Japonica Rice Cultivars Research Articles

Functional diversity of oxidosqualene cyclases in genus Oryza.

Triterpene skeletons, catalyzing by 2,3-oxidosqualene cyclases (OSCs), are essential for synthesis of steroids and triterpenoids. In japonica rice cultivars Zhonghua11, a total of 12 OsOSCs have been found. While the catalytic functions of OsOSC1, 3, 4, 9, and 10 remain unclear, the functions of the other OsOSCs have been well studied. In this study, we conducted a comprehensive analysis of 12 OSC genes within genus Oryza with the aid of 63 genomes from cultivated and wild rice. We found that OSC genes are relatively conserved within genus Oryza with a few exceptions. Collinearity analysis further suggested that, throughout the evolutionary history of genus Oryza, the OSC genes have not undergone significant rearrangements or losses. Further functional analysis of 5 uncharacterized OSCs revealed that OsOSC10 was a friedelin synthase, which affected the development of rice grains. Additionally, the reconstructed ancestral sequences of Oryza OSC3 and Oryza OSC9 had lupeol synthase and poaceatapetol synthase activity, respectively. The discovery of friedelin synthase in rice unlocks a new catalytic path and biological function of OsOSC10. The pan-genome analysis of OSCs within genus Oryza gives insights into the evolutionary trajectory and products diversity of Oryza OSCs.

Evolutionary Systems Biology Identifies Genetic Trade-offs In Rice Defense Against Above- and Belowground Attackers.

Like other plants, wild and domesticated rice species (Oryza nivara, O. rufipogon, and O. sativa) evolve in environments with various biotic and abiotic stresses that fluctuate in intensity through space and time. Microbial pathogens and invertebrate herbivores such as plant-parasitic nematodes and caterpillars show geographical and temporal variation in activity patterns and may respond differently to certain plant defensive mechanisms. As such, plant interactions with multiple community members may result in conflicting selection pressures on genetic polymorphisms. Here, through assays with different above- and belowground herbivores, the fall armyworm (Spodoptera frugiperda) and the southern root-knot nematode (Meloidogyne incognita), respectively, and comparison with rice responses to microbial pathogens, we identify potential genetic trade-offs at the KSL8 and MG1 loci on chromosome 11. KSL8 encodes the first committed step towards biosynthesis of either stemarane- or stemodane-type diterpenoids through the japonica (KSL8-jap) or indica (KSL8-ind) allele. Knocking out KSL8-jap and CPS4, encoding an enzyme that acts upstream in diterpenoid synthesis, in japonica rice cultivars increased resistance to S. frugiperda and decreased resistance to M. incognita. Furthermore, MG1 resides in a haplotype that provided resistance to M. incognita, while alternative haplotypes are involved in mediating resistance to the rice blast fungus Magnaporthe oryzae and other pests and pathogens. Finally, KSL8 and MG1 alleles are located within trans-species haplotypes and may be evolving under long-term balancing selection. Our data are consistent with a hypothesis that polymorphisms at KSL8 and MG1 may be maintained through complex and diffuse community interactions.

Genetic Diversity and Divergence between Southern Japonica and Northern Japonica Rice Varieties in China.

Given the notable ecological and breeding disparities between southern and northern rice regions, delving into the genetic diversity and divergence between southern and northern japonica rice contributes to enhancing the genetic pool for japonica rice breeding. In this study, we analyzed 90 southern and 51 northern japonica rice varieties, focusing on nucleotide diversity (Pi), agronomic trait variations, population structure, genetic divergence, and a neutral test. For genetic diversity, the results demonstrated higher Pi in northern japonica rice varieties (NJRVs) on Chr2, Chr5, Chr6, Chr8, and Chr10, whereas in southern japonica rice varieties (SJRVs) on Chr7 and Chr9. In addition, SJRVs exhibited higher grain width and thickness, whereas NJRVs featured a higher grain aspect ratio, filled grain number, and grain number per panicle. Regarding genetic divergence, geographic differentiation existed between NJRVs and SJRVs, with Chr5 exhibiting numerous higher genetic differentiation windows, including cloned grain shape-controlling genes RGA1 and SFD5, stemming from intensified selection pressure on SJRVs. In summary, SJRVs and NJRVs exhibited diversity differences and genetic differentiation. Hence, it was suggested to conduct genetic introgression between NJRVs and SJRVs to broaden the genetic basis of the local japonica rice germplasm. By exploiting their heterotic advantage, new japonica rice cultivars with superior comprehensive traits could be developed.

Cryptic cytoplasmic male sterility-causing gene in the mitochondrial genome of common japonica rice.

Cytoplasmic male sterility (CMS) is an agronomically significant trait that causes dysfunction in pollen and anther development. It is often observed during successive backcrossing between distantly related species. Here, we show that Asian japonica cultivars (Oryza sativa) exhibit CMS when the nucleus is replaced with that of the African rice Oryza glaberrima. The CMS line produced stunted anthers and did not set any seeds. Mitochondrial orf288 RNA was detected in the anthers of CMS lines but not in fertility restorer lines. The mitochondrial genome-edited japonica rice that was depleted of orf288 did not exhibit male sterility when backcrossed with O. glaberrima. These results demonstrate that orf288 is a CMS-causing gene. As orf288 commonly occurs in the mitochondrial genomes of japonica rice, these results indicate that common japonica rice cultivars possess a cryptic CMS-causing gene hidden in their mitochondrial genomes.

Straw type and nitrogen-water management balance rice yield and methane emissions by regulating rhizosphere microenvironment

Context or problemStraw incorporation improves soil fertility but also poses environmental challenges due to increasing methane (CH4) emissions in paddy fields. Whether nitrogen (N) and water management can balance rice yield and CH4 emissions under different crop straw incorporation is still not well-documented. ObjectiveA three-year field experiment was conducted to probe the comprehensive effects of N application ratios and irrigation regimes on rice yield, rhizosphere soil properties, and CH4 emissions, along with the underlying mechanisms of CH4 emission variations among different straw types. MethodsA two-factor randomized block design was used with two Japonica rice cultivars as materials in 2020 and 2021. The straw incorporation treatment included no straw incorporation (NS), wheat straw incorporation (WS), and rape straw incorporation (RS). The N fertilizer application treatments included local farmers' fertilizer practice (LFP) and increasing basal fertilizer rate (IBF). Two irrigation practices, continuously-flooded irrigation (CF) and alternate wetting and drying irrigation (AWD), were designed under the WS and RS treatments in 2022. Results1) WS-IBF and RS-IBF enhanced yield by 6.70∼9.03 % and 8.13∼9.50 % compared to WS-LFP and RS-LFP, respectively. AWD further increased yield by 6.28∼7.76 % compared to CF. 2) WS-IBF and RS-IBF enhanced dissolved organic carbon (DOC) content, synchronously boosted the methanogens (mcrA) and methanotrophs (pmoA) abundances, but decreased the pmoA/mcrA ratio, which significantly promoted CH4 emission flux in early growth stage. This resulted in a 5.04∼8.01 % and 4.60∼7.88 % increase in CH4 emissions compared to WS-LFP and RS-LFP, respectively, but a decrease in yield-scaled CH4 emissions. AWD reduced DOC content, facilitated the conversion of ammonium N to nitrate N, increased dissolved oxygen content, and hence decreased CH4 emissions by 23.41∼24.38 % compared to CF. 3) RS significantly increased microbial biomass C, N, and related metabolites, leading to a 1.29∼2.73 % increase in yield compared to WS. Meanwhile, RS promoted Nitrospira abundance as well as pterin and flavonoid metabolites associated with mcrA inhibition, while decreasing Anaeromyxobacter abundance, ammonium N, and DOC content, resulting in an increase in the pmoA/mcrA ratio and a noticeable drop in CH4 emissions compared to WS. ConclusionsRS combined with IBF and AWD is a more sustainable integrated practice in light of the synergistic improvement in rice production and environmental benefits. ImplicationsThe results reveal that optimizing N and water management can synergize high-yield and low-carbon by regulating rhizosphere microenvironment in rice production under crop straw incorporation.

Nitrogen topdressing at panicle initiation modulated nitrogen allocation between storage proteins and free nitrogenous compounds in grains of japonica rice

Nitrogen (N) topdressing often leads to a decline in cooking and eating quality due to alterations in grain chemical composition. Investigation into the response of N compounds in grains, such as protein-bound amino acids (PAA), free amino acids (FAA), and other residue N compounds (Nresidue), would be beneficial for understanding the impact of N on rice quality. The present study selected five japonica rice cultivars and conducted a two-year field experiment employing two N fertilization treatments: the CK treatment, where all N fertilizer was applied as basal fertilizer, and the topdressing treatment, where 50% of the total N was applied as topdressing at the panicle initiation stage. Averaging across years and cultivars, N topdressing resulted in a 16.3% increase in PAA and an 8.65% decrease in FAA, with no significant impact on Nresidue. N topdressing resulted in a decrease in the proportion of essential amino acids found in prolamins, as well as a reduction in the proportion of amino acids belonging to the aspartate family and arginine. Significant genotypic variations were observed in the response of nitrogen components to topdressing. Furthermore, elite cultivars with premium quality exhibited greater stability in amylose content compared to protein content.

Nitrogen and Potassium Application Effects on Grain-Filling and Rice Quality in Different Japonica Rice Cultivars

Nitrogen and Potassium Application Effects on Grain-Filling and Rice Quality in Different Japonica Rice Cultivars

Combination of Polymer-Coated Urea and Rapid-Release Urea Increases Grain Yield and Nitrogen Use Efficiency of Rice by Improving Root and Shoot Activities

The use of polymer-coated urea (PCU) can improve nitrogen use efficiency (NUE), compared to the application of rapid-release urea (RU). However, the effect of PCU-based nitrogen management on grain yield and the NUE of rice and its underlying mechanism remain unclear. A japonica rice cultivar Jinxiangyu 1 was grown in the field with four treatments including N omission (0N), split application of RU (Control), one-time application of 100% PCU (T1), and one-time application of 70% PCU + 30% RU (T2). Results showed that, compared to the control, the grain yield was significantly increased in the T2 treatment, while it was comparable in the T1 treatment. This was mainly due to increased total spikelets in the T2 treatment. Root oxidation activity (ROA) and root zeatin (Z) + zeatin riboside (ZR) content during booting were the distinct advantages of the T2 treatment, compared to either the control or T1 treatment, exhibiting significant or highly significant correlations with leaf photosynthesis. This process contributed significantly to total spikelets and total N uptake. Additionally, the T2 treatment absorbed more N than the control without reducing the internal N use efficiency (IEN), primarily due to its unchanged harvest index (HI) driven by comparable non-structural carbohydrate remobilization. In conclusion, combining PCU with RU can enhance the coordination of root and shoot traits during booting while maintaining a competitive HI at maturity, thereby significantly improving grain yield and achieving a balance in N uptake and utilization.

Co-mutation of OsLPR1/3/4/5 provides a promising strategy to minimize Cd contamination in rice grains

Minimizing cadmium (Cd) contamination in rice grains is crucial for ensuring food security and promoting sustainable agriculture. Utilizing genetic modification to generate rice varieties with low Cd accumulation is a promising strategy due to its cost-effectiveness and operational simplicity. Our study demonstrated that the CRISPR-Cas9-mediated quadruple mutation of the multicopper oxidase genes OsLPR1/3/4/5 in the japonica rice cultivar Tongjing 981 had little effect on yields. However, a notable increase was observed in the cell wall functional groups that bind with Cd. As a result, the quadruple mutation of OsLPR1/3/4/5 enhanced Cd sequestration within the cell wall while reducing Cd concentrations in both xylem and phloem sap, thereby inhibiting Cd transport from roots to shoots. Consequently, Cd concentrations in brown rice and husk in oslpr1/3/4/5 quadruple mutants (qm) decreased by 52% and 55%, respectively, compared to the wild-type. These findings illustrate that the quadruple mutation of OsLPR1/3/4/5 is an effective method for minimizing Cd contamination in rice grains without compromising yields. Therefore, the quadruple mutation of OsLPR1/3/4/5 via biotechnological pathways may represent a valuable strategy for the generation of new rice varieties with low Cd accumulation.

Transgenic early japonica rice: Integration and expression characterization of stem borer resistance Bt gene

BackgroundTransgenic insect-resistant rice offers an environmentally friendly approach to mitigate yield losses caused by lepidopteran pests, such as stem borers. Bt (Bacillus thuringiensis) genes encode insecticidal proteins and are widely used to confer insect resistance to genetically modified crops. This study investigated the integration, inheritance, and expression characteristics of codon-optimised synthetic Bt genes, cry1C* and cry2A*, in transgenic early japonica rice lines. MethodsThe early japonica rice cultivar, Songgeng 9 (Oryza sativa), was transformed with cry1C* or cry2A*, which are driven by the ubi promoter via Agrobacterium tumefaciens-mediated transformation. Molecular analyses, including quantitative PCR (qPCR), enzyme-linked immunosorbent assay (ELISA), and Southern blot analysis were performed to confirm transgene integration, inheritance, transcriptional levels, and protein expression patterns across different tissues and developmental stages. ResultsStable transgenic early japonica lines exhibiting single-copy transgene integration were established. Transcriptional analysis revealed variations in Bt gene expression among lines, tissues, and growth stages, with higher expression levels observed in leaves than in other organs. Notably, cry2A* exhibited consistently higher mRNA and protein levels than cry1C* across all examined tissues and developmental time points. Bt protein accumulation followed the trend of leaves > stem sheaths > young panicles > brown rice, with peak expression during the filling stage in the vegetative tissues. ConclusionsSynthetic cry2A* displayed markedly elevated transcription and translation compared to cry1C* in the transgenic early japonica rice lines examined. Distinct spatiotemporal patterns of Bt gene expression were elucidated, providing insights into the potential insect resistance conferred by these genes in rice. These findings will contribute to the development of insect-resistant japonica rice varieties and facilitate the rational deployment of Bt crops.

Phytotoxicity and Residual Effect of Some Herbicides on three Egyptian Rice Cultivars (Oryza sativa L.)

The current study was carried out at the experimental farm of Rice Research Department, Sakha Agricultural Research Station and Central Agricultural Pesticide Laboratory, ARC, Egypt during the summer season of 2021 and 2022 to study the sensitivity of three Egyptian rice cultivars against four common herbicides and its effect on physiological and agronomic characteristics of rice as well as herbicide residues in rice seeds. The field study was laid out in split plot design with three replications. Three rice cultivars (Giza 177, Giza 179 and Sakha 108) were randomly distributed in main plots, while weed control treatments included four herbicides at recommended doses (penoxsulam 2.5% OD, bispyribac-sodium 2% SL, cyhalofop-butyl 10% EC and fenoxaprop-p-ethyl 7.5% EW) as compared to hand weeding were devoted in sub plots. The obtained results revealed that Sakha 108 as Japonica rice cultivar was resistant against tested herbicides and recorded the highest values of rice dry weight, chlorophyll content, number of panicles m-2, panicle weight and grain yield, while scored the lowest values of total phenols. On the other hand, Giza 177 as short duration Japonica rice cultivar showed the higher sensitivity against tested herbicides and achieved the lowest values of abovementioned agronomic traits and chlorophyll content. Cyhalofop-butyl as ACCase herbicide was safer on studied rice cultivars and recorded the highest values of studied agronomic characteristics for rice as well as chlorophyll content, while recorded the lowest values of total phenols during the study. On the other hand, bispyribac-sodium 2% SL resulted in more toxic effect for rice cultivars and recorded the lowest values of studied agronomic traits for rice. fenoxaprop-p-ethyl 7.5% EW achieved the lowest chlorophyll content and highest values of total phenols during the study. Giza 179 as Indica-Japonica rice cultivar appeared high sensitivity when sprayed with bispyribac-sodium and fenoxaprop-ethyl but made recovery after 12 days from herbicidal application and recorded the highest chlorophyll content, dry weight, panicles m-2 and grain yield with no significant differences between Sakha 108 rice cultivar during the study. In respect of herbicide residues in rice seeds, all tested herbicides didn’t have residues in rice seeds at 100 days after treatment. It means that tested herbicides didn’t have any residues in rice seeds.

Effect of Cd(II) adsorption onto rice roots on its uptake by different indica and japonica rice varieties and toxicity effect of Cd(II) under acidic conditions.

The rapid increase in soil acidity coupled with the deleterious effects of cadmium (Cd) toxicity had led to a decline in worldwide agricultural production. Rice absorbs and accumulates Cd(II) from polluted paddy soils, increasing human health risks throughout the food chain. A 35-day hydroponic experiment with four japonica and four indica (two each of them tolerant and sensitive cultivars) was conducted in this study to investigate the adsorption and absorption of Cd(II) by rice roots as related with surface chemical properties of the roots. The results showed that the three chemical forms of exchangeable, complexed, and precipitated Cd(II) increased with the increase in Cd(II) concentration for all rice cultivars. The roots of indica rice cultivars carried more negative charges and had greater functional groups and thus adsorbed more exchangeable and complexed Cd(II) than those of japonica rice cultivars. This led to more absorption of Cd(II) by the roots and greater toxicity of Cd(II) to the roots of indica rice cultivars and more inhibition of Cd(II) stress on the growth of the roots and whole plants of indica rice cultivars compared with japonica rice cultivars, which was one of the main reasons for more declines in the biomass and length of indica rice roots and shoots than japonica rice cultivars. Cd(II) stress showed more toxicity to the sensitive rice cultivars and thus greater inhibition on the growth of the cultivars due to more exchangeable and complexed Cd(II) adsorbed by their roots induced by more negative charges and functional groups on the roots compared with tolerant rice cultivar for both indica and japonica, which resulted in greater decreases in the biomass and length of roots and shoots as well as chlorophyll contents of the sensitive cultivars than the tolerant cultivars. The roots of sensitive rice cultivars also absorbed more Cd(II) than tolerant rice cultivars due to the same reasons as above. These findings will provide useful references for the safe utilization and health risk prevention of Cd-contaminated paddy fields.

Moderate Salinity Stress Affects Rice Quality by Influencing Expression of Amylose- and Protein-Content-Associated Genes.

Salinity is an environmental stress that severely impacts rice grain yield and quality. However, limited information is available on the molecular mechanism by which salinity reduces grain quality. In this study, we investigated the milling, appearance, eating and cooking, and nutritional quality among three japonica rice cultivars grown either under moderate salinity with an electrical conductivity of 4 dS/m or under non-saline conditions in a paddy field in Dongying, Shandong, China. Moderate salinity affected rice appearance quality predominantly by increasing chalkiness rate and chalkiness degree and affected rice eating and cooking and nutritional quality predominantly by decreasing amylose content and increasing protein content. We compared the expression levels of genes determining grain chalkiness, amylose content, and protein content in developing seeds (0, 5, 10, 15, and 20 days after flowering) of plants grown under saline or non-saline conditions. The chalkiness-related gene Chalk5 was up-regulated and WHITE-CORE RATE 1 was repressed. The genes Nuclear factor Y and Wx, which determine amylose content, were downregulated, while protein-content-associated genes OsAAP6 and OsGluA2 were upregulated by salinity in the developing seeds. These findings suggest some target genes that may be utilized to improve the grain quality under salinity stress conditions via gene-pyramiding breeding approaches.

Controlling diurnal flower-opening time by manipulating the jasmonate pathway accelerates development of indica-japonica hybrid rice breeding.

Inter-subspecific indica-japonica hybrid rice (Oryza sativa) has the potential for increased yields over traditional indica intra-subspecies hybrid rice, but limited yield of F1 hybrid seed production (FHSP) hinders the development of indica-japonica hybrid rice breeding. Diurnal flower-opening time (DFOT) divergence between indica and japonica rice has been a major contributing factor to this issue, but few DFOT genes have been cloned. Here, we found that manipulating the expression of jasmonate (JA) pathway genes can effectively modulate DFOT to improve the yield of FHSP in rice. Treating japonica cultivar Zhonghua 11 (ZH11) with methyl jasmonate (MeJA) substantially advanced DFOT. Furthermore, overexpressing the JA biosynthesis gene OPDA REDUCTASE 7 (OsOPR7) and knocking out the JA inactivation gene CHILLING TOLERANCE 1 (OsHAN1) in ZH11 advanced DFOT by 1- and 2-h respectively; and knockout of the JA signal suppressor genes JASMONATE ZIM-DOMAIN PROTEIN 7 (OsJAZ7) and OsJAZ9 resulted in 50-min and 1.5-h earlier DFOT respectively. The yields of FHSP using japonica male-sterile lines GAZS with manipulated JA pathway genes were significantly higher than that of GAZS wildtype. Transcriptome analysis, cytological observations, measurements of elastic modulus and determination of cell wall components indicated that the JA pathway could affect the loosening of the lodicule cell walls by regulating their composition through controlling sugar metabolism, which in turn influences DFOT. This research has vital implications for breeding japonica rice cultivars with early DFOT to facilitate indica-japonica hybrid rice breeding.

Population Structure and Genetic Diversity of Shanlan Landrace Rice for GWAS of Cooking and Eating Quality Traits.

The Shanlan landrace rice in Hainan Province, China, is a unique upland rice germplasm that holds significant value as a genetic resource for rice breeding. However, its genetic diversity and its usefulness in rice breeding have not been fully explored. In this study, a total of eighty-four Shanlan rice, three typical japonica rice cultivars, and three typical indica rice cultivars were subjected to resequencing of their genomes. As a result, 11.2 million high-quality single nucleotide polymorphisms (SNPs) and 1.6 million insertion/deletions (InDels) were detected. Population structure analysis showed all the rice accessions could be divided into three main groups, i.e., Geng/japonica 1 (GJ1), GJ2, and Xian/indica (XI). However, the GJ1 group only had seven accessions including three typical japonica cultivars, indicating that most Shanlan landrace rice are different from the modern japonica rice. Principal component analysis (PCA) showed that the first three principal components explained 60.7% of the genetic variation. Wide genetic diversity in starch physicochemical parameters, such as apparent amylose content (AAC), pasting viscosity, texture properties, thermal properties, and retrogradation representing the cooking and eating quality was also revealed among all accessions. The genome-wide association study (GWAS) for these traits was conducted and identified 32 marker trait associations in the entire population. Notably, the well-known gene Waxy (Wx) was identified for AAC, breakdown viscosity, and gumminess of the gel texture, and SSIIa was identified for percentage of retrogradation and peak gelatinization temperature. Upon further analysis of nucleotide diversity in Wx, six different alleles, wx, Wxa, Wxb, Wxin, Wxla/mw, and Wxlv in Shanlan landrace rice were identified, indicating rich gene resources in Shanlan rice for quality rice breeding. These findings are expected to contribute to the development of new rice with premium quality.

Insights on the enhancement of chilling tolerance in Rice through over-expression and knock-out studies of OsRBCS3

ABSTRACT Chilling stress is an important environmental factor that affects rice (Oryza sativa L.) growth and yield, and the booting stage is the most sensitive stage of rice to chilling stress. In this study, we focused on OsRBCS3, a rice gene related to chilling tolerance at the booting stage, which encodes the key enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) small subunit in photosynthesis. The aim of this study was to elucidate the role and mechanism of OsRBCS3 in rice chilling tolerance at the booting stage. The expression levels of OsRBCS3 under chilling stress were compared in two japonica rice cultivars with different chilling tolerances: Kongyu131 (KY131) and Longjing11 (LJ11). A positive correlation was found between OsRBCS3 expression and chilling tolerance. Over-expression (OE) and knock-out (KO) lines of OsRBCS3 were constructed using over-expression and CRISPR/Cas9 technology, respectively, and their chilling tolerance was evaluated at the seedling and booting stages. The results showed that OE lines exhibited higher chilling tolerance than wild-type (WT) lines at both seedling and booting stages, while KO lines showed lower chilling tolerance than WT lines. Furthermore, the antioxidant enzyme activities, malondialdehyde (MDA) content and Rubisco activity of four rice lines under chilling stress were measured, and it was found that OE lines had stronger antioxidant and photosynthetic capacities, while KO lines had the opposite effects. This study validated that OsRBCS3 plays an important role in rice chilling tolerance at the booting stage, providing new molecular tools and a theoretical basis for rice chilling tolerance breeding.

A simplified and improved protocol of rice transformation to cater wide range of rice cultivars.

The latest CRISPR-Cas9-mediated genome editing technology is expected to bring about revolution in rice yield and quality improvement, and thus validation of rice transformation protocols using CRISPR-Cas9-gRNA constructs is the need of the hour. Moreover, regeneration of more number of transgenic rice plants is prerequisite for developing genome-edited rice lines, as recalcitrant rice varieties were shown to have lower editing efficiencies which necessities screening of large number of transgenic plants to find the suitable edits. In the present study, we have simplified the Agrobacterium-mediated rice transformation protocol for both Indica and Japonica rice cultivars using CRISPR/Cas9 empty vector construct, and the protocols have been suitably optimized for getting large numbers of the regenerated plantlets within the shortest possible time. The Japonica transgenic lines were obtained within 65days and for the Indica cultivars, it took about 76-78days. We also obtained about 90% regeneration efficiency for both Japonica and Indica cultivars. The transformation efficiency was about 97% in the case of Japonica and 69-83% in the case of Indica rice cultivars. Furthermore, we screened the OsWRKY24 gene editing efficiency by transforming rice cultivars with CRISPR/Cas9 construct harbouring sgRNA against OsWRKY24 gene and found about 90% editing efficiency in Japonica rice cultivars, while 30% of the transformed Indica cultivars were found to be edited. This implicated the presence of a robust repair mechanism in the Indica rice cultivars.

A Preliminary Study of the Impacts of Duckweed Coverage during Rice Growth on Grain Yield and Quality.

The overuse and misuse of fertilizers have been causing duckweed outbreaks in irrigation ditches and paddy fields in many rice-growing areas. However, how duckweed coverage in a paddy field affects the rice yield and grain quality is under debate because duckweed may act as either a weed, competing with rice for mineral nutrients, or a "nutrient buffer", providing significant ecological and economic benefits. To understand the effects of duckweed coverage throughout rice growth on the yield and quality of rice grains, an experiment with three Japonica rice cultivars was conducted with fertile lotus-pond bottom soil as a growth medium to provide sufficient mineral nutrients for both the duckweed and rice. Averaged across three rice cultivars, duckweed coverage decreased the panicle density but increased the spikelet density and grain weight, resulting in no significant change in the rice yield. Duckweed coverage had no impact on the processing and appearance quality in general, but significant duckweed-by-cultivar interactions were detected in the head rice percentage and grain chalkiness, indicating different sensitivities of different cultivars in response to the duckweed treatment. The decrease in breakdown and increase in setback values in the rapid visco analyzer (RVA) profile of rice flour suggested that duckweed coverage during rice growth worsened the cooking quality of the rice. However, no significant change in the palatability of the cooked rice was found. The most profound change induced by the duckweed was the nutritional quality; duckweed coverage increased the protein concentration but decreased the concentrations of Mg, Mn, Cu, and Zn in rice grains. This preliminary study suggested that duckweed coverage during rice growth has profound effects on the rice nutrient uptake and grain nutritional quality under the circumstances, and further research on the responses of the rice quality to the duckweed coverage in paddy fields in multiple locations and years is needed.

Wetting alternating with partial drying during grain filling increases lysine biosynthesis in inferior rice grain

Lysine content is a criterion of the nutritional quality of rice. Understanding the process of lysine biosynthesis in early-flowering superior grain (SG) and late-flowering inferior grain (IG) of rice would advance breeding and cultivation to improve nutritional quality. However, little information is available on differences in lysine anabolism between SG and IG and the underlying mechanism, and whether and how irrigation regimes affect lysine anabolism in these grains. A japonica rice cultivar was grown in the field and two irrigation regimes, continuous flooding (CF) and wetting alternating with partial drying (WAPD), were imposed from heading to the mature stage. Lysine content and activities of key enzymes of lysine biosynthesis, and levels of brassinosteroids (BRs) were lower in the IG than in the SG at the early grain-filling stage but higher at middle and late grain-filling stages. WAPD increased activities of these key enzymes, BR levels, and contents of lysine and total amino acids in IG, but not SG relative to CF. Application of 2,4-epibrassinolide to rice panicles in CF during early grain filling reproduced the effects of WAPD, but neither treatment altered the activities of enzymes responsible for lysine catabolism in either SG or IG. WAPD and elevated BR levels during grain filling increased lysine biosynthesis in IG. Improvement in lysine biosynthesis in rice should focus on IG.

High canopy photosynthesis before anthesis explains the outstanding yield performance of rice cultivars with ideal plant architecture

ContextThe majority of investigations on mechanisms underlying yield difference between superior and conventional cultivars has focused on the grain-filling stage after anthesis. However, the physiological process before anthesis has been less explored. ObjectiveWe aimed to quantify the contribution of canopy photosynthesis at pre-anthesis stage to grain yield formation and clarify the mechanism underlying the yield advantage of the superior cultivars. MethodsThis study selected representative cultivars of indica and japonica rice, each type having a pair of cultivars with ideal plant architecture (IPA) and conventional architecture. Field experiments over four years (2019–2022) were conducted to compare the relevance of canopy photosynthesis to yield formation between the phases before and after anthesis. ResultsThe two IPA cultivars, JYZK-6 and ZZY-1, produced higher grain yield than the two conventional cultivars, NJ-5055 and HHZ. JYZK-6 and ZZY-1 also had good distribution of light interception inside the canopy, with the middle and lower part receiving more light than NJ-5055 and HHZ. At leaf level, there was no significant difference in photosynthetic rate between the two types of cultivars. At canopy level, the IPA cultivars had stronger photosynthetic activities before anthesis, outperforming NJ-5055 by 35.56% in averaged canopy photosynthesis, and by 46.65% in averaged daily net photosynthetic accumulation in 2021–2022. By contrast, canopy photosynthesis after anthesis was not significantly different between the two types. The IPA cultivars stored heavier biomass in internodes and sheaths before anthesis and remobilized more reserves to the developing grains, with the remobilization rate being 16.42% (internodes) and 35.34% (sheaths) higher than NJ-5055 and HHZ. ConclusionThe higher canopy productivity before anthesis is one of the main reasons accounting for the superior IPA cultivars, suggesting the necessity of optimizing plant growth and development during the vegetative and reproductive phases.