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Improving the efficiency of germplasm innovation has always been the aim of rice breeders. Traditional hybrid breeding methods for variety selection rarely meet the practical needs of rice production. The emergence of genome-editing technologies, such as CRISPR/Cas9, provides a new approach to the genetic improvement of crops such as rice. The number of published scientific papers related to "gene editing" and "CRISPR/Cas9" retrievable on websites both from China and other countries exhibited an increasing trend, year by year, from 2014 to 2023. Research related to gene editing in rice accounts for 33.4% and 12.3% of all the literature on gene editing published in China and other countries, respectively, much higher than that on maize and wheat. This article reviews recent research on CRISPR/Cas9 gene-editing technology in rice, especially germplasm innovation and genetic improvement of commercially promoted varieties with improved traits such as disease, insect, and herbicide resistance, salt tolerance, quality, nutrition, and safety. The aim is to provide a reference for the precise and efficient development of new rice cultivars that meet market demand. [ABSTRACT FROM AUTHOR]

Rice, a globally important food crop, faces significant challenges due to salt and drought stress. These abiotic stresses severely impact rice growth and yield, manifesting as reduced plant height, decreased tillering, reduced biomass, and poor leaf development. Recent advances in molecular biology and genomics have uncovered key physiological and molecular mechanisms that rice employs to cope with these stresses, including osmotic regulation, ion balance, antioxidant responses, signal transduction, and gene expression regulation. Transcription factors such as DREB, NAC, and bZIP, as well as plant hormones like ABA and GA, have been identified as crucial regulators. Utilizing CRISPR/Cas9 technology for gene editing holds promise for significantly enhancing rice stress tolerance. Future research should integrate multi-omics approaches and smart agriculture technologies to develop rice varieties with enhanced stress resistance, ensuring food security and sustainable agriculture in the face of global environmental changes. [ABSTRACT FROM AUTHOR]

Through backcrossing and marker-assisted selection, gene Pi9 for resistance to rice blast was introduced into the water-saving and drought-resistant rice variety, Hanhui 3. The genetic background identity between Hanhui 8200 and Hanhui 3 was 91.4%. The drought resistance and drought avoidance abilities of Hanhui 8200 were equivalent to those of Hanhui 3. The resistance to rice blast was improved from grade 7 to grade 1. The rice quality of Hanhui 8200 meets the Ministry of Agriculture's grade 3 rice standards. The two-line and three-line hybrids formulated with Hanhui 8200 have high yield potential. Among them, the three-line hybrid Hanyou 8200 (Approval No.: Evaluated Rice 20210073), formulated with Huhan 7A, passed the Hubei Provincial approval in 2021, and the two-line hybrid Hanyouliangyou 8200 (Approval No.: Nationally Validated Rice 20210448), formulated with Huhan 82S, passed the national variety approval in 2021. Both hybrids demonstrated strong resistance to rice blast, moderate resistance to bacterial leaf blight, strong drought resistance, high quality, and high yield. [ABSTRACT FROM AUTHOR]

Life cycle (annual vs perennial) and leaf venation pattern (parallel and reticular) are known to be related to water use strategies in herb species and critical adaptation to certain climatic conditions. However, the effect of these two traits and how they influence the coordination between vein density (vein length per area, VLA) and stomatal density (SD) remains unclear. In this study, we examined the leaves of 53 herb species from a subtropical botanical garden in Guangdong Province, China, including herbs with different life cycles and leaf venation patterns. We assessed 21 leaf water-related functional traits for all species, including leaf area (LA), major and minor VLA, major and minor vein diameter (VD), SD and stomatal length (SL). The results showed no significant differences in mean SD and SL between either functional group (parallel venation vs reticular venation and annual vs perennial). However, parallel vein herbs and perennial herbs displayed a significantly higher mean LA and minor VD, and lower minor VLA compared to reticular vein herbs and annual herbs, respectively. There was a linear correlation between total VLA and SD in perennial and reticular vein herbs, but this kind of correlation was not found in annual and parallel vein herbs. The major VLA and minor VD were significantly affected by the interaction between life cycle and leaf venation pattern. Our findings suggested that VLA, rather than SD, may serve as a more adaptable structure regulated by herbaceous plants to support the coordination between leaf water supply and demand in the context of different life cycles and leaf venation patterns. The results of the present study provide mechanistic understandings of functional advantages of different leaf types, which may involve in species fitness in community assembly and divergent responses to climate changes. [ABSTRACT FROM AUTHOR]

Plant architecture is a complex trait and has a profound impact on crop performance and productivity. We applied the CRISPR/Cas9 system to mutate the DEP1 gene, which has been reported to function as regulator of plant architecture, in elite Xian cultivar 'Huhan1509'. Sequencing analysis of T0 transformed plants showed that the CRISPR/Cas9 system was highly efficient in mutagenesis of targeted DEP1 gene, with 30% of the homozygous mutations and 70% of the heterozygous mutations. T2 homozygous mutants without T‐DNA were further examined for the agronomic traits. The DEP1 mutants exhibited an altered plant architecture along with a shorter plant height and grain size and increased spikelets and grain density. Furthermore, phenotypes of raising primary branches and stem diameters were observed in the DEP1 mutants. Our results demonstrate that favourable alleles of the DEP1 gene, developed by CRISPR/Cas9 system, could be used to improve plant architecture in Xian rice. [ABSTRACT FROM AUTHOR]

Of all the crops, rice is the one that consumes the most water. Rice yields and quality are significantly influenced by irrigation. However, it is still unknown how different irrigation practices would affect the grain yield and quality of water-saving and drought-resistant rice. Hyou 518 (high-yielding rice variety) and Hanyou 73 (water-saving and drought-resistant rice variety) were employed as materials. Three irrigation regimes were set up in the field: conventional flooding irrigation (CF), alternate wetting and moderate soil drying irrigation (AWD), and dry cultivation (D). It was investigated how various irrigation regimes affected the two varieties' yield and quality. The results revealed the following: 1. D considerably increased water-use efficiency while drastically reducing the yield, compared to CF and AWD. In comparison to other irrigation regimes, the grain yield and water use efficiency of Hanyou 73 enhanced synergistically under AWD treatment. 2. In contrast to CF treatment, AWD and D (especially) treatments decreased perfect rice kernel, total starch content, amylose content, amylopectin content, amylose/amylopectin, gel consistency, and breakdown, but increased green rice kernel, chalky kernel, protein content, and setback. 3. After heading, AWD and D lowered, and D treatment decreased more, the activities of ADP-glucose pyrophosphorylase (AGP), soluble starch synthase (SSS), and starch branching enzyme (SBE). AGP, SSS, and SBE were strongly inversely linked with perfect rice kernel, amylopectin content, gel consistency, and breakdown, but significantly negatively correlated with green rice kernel, chalky kernel, protein content, and setback. The results indicate that with AWD treatment, Hanyou 73 might provide a synergistic boost grain production, water-use efficiency, and quality. D treatment could significantly improve water-use efficiency. Compared with Hyou518, Hanyou 73 could maintain higher AGP, SSS, and SBE activities, head milled rice, perfect rice kernel, amylopectin content, and gel consistency under AWD and D treatment. [ABSTRACT FROM AUTHOR]

Rice production is often affected by biotic and abiotic stressors. The breeding of resistant cultivars is a cost-cutting and environmentally friendly strategy to maintain a sustainable high production level. An elite water-saving and drought-resistant rice (WDR), Hanhui3, is susceptible to blast and bacterial blight (BB). This study was conducted to introgress three resistance genes (Pi2, xa5, and Xa23) for blast and BB into Hanhui3, using marker-assisted selection (MAS) for the foreground selection and a whole-genome single-nucleotide polymorphism (SNP) array for the background selection. As revealed by the whole-genome SNP array, the recurrent parent genome (RPG) recovery of the improved NIL was 94.2%. The resistance levels to blast and BB of the improved NIL and its derived hybrids were higher than that of the controls. In addition, the improved NIL and its derived hybrids retained the desired agronomic traits from Hanhui3, such as yield. The improved NIL could be useful to enhance resistance against biotic stressors and produce stable grain yields in Oryza sativa subspecies indica rice breeding programs. [ABSTRACT FROM AUTHOR]

Eukaryotic translation initiation factor 4E1 (eIF4E1) is required for the initiation of protein synthesis. The biological function of eIF4E1 in plant–potyvirus interactions has been extensively studied. However, the role of eIF4E1 in Arabidopsis development remains unclear. In this study, we show that eIF4E1 is highly expressed in the embryo and root apical meristem. In addition, eIF4E1 expression is induced by auxin. eIF4E1 mutants show embryonic cell division defects and short primary roots, a result of reduced cell divisions. Furthermore, our results show that mutation in eIF4E1 severely reduces the accumulation of PIN-FORMED (PIN) proteins and decreases auxin-responsive gene expression at the root tip. Yeast two-hybrid assays identified that eIF4E1 interacts with an RAC/ROP GTPase activator, RopGEF7, which has been previously reported to be involved in the maintenance of the root apical meristem. The interaction between eIF4E1 and RopGEF7 is confirmed by protein pull-down and bimolecular fluorescent complementation assays in plant cells. Taken together, our results demonstrated that eIF4E1 is important for auxin-regulated embryo development and root growth. The eIF4E1–RopGEF7 interaction suggests that eIF4E1 may act through ROP signaling to regulate auxin transport, thus regulating auxin-dependent patterning. [ABSTRACT FROM AUTHOR]

Introduction: Transcriptomic divergence drives plant ecological adaptation. Upland rice is differentiated in drought tolerance from lowland rice during its adaptation to the drought‐prone environment. They provide a good system to learn the evolution of drought tolerance in rice. Methods and Results: We estimate morphological differences between the two rice ecotypes under well‐watered and drought conditions, as well as their genetic and transcriptomic divergences by the high‐throughput sequencing. Upland rice possesses higher expression diversity than lowland rice does. Thousands of genes exhibit expression divergences between the two rice ecotypes, which contributes to their morphological differences in drought tolerance. These transcriptomic divergences contribute to drought adaptation of upland rice during its domestication. Mutations in transcriptional regulatory regions, which cause presence and absence of cis‐elements, are the cause of expression divergence. About 15.3% transcriptionally selected genes also receive sequence‐based selection in upland or lowland ecotype. Some highly differentiated genes promote the transcriptomic divergence between rice ecotypes via gene co‐expression network. In addition, we also detected transcriptomic trade‐offs between drought tolerance and productivity. Discussion: Many key genes, which promote transcriptomic adaptation to drought in upland rice, have great prospective in breeding water‐saving and drought‐resistant rice. Meanwhile, appropriate strategies are required in breeding to overcome the potential transcriptomic trade‐off. [ABSTRACT FROM AUTHOR]

Emergent aquatic plants mostly occur in shallow waters and root in bottom substrates, but their leaves emerge from the water surface and are thus exposed to air, similar to the leaves of terrestrial plants. Previous studies have found coordination between leaf water supply and demand in terrestrial plants; however, whether such a coordination exists in emergent aquatic plants remains unknown. In this study, we analysed leaf veins and stomatal characteristics of 14 emergent aquatic and 13 terrestrial monocotyledonous herb species (EMH and TMH), with 5 EMH and 8 TMH belonging to Poaceae. We found that EMH had significantly higher mean leaf area, leaf thickness, stomatal density, stomatal number per vein length and major vein diameter, but lower mean major vein length per area (VLA) and total VLA than TMH. There was no significant difference in stomatal length, minor VLA and minor vein diameter between the two groups. Stomatal density and total VLA were positively correlated among the EMH, TMH, as well as the 8 Poaceae TMH species, but this correlation became non-significant when data from both the groups were pooled. Our results showed that the differences in water supply between emergent aquatic and terrestrial plants modify the coordination of their leaf veins and stomatal traits. [ABSTRACT FROM AUTHOR]

Summary: The growth and development of roots in plants depends on the specification and maintenance of the root apical meristem. Here, we report the identification of CBL, a gene required for embryo and root development in Arabidopsis, and encodes cystathionine beta‐lyase (CBL), which catalyzes the penultimate step in methionine (Met) biosynthesis, and which also led to the discovery of a previous unknown, but crucial, metabolic contribution by the Met biosynthesis pathway. CBL is expressed in embryos and shows quiescent center (QC)‐enriched expression pattern in the root. cbl mutant has impaired embryo patterning, defective root stem cell niche, stunted root growth, and reduces accumulation of the root master regulators PLETHORA1 (PLT1) and PLT2. Furthermore, mutation in CBL severely decreases abundance of several PIN‐FORMED (PIN) proteins and impairs auxin‐responsive gene expression in the root tip. cbl seedlings also exhibit global reduction in histone H3 Lys‐4 trimethylation (H3K4me3) and DNA methylation. Importantly, mutation in CBL reduces the abundance of H3K4me3 modification in PLT1/2 genes and downregulates their expression. Overexpression of PLT2 partially rescues cbl root meristem defect, suggesting that CBL acts in part through PLT1/2. Moreover, exogenous supplementation of Met also restores the impaired QC activity and the root growth defects of cbl. Taken together, our results highlight the unique role of CBL to maintain the root stem cell niche by cooperative actions between Met biosynthesis and epigenetic modification of key developmental regulators. Significance Statement: Growth and development depend on metabolic activities that properly support fundamental cellular functions, but mechanistic links are scarce. Here, we demonstrate that a quiescent center‐enriched CBL gene, encoding the second committed enzyme in methionine (Met) biosynthesis, plays a crucial role in root apical meristem maintenance by epigenetically influencing expression of the root master regulators PLETHORA (PTL1/2), elucidating a functional link between Met metabolic contribution and auxin‐dependent PLT‐mediated maintenance of root stem cell niche. [ABSTRACT FROM AUTHOR]

Soil relative water content and seed plumpness have been shown to be the key factors affecting seed germination and seedling growth of rice under direct drought cropping. It remains to be determined whether seed germination and seedling growth of water-saving and drought-resistant rice (WDR) and conventional rice with the same proportion of rice seed have the same response to soil moisture changes. The purpose of this study was to investigate the seed germination and physiological characteristics of the rice cultivars Guangliangyou 1813 (GLY-1813,indica hybrid rice) and Hanyou 73 ((HY-73), WDR) with four different specific gravities (T1, T2, T3, and T4; the rice seeds were divided into four specific gravity levels by weight using saline water, the representative specific gravities were <1.0, 1.0–1.1, 1.1–1.2 and >1.2 kg m−3, respectively), at five soil moisture content gradients (soil relative water contents of 10–20%, 20–40%, 40–60%, 60–80%, and 80–100%), under dry direct seeding conditions. The results showed that GLY-1813 had a higher germination potential, germination and seedling emergence rates, greater root dry weight, seedling dry weight, root oxidation activity, and chlorophyll content, and lower malondialdehyde (MDA) content when the soil relative water content was 20–40% or 40–60%. Cultivar HY-73 had the highest germination rate and seedling physiological activity at 20–40% relative water content; its growth vigor was better than that of GLY-1813 at the same soil moisture level. In conclusion, the soil relative water content for seed germination of HY-73 was 20–40%, which was less than that of GLY-1813. When soil relative water content was sufficient for seed germination and growth, the higher the plumpness of the rice seed, the easier it was to resist the negative effects of an adverse growth environment. [ABSTRACT FROM AUTHOR]

Leaf phenology (evergreen vs. deciduous) and morphology (simple vs. compound) are known to be related to water use strategies in tree species and critical adaptation to certain climatic conditions. However, the effect of these two traits and their interactions on the coordination between minor vein density (MVD) and stomatal density (SD) remains unclear. In this study, we examined the leaves of 108 tree species from plots in a primary subtropical forest in southern China, including tree species with different leaf morphologies and phenologies. We assessed nine leaf water-related functional traits for all species, including MVD, SD, leaf area (LA), minor vein thickness (MVT), and stomatal length (SL). The results showed no significant differences in mean LA and SD between either functional group (simple vs. compound and evergreen vs. deciduous). However, deciduous trees displayed a significantly higher mean MVD compared to evergreen trees. Similarly, compound-leaved trees have a higher (marginally significant) MVD than simple-leaved trees. Furthermore, we found that leaf morphology and phenology have significantly interactive effects on SL, and the compound-leafed deciduous trees exhibited the largest average SL among the four groups. There were significant correlations between the MVD and SD in all different tree groups; however, the slopes and interceptions differed within both morphology and phenology. Our results indicate that MVD, rather than SD, may be the more flexible structure for supporting the coordination between leaf water supply and demand in different leaf morphologies and phenologies. The results of the present study provide mechanistic understandings of the functional advantages of different leaf types, which may involve species fitness in community assembly and divergent responses to climate changes., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The reviewer YN declared a shared affiliation with authors WZ, PF, QM, and PZ to the handling editor at the time of review., (Copyright © 2023 Zhao, Fu, Mao, Liu, Li, Xia and Zhao.)

Rice is one of the most economically important staple food crops in the world. Soil salinization and drought seriously restrict sustainable rice production. Drought aggravates the degree of soil salinization, and, at the same time, increased soil salinity also inhibits water absorption, resulting in physiological drought stress. Salt tolerance in rice is a complex quantitative trait controlled by multiple genes. This review presents and discusses the recent research developments on salt stress impact on rice growth, rice salt tolerance mechanisms, the identification and selection of salt-tolerant rice resources, and strategies to improve rice salt tolerance. In recent years, the increased cultivation of water-saving and drought-resistance rice (WDR) has shown great application potential in alleviating the water resource crisis and ensuring food and ecological security. Here, we present an innovative germplasm selection strategy of salt-tolerant WDR, using a population that is developed by recurrent selection based on dominant genic male sterility. We aim to provide a reference for efficient genetic improvement and germplasm innovation of complex traits (drought and salt tolerance) that can be translated into breeding all economically important cereal crops.

The commercialization of hybrid rice has greatly contributed to the increase in rice yield, with the improvement of its seed production capacity having played an important role. The stigma exsertion rate (SER) is a key factor for improving the outcrossing of the sterile line and the hybrid rice seed production. We used the Zhenshan 97B × IRAT109 recombinant inbred population comprising 163 lines and a natural population of 138 accessions to decipher the genetic foundation of SER over 2 years in three environments. Additionally, we detected eight QTLs for SER on chromosomes 1, 2, and 8 via linkage mapping. We also identified seven and 19 significant associations for SER using genome-wide association study in 2016 and 2017, respectively. Interestingly, we located two lead SNPs (sf0803343504 and sf083344610) on chromosome 8 in the qTSE8 QTL region that were significantly associated with total SER. After transcriptomic analysis, quantitative real-time PCR, and haplotype analysis, we found 13 genes within this reliable region as important candidate genes. Our study results will be beneficial to molecular marker-assisted selection of rice lines with high outcrossing rate, thereby improving the efficiency of hybrid seed production., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Liu, Fu, Kong, Ma, Zhang, Wang, Wang, Xia, Liu, Yu and Luo.)

Low amylose content (AC) is a desirable trait for rice ( Oryza sativa L.) cooking quality and is selected in soft rice breeding. The Wx hp allele was derived from a Yunnan rice landrace in China, Haopi, with low AC. To efficiently and rapidly utilize the low amylose content-associated gene Wx hp in rice molecular breeding programs, we developed a tetra-primer amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) method, according to the single-nucleotide variation of the Wx hp . Four Wx hp -specific primers were used to perform PCR assays using genomic DNA extracted from several rice varieties. Based on the band pattern of the amplified products after electrophoresis, this method can accurately distinguish three Wx hp -related genotypes (i.e., Wx hp homozygotes, Wx hp heterozygotes, and wild-type), and the genotypes completely correspond to the appearance of mature endosperm. This method represents a novel approach that is both inexpensive and highly efficient and can be widely used for genotyping Wx hp alleles in rice germplasm collections and may aid breeding programs with marker-assisted selection (MAS)., (Copyright © 2019 by JAPANESE SOCIETY OF BREEDING.)

RNA editing of mitochondrial gene transcripts plays a central role during plant development and evolutionary adaptation. RNA editing has previously been reported to differ between the rice cytoplasmic male sterile (CMS) line and its maintainer line, which has been suggested as a cause for their different performances under environmental stress. To specifically test this hypothesis, a wild abortive (WA) CMS line (Huhan-1A) and its maintainer line (Huhan-1B) were utilized to investigate performances in response to oxidative stress, as well as RNA editing efficiencies on transcripts of six selected mitochondrial genes. Compared to the maintainer line, Huhan-1A represented both lower plant height and total antioxidant capacity, possessed higher total soluble protein and chlorophyll contents, accumulated less H 2 O 2 content on the 3rd day after treatment (DAT), and exhibited higher survival ratio after re-watering. Furthermore, a total of 90 editing sites were detected on transcripts of six mitochondrial genes ( atp9, nad2, nad7, nad9, ccmB , and ccmC ) in both Huhan-1A and Huhan-1B on the 0, 1st, and 3rd DAT. Forty-eight sites were furthermore determined as stress-responsive sites (SRS). Generally, in response to oxidative stress, SRS in Huhan-1A increased the resulting editing efficiencies, while SRS in Huhan-1B decreased the resulting editing efficiencies. In addition, 33 and 22 sites at ccmB and ccmC were differentially edited between Huhan-1A and Huhan-1B, respectively, on the 0, 1st, and 3rd DAT. Editing efficiencies of ccmB and ccmC were generally lower in Huhan-1A ( ccmB , 37.3-47.8%; ccmC , 41.2-52.3%) than those in Huhan-1B ( ccmB , 82.6-86.5%; ccmC , 81.0-82.9%). Deficiencies of RNA editing in Huhan-1A at ccmB and ccmC could lead to the loss of transmembrane domains in their protein structures. Consequently, differences in RNA editing at ccmB and ccmC between the WA-CMS line and its maintainer line partially explained their different performances under stress. Moreover, we detected differences in expressions of pentatricopeptide repeat (PPR) genes between both lines, as well as significant correlations with RNA editing. Our study indicated potential associations of RNA editing and PPR genes in rice tolerance to abiotic stresses. However, the underlying molecular mechanisms of stress-adaptation, which are attributed to RNA editing on transcripts of mitochondrial genes, require further investigation.