels and PH variations, and MADS56 was shown to physically interact with MADS57 to coregulate expression of gibberellin (GA) metabolic genes OsGA2ox3 and Elongated Uppermost Internode1 (EUI1). Our study uncovered the multifaceted genetic architectures of rice PH, and provided novel and abundant genetic resources for breeding semi-dwarf rice and new candidates for further mechanistic studies on regulation of PH in rice.

Brassinosteroids (BRs) regulate a diverse spectrum of processes during plant growth and development and modulate plant physiology in response to environmental fluctuations and stress factors. Thus, the BR signaling regulators have the potential to be targeted for gene editing to optimize the architecture of plants and make them more resilient to environmental stress. Our understanding of the BR signaling mechanism in monocot crop species is limited compared to our knowledge of this process accumulated in the model dicot species - Arabidopsis thaliana. A deeper understanding of the BR signaling and response during plant growth and adaptation to continually changing environmental conditions will provide insight into mechanisms that govern the coordinated expression of the BR signaling genes in rice (Oryza sativa) which is a model for cereal crops. Therefore, in this study a comprehensive and detailed in silico analysis of promoter sequences of rice BR signaling genes was performed. Moreover, expression profiles of these genes during various developmental stages and reactions to several stress conditions were analyzed. Additionally, a model of interactions between the encoded proteins was also established. The obtained results revealed that promoters of the 39 BR signaling genes are involved in various regulatory mechanisms and interdependent processes that influence growth, development, and stress response in rice. Different transcription factor-binding sites and cis-regulatory elements in the gene promoters were identified which are involved in regulation of the genes\' expression during plant development and reactions to stress conditions. The in-silico analysis of BR signaling genes in O. sativa provides information about mechanisms which regulate the coordinated expression of these genes during rice development and in response to other phytohormones and environmental factors. Since rice is both an important crop and the model species for other cereals, this information may be important for understanding the regulatory mechanisms that modulate the BR signaling in monocot species. It can also provide new ways for the plant genetic engineering technology by providing novel potential targets, either cis-elements or transcriptional factors, to create elite genotypes with desirable traits.

The calmodulin-binding transcription activator (CAMTA) is a family of transcriptional factors containing a cluster of calmodulin-binding proteins that can activate gene regulation in response to stresses. The presence of this family of genes has been reported earlier, though, the comprehensive analyses of rice CAMTA (OsCAMTA) genes, their promoter regions, and the proteins were not deliberated till date. The present report revealed the existence of seven CAMTA genes along with their alternate transcripts in five chromosomes of rice (Oryza sativa) genome. Phylogenetic trees classified seven CAMTA genes into three clades indicating the evolutionary conservation in gene structure and their association with other plant species. The in silico study was carried out considering 2 kilobases (kb) promoter regions of seven OsCAMTA genes regarding the distribution of transcription factor binding sites (TFbs) of major and plant-specific transcription factors whereas OsCAMTA7a was identified with highest number of TFbs, while OsCAMTA4 had the lowest. Comparative modelling, i.e., homology modelling, and molecular docking of the CAMTA proteins contributed the thoughtful comprehension of protein 3D structures and protein-protein interaction with probable partners. Gene ontology annotation identified the involvement of the proteins in biological processes, molecular functions, and localization in cellular components. Differential gene expression study gave an insight on functional multiplicity to showcase OsCAMTA3b as most upregulated stress-responsive gene. Summarization of the present findings can be interpreted that OsCAMTA gene duplication, variation in TFbs available in the promoters, and interactions of OsCAMTA proteins with their binding partners might be linked to tolerance against multiple biotic and abiotic cues.

Evaluating the impact of increasing CO2 on rice quality is becoming a global concern. However, whether adjusting the source-sink ratio will affect the response of rice grain quality to elevated CO2 concentrations remains unknown. In 2016-2018, we conducted a free-air CO2 enrichment experiment using a popular japonica cultivar grown at ambient and elevated CO2 levels (eCO2, increased by 200 ppm), reducing the source-sink ratio via cutting leaves (LC) at the heading stage, to investigate the effects of eCO2 and LC and their interactions on rice processing, appearance, nutrition, and eating quality. Averaged across 3 years, eCO2 significantly decreased brown rice percentage (-0.5%), milled rice percentage (-2.1%), and head rice percentage (-4.2%) but increased chalky grain percentage (+ 22.3%) and chalkiness degree (+ 26.3%). Markedly, eCO2 increased peak viscosity (+ 2.9%) and minimum viscosity (+ 3.8%) but decreased setback (-96.1%) of powder rice and increased the appearance (+ 4.5%), stickiness (+ 3.5%) and balance degree (+ 4.8%) of cooked rice, while decreasing the hardness (-6.7%), resulting in better palatability (+ 4.0%). Further, eCO2 significantly decreased the concentrations of protein, Ca, S, and Cu by 5.3, 4.7, 2.2, and 9.6%, respectively, but increased K concentration by 3.9%. Responses of nutritional quality in different grain positions (brown and milled rice) to eCO2 showed the same trend. Compared with control treatment, LC significantly increased chalky grain percentage, chalkiness degree, protein concentration, mineral element levels (except for B and Mn), and phytic acid concentration. Our results indicate that eCO2 reduced rice processing suitability, appearance, and nutritional quality but improved the eating quality. Rice quality varied significantly among years; however, few CO2 by year, CO2 by LC, or CO2 by grain position interactions were detected, indicating that the effects of eCO2 on rice quality varied little with the growing seasons, the decrease in the source-sink ratios or the different grain positions.

Excessive Cd content and high Cd/Zn ratio in rice grains threaten human health. To study the reduction effects of combined soil amendments on Cd content and Cd/Zn ratio in rice planting in soils with different Cd contamination levels, we conducted field trials in three regions of Hunan province, China. Six field treatments were designed in each study area, including control (CK), lime alone (L), lime combined with sepiolite (LS), phosphate fertilizer (LP), organic fertilizer (LO) and phosphate fertilizer + organic fertilizer (LPO). The application of the combined amendments reduced the Cd content in rice grains to less than the Food Health Standard of China (0.2 mg/kg) and the Cd/Zn ratio to less than the safety threshold of 0.015. The average reduction rates of grain Cd content under the combined treatments among the three regions increased with the increase in Cd content in the soil. Meanwhile, the amendments also decreased the soil available Cd and Zn concentration significantly. The LO had the highest efficiency on decreasing Cd content in rice grains among these amendments, which is ranged from 44.6% to 52.8% in the three regions compared with CK. Similarly, high reduction rates of Cd/Zn ratio were found in the LO treatment, with an average value of 57.3% among the three regions. The grain Cd contents and Cd/Zn ratios were significantly correlated with the soil available Cd concentrations, plant uptake factor and the straw to rice grain translocation factor (TFgs) (P < 0.05). The results indicated that the combined soil amendments, especially lime combined with organic fertilizer, would be an effective way to control Cd content in rice.

To understand the effects of source-sink relationships on rice yield response to elevated CO2 levels (eCO2), we conducted a field study using a popular japonica cultivar grown in a free-air CO2 enrichment environment in 2017-2018. The source-sink ratio of rice was set artificially via source-sink treatments (SSTs) at the heading stage. Five SSTs were performed in 2017 (EXP1): cutting off the flag leaf (LC1) and the top three functional leaves (LC3), removing one branch in every three branches of a panicle (SR1/3) and one branch in every two branches of a panicle (SR1/2), and the control (CK) without any leaf cutting or spikelet removal. The eCO2 significantly increased grain yield by 15.7% on average over all treatments; it significantly increased grain yield of CK, LC1, LC3, SR1/3, and SR1/2 crops by 13.9, 18.1, 25.3, 12.0, and 10.9%, respectively. The yield response to eCO2 was associated with a significant increase of panicle number and fully-filled grain percentage (FGP), and the response of crops under different SSTs was significantly positively correlated with FGP and the average grain weight of the seeds. Two SSTs (CK and LC3) were performed in 2018 (EXP2), which confirmed that the yield response of LC3 crops (25.1%) to eCO2 was significantly higher than that of CK (15.9%). Among the different grain positions, yield response to eCO2 of grains attached to the lower secondary rachis was greater than that of grains attached to the upper primary rachis. Reducing the source-sink ratio via leaf-cutting enhanced the net photosynthetic rate response of the remaining leaves to eCO2 and increased the grain filling ability. Conversely, spikelet removal increased the non-structural carbohydrate (NSC) content of the stem, causing feedback inhibition and photosynthetic down-regulation. This study suggests that reducing the source-sink ratio by adopting appropriate management measures can increase the response of rice to eCO2.

Species interactions and mechanisms affect plant coexistence and community assembly. Despite increasing knowledge of kin recognition and allelopathy in regulating inter-specific and intra-specific interactions among plants, little is known about whether kin recognition mediates allelopathic interference. We used allelopathic rice cultivars with the ability for kin recognition grown in kin versus non-kin mixtures to determine their impacts on paddy weeds in field trials and a series of controlled experiments. We experimentally tested potential mechanisms of the interaction via altered root behaviour, allelochemical production and resource partitioning in the dominant weed competitor, as well as soil microbial communities. We consistently found that the establishment and growth of paddy weeds were more inhibited by kin mixtures compared to non-kin mixtures. The effect was driven by kin recognition that induced changes in root placement, altered weed carbon and nitrogen partitioning, but was associated with similar soil microbial communities. Importantly, genetic relatedness enhanced the production of intrusive roots towards weeds and reduced the production of rice allelochemicals. These findings suggest that relatedness allows allelopathic plants to discriminate their neighbouring collaborators (kin) or competitors and adjust their growth, competitiveness and chemical defense accordingly.