Irrigation regimes modulate non-structural carbohydrate remobilization and improve grain filling in rice(Oryza sativa L.)by regulating starch metabolism

Recently developed ‘super’ rice cultivars with greater yield potentials often suffer from the problem of poor grain filling, especially in inferior spikelets. Here, we studied the activities of enzymes related to starch metabolism in rice stems and grains, and the microstructures related to carbohydrate accumulation and transportation to investigate the effects of different water regimes on grain filling. Two ‘super’ rice cultivars were grown under two irrigation regimes of well-watered(WW) and alternate wetting and moderate soil drying(AWMD). Compared with the WW treatment,the activities of ADP glucose pyrophosphorylase(AGPase), starch synthase(StSase) and starch branching enzyme(SBE), and the accumulation of non-structural carbohydrates(NSCs) in the stems before heading were significantly improved, and more starch granules were stored in the stems in the AWMD treatment. After heading, the activities of α-amylase, β-amylase, sucrose phosphate synthase(SPS) and sucrose synthase in the synthetic direction(SSs)were increased in the stems to promote the remobilization of NSCs for grain filling under AWMD. During grain filling, the enzymatic activities of sucrose synthase in the cleavage direction(SSc), AGPase, StSase and SBE in the inferior spikelets were increased, which promoted grain filling, especially for the inferior spikelets under AWMD.However, there were no significant differences in vascular microstructures. The grain yield and grain weight could be improved by 13.1 and 7.5%, respectively, by optimizing of the irrigation regime. We concluded that the low activities of key enzymes in carbon metabolism is the key limitation for the poor grain filling, as opposed to the vascular microstructures, and AWMD can increase the amount of NSC accumulation in the stems before heading, improve the utilization rate of NSCs after heading, and increase the grain filling, especially in the inferior spikelets, by altering the activities of key enzymes in carbon metabolism.

The auxin transporter OsAUX1 regulates tillering in rice(Oryza sativa)

Tillering is an important agronomic trait of rice(Oryza sativa)that affects the number of effective panicles,thereby affecting yields.The phytohormone auxin plays a key role in tillering.Here we identified the high tillering and semi-dwarf 1(htsd1)mutant with auxin-deficiency root characteristics,such as shortened lateral roots,reduced lateral root density,and enlarged root angles.htsd1 showed reduced sensitivity to auxin,but the external application of indole-3-acetic acid(IAA)inhibited its tillering.We identified the mutated gene in htsd1 as AUXIN1(OsAUX1,LOC_Os01g63770),which encodes an auxin influx transporter.The promoter sequence of OsAUX1 contains many SQUAMOSA PROMOTER BINDING PROTEIN-LIKE(SPL)binding sites,and we demonstrated that SPL7 binds to the OsAUX1 promoter.TEOSINTE BRANCHED1(OsTB1),a key gene that negatively regulates tillering,was significantly downregulated in htsd1.Tillering was enhanced in the OsTB1 knockout mutant,and the external application of IAA inhibited tiller elongation in this mutant.Overexpressing OsTB1 restored the multi-tiller phenotype of htsd1.These results suggest that SPL7 directly binds to the OsAUX1 promoter and regulates tillering in rice by altering OsTB1 expression to modulate auxin signaling.

Effects of Paclobutrazol Seed Priming on Seedlings Quality,Physiological and Bakanae Disease Index Characteristics of Rice(Oryza sativa L.)

Rice(Oryza sativa L.)is one of the most important cereal crops in the world.Bakanae disease is a significant rice disease widely distributed in rice-growing regions worldwide.Therefore,the present investigation aimed to assess the optimal concentrations of paclobutrazol(PBZ)as a treatment for rice grains(cv.Sakha 108)to control bakanae disease,also evaluating its impact on grain germination,seedling growth parameters as well as disease index.Paclobutrazol concentrations had no significant impact on seed germination,regardless of whether the seeds were incubated with Fusarium fujikuroi or not.Application of PBZ,either alone or in combination with fungal pathogens,negatively impacted the rice seedlings’height.Paclobutrazol at 25,50 and 100 mg/L,combined with the fungal pathogen positively impacted root length.Paclobutrazol at 3 and 6 mg/L mitigated the adverse impact on chlorophyll pigments content in infected seedlings.The highest proline contents were achieved by 100 mg/L PBZ alone or in combination with fungal pathogens.It has been observed that the application of PBZ,either alone or in combination with a fungal pathogen,leads to the enhancement of catalase,peroxidase,and polyphenol oxidase activities.The median lethal concentration of PBZ was 0.874 mg/L;applying low concentrations of paclobutrazol effectively increased the percentage of fungal growth suppression.Application of PBZ,at higher concentrations(50 and 100 mg/L),decreased infection percentage and disease severity index(DSI)significantly.These findings suggest that PBZ can be an effective treatment for controlling bakanae disease and enhancing resistance in rice plants.

The Magnaporthe oryzae effector Avr-PikD suppresses rice immunity by inhibiting an LSD1-like transcriptional activator

Avirulence effectors(Avrs),encoded by plant pathogens,can be recognized by plants harboring the corresponding resistance proteins,thereby initiating effector-triggered immunity(ETI).In susceptible plants,however,Avrs can function as effectors,facilitating infection via effector-triggered susceptibility(ETS).Mechanisms of Avr-mediated ETS remain largely unexplored.Here we report that the Magnaporthe oryzae effector Avr-PikD enters rice cells via the canonical cytoplasmic secretion pathway and suppresses rice basal defense.Avr-PikD interacts with an LSD1-like transcriptional activator AKIP30 of rice,and AKIP30 is also a positive regulator of rice immunity,whereas Avr-PikD impedes its nuclear localization and suppresses its transcriptional activity.In summary,M.oryzae delivers Avr-PikD into rice cells to facilitate ETS by inhibiting AKIP30-mediated transcriptional regulation of immune response against M.oryzae.

DDG1 and G Protein α Subunit RGA1 Interaction Regulates Plant Height and Senescence in Rice(Oryza sativa)

Many studies have already shown that dwarfism and moderate delayed leaf senescence positively impact rice yield,but the underlying molecular mechanism of dwarfism and leaf senescence remains largely unknown.Here,using map-based cloning,we identified an allele of DEP2,DDG1,which controls plant height and leaf senescence in rice.The ddg1 mutant displayed dwarfism,short panicles,and delayed leaf senescence.Compared with the wild-type,ddg1 was insensitive to exogenous gibberellins(GA)and brassinolide(BR).DDG1 is expressed in various organs,especially in stems and panicles.Yeast two-hybrid assay,bimolecular fluorescent complementation and luciferase complementation image assay showed that DDG1 interacts with theα-subunit of the heterotrimeric G protein.Disruption of RGA1 resulted in dwarfism,short panicles,and darker-green leaves.Furthermore,we found that ddg1 and the RGA1 mutant was more sensitive to salt treatment,suggesting that DDG1 and RGA1 are involved in regulating salt stress response in rice.Our results show that DDG1/DEP2 regulates plant height and leaf senescence through interacting with RGA1.

Conventional Breeding and Molecular Markers for Blast Disease Resistance in Rice (Oryza sativa L.)

Monogenic lines,which carried 23 genes for blast resistance were tested and used donors to transfer resistance genes by crossing method.The results under blast nursery revealed that 9 genes from 23 genes were susceptible to highly susceptible under the three locations(Sakha,Gemmeza,and Zarzoura in Egypt);Pia,Pik,Pik-p,Piz-t,Pita,Pi b,Pi,Pi 19 and Pi 20.While,the genes Pii,Pik-s,Pik-h,Pi z,Piz-5,Pi sh,Pi 3,Pi 1,Pi 5,Pi 7,Pi 9,Pi 12,Pikm and Pita-2 were highly resistant at the same locations.Clustering analysis confirmed the results,which divided into two groups;the first one included all the susceptible genes,while the second one included the resistance genes.In the greenhouse test,the reaction pattern of five races produced 100%resistance under artificial inoculation with eight genes showing complete resistance to all isolates.The completely resistant genes:Pii,Pik-s,Piz,Piz-5(=bi2)(t),Pita(=Pi4)(t),Pita,Pi b and Pi1 as well as clustering analysis confirmed the results.In the F1 crosses,the results showed all the 25 crosses were resistant for leaf blast disease under field conditions.While,the results in F2 population showed seven crosses with segregation ratio of 15(R):1(S),two cross gave segregated ratio of 3 R:1 S and one gave 13:3.For the identification of blast resistance genes in the parental lines,the marker K3959,linked to Pik-s gene and the variety IRBLKS-F5 carry this gene,which was from the monogenic line.The results showed that four genotypes;Sakha 105,Sakha 103,Sakha 106 and IRBLKS-F5 were carrying Pik-s gene,while was absent in the Sakha 101,Sakha 104,IRBL5-M,IRBL9-W,IRBLTACP1 and IRBL9-W(R)genotypes.As for Pi 5 gene,the results showed that it was present in Sakha 103 and Sakha 104 varieties and absent in the rest of the genotypes.In addition,Pita-Pita-2 gene was found in the three Egyptian genotypes(Sakha 105,Sakha 101 and Sakha 104)plus IRBLTACP1 monogenetic.In F2 generation,six populations were used to study the inheritance of blast resistance and specific primers to confirm the ratio and identify the resistance genes.However,the ratios in molecular markers were the same of the ratio under field evaluation in the most population studies.These findings would facilitate in breeding programs for gene pyramiding and gene accumulation to produce durable resistance for blast using those genotypes.

A Farmer’s Approach to Detecting Photoperiod Sensitivity in Rice (Oryza sativa ssp. indica) Landraces

Most indigenous rice landraces are sensitive to photoperiod during short day seasons,and this sensitivity is more pronounced in indica than in japonica landraces.Attempts to identify photoperiod sensitive(PPS)cultivars based on the life history stages of the rice plant,and several models and indices based on phenology and day length have not been precise,and in some cases yield counterfactual inferences.Following the empirical method of traditional Asian rice farmers,the author has developed a robust index,based on the sowing and flowering dates of a large number of landraces grown in different seasons from 2020 to 2023,to contradistinguish PPS from photoperiod insensitive cultivars.Unlike other indices and models of photoperiod sensitivity,the index does not require the presumed duration of different life history stages of the rice plant but relies only on the flowering dates and the number of days till flowering of a rice cultivar sown on different dates to consistently identify photoperiod sensitive cultivars.

Influence of Epistasis and QTL×Environment Interaction on Heading Date of Rice(Oryza sativa L.)

QTLs for heading date of rice （Oryza sativa L.） with additive, epistatic, and QTL × environment （QE） interaction effects were studied using a mixed-model-based composite interval mapping （MCIM） method and a double haploid （DH） population derived from IR64/Azucena in two crop seasons. Fourteen QTLs conferring heading date in rice, which were distributed on ten chromosomes except for chromosomes 5 and 9, were detected. Among these QTLs, eight had single-locus effects, five pairs had double-locus interaction effects, and two single-loci and one pair of double-loci showed QTL × environment interaction effects. All predicted values of QTL effects varied from 1.179 days to 2.549 days, with corresponding contribution ratios of 1.04%-4.84%. On the basis of the effects of the QTLs, the total genetic effects on rice heading date for the two parents and the two superior lines were predicted, and the putative reasons for discrepancies between predicted values and observed values, and the genetic potentiality in the DH population for improvement of heading date were discussed. These results are in agreement with previous results for heading date in rice, and the results provide further information, which indicate that both epistasis and QE interaction are important genetic basis for determining heading date in rice.

Advances in Physiological-biochemical and Genetic Mechanisms of Seed Aging in Rice(Oryza sativa L.)

Reasons causing or accelerating seed aging are mainly damage of mem- branes, DNA and proteins, decline of protein synthesis capacity and excessive ac- cumulation of reactive oxygen species. With the application of natural aging or artifi- cial aging methods, it was reported that quantitative trait loci （QTLs） of seed stora- bility in rice were widely distributed on the chromosomes except the 10th chromo- some. In this paper, we reviewed the progresses in the research on physiological- biochemical and genetic mechanisms of seed aging, and analyzed the existing problems and developing prospect in molecular breeding of rice with improved seed storability, in order to provide reference for the basic research and genetic improve- ment of rice seed storabUity.

Comparative Studies on the Changes of Microtubule Distribution and Reorganization During the Meiotic Stages of Development in Normal (IR36) and a Temperature/photoperiod Sensitive Male Sterile Line (Peiai 64S) of Rice ( Oryza sativa )

Changes in the pattern of organization of microtubules in the meiotic stages of development of pollen (i.e. from pre-meiotic interphase to more or less metaphase I) of a normal (IR36) and a temperature/photoperiod sensitive male sterile line (Peiai 64S) of rice were studied using immunofluorescence confocal microscopy. In IR36, from pre-meiotic interphase to metaphase I, the pattern of microtubule distribution in the meiocytes underwent a series of changes. Some new organizational patterns of microtubules (that have not been described before) were observed during microsporogenesis, including the existence of a broad band of perinuclear microtubules at the diakinesis stage of development. The pattern of microtubule distribution in the meiocytes of the male sterile line, Peiai 64S, was quite different front that seen in IR36. In Peiai 64S, the microtubules showed abnormal patterns of distribution from pre-meiotic interphase to metaphase I. For example the broad band of perinuclear microtubules seen at diakinesis in IR36 was much disorganized and loosened in Peiai 64S. The spindles formed were also very abnormal and different from the normal spindle. The appearance of abnormal microtubule distribution in the early stages of microsporogenesis may contribute to the malformation and ultimate abortion of pollen in Peiai 64S.

Novel QTLs from Wild Rice Oryza longistaminata Confer Strong Tolerance to High Temperature at Seedling Stage

Global warming poses a threat to rice production.Breeding heat-tolerant rice is an effective and economical approach to address this challenge.African rice is a valuable genetic resource for developing heat-tolerant crops due to its intricate mechanism for adapting to high temperatures.Oryza longistaminata,a widely distributed wild rice species in Africa,may harbor an even richer gene pool for heat tolerance,which remains untapped.In this study,we identified three heat tolerance QTLs from O.longistaminata at the seedling stage,including novel heat tolerance loci qTT4 and qTT5.Our findings demonstrated that the O.longistaminata alleles for these two QTLs can enhance the heat tolerance of rice seedlings.Remarkably,qTT5 was mapped to a region spanning approximately 287.2 kb,which contains 46 expressing genes.Through the analysis of Gene Ontology and expression differences under heat induction,we identified four candidate genes.Our results lay the foundation for discovering heat tolerance genes underlying O.longistaminata and developing new genetic resources for heat-tolerant rice breeding.

Calcineurin B-like interacting protein kinase OsCIPK23 functions in pollination and drought stress responses in rice (Oryza sativa L.)

Drought is very harmful to grain yield due to its adverse effect on reproduction, especially on pollination process in rice. However, the molecular basis of such an effect still remains largely unknown. Here, we report the role of a member of CBL （Calcineurin B-Like） Interacting Protein Kinase （CIPK） family, OsCIPK23, in pollination and stress responses in rice. Molecular analyses revealed that it is mainly expressed in pistil and anther but up-regulated by pollination, as well as by treatments of various abiotic stresses and phytohormones. RNA interference-mediated suppression of OsCIPK23 expression significantly reduced seed set and conferred a hypersensitive response to drought stress, indicating its possible roles in pollination and drought stress. In consistent, overexpression of OsCIPK23 induced the expression of several drought tolerance related genes. Taken together, these results indicate that OsCIPK23 is a multistress induced gene and likely mediates a signaling pathway commonly shared by both pollination and drought stress responses in rice.

Bulked Segregant Analysis to Detect QTL Related to Heat Tolerance in Rice(Oryza sativa L.) Using SSR Markers

The study was undertaken to assess the genetic effect of quantitative trait loci （QTLs） conferring heat tolerance at flowering stage in rice. A population consisting of 279 F2 individuals from the cross between 996, a heat tolerant cultivar and 4628, a heat-sensitive cultivar, was analyzed for their segregation pattern of the difference of seed set rate under optimal temperature condition and high temperature condition. The difference of seed set rate under optimal temperature condition and high temperature condition showed normal distribution, indicating the polygenic control over the trait. To identify main effect of QTL for heat tolerance, the parents were surveyed with 200 primer pairs of simple sequence repeats （SSR）. The parental survey revealed 30% polymorphism between parents. In order to detect the main QTL association with heat tolerance, a strategy of combining the DNA pooling from selected segregants and genotyping was adopted. The association of putative markers identified based on DNA pooling from selected segregants was established by single marker analysis （SMA）. The results of SMA revealed that SSR markers, RM3735 on chromosome 4 and RM3586 on chromosome 3 showed significant association with heat tolerance respectively, accounted for 17 and 3% of the total variation respectively. The heat tolerance during flowering stage in rice was controlled by multiple gene. The SSR markers, RM3735 on chromosome 4 and RM3586 on chromosome 3 showed significant association with heat tolerance respectively, accounted for 17 and 3% of the total variation respectively. The two genetic loci, especially for RM3735 on chromosome 4, can be used in marker-assistant-selected method in heat tolerance breeding in rice.

Studies on the Mechanism of Single Basal Application of Controlled-Release Fertilizers for Increasing Yield of Rice (Oryza sativa L.)

This paper was to explore the mechanism of single basal application of controlled-release fertilizers for increasing yield of rice （Oryza sativa L.）. Pot trials and cylinder trials were carried out from 2002 to 2005 to study the influences of single basal application of 3 controlled-release fertilizers on the changes of soil available N, root development, senescence and lodging resistance at late growth stages. Results showed that at 30 days after fertilization, single basal application of controlled-release fertilizers coated with vegetal-substance （CRF1） and polymer materials （CRF3） increased soil available N to 12.0 and 147.9%, respectively, in comparison to split fertilization of rice-specific fertilizer （RSF1）. Treatments of the two CRFs obviously benefited the development of root system, resulting in greater rice root weights with extensive distribution and higher root activity. In addition, the two CRF treatments, in comparison to RSF1, enhanced chlorophyll consents of the flag leaves to 9.5 and 15.5%, and soluble protein up to 89.7 and 108.0% respectively. Application of the two CRFs also made the base of rice stems strong and large, declined the proportion of shoot and root, increased root depth index. Though relatively low K rate, single basal application of the CRF3 coated with NH4MgPO4 could also promote the development of root system, enhance root activity and some physiological functions of flag leaves. Based on these results, it was concluded that major mechanisms for increasing rice yield by single basal application of the CRFs should be attributed to grater soil available N supply, superior development of root systems, better nutrient absorption capacity, slower senescence and enhancement of lodging resistance at late stages.

Genotypic Difference in the Responses of Seedling Growth and Cd Toxicity in Rice(Oryza sativa L.)

The experiment was carried out to study the genotypic difference in the responses of seed germination, growth and physiological characters of rice seedlings to Cd toxicity. The result showed that the germination was slightly stimulated under low Cd concentration （0.01-1.5 mM Cd）, while severely depressed under higher Cd concentration （2.0 mM）. Rice seedlings exposed to 0.01 mM Cd showed slight increases in plant height, root volume, biomass and chlorophyll concentration. These parameters were significantly reduced when Cd level in the medium was increased to 0.5 mM, and meanwhile corresponding increase in superoxide dismutase （SOD） and peroxidase （POD） activities, and MDA （malondialdehyde） content was observed. However, SOD and POD activities declined when plants were exposed to 1 mM Cd when compared with those under 0.5 mM Cd. Cadmium addition lowered Fe, Cu and Mn concentrations in roots and shoots. There was significant genotypic difference in the response of these parameters to Cd stress. Under Cd stress, Xiushui 110 had the least inhibition of growth and increase in MDA content, higher shoot Cd concentration, and greatest increase in POD and SOD activities, indicating its higher tolerance to Cd toxicity, while Bing 9914 had the greatest reduction of growth, and Zn, Cu, Fe, and Mn contents, but greatest increase in MDA content, and least increase in activities of antioxidative enzymes, indicating its sensitivity to Cd toxicity.

Biomass-Based Rice (Oryza sativa L.) Aboveground Architectural Parameter Models

To quantify the relationships between rice plant architecture parameters and the corresponding organ biomass, and to research on functional structural plant models of rice plant, this paper presented a biomass-based model of aboveground architectural parameters of rice （Oryza sativa L.） in the young seedling stage, designed to explain effects of cultivars and environmental conditions on rice aboveground morphogenesis at the individual leaf level. Various model variables, including biomass of blade and blade length, were parameterized for rice based on data derived from an outdoor experiment with rice cv. Liangyou 108, 86You 8, Nanjing 43, and Yangdao 6. The organ dimensions of rice aboveground were modelled taking corresponding organ biomass as an independent variable. Various variables in rice showed marked consistency in observation and simulation, suggesting possibilities for a general rice architectural model in the young seedling stage. Our descriptive model was suitable for our objective. However, they can set the stage for connection to physiological model via biomass and development of functional structural rice models （FSRM）, and start with the localized production and partitioning of assimilates as affected by abiotic growth factors. The finding of biomass-based rice architectural parameter models also can be used in morphological models of blade, sheath, and tiller of the other stages in rice life.

Genetic Analysis and Primary Mapping of pms4, a Photoperiod-Sensitive Genic Male Sterility Gene in Rice (Oryza sativa)

To understand the genetic characteristics of a new photoperiod-sensitive genic male sterile line Mian 9S, some reciprocal crosses were made between Mian 9S and six indica rice materials, Yangdao 6, Luhui 602, Shuihui 527, Mianhui 725, Fuhui 838 and Yixiang 1B. Genetic analysis results suggested that the photoperiod-sensitive genic male sterility （PGMS） of Mian 9S was controlled by a single recessive nuclear gene. Thus, the F2 population derived from the cross of Yangdao 6/Mian 9S was used to map the PGMS gene in Mian 9S. By using SSR markers, the PGMS gene of Mian 9S was mapped on one side of the markers, RM6659 and RM1305, on rice chromosome 4, with the genetic distances of 3.0 cM and 3.5 cM, respectively. The gene was a novel PGMS gene and designated tentatively as pms4. In addition, the application of the pms4 gene was discussed.

Validation of qGSIO, a quantitative trait locus for grain size on the long arm of chromosome 10 in rice (Oryza sativa L.)

Grain size is a major determinant of grain weight and a trait having important impact on grain quality in rice. The objective of this study is to detect QTLs for grain size in rice and identify important QTLs that have not been well characterized before. The QTL mapping was first performed using three recombinant inbred line populations derived from indica rice crosses Teqing/IRBB lines, Zhenshan 97/Milyang 46, Xieqingzao/Milyang 46. Fourteen QTLs for grain length and 10 QTLs for grain width were detected, including seven shared by two populations and 17 found in one population. Three of the seven com- mon QTLs were found to coincide in position with those that have been cloned and the four others remained to be clarified. One of them, qGSIO located in the interval RM6100-RM228 on the long arm of chromosome 10, was validated using F2：3 populations and near isogenic lines derived from residual heterozygotes for the interval RM6100-RM228. The QTL was found to have a considerable effect on grain size and grain weight, and a small effect on grain number. This region was also previously detected for quality traits in rice in a number of studies, providing a good candidate for functional analysis and breeding utilization.

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.

Enclosed stigma contributes to higher spikelet fertility for rice(Oryza sativa L.) subjected to heat stress

With global warming, rice plants may be subjected to heat stress more regularly during the heatsensitive flowering stage, causing spikelet sterility and grain yield loss.Stigma exsertion is considered to increase pollen reception and promote female reproductive success.The aim of this study was to investigate the role of stigma exsertion on spikelet fertility at high temperatures.Five rice cultivars(Liangyoupeijiu, Shanyou 63, Huanghuazhan, Nagina 22, and IR64) with differing degrees of stigma exsertion were cultivated and exposed to high temperature at anthesis.Heat-tolerant cultivars did not always show a high percentage of spikelets with exserted stigmas, and vice versa.Irrespective of the presence of more pollen grains on exserted stigmas, spikelets with exserted stigmas did not show greater spikelet fertility than spikelets with fewer exserted stigmas or hidden stigmas under heat stress.GA3 application augmented the percentage of spikelets with exserted stigmas;however, it did not increase spikelet fertility under heat stress.Spikelet fertility of whole panicles was negatively correlated with the percentage of spikelets with exserted stigmas, but positively with that with hidden stigmas.Viability of the hidden stigmas was less reduced than that of exserted stigmas under heat stress, suggesting that hidden stigmas have an advantage in maintaining viability.Heat stress delayed anther dehiscence and reduced the viabilities of both exserted stigmas and pollens, thereby causing low spikelet fertility.Together, these results suggest that high spikelet fertility does not depend on stigma exsertion and that enclosed stigma generally contributes to higher spikelet fertility and heat tolerance under high-temperature conditions during flowering in rice.