qGW11a/OsCAT8,encoding an amino acid permease,negatively regulates grain size and weight in rice

Grain size is a key factor influencing grain weight in rice.In this study,a chromosome segment substitution line(CSSL9-17)was identified,that exhibits a significant reduction in both grain size and weight compared to its donor parent 93-11.Further investigation identified two quantitative trait loci(QTL)on chromosome 11,designated qGW11a and qGW11b,which contribute to 1000-grain weight with an additive effect.LOC_Os11g05690,encoding the amino acid permease OsCAT8,is the target gene of qGW11a.Overexpression of OsCAT8 resulted in decreased grain weight,while OsCAT8 knockout mutants exhibited increased grain weight.The 287-bp located within the OsCAT8 promoter region of 93-11 negatively regulates its activity,which is subsequently correlated with an increase in grain size and weight.These results suggest that OsCAT8 functions as a negative regulator of grain size and grain weight in rice.

Transcriptomic and metabolomic analysis of changes in grain weight potential induced by water stress in wheat

The sink strength of developing ovaries in wheat determines the grain weight potential.The period from booting to the grain setting stage is critical for ovary growth and development and potential sink capacity determination.However,the underlying regulatory mechanism during this period by which the wheat plant balances and coordinates the floret number and ovary/grain weight under water stress has not been clarified.Therefore,we designed two irrigation treatments of W0(no seasonal irrigation)and W1(additional 75 mm of irrigation at the jointing stage)and analyzed the responses of the ovary/grain weight to water stress at the phenotypic,metabolomic,and transcriptomic levels.The results showed that the W0 irrigation treatment reduced the soil water content,plant height,and green area of the flag leaf,thus reducing grain number,especially for the inferior grains.However,it improved the grain weight of the superior and inferior grains as well as average grain weight at maturity,while the average ovary/grain weight and volume during–3 to 10 days after anthesis(DAA)also increased.Transcriptomic analysis indicated that the genes involved in both sucrose metabolism and phytohormone signal transduction were prominently accelerated by the W0 treatment,accompanied by greater enzymatic activities of soluble acid invertase(SAI)and sucrose synthase(Sus)and elevated abscisic acid(ABA)and indole-3-acetic acid(IAA)levels.Thus,the sucrose content decreased,while the glucose and fructose contents increased.In addition,several TaTPP genes(especially TaTPP-6)were down-regulated and the IAA biosynthesis genes TaTAR1 and TaTAR2 were up-regulated under the W0 treatment before anthesis,which further increased the IAA level.Collectively,water stress reduced the growth of vegetative organs and eliminated most of the inferior grains,but increased the ABA and IAA levels of the surviving ovaries/grains,promoting the enzymatic activity of Sus and degrading sucrose into glucose and fructose.As a result,the strong sucrose utilization ability,the enhanced enzymatic activity of SAI and the ABA-and IAA-mediated signaling jointly increased the weight and volume of the surviving ovaries/grains,and ultimately achieved the tradeoff between ovary/grain weight and number in wheat under water stress.

GSW3.1,a novel gene controlling grain size and weight in rice

Grain size and weight are closely related traits determining yield in rice(Oryza sativa L.).Since indica and japonica rice varieties differ significantly in multiple traits,a high-generation recombinant inbred line(RIL)population derived from the crossing LH9(indica)and RPY(japonica)was used to map grainrelated traits in six environments.Pyramiding of the quantitative trait loci(QTL)for thousand-grain weight showed that combinations of multiple QTL significantly increased the phenotypic effect.A novel gene named GSW3.1 controlling grain size and weight was discovered using the major QTL for the colocalization of grain width and thousand-grain weight on chromosome 3.Gene editing revealed that GSW3.1(LOC_Os03g16850)was pleiotropic,positively regulating grain size and weight while affecting several other agronomic traits.Haplotype analysis indicated that some traits,including grain width and weight,were highly correlated with indica-japonica differentiation.

Effect of delayed sowing on grain number, grain weight, and protein concentration of wheat grains at specific positions within spikes

Delays in sowing have significant effects on the grain yield,yield components,and grain protein concentrations of winter wheat.However,little is known about how delayed sowing affects these characteristics at different positions in the wheat spikes.In this study,the effects of sowing date were investigated in a winter wheat cultivar,Shannong 30,which was sown in 2019 and 2020 on October 8(normal sowing)and October 22(late sowing)under field conditions.Delayed sowing increased the partitioning of ^(13)C-assimilates to spikes,particularly to florets at the apical section of a spike and those occupying distal positions on the same spikelet.Consequently,the increase in grain number was the greatest for the apical sections,followed by the basal and central sections.No significant differences were observed between sowing dates in the superior grain number in the basal and central sections,while the number in apical sections was significantly different.The number of inferior grains in each section also increased substantially in response to delayed sowing.The average grain weights in all sections remained unchanged under delayed sowing because there were parallel increases in grain number and ^(13)C-assimilate partitioning to grains at specific positions in the spikes.Increases in grain number m^(–2) resulted in reduced grain protein concentrations as the limited nitrogen supply was diluted into more grains.Delayed sowing caused the greatest reduction in grain protein concentration in the basal sections,followed by the central and apical sections.No significant differences in the reduction of the grain protein concentration were observed between the inferior and superior grains under delayed sowing.In conclusion,a 2-week delay in sowing improved grain yield through increased grain number per spike,which originated principally from an increased grain number in the apical sections of spikes and in distal positions on the same spikelet.However,grain protein concentrations declined in each section because of the increased grain number and reduced N uptake.

Identification and validation of major QTL for grain size and weight in bread wheat(Triticum aestivum L.)

Grain size and weight are key components of wheat yield.Exploitation of major underlying quantitative trait loci(QTL)can improve yield potential in wheat breeding.A recombinant inbred line(RIL)population was constructed to detect QTL for thousand-grain weight(TGW),grain length(GL)and grain width(GW)across eight environments.Genomic regions associated with grain size and grain weight were identified on chromosomes 4A and 6A using bulked segregant exome sequencing(BSE-Seq)analysis.After constructing genetic maps,six major QTL detected in at least four individual environments and in best linear unbiased estimator(BLUE)datasets,explained 7.50%-23.45%of the phenotypic variation.Except for QGl.cib-4A,the other five QTL were co-located in two regions,namely QTgw/Gw.cib-4A and QTgw/Gw/Gl.cib-6A.Interactions of these QTL were analyzed.Unlike QTgw/Gw/Gl.cib-6A,QTgw/Gw.cib-4A and QGl.cib-4A had no effect on grain number per spike(GNS).The QTL were validated in a second cross using Kompetitive Allele Specific PCR(KASP)markers.Since QTgw/Gw.cib-4A was probably a novel locus,it and the KASP markers reported here can be used in wheat breeding.TraesCS4A03G0191200 was predicted to be potential candidate gene for QTgw/Gw.cib-4A based on the sequence differences,spatiotemporal expression patterns,gene annotation and haplotype analysis.Our findings will be useful for fine mapping and for marker-assisted selection in wheat grain yield improvement.

Introgression of chromosome 6PL terminal segment from Agropyron cristatum to increase both grain number and grain weight in wheat

Agropyron cristatum(2n=4x=28,PPPP),which harbours many high-yield and disease-resistance genes,is a promising donor for wheat improvement.Narrow genetic diversity and the trade-off between grain weight and grain number have become bottlenecks for increasing grain yield in wheat.In this study,a novel translocation line,WAT650l,was derived from the chromosome 6P addition line 4844–12,which can simultaneously increase both grain number per spike(GNS)and thousand-grain weight(TGW).Cytological analysis and molecular marker analysis revealed that WAT650l was a 5BL.5BS-6PL(bin 12–17)translocation line.Assessment of agronomic traits and analysis of the BC4F2 and BC5F2 populations suggested that the 6PL terminal chromosome segment in WAT650l resulted in increased grain number per spike(average increased by 14.07 grains),thousand-grain weight(average increased by 4.31 g),flag leaf length,plant height,spikelet number per spike and kernel number per spikelet during the two growing seasons of 2020–2021 and 2021–2022.Additionally,the increased GNS locus and high-TGW locus of WAT650l were mapped to the bins 16–17 and 12–13,respectively,on chromosome 6PL by genetic population analysis of three translocation lines.In summary,we provide a valuable germplasm resource for broadening the genetic base of wheat and overcoming the negative relationship between GNS and TGW in wheat breeding.

The 218th amino acid change of Ser to Ala in TaAGPS-7A increases enzyme activity and grain weight in bread wheat

ADP-glucose pyrophosphorylase(AGPase)influences cereal productivity.There are few reports on the function of cytosolic AGPase small subunit in bread wheat(TaAGPS).In the present study,TaAGPS was preferentially expressed in developing endosperm during grain-filling stages in bread wheat.TaAGPS allelic variations were characterized in 143 wheat accessions by PacBio RS II sequencing.Two haplotypes(TaAGPS-7A-TG and TaAGPS-7A-CT)of TaAGPS-7A were identified and corresponding functional markers were developed,whereas no variants of TaAGPS-7B and TaAGPS-7D were detected.TaAGPS-7A was associated with thousand-kernel weight(TKW)by haplotype–trait association analysis in two populations.Near-isogenic lines(NILs)with TaAGPS-7A-TG showed higher TKW and total kernel starch content than those with TaAGPS-7A-CT,owing to the higher AGPase activity of TaAGPS-7A-TG than TaAGPS-7A-CT both in vitro and in vivo.Overexpression of TaAGPS-7A-TG in bread wheat doubled the transcription levels of TaAGPS and increased AGPase activity by 55.7%,resulting in a 3.0-g higher TKW than in the wild type(WT).Knockdown of TaAGPS led to reduced expression of TaAGPS,AGPase activity,and TKW than in the WT.Thus,owing to the 218th amino acid change of Ser to Ala in TaAGPS-7A,the favorable haplotype TaAGPS-7A-TG showed higher AGPase activity,resulting in higher kernel starch content and grain weight.This finding could be applied to increasing starch content and grain weight in bread wheat.

Effects of Spraying Nitrogen and Zinc Fertilizers after Flowering on Grain Weight and Nutritional Quality of Winter Wheat

In order to explore the effect of spraying fertilizers after flowering on grain weight and nutritional quality of different winter wheat cultivars, twenty wheat culti- vars in wheat zones along Yellow River and Huai River, were used to investigate the change of thousand kernel weight （TKW）, phytic acid content （PAC）, contents of Fe, Zn and Mg and bioavailability by spraying zinc and nitrogen fertilizers after flowering. The results showed that both of TKW and protein content in grains in- creased by spraying Zn and N fertilizers, and the effect of N fertilizer proved more significant. The PAC decreased significantly after Zn treatment, for example, PAC of ＂955159＂, a wheat cultivar, decreased by 27.95%. However, PAC was in- significantly influenced by N fertilizer. On the other hand, Zn content in wheat grains were improved after spraying Zn fertilizer and the increase aveaged 54.38%. Furthermore, Fe and Zn contents grew significantly after spraying N fertilizer, and the increase averaged 36.88% and 11.25% respectively. However, Mg content in grains declined by N fertilizer. Still, the bioavailability of zinc in grains was enhanced after spraying Zn fertilizer, and of Fe was also increased greatly after spraying N fertilizer. In conclusion, grain weight, and content and effectiveness of mineral ele-ments can be improved simultaneously by rational spraying of N and Zn fertilizers to improve quality of grains.

Effect of Enhanced UV-B Radiation on Seed Setting Rate and 1 000-grain Weight of Indica Hybrid Rice Restorer Lines

[Objective] This study was to investigate the effect of enhanced UV-B radiation on seed setting rate and 1 000-grain weight of hybrid rice combinations. [Method] The seed setting rate and 1 000-grain weight of 10 new sterile indica restorer lines planted in pots under enhanced UV-B radiation and fluorescent lamps （control） were respectively measured, and the differences were compared. [Result] The enhanced UV-B radiation significantly reduced the seed setting rate of indica restorer lines, and the differences between that UV-B radiation treatment and control all reached extremely significant level. In addition, the enhanced UV-B radiation reduced the 1 000-grain weight of most indica restorer lines, and compared with that of control the difference achieved significant or very significant level. However, the effect of enhanced UV-B radiation on seed setting rate and 1 000-grain weight differed to different indica restorer lines, and the differences among restorer lines tested were significant or very significant, which indicated the possibility to screen antiUV-B radiation rice materials and combinations. Finally, the indica restorer lines 09R-14, Luhui 37 and 10R-7703 which were strongly resistant to UV-B radiation were screened out. [Conclusion] This study laid foundation for breeding hybrid rice varieties resistance to UV-B radiation.

Isolation and initial characterization of GW5, a major QTL associated with rice grain width and weight

Grain weight is a major determinant of crop grain yield and is controlled by naturally occurring quantitative trait loci （QTLs）. We earlier identified a major QTL that controls rice grain width and weight, GW5, which was mapped to a recombination hotspot on rice chromosome 5. To gain a better understanding of how GW5 controls rice grain width, we conducted fine mapping of this locus and uncovered a 1 212-bp deletion associated with the increased grain width in the rice cultivar Asominori, in comparison with the slender grain rice IR24. In addition, genotyping analyses of 46 rice cultivars revealed that this deletion is highly correlated with the grain-width phenotype, suggesting that the GW5 deletion might have been selected during rice domestication. GW5 encodes a novel nuclear protein of 144 amino acids that is localized to the nucleus. Furthermore, we show that GW5 physically interacts with polyubiquitin in a yeast two-hybrid assay. Together, our results suggest that GW5 represents a major QTL underlying rice width and weight, and that it likely acts in the ubiquitin-proteasome pathway to regulate cell division during seed development. This study provides novel insights into the molecular mechanisms controlling rice grain development and suggests that GW5 could serve as a potential tool for high-yield breeding of crops.

Fine-Mapping of qTGW1.2a, a Quantitative Trait Locus for 1000-Grain Weight in Rice

Thousand-grain weight (TGW) is a key component of grain yield in rice. This study was conducted to validate and fine-map qTGW1.2a, a quantitative trait locus for grain weight and grain size previously located in a 933.6-kb region on the long arm of rice chromosome 1. Firstly, three residual heterozygotes (RHs) were selected from a BC2F11 population of the indica rice cross Zhenshan 97 (ZS97)///ZS97//ZS97/Milyang 46. The heterozygous segments in these RHs were arranged successively in physical positions, forming one set of sequential residual heterozygotes (SeqRHs). In each of the populations derived, non-recombinant homozygotes were identified to produce near isogenic lines (NILs) comprising the two homozygous genotypes. The NILs were tested for grain weight, grain length and grain width. QTL analyses for the three traits were performed. Then, the updated QTL location was followed for a new run of SeqRHs identification-NIL development-QTL mapping. Altogether, 11 NIL populations derived from four sets of SeqRHs were developed and used. qTGW1.2a was finally delimitated into a 77.5-kb region containing 13 annotated genes. In the six populations segregating this QTL, which were in four generations and were tested across four years, the allelic direction of qTGW1.2a remained consistent and the genetic effects were stable. For TGW, the additive effects ranged from 0.23 to 0.38 g and the proportions of phenotypic variance explained ranged from 26.15% to 41.65%. These results provide a good foundation for the cloning and functional analysis of qTGW1.2a.

Dissection of two quantitative trait loci for grain weight linked in repulsion on the long arm of chromosome 1 of rice(Oryza sativa L.)

Grain weight is a key determinant of grain yield in rice. Three sets of rice populations with overlapping segregating regions in isogenic backgrounds were established in the generations of BC2 F5, BC2 F6 and BC2 F7, derived from Zhenshan 97 and Milyang 46, and used for dissection of quantitative trait loci(QTL) for grain weight. Two QTL linked in repulsion phase on the long arm of chromosome 1 were separated. One was located between simple sequence repeat(SSR) markers RM11437 and RM11615, having a smaller additive effect with the enhancing allele from the maintainer line Zhenshan 97 and a partially dominant effect for increasing grain weight. The other was located between SSR markers RM11615 and RM11800, having a larger additive effect with the enhancing allele from the restorer line Milyang 46 and a partially dominant effect for increasing grain weight. When the two QTL segregated simultaneously, a residual additive effect with the enhancing allele from Milyang 46 and an over-dominance effect for increasing grain weight were detected. This suggests that dominant QTL linked in repulsion phase might play an important role in heterosis in rice. Our study also indicates that the use of populations with overlapping segregating regions in isogenic backgrounds is helpful for the dissection of minor linked QTL.

Fine mapping of qTGW10-20.8, a QTL having important contribution to grain weight variation in rice

Grain weight is one of themost important determinants of grain yield in rice.In this study,QTL analysis for grain weight,grain length,and grainwidthwas performed using populations derived from crosses between major parental lines of three-line indica hybrid rice.A total of 27 QTL for grain weight were detected using three recombinant inbred line populations derived from the crosses Teqing/IRBB lines,Zhenshan 97/Milyang 46,and Xieqingzao/Milyang 46.Of these,10 were found in only a single population and the other 17 in two or all three populations.Nine of the 17 common QTL were located in regions where no QTL associated with grain weight have been cloned and onewas selected for fine-mapping.Eight populations segregating in an isogenic background were derived from one F7 residual heterozygote of Teqing/IRBB52.The target QTL,qTGW10-20.8 controlling grain weight,grain length,and grain width,was localized to a 70.7-kb region flanked by InDel markers Te20811 and Te20882 on the long arm of chromosome 10.The QTL region contains seven annotated genes,ofwhich six encode proteins with known functional domains and one encodes a hypothetical protein.One of the genes,Os10g0536100 encoding the MIKC-type MADS-box protein OsMADS56,is the most likely candidate for qTGW10-20.8.These results provide a basis for cloning qTGW10-20.8,which has an important contribution to grain weight variation in rice.

QTL-Seq Identified a Major QTL for Grain Length and Weight in Rice Using Near Isogenic F_2 Population

Mapping and isolation of quantitative trait loci(QTLs)or genes controlling grain size or weight is very important to uncover the molecular mechanisms of seed development and crop breeding.To identify the QTLs controlling grain size and weight,we developed a near isogenic line F_2(NIL-F_2)population,which was derived from a residual heterozygous plant in an F_7 generation of recombinant inbred line(RIL).With the completion of more than 30×whole genome re-sequencing of the parents,two DNA bulks for large and small grains,a total of 58.94 Gb clean nucleotide data were generated.A total of455 262 single nucleotide polymorphisms(SNPs)between the parents were identified to perform bulked QTL-seq.A candidate genomic region containing SNPs strongly associated with grain length and weight was identified from 15 to 20 Mb on chromosome 5.We designated the major QTL in the candidate region as q TGW5.3.Then,q TGW5.3 was further validated with PCR-based conventional QTL mapping method through developing simple sequence repeat and Insertion/Deletion markers in the F_2 population.Furthermore,recombinants and the progeny tests delimited the candidate region of q TGW5.3 to 1.13 Mb,flanked by HX5009(15.15 Mb)and HX5003(16.28 Mb).A set of NILs,selected from the F_2 population,was developed to evaluate the genetic effect of q TGW5.3.Significant QTL effects were detected on grain length,grain width and 1000-grain weight of H12-29 allele with 1.14 mm,-0.11 mm and 3.11 g,which explained 99.64%,95.51%and 97.32%of the phenotypic variations,respectively.

Weight Determining of Factors Influencing Grain Output Based on Entropy Weight Method

This article selects 8 main factors(the number of rural employees,total power of agricultural machinery,effective irrigation area of crops,growing area of grain crops,fertilizer consumption,electricity consumption in rural areas,area affected and area covered) as the factors influencing grain output,and offers the method of determining weight of factors influencing grain output using entropy weight method.According to the relevant data in the period 1985-2005,we analyze the weight of factors influencing grain output in China by example.The results show that the electricity consumption in rural areas has the greatest impact on grain output,followed by total power of agricultural machinery,fertilizer consumption and area covered.To increase grain output,we must enhance the degree of mechanization,free people from the former process of direct cultivation,strengthen water conservancy construction,and do a good job in disaster prevention and mitigation.

Correlation Analysis of Rice Seed Setting Rate and Weight of 1 000-Grain and Agro-Meteorology over the Middle and Lower Reaches of the Yangtze River,China

The purpose of this study is to reveal the effects of historic climate change on rice yield over the middle and lower reaches of the Yangtze River, China, and to better adapt to climate change in the future. This study presents the relation of temperature and precipitation and rice components from 1981 to 2003 at 48 early rice stations and 30 middle rice stations. It focuses on an analysis of three stages： flowering, pre-milk, and late milk. The results show that mean maximum temperature and mean daily precipitation at the stages of flowering and pre-milk are most related to early rice yield. Yield change of middle rice is mainly because of mean precipitation change at the flowering stage. Furthermore, percentage of undeveloped grain increases as mean maximum temperature rises at the flowering stage. Over-precipitation in the reproductive stage is a major reason for reduction in yield of early rice. Consecutive rainfall and continuous high temperature can have negative effects on middle rice yield. Global warming would affect middle rice more seriously than early rice.

Poaceae Orthologs of Rice OsSGL,DUF1645 Domain-Containing Genes,Positively Regulate Drought Tolerance,Grain Length and Weight in Rice

Grain yield is a polygenic trait that can be influenced by environmental factors and genetic compositions at all plant growth stages.Currently,the molecular mechanisms behind the coordination of the interaction between grain yield-related traits remain unknown.In this study,we characterized the function of four STRESS_tolerance and GRAIN_LENGTH(Os SGL)Poaceae ortholog genes that are transcribed into DUF1645 domain-containing proteins in relation to the grain length,grain weight,and drought stress-tolerance of rice.The transgenic plants with overexpressing or heterologous high levels of Poaceae OsSGL ortholog genes exhibited longer grain size than the wild type plants.Larger cells were seen in panicles of the four transgenic lines with paraffin sectioning and scanning electron microscopy analyses.In addition,four Poaceae OsSGL ortholog genes positively affected the drought tolerance of rice.Four transgenic plants displayed higher resistance to drought stress at the seedling and vegetative stages.RNA-sequencing and qRT-PCR results indicated that over-or heterologous-expression of four Poaceae OsSGL ortholog genes also affected the transcriptome of rice plants.These genes may play a role in auxin and cytokinin biosynthesis and their transduction pathways.Taken together,these results suggested that the four OsSGL orthologs have a conserved function in the regulation of stress-tolerance and cell growth by modulating hormonal biosynthesis and signaling.

Fine Mapping of qTGW3-1,a QTL for 1000-Grain Weight on Chromosome 3 in Rice

The QTL qTGW3-1 was located on chromosome 3 of rice （Oryza sativa L.） and associated with the 1 000-grain weight （TGW） according to the result of our earlier study. With the objective of fine mapping of this locus, we developed a F2 population consisting of 3 428 plants derived from the cross between TGW-related near isogenic line DL017 （BC3F4 generation of GSL 156×Nipponbare） and the recurrent parent Nipponbare. Using six microsatellites, this QTL was delimited between RM5477 and RM6417. Markers MM 1455 and MM 1456 within this region were used for further mapping of this QTL. Finally, qTGW3-1 was fine-mapped into a 89-kb interval between RM5477 and MM1456, which locates in the BAC clone AC107226 harboring five putative candidate genes.

QTL mapping of grain appearance quality traits and grain weight using a recombinant inbred population in rice(Oryza sativa L.)

supported by a grant from the National High-Tech R＆D Program of China （2014AA10A603, 2014AA10A604）;a grant from the Youth Foundation in Sichuan, China （2011JTD0022）;the special fund for China Agricultural Research System （CARS-01-08）;the Provincial Specialized Funds for Innovation Ability Promotion in Sichuan, China （2013GXJS005）

TaABI19 positively regulates grain development in wheat

Starch is the most important component in the endosperm, and its synthesis is regulated by multiple transcription factors(TFs) in cereals. However, whether the functions of these TFs are conserved among cereals remains unclear. In this study,we cloned a B3 family TF in wheat, named TaABI19, based on its orthologous sequence in maize(Zea mays L.). Alignment of the DNA and protein sequences showed that ABI19 is conserved in maize and wheat(Triticum aestivum L.). We found that TaABI19 is highly expressed in young spikes and developing grains, and encodes a nucleus-localized transcriptional activator in wheat. The taabi19-b1 null mutants obtained by EMS exhibited a down-regulation of starch synthesis, shorter grain length and lower thousand-grain weight(TGW). Furthermore, we proved that TaABI19 could bind to the promoters of TaPBF homologous genes and enhance their expression. Haplotype association showed that TaABI19-B1 is significantly associated with TGW. We found that Hap2 and Hap3 were favored and had undergone positive selection in China’s wheat breeding programs. Less than 50% of the modern cultivars convey the favored haplotypes, indicating that TaABI19 still can be considered as a target locus for marker-assisted selection breeding to increase TGW in China.