A Rapid and Accurate Quantitative Analysis of Cellulose in the Rice Bran Layer Based on Near-Infrared Spectroscopy.

Cultivating rice varieties with lower cellulose content in the bran layer has the potential to enhance both the nutritional value and texture of brown rice. This study aims to establish a rapid and accurate method to quantify cellulose content in the bran layer utilizing near-infrared spectroscopy (NIRS), thereby providing a technical foundation for the selection, screening, and breeding of rice germplasm cultivars characterized by a low cellulose content in the bran layer. To ensure the accuracy of the NIR spectroscopic analysis, the potassium dichromate oxidation (PDO) method was improved and then used as a reference method. Using 141 samples of rice bran layer (rice bran without germ), near-infrared diffuse reflectance (NIRdr) spectra, near-infrared diffuse transmittance (NIRdt) spectra, and fusion spectra of NIRdr and NIRdt were used to establish cellulose quantitative analysis models, followed by a comparative evaluation of these models' predictive performance. Results indicate that the optimized PDO method demonstrates superior precision compared to the original PDO method. Upon examining the established models, their predictive capabilities were ranked in the following order: the fusion model outperforms the NIRdt model, which in turn surpasses the NIRdr model. Of all the fusion models developed, the model exhibiting the highest predictive accuracy utilized fusion spectra (NIRdr-NIRdt (1st der)) derived from preprocessed (first derivative) diffuse reflectance and transmittance spectra. This model achieved an external predictive R2p of 0.903 and an RMSEP of 0.213%. Using this specific model, the rice mutant O2 was successfully identified, which displayed a cellulose content in the bran layer of 3.28%, representing a 0.86% decrease compared to the wild type (W7). The utilization of NIRS enables quantitative analysis of the cellulose content within the rice bran layer, thereby providing essential technical support for the selection of rice varieties characterized by lower cellulose content in the bran layer.

CEF3 is involved in membrane trafficking and essential for secondary cell wall biosynthesis and its mutation enhanced biomass enzymatic saccharification in rice.

BACKGROUND: As one of the most important staple food crops, rice produces large of agronomic biomass residues that contain lots of secondary cell walls (SCWs). Membrane trafficking plays key roles in SCWs biosynthesis, but information association membrane trafficking and SCWs formation in plants is limited. RESULTS: In this study, we report the function characterization of a rice mutant, culm easily fragile 3 (cef3), that exhibits growth retardation and fragile culm phenotype with significantly altered cell wall composition and reduced secondary wall thickness. Map-based cloning revealed that CEF3 encodes a homologous protein of Arabidopsis STOMATAL CYTOKINESIS DEFECTIVE2 (SCD2). The saccharification assays revealed that CEF3 mutation can improve biomass enzymatic saccharification. Expression pattern analysis indicated that CEF3 is ubiquitously expressed in many organs at different developmental stages. Subcellular localization revealed that CEF3 is a Golgi-localized protein. The FM4-64 uptake assay revealed CEF3 is involved in endocytosis. Furthermore, mutation of CEF3 not only affected cellulose synthesis-related genes expression, but also altered the abundance of cellulose synthase catalytic subunit 9 (OsCESA9) in the PM and in the endomembrane systems. CONCLUSIONS: This study has demonstrated that CEF3 participates in the membrane trafficking that is essential for normal cellulose and other polysaccharides biosynthesis of the secondary cell wall, thereby manipulation of CEF3 could alter cellulose content and enhance biomass enzymatic saccharification in rice plants. Therefore, the study of the function of CEF3 can provide a strategy for genetic modification of SCWs in bioenergy crops.

Low grain weight, a new allele of BRITTLE CULM12, affects grain size through regulating GW7 expression in rice.

Grain weight is a major determinant in rice yield, which is tightly associated with grain size. However, the underlying molecular mechanisms that control this trait remain unclear. Here, we report a rice (Oryza sativa) mutant, low grain weight (lgw), which shows that reduced grain length is caused by decreased cell elongation and proliferation. Map-based cloning revealed that all mutant phenotypes resulted from a nine-base pair (bp) deletion in LGW, which encodes the kinesin-like protein BRITTLE CULM12 (BC12). Protein sequence alignment analysis revealed that the mutation site was located at the nuclear localization signal (NLS) of LGW/BC12, resulting in the lgw protein not being located in the nucleus. LGW is preferentially expressed in both culms and roots, as well as in the early developing panicles. Overexpression of LGW increased the grain length, indicating that LGW is a positive regulator for regulating grain length. In addition, LGW/BC12 is directly bound to the promoter of GW7 and activates its expression. Elevating the GW7 expression levels in lgw plants rescued the small grain size phenotype. We conclude that LGW regulates grain development by directly binding to the GW7 promoter and activating its expression. Our findings revealed that LGW plays an important role in regulating grain size, and manipulation of this gene provides a new strategy for regulating grain weight in rice.

A Semi-Dominant Mutation in OsCESA9 Improves Salt Tolerance and Favors Field Straw Decay Traits by Altering Cell Wall Properties in Rice.

BACKGROUND: Cellulose synthase (CESA) mutants have potential use in straw processing due to their lower cellulose content, but almost all of the mutants exhibit defective phenotypes in plant growth and development. Balancing normal plant growth with reduced cellulose content remains a challenge, as cellulose content and normal plant growth are typically negatively correlated with one another. RESULT: Here, the rice (Oryza sativa) semi-dominant brittle culm (sdbc) mutant Sdbc1, which harbors a substitution (D387N) at the first conserved aspartic acid residue of OsCESA9, exhibits lower cellulose content and reduced secondary wall thickness as well as enhanced biomass enzymatic saccharification compared with the wild type (WT). Further experiments indicated that the OsCESA9D387N mutation may compete with the wild-type OsCESA9 for interacting with OsCESA4 and OsCESA7, further forming non-functional or partially functional CSCs. The OsCESA9/OsCESA9D387N heterozygous plants increase salt tolerance through scavenging and detoxification of ROS and indirectly affecting related gene expression. They also improve rice straw return to the field due to their brittle culms and lower cellulose content without any negative effects in grain yield and lodging. CONCLUSION: Hence, OsCESA9D387N allele can improve rice salt tolerance and provide the prospect of the rice straw for biofuels and bioproducts due to its improved enzymatic saccharification.

Differences in cadmium accumulation between indica and japonica rice cultivars in the reproductive stage.

Excessive cadmium (Cd) in rice grains is of great concern worldwide, particularly in southern China where heavy metal pollution in the soil is widespread. Much work has been done regarding the key genes responsible for Cd absorption, transport, and accumulation in rice, but little is known about the differences of Cd accumulation between indica and japonica rice cultivars during the reproductive stage. Furthermore, physiological parameters, such as nonstructural carbohydrate content, involved in Cd accumulation have not been fully elucidated. We studied several indica and japonica cultivars under three different Cd treatment levels and harvested them at different periods after heading. Differences in Cd accumulation between subspecies mainly were generated during the reproductive stage. An increase in the Cd pollution level caused the average absorption rate of Cd in the aerial parts of the indica cultivars in the reproductive stage to be 6.17, 4.52, and 3.89 times greater than that of the japonica cultivars across the three Cd treatments. The contribution of Cd absorption by shoots to Cd accumulation at the pre- or postheading stages was 33.8% and 66.2% in indica, and 44.9% and 55.1% in japonica. We found a significant negative correlation between Cd content in the rice grains and the content of nonstructural carbohydrates in the sheath (P < 0.05). Cd translocation from sheath to grain occurred along with sugar transfer in the indica cultivars. The Cd content of the indica cultivar grain was 1.84-4.14 times higher than that of the japonica cultivars (P < 0.05). The japonica cultivars thus met the cereal Cd limits of China (0.2 mg kg-1) under low and moderate soil Cd pollution. These findings are helpful for the selection of proper cultivars and field management practices to alleviate Cd exposure risk in rice production.

A calibration transfer optimized single kernel near-infrared spectroscopic method.

Single kernel near-infrared spectroscopy (SKNIRS) could aid in the quality screening of early-generation seeds, to improve the efficiency of seed breeding. However, the application of SKNIRS is limited due to the irregular physical characteristics, the heterogeneous constituent distributions of individual seeds, and the insufficient detection accuracy of the reference method. The reported near-infrared detection results of single seeds are often less accurate than those of dehusked seeds and seed flour. In this paper, a calibration transfer-optimized single kernel near-infrared spectroscopic method is proposed. This method aims to accurately detect the chemical composition of single seeds by using the calibration model of the corresponding dehusked seeds or seed flour. The proposed method was applied to the analysis of the protein content of a single rice kernel. The near-infrared transmission spectra of three forms of rice (single rice kernel (SRK), single brown rice kernel (SBK) and rice flour (RF)) of 201 individual rice seeds and the corresponding protein content values were obtained. By comparing different pretreatment methods and spectral ranges, the spectral range 950-1250 nm, the standard normal variate transformation (SNV) pretreatment, and 9 PLS factors were selected to construct the optimal partial least squares (PLS) regression models. Then, the protein content of single rice kernels were determined through two different methods: (i) the direct method, in which single rice kernels were analyzed using the single rice kernel model directly; and (ii) the proposed method, in which the spectra of single rice kernels were transferred into the spectra of single brown rice kernels and rice flours with a calibration transfer algorithm, spectral space transformation (SST), and were analyzed by the respective calibration models. The external validation coefficient correlation (R) value of the direct method was 0.971, and the R values of the proposed method were 0.962 (SBK) and 0.975 (RF). The root mean square error of prediction (RMSEP) value of the direct method was 0.423, and the RMSEP of the proposed method were 0.480 (SBK) and 0.401 (RF). In addition, the transfer results among the spectra of three forms of rice were compared. By comparison, the results of the proposed method are fairly close to the results of the direct method. The results indicate that the spectra generated from one individual rice seed can be transferred freely among the three forms by means of calibration transfer. The proposed method is a promising way to overcome the challenges associated with analyzing individual seeds and to improve SKNIRS.