Background Rice farming faces a significant challenge from the brown planthopper (BPH), a destructive pest that threatens crop yields. Developing BPH-resistant rice varieties is critical for ensuring food security. The pyramiding of BPH resistance genes, BPH14 and BPH15, has proven effective in providing protection in elite rice strains. MicroRNAs (miRNAs) play a pivotal role in plant defense by fine-tuning resistance responses through the modulation of genes involved in various signaling pathways and metabolic processes. B1415, the pyramiding line containing BPH14 and BPH15, exhibits stronger BPH resistance compared to its recurrent parent, Wushansimiao, without affecting other important agronomic traits. However, the molecular basis underlying the resistance conferred by the BPH14/BPH15 pyramiding rice remains largely unclear, particularly with respect to the potential regulatory role of miRNAs. Studying miRNAs in resistance gene pyramiding lines like B1415 is essential for advancing rice breeding efforts, as such research can uncover regulatory networks that enhance pest resistance and identify miRNA-mRNA interactions as potential targets for genetic manipulation. Results The study investigated miRNA levels in B1415 and their recurrent parent (RP) under BPH infestation employing high-throughput sequencing and revealed 136 differentially expressed miRNAs (DEMs) among 550 known miRNAs. An integrated analysis highlighted that 587 miRNA-mRNA pairs linking 95 DEMs to 537 targeted genes were enriched in phenylpropanoid and lignin metabolism, circadian rhythms, and amino acid metabolism. The candidate DEMs, miR172d-3p, and miR396 family members were identified as negative regulators to decrease their target genes Os06g0708700 (encoding a nodulin-like protein) and Os11g0129700 (encoding an AP2 domain transcription factor), suggesting their key roles in rice against BPH. Conclusions Our investigation provides the first insights into miRNA-mediated defense mechanisms in the B1415. Identifying miRNAs and their target mRNAs in BPH resistance opens a new avenue for rice breeding programs, offering potential targets for improving pest resistance. Understanding these molecular interactions paves the way for developing more resistant rice cultivars, thereby contributing to sustainable rice production and food security.

High temperature is a major environmental constraint that severely limits rice (Oryza sativa) growth, yield potential, and geographical adaptability. The molecular mechanisms underlying rice adaptation to warm climates remain poorly understood. By integrating population genomic data with multi-scale phenotyping, we show that a single nucleotide polymorphism (SNP-1456) in the promoter of the defense transcription factor WRKY53 determines rice thermotolerance and grain yield. The thermotolerant allele strengthens the binding of the membrane-tethered NAC factor OsNTL3, which represses WRKY53 expression. Low WRKY53 derepresses catalase A (CatA), thereby lowering hydrogen peroxide (H2O2) levels. CRISPR knockout of WRKY53 boosts seed set and increases yield under field heat waves. Environmental-cline analysis of 4315 accessions reveals that the thermotolerant allele of WRKY53 is predominantly distributed in low-latitude regions with warm climates. Our findings reveal a previously uncharacterized WRKY53-centered regulatory network that mediates thermotolerance and facilitates rice adaptation to warm environments, offering promising targets for molecular breeding of climate-resilient rice varieties.

IntroductionThe application of genetically engineered (GE) crops in pest management raises biosafety concerns among governments, the scientific community, and the public, especially with the emergence of RNA interference (RNAi)-based crops expressing insecticidal double-stranded RNA (dsRNA). These crops may pose challenges to public health, agriculture, and conservation, and they could also present risks to non-target organisms, including beneficial natural enemies of pests. Natural enemies of insects are a significant component of global biodiversity and play a crucial role in managing insect pests within agroecosystems. This study addresses the biosafety concerns associated with insect-resistant transgenic dsRNA-expressing crops, focusing on their potential unintended effects on non-target organisms, particularly natural enemies.MethodsWe combined biological and bioinformatic approaches, utilizing both food-chain delivery and animal-feeding systems, to comprehensively evaluate the potential unintended effects of exogenous insecticidal dsRNA expressed by dsAllim cotton on the biological parameters and transcriptome of the cotton-field predatory natural enemy, Orius similis.ResultsThe findings indicate that dsAllim cotton had no adverse effects on O. similis, suggesting its potential safety for non-target beneficial insects. At both developmental and transcriptomic levels, dsAllim cotton showed no significant impact on O. similis.DiscussionThese results support the use of dsAllim cotton as a reference in developing regulatory frameworks for the risk assessment of RNAi crops. Together with previous research, our findings underscore the importance of conducting RNAi crop safety evaluations for non-target organisms on a case-by-case basis, with particular attention to potential off-target effects.

Polyploidization drives evolutionary innovation and crop domestication by leveraging novel gene combinations and increased genetic variation. Although tetraploid rice shows promise for yield enhancement due to its larger grains, the underlying regulatory mechanisms remain unclear. In this study, using tetraploid rice CX35-4x and its diploid counterpart CX35-2x, we constructed the first comprehensive mRNA and microRNA atlas during grain development. Transcriptomic analysis revealed that during key stages of hull development, transcription factors and genes involved in plant hormone signal transduction exhibited pronounced dose-sensitive effects in tetraploid rice, forming a core regulatory network for polyploid grain development. Integrated analysis revealed that dose-sensitive miRNAs regulate these key genes, enabling construction of a miRNA-target network controlling grain size. Functional validation of key dose-sensitive miR164e and WRKY50 transcription factors confirmed that miR164e inhibit WRKY50 expression via 3'UTR targeting, negatively regulating grain development. miR164e mutation increased thousand-grain weight by 25%, while WRKY50 mutation caused 16% reduction. Further DAP-seq analysis identified JAZ7 as direct WRKY50 targets. As dose-sensitive hormone signaling components, JAZ7 expression is activated by WRKY50, and JAZ7 loss-of-function reduced thousand-grain weight by 18%. Our study systematically reveals the dose-sensitive regulatory network controlling polyploid rice grain size, and for the first time, establishes the novel miR164e-WRKY50-JAZ7 dose-sensitive module as a key regulatory mechanism. These findings provide new insights into polyploidy advantage and valuable genetic resources for further rice yield improvement.

Enhancing crop resilience to guarantee stable, high yields under adverse conditions has long been a central goal of rice breeding, but it remains challenging because of inherent trade-offs. Here, we show that the transcriptional activity of the aldehyde-dehydrogenase OsALDH2B1 simultaneously increases grain length and alkaline tolerance. Upon alkali stress, the plasma-membrane leucine-rich-repeat receptor-like kinase SERL1 phosphorylates OsALDH2B1 at Thr-481, thereby blocking its 26S-proteasome-mediated degradation. Stabilized OsALDH2B1 directly represses GS3, a negative regulator of both grain size and alkaline tolerance, and activates all three catalase genes, leading to reduced hydrogen-peroxide (H2O2) accumulation. OsALDH2B1 over-expression enlarged grains and raised grain yield under field alkalinity, whereas osaldh2b1 mutants were hypersensitive. CRISPR knockouts of SERL1 or the catalase genes phenocopied these defects, confirming that the SERL1-OsALDH2B1 module constitutes an integrated signaling axis that links membrane perception to nuclear reprogramming through GS3 repression and catalase activation. This dual-purpose circuit provides an immediate, breeder-friendly target for high-yield, alkaline-resilient rice.

Insect pests and weeds are the two major biotic factors affecting crop yield in the modern agricultural system. In this study, a brown planthopper (BPH) resistance gene (BPH9) and glufosinate tolerance gene (bar) were stacked into a single T-DNA cassette and transformed into an indica rice (Oryza sativa L.) line H23. The present study employed a gene stacking process that combines more than one gene/trait into an individual transgenic plant to meet the increasing cropping demands under complex conditions. The transgenic rice H23 (H23R) co-expressing bar and BPH9 genes demonstrated both glufosinate tolerance and BPH resistance. We utilized transcriptome data to reveal the mechanism of BPH9-mediated brown planthopper resistance and to analyze the impact of exogenous transgenic fragments on upstream and downstream genes at insertion sites. The evaluation of insect resistance and glufosinate tolerance confirmed H23R as an excellent double-resistant transgenic rice. These findings indicate that H23R can satisfy insect management and weed control in the modern rice agricultural system. However, a deregulation study will help with prospective commercial planting.

Rice yield remains limited by trade-offs between effective panicle number, grain number per panicle, and grain weight. However, the molecular mechanisms linking auxin transport to panicle formation in rice remain largely unknown. In this study, we conducted genome-wide association studies and identified Suppressor of Effective Panicle 1 (SEP1), which encodes a basic helix-loop-helix transcription factor that negatively regulates effective panicle number and yield. SEP1 directly activates OsPIN1a and OsPIN1b, two auxin efflux carriers that modulate auxin transport and distribution in tiller buds. Natural variation in SEP1 alters the transcriptional activation capacity of SEP1, and the SEP1Hap2 allelic variant exhibits weaker transcriptional activation of OsPIN1a and OsPIN1b, correlating with its prevalence in Xian cultivars with relatively higher panicle numbers. Furthermore, we discovered that Gnp4/LAX PANICLE 2, a RING finger and WD40-associated ubiquitin-like domain-containing protein, destabilizes SEP1 via ubiquitin-proteasome degradation, fine-tuning auxin transport and tiller bud elongation. Notably, knockout of SEP1 in Xian/Geng cultivars significantly increases yield in field trials. Collectively, our study reveals a molecular mechanism for regulating rice yield and provides a practical strategy for breeding high-yield rice.

Cadmium (Cd) pollution in rice crops is a global environmental challenge, endangering food security and sustainable agricultural development. Cd ions are highly dynamic and toxic and can easily accumulate in rice grains, resulting in adverse consequences on human health and ecological safety. With accelerated industrialization and abundant agricultural activities, Cd enters paddy soils through multiple pathways, leading to increasingly complex processes of migration and transformation of Cd in the soil–rice ecosystem. Although recent studies have substantially advanced our comprehension of the pathways promoting the uptake, transport, and accumulation of Cd in rice, this information is scattered and lacks systematic integration, leading to an incomplete understanding of the entire contamination process. This review adopts a rigorous perspective spanning from soil input to grain accumulation and comprehensively summarizes the absorption pathways, translocation mechanisms, and remediation strategies for Cd pollution in rice. The effects of phytotoxicity induced by Cd on rice growth are thoroughly analyzed, and recent advances in various mitigation strategies are highlighted, including agronomic management, cultivar improvement, bioremediation, and signal regulation. By integrating the findings of latest research, this review (i) proposes a mechanistic network of Cd contamination occurrence and control in rice; (ii) elucidates critical regulatory nodes; and (iii) offers a theoretical framework for growing rice cultivars with a low Cd content, remediating Cd-contaminated farmlands, and ensuring food safety.

This study investigated the effects of high-temperature (HT) storage on the quality of three indica rice varieties (Huang Huazhan, Quan You 607, and E Zhong 5). Changes in rice nutrients, gamma-oryzanol content, enzyme activities, oxidative stress, processing quality, and palatability were examined under HT storage. The results indicated that HT storage accelerated decline in peroxidase (POD) activity, palatability, and head rice recovery, but increased fatty acid value, and malondialdehyde content, meanwhile altering moisture and fat contents, lipase (LPS), and lipoxygenase (LOX) activities, ferric reducing antioxidant power, and gamma-oryzanol content. During HT storage, gamma-oryzanol and its four monomer contents exhibited a trend of first significant increase and then a decrease. Huang Huazhan and Quan You 607 varieties exhibited significant H2O2 accumulation from day 20 onward under HT. Correlation analysis revealed that temperature and storage time were key factors deteriorating rice quality. Mechanistically, elevated temperature and prolonged storage increased LOX and LPS activities, which accelerated lipid peroxidation and H2O2 accumulation, aggravated lipid oxidative damage, eventually led to grain quality deterioration. Additionally, HT-induced oxidative stress activated antioxidant defense mechanisms in rice grain, enhancing gamma-oryzanol biosynthesis, whereas POD activity decreased. These findings elucidated the adaptive mechanism of indica rice under HT storage stress.

Grain size, which encompasses grain length, width, and thickness, is a critical determinant of both grain weight and quality in rice. Despite the extensive regulatory networks known to determine grain length and width, the pathway(s) that regulate grain thickness remain to be clarified. Here, we present the map- based cloning and characterization of qGT3, a major quantitative trait locus for grain thickness in rice that encodes the MADS-domain transcription factor OsMADS1. Our findings demonstrate that OsMADS1 regulates grain thickness by affecting sugar delivery during grain filling, and we show that OsMADS1 modulates expression of the downstream monosaccharide transporter gene MST4. A natural variant leads to alternative splicing and thus to a truncated OsMADS1 protein with attenuated transcriptional repressor activity. The truncated OsMADS1 protein results in increased expression of MST4, leading to enhanced loading of monosaccharides into the developing endosperm and thereby increasing grain thickness and improving grain quality. In addition, our results reveal that NF-YB1 and NF-YC12 interact directly with OsMADS1, acting as cofactors to enhance its transcriptional activity toward MST4. Collectively, these findings reveal a novel molecular mechanism underlying grain thickness regulation that is controlled by the OsMADS1-NF-YB1-YC12 complex and has great potential for synergistic improvement of grain yield and quality in rice.

Grain shape and tiller angle are two important agronomic traits influencing grain yield and quality in rice. Herein, we map-based cloned a grain shape gene GLW9 (Grain Length and Width on chromosome 9), which encodes a DNA binding with one finger (DOF) family transcription factor OsDOF25. GLW9 positively regulates grain length and negatively regulates grain width, consequently improving grain length-to-width ratio and appearance quality. GLW9 binds to the EXPA6 promotor to upregulate its expression, thereby positively regulating cell expansion and grain shape. On the other hand, GLW9 directly upregulates the expression of OsPIN1b to reduce tiller angle. This study elucidates the mechanism by which GLW9 coordinately regulates grain shape and tiller angle, providing theoretical reference and gene resources for the improvement of grain shape and tiller angle in rice.

The anther culture-based breeding of rice is a plant tissue culture technique that utilizes rice pollen to rapidly obtain haploid plants. In comparison with traditional breeding methods, this technique shortens the breeding cycle and enables the quick generation of homozygous plants, which is of great significance for the development of new rice varieties and the expansion of germplasm resources. With the advancement of technologies, the use of the anther culture technique in rice breeding has matured and has been applied to the development and utilization of new varieties with high yield, multiple resistances, and superior quality, in combination with other breeding methods. This technique has gained widespread attention globally, with many countries adopting it to create new germplasm resources. This study reviews advances in the rice anther culture technique, the factors influencing anther culture efficiency, and the progress in breeding rice varieties using this technique, as well as analyzes the current challenges and future prospects of anther culture breeding.

Rice, a vital crop, faces significant threats from the brown planthopper (BPH), which impacts plant growth and yield. Pyramiding the BPH resistance genes BPH14 and BPH15 provides rice crops with reliable and lasting protection against BPH. Nonetheless, current research lacks clarity on the molecular processes responsible for BPH14/BPH15-mediated resistance to BPH. In this study, utilizing high-throughput metabolomics and integrating transcriptomic data, we investigated the metabolic adaptations of the BPH14/BPH15 pyramiding line (B1415) and its recurrent parent (RP) during early and late infestation stages. The analysis identified 1007 metabolites, mainly consisting of lipids and lipid-like molecules, together with phenylpropanoid and polyketide classes. Differentially accumulated metabolites (DAMs) displayed different patterns in B1415 and RP, particularly in flavonoid and phenylpropanoid biosynthesis pathways, which were more pronounced in the resistant B1415. Furthermore, ferulic acid (FA) was found to negatively regulate BPH resistance. These findings elucidate critical metabolic pathways involved in rice defense mechanisms and underscore the potential of B1415’s enhanced metabolic responses in conferring durable resistance against BPH.

The brown planthopper (Nilaparvata lugens Stål, BPH) is a destructive pest of rice. Non-coding RNAs (ncRNAs) regulate the defense mechanisms in rice and the adaptive strategies of BPHs. In rice, ncRNAs modulate key resistance pathways such as jasmonic acid biosynthesis, flavonoid production, and phenylpropanoid metabolism, which increases BPH resistance. In BPHs, ncRNAs regulate processes such as reproduction, metabolism, and wing polyphenism, which facilitate adaptation and virulence. Cross-kingdom interactions between rice and BPHs reveal the dynamic molecular interplay that underpins this pest–host relationship. These new insights into ncRNA functions will help improve innovative pest management strategies and equip rice varieties with enhanced BPH resistance.

Grain chalkiness is an undesirable agronomic trait that negatively affects both the yield and quality of rice (Oryza sativa). The molecular mechanisms underlying chalky grain phenotype have remained largely unclear. In this study, we selected the rice variety HK300 with a high chalkiness, and ZR24D with a low chalkiness, as experimental materials and systematically characterized the reasons of grain chalkiness formation at molecular level by means of RNA-seq analysis. Analysis results revealed that the differentially expressed genes (DEGs) in these two rice varieties were significantly enriched in transcriptional regulation, sucrose and starch metabolism, and phytohormone signal transduction. Moreover, we found the expression of 13 genes related to trehalose pathway (4 out of 14 TPS genes and 9 out of 13 TPP genes in rice genome) were significantly different between the two varieties, indicating trehalose synthesis pathways may contribute to the increased chalkiness formation. Notably, the number of DEGs associated with the signal transduction pathway for indole-3-acetic acid (IAA), which has been rarely studied for its involvement in chalkiness formation, was the highest among those associated with plant hormone signal transduction. Among them, the expression of two IAA receptor genes, OsAFB3 and OsAFB5, were significantly lower in HK300 than that in ZR24D through RNA-seq and qRT-PCR. Furthermore, we newly validated the two genes negatively regulated the formation of chalkiness through gene knockout. Our findings provided the theoretical basis and novel gene resources for molecular breeding aimed at improving rice quality.

RH (Rathu Heenati), an indica rice cultivar from Sri Lanka, is highly resistant to several different biotypes of brown planthopper (Nilaparvata lugens Stål, BPH). In contrast, another indica rice cultivar, TN1 (Taichung Native 1), is highly susceptible to all biotypes of BPH. To reveal the molecular biological mechanism of biotic and abiotic stress mediated by BPH suction in two rice cultivars, RH and TN1, the genome-wide gene expressional profiling of these two rice varieties at 6 h after BPH infestation and needle puncturing was conducted by performing transcriptome analysis. In total, six samples (T6C, T6N, T6B, R6C, R6N, and R6B) were collected after performing different treatments for this expressional analysis. The results provide important information for the further isolation of BPH-resistance genes from RH as well as research on the mechanism of BPH resistance in RH. The transcriptome results revealed that TN1 (a BPH-susceptible rice variety) and RH (a BPH-resistant rice variety) harbor different pathways that respond to the physical injury of acupuncture and BPH infestation. The susceptible variety TN1 is more sensitive to the physical damage caused by acupuncture, whereas in the resistant variety RH, the response to insect damage is more rapid and direct. By comparing the gene changes of various plant hormones between the two varieties under different stresses, it was found that, in addition to the traditional SA and ET pathways related to the feeding induction of piercing-sucking pests, the expression patterns of JAZ-related genes in the JA signaling pathway also changed under two stresses (acupuncture induction and brown planthopper feeding induction), while the IAA hormone pathway-related genes in the susceptible variety TN1 also changed significantly after brown planthopper feeding. Furthermore, the results of this study help us to understand the biological process of rice responses to BPH and provide a promising direction to identify BPH-resistance-related genes in RH.

Approximately one-third of the total annual food production in the world is lost owing to pests, diseases and weeds. Therefore, the challenges posed by crop losses and population growth have emphasized the need for better breeding techniques (FAO et al., 2023). Practical experience has demonstrated that the utilization of existing resistance genes to breed and cultivate herbicide- and pest-resistant rice cultivars is the most economical, safe and effective method for preventing and controlling weeds and pests (Zhang, 2007). The incorporation of a single or few resistance genes during rice breeding is no longer adequate for in-demand production. In addition, hybridization and backcrossing involve a long breeding cycle, and the issue of linkage drag may occur. The multi-gene transformation strategy can be utilized for the rapid and accurate incorporation of multiple resistance genes into rice (Zhu et al., 2017). The fact that a trade-off between growth and defence generally exists in crops is universally accepted. Therefore, the overexpression of multi-resistance genes in rice causes considerable changes to the agronomic traits of crops, especially yield. The crop yield is positively correlated with the flowering stage within a certain range. For example, editing Ehd1 or overexpressing Ghd7 to appropriately extend the basic vegetative growth period of rice may be possible, and ultimately promote rice yield and quality (Eshed and Lippman, 2019; Zhou et al., 2023). This strategy is more effective for rice varieties with shorter growth periods. For some rice varieties with longer growth periods, we can use editing other yield related genes (grain type or grain weight), such as GS3 and GS5 (Ren et al., 2023). The herbicide resistance gene I. variabilis-EPSPS*, brown planthopper resistance genes Bph14* and OsLecRK1*, borer resistance gene Cry1C*, bacterial blight resistance gene Xa23* and blast resistance gene Pi9* are resistance gene resources in rice that have been extensively validated for use in rice breeding (Appendix S1). In our work, a highly efficient transgene system was used to construct an assembly of six resistance genes (about 26 Kb) mentioned earlier (380-6G) and Ehd1 CRISPR/Cas9 editing vector (Cas9-Ehd1) (Figure 1a; Appendix S2 and S3). We expect to extend the basic vegetative growth period of multi-resistance gene transgenic rice by editing Ehd1 to improve the agronomic traits (especially yield) and obtain a new multi-resistance and high-yield rice germplasm resource, termed MR&HY rice. We transformed two vectors, 380-6G and Cas9-Ehd1, into ZH11 rice varieties using Agrobacterium-mediated dual-strain transformation and screened using glyphosate and hygromycin simultaneously. When T0 transgenic plants were obtained, single-copy families with correct expression of resistance genes and correct editing of Ehd1 were screened out. Subsequently, the lines with homozygous single copy of multi-resistance genes and the Cas9-free family with Ehd1 mutation were screened in the T1 generation for further study (Figures 1b and S2). According to the process shown in Figure 1b, we obtained MR&HY-3 and MR&HY-5 with six resistance genes single copy homozygous, Ehd1-editing and Cas9-free. In MR&HY-3 and MR&HY-5 T2 generation, all six resistance genes were expressed normally and Ehd1 was mutated as expected (Figures 1c–g and S2). The MR&HY rice not only possessed resistance to herbicide (glyphosate), pests (brown planthopper and stem borer) and diseases (bacterial blight and blast), but also exhibited a considerable increase in yield (Figure 1h–m). In addition, the resistance rice transformed with multiple resistance genes had substantially better resistance to specific diseases and pests than single gene effects. In field experiments, although the MR&HY rice was cultivated without pesticide throughout the entire growth period, as compared with the ZH11 with pesticide, the yield increased by 20%; compared with ZH11 without pesticide throughout the entire growth period, the yield had almost increased by three times (Figure 1p,q). Although the growth period of MR&HY rice was extended by approximately 13 days, its yield and even quality had improved (Li et al., 2022a). Improving yield and disease resistance are important indicators of breeding, but the signals regulating yield and disease resistance often contrast each other. Currently, it is reported that only a few genes can simultaneously promote growth and resistance (Li et al., 2020, 2022b; Sun et al., 2019). In this study, we used multigene transformation techniques to enhance rice resistance against diseases, pests and herbicides and simultaneously performed CRISPR/Cas9 gene editing to adjust the heading stage of rice. This approach successfully balanced the tradeoff between rice growth and defence, and created rice germplasm with resistance to diseases, pests and weeds as well as increased yield. The multiple resistance of the resultant rice germplasm decreases the use of pesticide, which lowers rice production cost, reduces environmental pollution, enhances rice quality and renders the rice safe for human consumption. Its high yield effectively increases grain production, thereby addressing the global food crisis that has become increasingly severe with continuous population growth. This research was supported by the Biological Breeding-Major Projects (2023ZD04074), the Hubei Province Outstanding Youth Project (2024AFA088) and the Science and Technology Major Program of Hubei Province (2022ABA001 and 2021ABA011). The authors declare no competing interests. A.Q.Y., Y.J.L., L.Z. and C.Y.L. designed the research; C.Y.L., Z.H.Z., X.Z.X., C.X.L., C.H.L., E.-L. and J.Y.W. performed the research; C.Y.L, H.C., W.Z. and B.W. analysed the data; C.Y.L., L.Z., Y.J.L. and A.Q.Y. wrote the paper. The data that support the findings of this study are available in TIGR at http://rice.uga.edu/analyses_search_locus.shtml. These data were derived from the following resources available in the public domain: – LOC_Os03g63150, http://rice.uga.edu/cgi-bin/sequence_display.cgi?orf=LOC_Os03g63150.2 – LOC_Os04g12540, http://rice.uga.edu/cgi-bin/sequence_display.cgi?orf=LOC_Os04g12540.1 – LOC_Os11g37620, http://rice.uga.edu/cgi-bin/sequence_display.cgi?orf=LOC_Os11g37620.1 – LOC_Os06g17900, http://rice.uga.edu/cgi-bin/sequence_display.cgi?orf=LOC_Os06g17900.1. Appendix S1 Six resistance gene sequences optimized for codons. Appendix S2 The process of obtaining Multi-resistance and high-yield rice. Appendix S3 Primers used in this study. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.