Rice Improvement Research Articles

The Landscape of Presence/Absence Variations during the Improvement of Rice

Rice is one of the most important staple crops in the world; therefore, the improvement of rice holds great significance for enhancing agricultural production and addressing food security challenges. Although there have been numerous studies on the role of single-nucleotide polymorphisms (SNPs) in rice improvement with the development of next-generation sequencing technologies, research on the role of presence/absence variations (PAVs) in the improvement of rice is limited. In particular, there is a scarcity of studies exploring the traits and genes that may be affected by PAVs in rice. Here, we extracted PAVs utilizing resequencing data from 148 improved rice varieties distributed in Asia. We detected a total of 33,220 PAVs and found that the number of variations decreased gradually as the length of the PAVs increased. The number of PAVs was the highest on chromosome 1. Furthermore, we identified a 6 Mb hotspot region on chromosome 11 containing 1091 PAVs in which there were 29 genes related to defense responses. By conducting a genome-wide association study (GWAS) using PAV variation data and phenotypic data for five traits (flowering time, plant height, flag leaf length, flag leaf width, and panicle number) across all materials, we identified 186 significantly associated PAVs involving 20 cloned genes. A haplotype analysis and expression analysis of candidate genes revealed that important genes might be affected by PAVs, such as the flowering time gene OsSFL1 and the flag leaf width gene NAL1. Our work investigated the pattern in PAVs and explored important PAV key functional genes associated with agronomic traits. Consequently, these results provide potential and exploitable genetic resources for rice breeding.

OsLSC6 Regulates Leaf Sheath Color and Cold Tolerance in Rice Revealed by Metabolite Genome Wide Association Study

Plant metabolites including anthocyanins play an important role in the growth of plants, as well as in regulating biotic and abiotic stress responses to the environment. Here we report comprehensive profiling of 3315 metabolites and a further metabolic-based genome-wide association study (mGWAS) based on 292,485 SNPs obtained from 311 rice accessions, including 160 wild and 151 cultivars. We identified hundreds of common variants affecting a large number of secondary metabolites with large effects at high throughput. Finally, we identified a novel gene namely OsLSC6 (Oryza sativa leaf sheath color 6), which encoded a UDP 3-O-glucosyltransferase and involved in the anthocyanin biosynthesis of Cyanidin-3-Galc (sd1825) responsible for leaf sheath color, and resulted in significant different accumulation of sd1825 between wild (purple) and cultivars (green). The results of knockout transgenic experiments showed that OsLSC6 regulated the biosynthesis and accumulation of sd1825, controlled the purple leaf sheath. Our further research revealed that OsLSC6 also confers resistance to cold stress during the seedling stage in rice. And we identified that a SNP in OsLSC6 was responsible for the leaf sheath color and chilling tolerance, supporting the importance of OsLSC6 in plant adaption. Our study could not only demonstrate that OsLSC6 is a vital regulator during anthocyanin biosynthesis and abiotic stress responses, but also provide a powerful complementary tool based on metabolites-to-genes analysis by mGWAS for functional gene identification andpromising candidate in future rice breeding and improvement.

Improvement of rice yields in the Mancherial district of Telangana with the use of the STCR targeted yield strategy

Improvement of rice yields in the Mancherial district of Telangana with the use of the STCR targeted yield strategy

Comparative miRNome and transcriptome analyses reveal the expression of novel miRNAs in the panicle of rice implicated in sustained agronomic performance under terminal drought stress.

Differential expression of 128 known and 111 novel miRNAs in the panicle of Nagina 22 under terminal drought stress targeting transcription factors, stress-associated genes, etc., enhances drought tolerance and helps sustain agronomic performance under terminal drought stress. Drought tolerance is a complex multigenic trait, wherein the genes are fine-tuned by coding and non-coding components in mitigating deleterious effects. MicroRNA (miRNA) controls gene expression at post-transcriptional level either by cleaving mRNA (transcript) or by suppressing its translation. miRNAs are known to control developmental processes and abiotic stress tolerance in plants. To identify terminal drought-responsive novel miRNA in contrasting rice cultivars, we constructed small RNA (sRNA) libraries from immature panicles of drought-tolerant rice [Nagina 22 (N 22)] and drought-sensitive (IR 64) cultivars grown under control and terminal drought stress. Our analysis of sRNA-seq data resulted in the identification of 169 known and 148 novel miRNAs in the rice cultivars. Among the novel miRNAs, 68 were up-regulated while 43 were down-regulated in the panicle of N 22 under stress. Interestingly, 31 novel miRNAs up-regulated in N 22 were down-regulated in IR 64, whereas 4 miRNAs down-regulated in N 22 were up-regulated in IR 64 under stress. To detect the effects of miRNA on mRNA expression level, transcriptome analysis was performed, while differential expression of miRNAs and their target genes was validated by RT-qPCR. Targets of the differentially expressed miRNAs include transcription factors and stress-associated genes involved in cellular/metabolic/developmental processes, response to abiotic stress, programmed cell death, photosynthesis, panicle/seed development, and grain yield. Differential expression of the miRNAs could be validated in an independent set of the samples. The findings might be useful in genetic improvement of drought-tolerant rice.

Improvement of Broad-Spectrum Disease-Resistant Rice by the Overexpression of BSR1 via a Moderate-Strength Constitutive Promoter and a Pathogen-Inducible Promoter.

Conferring crops with resistance to multiple diseases is crucial for stable food production. Genetic engineering is an effective means of achieving this. The rice receptor-like cytoplasmic kinase BSR1 mediates microbe-associated molecular pattern-induced immunity. In our previous study, we demonstrated that rice lines overexpressing BSR1 under the control of the maize ubiquitin promoter exhibited broad-spectrum resistance to rice blast, brown spot, leaf blight, and bacterial seedling rot. However, unfavorable phenotypes were observed, such as a decreased seed germination rate and a partial darkening of husked rice. Herein, we present a strategy to address these unfavorable phenotypes using an OsUbi7 constitutive promoter with moderate expression levels and a pathogen-inducible PR1b promoter. Rice lines expressing BSR1 under the influence of both promoters maintained broad-spectrum disease resistance. The seed germination rate and coloration of husked rice were similar to those of the wild-type rice.

Functional differentiation and genetic diversity of rice cation exchanger (CAX) genes and their potential use in rice improvement

Cation exchanger (CAX) genes play an important role in plant growth/development and response to biotic and abiotic stresses. Here, we tried to obtain important information on the functionalities and phenotypic effects of CAX gene family by systematic analyses of their expression patterns, genetic diversity (gene CDS haplotypes, structural variations, gene presence/absence variations) in 3010 rice genomes and nine parents of 496 Huanghuazhan introgression lines, the frequency shifts of the predominant gcHaps at these loci to artificial selection during modern breeding, and their association with tolerances to several abiotic stresses. Significant amounts of variation also exist in the cis-regulatory elements (CREs) of the OsCAX gene promoters in 50 high-quality rice genomes. The functional differentiation of OsCAX gene family were reflected primarily by their tissue and development specific expression patterns and in varied responses to different treatments, by unique sets of CREs in their promoters and their associations with specific agronomic traits/abiotic stress tolerances. Our results indicated that OsCAX1a and OsCAX2 as general signal transporters were in many processes of rice growth/development and responses to diverse environments, but they might be of less value in rice improvement. OsCAX1b, OsCAX1c, OsCAX3 and OsCAX4 was expected to be of potential value in rice improvement because of their associations with specific traits, responsiveness to specific abiotic stresses or phytohormones, and relatively high gcHap and CRE diversity. Our strategy was demonstrated to be highly efficient to obtain important genetic information on genes/alleles of specific gene family and can be used to systematically characterize the other rice gene families.

SCREENING OF RICE VARIETIES TOLERANT AND SUSCEPTIBLE TO IRON TOXICITY IN INLAND VALLEY SWAMPS (IVS) IN SIERRA LEONE

Iron toxicity is a nutrient disorder, which is brought about by the uptake of Fe (II) in amounts that disrupt or over-expressseveral metabolic processes, resulting in damage to the rice plant. A pot experiment was conducted in a greenhouse at Sierra Leone Agricultural Business Initiatives (SABI) Center Newton to enable the screening of many different varieties thereby resulting in the identification of both tolerant and susceptible varieties that could be used as breeding materials for rice improvement programs in the tropics. The hot spot iron toxic soils used in the pot were weighed at a uniform rate The parameters or items of measurement to be looked at could be the dry weight of root, dry weight of stem, dry weight of filled grain, dry weight of unfilled grains, plant height (cm) of the genotypes, tiller number of the genotypes, leaf area (cm2)of the genotypes, phenotypic acceptability of the genotypes, Iron-toxicity score of the genotypes Based on their performance in terms of toxicity scores, dry root weight, stem weight, leaf area, unfilled grains, filled grains, tiller count, plant height, and phenotypic acceptability the following varieties have been identified as iron toxicity tolerant with high yield potential Gbassay, Rok-24, Chim, kiamp.

Cereals can trap endophytic bacteria with potential beneficial traits when grown ex-situ in harsh soils.

Microbial communities associated with plants growing in harsh conditions, including salinity and water deficiency, have developed adaptive features which permit them to grow and survive under extreme environmental conditions. In the present study, an ex-situ plant trapping method has been applied to collect the culturable microbial diversity associated with the soil from harsh and remote areas. Oryza sativa cv. Baldo and Triticum durum Primadur plants were used as recruiters, while the soil surrounding the roots of Oryza glaberrima plants from remote regions of Mali (West Africa) was used as substrate for their growth. The endophytic communities recruited by the two plant species belonged to Proteobacteria and Firmicutes, and the dominant genera were Bacillus, Kosakonia, and Enterobacter. These endophytes were characterized by analyzing some of the most common plant growth promoting traits. Halotolerant, inorganic phosphate-solubilizing and N-fixing strains were found, and some of them simultaneously showing these three traits. We verified that 'Baldo' recruited mostly halotolerant and P-solubilizers endophytes, while the endophytes selected by 'Primadur' were mainly N-fixers. The applied ex-situ plant trapping method allowed to isolate endophytes with potential beneficial traits that could be applied for the improvement of rice and wheat growth under adverse environmental conditions.

Physio-biochemical responses of rice to smart nitrogen fertilizer nano-hydrogel under drought stress and flooding alternation: An eco-friendly innovative approach to enhance the grain quality

Fertilizer nanohydrogels have been newly introduced to agriculture. Herein, we investigated the effects of smart nitrogen (N) fertilizer nano-hydrogel on rice's physio-biochemical traits and seed quality under different drought stress and flooding conditions. Therefore, a pot experiment was conducted with no N fertilizer delivery 0 g N/kg (NND), with conventional N fertilizer delivery 0.175 g N/kg (CND), and smart N fertilizer delivery 0.175 g N/kg (SND) using three irrigation options: flooding/moderate dryness (FMD, 90% saturation, 30 mm submerged), flooding/sharp dryness (FSD, 70% saturation, 30 mm submerged), and flooding/extreme dryness (FED, 50% saturation, 30 mm submerged). Results showed that rice root's active acquisition area and enzymatic activity were larger in FMD × SND than in FED × NND, as the former combination increased NH4+, NO3−, and TN contents in the soil while the latter reduced their availability. The optimal N nutrition improved the plant cell ultrastructure, and the catalase activity was higher in the FMD × SND (42.0 µg/min) compared to the FED × NND (25.2 µg/min), leading to the alleviation of oxidative cell injury, as verified by lesser radical oxidative sequences and lipid peroxidation. The leaf soluble protein (8.8 mg/g) and sugar (36.9 mg/g) were greater in FMD × SND than in FED × NND (5.3 mg/g), (10.4 mg/g), and (8.3 µg/g h), respectively. Moreover, α-amylase and glutamine synthetase activities were larger in the FMD × SND than in FED × NND. Meanwhile, the FMD × SND resulted in the highest milled rice (87.3%), amylose (17.6%), protein (12.4%), and fatty acids (11.1%). However, the combination of FED × NND led to the lowest milled rice (70.0%), amylose (5.6%), protein (5.3%), and fatty acids (6.6%). This study suggests that the FMD × SND can guarantee physio-biochemical improvements in rice, thereby increasing grain quality.

Molecular basis of distinct responses to drought between rice and wheat genotypes

The water-intensive cultivation of rice in flooded conditions poses a great challenge to global water resources, particularly in the face of climate change. Wheat requires significantly less water than rice with fairly high yields. A main research question is whether there are any drought adaptive mechanisms in rice similar to those existing in wheat. We report a study, where precise moisture stress was imposed at the vegetative stage using a high-throughput drought simulator phenomics facility. Variability in stress response was captured through untargeted metabolome analysis. In the first experiment, using two rice cultivars with contrasting drought responses (IR64 and Apo) and a wheat cultivar, Weebill, we found distinct variations in regulation of metabolite levels under well-watered or water-limited conditions, both between and within species. Several biosynthetic pathways, including those of polyamines, flavonoids, flavones, and specific metabolites like sphinganine-1-phosphate and anthranilate were enhanced under water-limited conditions. These metabolites were more abundant in wheat than in rice. Among the rice cultivars, Apo had higher abundance of metabolites than IR64. Regulatory patterns governing these metabolic pathways provided basis for stress responses of the tolerant rice cultivar and wheat. A second experiment was conducted to screen rice germplasm lines for acquired tolerance traits contrasting for drought responsiveness. Eight genotypes (four sensitive and four tolerant ones) were selected to validate the candidate metabolites from the first experiment. In the third experiment, an RT-PCR analysis of the key regulatory genes governing the metabolic pathways was conducted, using plant materials from both experiments. The results of this third experiment provided a strong validation for the relevance of these metabolic pathways under stress in rice. We identified promising rice genotypes that exhibited fold change levels of stress resistance metabolites comparable to those of the wheat cultivar Weebill. This study helped identify key candidate metabolites that can be either developed as markers or as elicitors, and genotypes that can be used as trait donor lines for rice improvement.

Influence of nitrogen substitution at an equivalent total nitrogen level on bacterial and fungal communities, as well as enzyme activities of the ditch bottom soil in a rice-fish coculture system.

Rice-fish coculture system (RFS) operates through effectively utilizing water and land resources in a complementary form, but it requires more efficient utilization of fertilizer and feed without compromising rice yield. However, the knowledge of how to regulate the proportion of nitrogen (N) supplied from fertilizer and feed at an equivalent total N levelto improve the benefits of RFS remains limited. Therefore, four treatments (S0: RFS with 0% N fromfertilizer and 100% N from feed; S25: RFS with 25% N from fertilizer and 75% N from feed; S50: RFS with 50% N from fertilizer and 50% N from feed; S75: RFS with 75% N from fertilizer and 25% N from feed) were conducted to assess the variation of ditch bottom soil properties, microbial communities and enzyme activities, as well as to obtain the optimal ratio of N supplied from fish feed and fertilizer. The experiments showed that the contents of soil organic matter, total carbon and total N, and the activities of urease, N-acetyl-β-D-glucosaminidase, protease, β-1,4-glucosidase and catalase in the ditch bottom soil significantly reduced in S25 treatment, compared with the other three treatments. Ammonium N content decreased with increasing percentage of the basal fertilizer, whereas nitrate N content and pH value showed an adverse trend. However, the bacterial and fungal communities were unaffected by the ratio shifts between fertilizer-N and feed-N, but their dominant phyla were influenced by the ditch bottom soil N level. Moreover, the bacterial community composition was positively related to nitrate N, whereas fungal diversity was positively correlated with pH, ammonium N and nitrate N, and urease. We also found that the treatment of N input with 25% N from fertilizer and 75% N from feed can reduce N deposition in the ditch bottom soil in the rice-fish coculture system. Our findings indicate that under the equivalent total N input level, the relative higher ratio of N from fish feed increased (S0 treatment) or reduced (S25 treatment) the deposition of N in the ditch bottom soil, and improved fish production, but decreased rice yield; while the higher ratio of N from basal fertilizer increased the transportation of nutrients into the ditch bottom soil and rice yield, but reduced fish production. So when considering multi-balance and multiple benefits, we recommend that a selective substitution ratio within 50% ~ 75% from fish feed to substitute for the basal fertilizer under the equivalent total N input may achieve a good balance of rice and fish production improvement, and reduce nutrients wastage to the ditch bottom, as well as alleviate the potential of non-point source pollution. This study also provides an evidence for regulating and optimizing the ratio of N supplied from fertilizer and fish feed at an equivalent total N level through monitoring the nutrient accumulation in ditch bottom soil in the rice-fish coculture system. © 2024 Society of Chemical Industry.

Nano zero-valent iron and melatonin synergistically alters uptake and translocation of Cd and As in soil-rice system and mechanism in soil chemistry and microbiology

Nanoscale zero-valent iron (Fe) is a promising nanomaterial for remediating heavy metal-contaminated soils. Melatonin (MT) is essential to alleviate environmental stress in plants. However, the conjunction effects of Fe and MT (FeMT) on rice Cd, As accumulation and the mechanism of soil chemical and microbial factors interaction are unclear. Here, a pot experiment was conducted to evaluated the effects of the FeMT for rice Cd, As accumulation and underlying mechanisms. The findings showed that FeMT significantly reduced grains Cd by 92%–87% and As by over 90%, whereas improving grains Fe by over 213%. Soil available-Cd and iron plaques-Cd (extracted by dithionite-citrate-bicarbonate solution, DCB-Cd) significantly regulated roots Cd, thus affected Cd transport to grains. Soil pH significantly affected soil As and DCB-As, which further influenced roots As uptake and the transport to shoots and grains. The interactions between the soil bacterial community and soil Fe, available Fe, and DCB-Fe together affected root Fe absorption and transportation in rice. FeMT significantly influenced rhizosphere soil bacterial α- and β-diversity. Firmicutes as the dominant phylum exhibited a significant positive response to FeMT measure, and acted a key role in reducing soil Cd and As availability mainly by improving iron-manganese plaques. The increase of soil pH caused by FeMT was beneficial only for Actinobacteriota growth, which reduced Cd, As availability probably through complexation and adsorption. FeMT also showed greater potential in reducing human health and ecological risks by rice consumption and straw returning. These results showed the important role of both soil chemical and microbial factors in FeMT-mediated rice Cd, As reduction efficiency. This study opens a novel strategy for safe rice production and improvement of rice iron nutrition level in heavy-metals polluted soil, but also provides new insights into the intricate regulatory relationships among soil biochemistry, toxic elements, microorganism, and plants.

Linkage map construction and QTL identification for yield related traits of rice (Oryza sativa L.)

The study was undertaken to construct a linkage map and to identify the QTLs for heading date, plant height, and culm strength using a F2 population derived from Ayeyarmin/IR 64, two varieties distinctly different in heading date and plant height. A total of 233 F2 individuals were genotyped using 70 SSR primers, and phenotypic data were collected for heading date, plant height and culm strength traits. Fifty-four out of 70 marker loci were mapped across twelve rice chromosomes spanning a total map length of 1592.30 cM with an average distance of 29.49 cM between adjacent marker loci using MAPMAKER/EXP 3. Composite interval mapping for QTL analysis was conducted with 1000 permutation time at 0.05 probability level using WinQTL Cartographer version 2.5. One major QTL for heading date was mapped between marker RM204 and RM50 on chromosome 6. One major QTL for culm strength was mapped between marker RM1003-RM5931, and two major QTLs for plant height were mapped between RM84-RM1003 and RM1003-RM5931 on chromosome 1. The molecular markers linked to the major QTLs could be potentially used in marker-assisted selection for the rainfed rice improvement breeding program.

Morpho-molecular and nutritional profiling for yield improvement and value addition of indigenous aromatic Joha rice of Assam

Short-grain aromatic Joha rice of Assam is a unique class of specialty rice having tremendous potential in domestic and international markets. The poor yielding ability of Assam's Joha rice demands its systematic characterization for an effective breeding program. This study investigates the morphological, molecular and biochemical profiles of twenty popular Joha (aromatic) rice cultivars indigenous to Assam. Distinctiveness, Uniformity and Stability (DUS) characterization of the cultivars revealed polymorphism in thirty-seven traits, establishing distinctiveness for their utilization in breeding programs. Unweighted Neighbor Joining (UNJ) clustering based on usual Euclidean distances for the polymorphic morphological markers grouped the cultivars into three clusters with eight, eleven, and one genotypes. The Joha rice cultivars showed significant differences for all the quantitative traits except for panicle length. The genotypic and phenotypic coefficients of variability (GCV & PCV) were high for grain yield ha−1 (24.62 & 24.85%) and filled grains panicle−1 (23.69 & 25.02%). Mahalanobis D2 analysis revealed three multi-genotypic and four mono-genotypic clusters of the cultivars. The first five principal components explain 85.87% of the variation among the cultivars for the traits under study; filled grain panicle−1 (0.91) and stem thickness (0.55) positively contributed to the first PC. The cultivars' average polyunsaturated fatty acids were 37.9% oleic acid, 39.22% linoleic acid, and 0.5% linolenic acid. Kon Joha 4 and Ronga Joha contained the highest iron (82.88 mg kg−1) and zinc (47.39 mg kg−1), respectively. Kalijeera, Kunkuni Joha, Kon Joha-5, Manimuni Joha and Kon Joha-2 accorded a strong aroma. PCR amplified 174 alleles with a mean value 2.64 across the 66 polymorphic SSR markers. PIC values ranged from 0.091 to 0.698, with an average of 0.326. The highly informative (PIC > 0.50) markers were RM316, RM283, RM585, RM1388, RM3562, RM171, R1M30, RM118, RM11and RM29 for identification of the twenty aromatic rice cultivars. PCR amplification of 27 SSR markers identified 28 unique alleles (97–362 bp) in 13 Joha rice cultivars, which can help their identification/DNA fingerprinting. The UNJ clustering based on Jaccard's coefficients classified the cultivars into three distinct clusters with eight, ten, and two genotypes. Our study revealed the nutritional richness of these specialty Joha rice cultivars and sufficient scope for yield enhancement through their interbreeding to keep quality intact.

A mediator of OsbZIP46 deactivation and degradation negatively regulates seed dormancy in rice

Preharvest sprouting (PHS) is a deleterious phenotype that occurs frequently in rice-growing regions where the temperature and precipitation are high. It negatively affects yield, quality, and downstream grain processing. Seed dormancy is a trait related to PHS. Longer seed dormancy is preferred for rice production as it can prevent PHS. Here, we map QTLs associated with rice seed dormancy and clone Seed Dormancy 3.1 (SDR3.1) underlying one major QTL. SDR3.1 encodes a mediator of OsbZIP46 deactivation and degradation (MODD). We show that SDR3.1 negatively regulates seed dormancy by inhibiting the transcriptional activity of ABIs. In addition, we reveal two critical amino acids of SDR3.1 that are critical for the differences in seed dormancy between the Xian/indica and Geng/japonica cultivars. Further, SDR3.1 has been artificially selected during rice domestication. We propose a two-line model for the process of rice seed dormancy domestication from wild rice to modern cultivars. We believe the candidate gene and germplasm studied in this study would be beneficial for the genetic improvement of rice seed dormancy.

Yield gap variation in rice cultivation in Indonesia

Abstract The rice yield gap (YG) is a global concern, requiring more detailed studies spatially and temporally. As a staple food in Indonesia, rice was produced from 7.4 Mha paddy fields in 2019. Better insight into the YG helps assess measures to boost rice production. However, the information on YG variation among regions scale is limited. This study aimed to identify the rice YG based on 295 historical trial datasets from 23 provinces in Indonesia. We surveyed published trial results from 2012 to 2022 and analyzed YGs, expressed as the percentage of farmer yield (FY). The potential yield (PY) was estimated from field trial results using introduced rice cultivation technology package, whereas FY from results using existing farmer practices. Our study showed that the average YG was 62% in rainfed, 54% in tidal, and 32% in irrigated paddy fields. The YG was significantly high in the paddy fields of Kalimantan (74%) and Maluku-Papua (49%), while the lowest was in Sulawesi (27%) and Java (31%). The YG varied significantly with geo-regions, rice varieties, and cultivation technology packages. Closing the YG and ensuring sustainable rice production requires the implementation of sustainable intensification through applying site-specific technology packages, reallocation of agricultural interventions to a higher YG region, and rice variety improvement to increase PY.

Rapid Visual Detection of Elite Erect Panicle Dense and Erect Panicle 1 Allele for Marker-Assisted Improvement in Rice (Oryza sativa L.) Using the Loop-Mediated Isothermal Amplification Method.

Molecular-assisted breeding is an effective way to improve targeted agronomic traits. dep1 (dense and erect panicle 1) is a pleiotropic gene that regulates yield, quality, disease resistance, and stress tolerance, traits that are of great value in rice (Oryza sativa L.) breeding. In this study, a colorimetric LAMP (loop-mediated isothermal amplification) assay was developed for the detection of the dep1 allele and tested for the screening and selection of the heavy-panicle hybrid rice elite restorer line SHUHUI498, modified with the allele. InDel (Insertion and Deletion) primers (DEP1_F and DEP1_R) and LAMP primers (F3, B3, FIP, and BIP) for genotyping were designed using the Primer3 Plus (version 3.3.0) and PrimerExplore (version 5) software. Our results showed that both InDel and LAMP markers could be used for accurate genotyping. After incubation at a constant temperature of 65 °C for 60 min with hydroxynaphthol blue (HNB) as a color indicator, the color of the LAMP assay containing the dep1 allele changed to sky blue. The SHUHUI498 rice line that was detected in our LAMP assay displayed phenotypes consistent with the dep1 allele such as having a more compact plant architecture, straight stems and leaves, and a significant increase in the number of effective panicles and spikelets, demonstrating the effectiveness of our method in screening for the dep1 allele in rice breeding.

Genome-wide identification of rice CXE gene family and mining of alleles for potential application in rice improvement.

Carboxylesterases (CXE, EC 3.1.1.1), a class of hydrolases with an α/β folding domain, play important roles in plant growth and development and stress response. Here, we identified 32, 63, 41, and 45 CXE genes in Oryza sativa Japonica (Nipponbare), Oryza sativa Indica (93-11), Oryza sativa Indica (Xian-1B1 var.IR64), and Oryza sativa Japonica (Geng-sbtrp var.ChaoMeo), respectively. Then, we analyzed the chromosomal location, physical and chemical properties, subcellular localization, collinearity, and selection pressure of CXE genes in four rice varieties. We also analyzed the functional interaction network, cis-regulatory elements, evolutionary relationship, and protein tertiary structure, and performed gene expression profiling and qPCR verification under abiotic stress, as well as diversity analysis of 3010 gene-CDS-haplotype (gcHap) rice samples, aiming to understand the potential function of the 32 OsCXE genes. Our results indicated that fragment replication is the main reason for amplification of the CXE gene family in rice, and the gene family has undergone strong purification selection. OsCXE3.1, OsCXE3.2, OsCXE3.3, OsCXE5.1, and OsCXE7.3 may be used to improve the tolerance of rice to abiotic stress. OsCXE play important roles in rice population differentiation and improvement, and the major gcHaps at most OsCXE locus are significantly associated with yield traits. Therefore, natural variations of most OsCXE locus have great potential value for improvement of rice productivity.

Development of CRISPR-Cas9 Multiple Editing System for Genetic Improvement of Rice

Development of CRISPR-Cas9 Multiple Editing System for Genetic Improvement of Rice

Strategies for Rice Improvement: Utilizing Genetic Resources from Wild and Cultivated Oryza Species

Strategies for Rice Improvement: Utilizing Genetic Resources from Wild and Cultivated Oryza Species