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.

Genetic Analysis of Cold Tolerance at Seedling Stage and Heat Tolerance at Anthesis in Rice (Oryza sativa L.)

A set of 240 introgression lines derived from the advanced backcross population of a cross between a japonica cultivar,Xiushui 09,and an indica breeding line,IR2061,was developed to dissect QTLs affecting cold tolerance （CT） at seedling stage and heat tolerance （HT） at anthesis.Survival rate of seedlings （SRS） and spikelet fertility （SF）,the index traits of CT and HT,showed significant differences between the two parents under stresses.A total of four QTLs （qSRS1,qSRS7,qSRS11a and qSRS11b） for CT were identified on chromosomes 1,7,11,and the Xiushui 09 alleles increased SRS at all loci except qSRS7.Four QTLs for SF were identified on chromosomes 4,5,6,and 11.These QTLs could be classified into two major types based on their behaviors under normal and stress conditions.The first was QTL expressed only under normal condition;and the second QTL was apparently stress induced and only expressed under stress.Among them,two QTLs （qSF4 and qSF6） which reduced the trait difference between heat stress and normal conditions must have contributed to HT because of their obvious contribution to trait stability,and the IR2061 allele at the qSF6 and the Xiushui 09 allele at the qSF4 improved HT,respectively.No similar QTL was found between CT at seedling stage and HT at anthesis.Therefore,it is possible to breed a new variety with CT and HT by pyramiding the favorable CT-and HT-improved alleles at above loci from Xiushui 09 and IR2061,respectively,through marker-assisted selection （MAS）.

Identification for Heat Tolerance in Backcross Recombinant Lines and Screening of Backcross Introgression Lines with Heat Tolerance at Milky Stage in Rice

The present study aimed at breeding new rice germplasms with similar genome but significantly differed in heat tolerance during the grain filling stage.A total of 791 BC1F8 backcross recombinant lines,derived from the cross of Xieqingzao B /N22 //Xieqingzao B,were used as materials.Each rice line was separated evenly into two groups,and the heat tolerance of all rice lines were evaluated at natural high temperature in fields.The rice lines with heat tolerant index higher than 90% or lower than 40% were selected to compare the phenotypic characters and further identify heat tolerance at the early milky stage in a phytotron.Rice lines with similar phenotypic characters but significantly differed in heat tolerance at the milky stage were analyzed by 887 simple sequence repeat markers that were evenly distributed on the 12 rice chromosomes.In the result,12(6 pairs) rice lines with similar phenotypic characters but significantly differed in heat tolerance at the milky stage were obtained.Molecular marker analysis indicated that the genomic polymorphism between 703T and 704S was the smallest in the 6 pairs of rice lines,with only 16 polymorphic sites,including 22 different alleles.The application of these two backcross introgression rice lines for future study on the mechanisms of heat tolerance in rice at the milky stage will be theoretically beneficial in reducing the interference caused by genetic differences from experimental materials.

Germplasm Innovation of Heat Tolerance in Rice for Irrigated Lowland Conditions in the Philippines

Heat-tolerant varieties, such as N22 and Dular, which were used in this study, usually have low yield potential and undesirable plant characteristics but combining them with high yielding and improved rice varieties, new heat-tolerant rice genotypes with high yield potential can be achieved. In this study, phenotyping and selecting desirable materials from various crosses were performed under high temperature conditions during the reproductive stage. Screening was performed in the field and glasshouse to select individuals with heat tolerance and high yield potential. Several advanced breeding lines from Gayabyeo/N22 cross produced desirable individuals with heat tolerance, resistance to pests and diseases, and high yield potential. The genetic variation in percent sterility among the selected backcross populations grown in high temperature environments showed that large number of plants can be identified and selected with lower percent sterility.

Identification of Heat Tolerance Linked Molecular Markers of Chinese Cabbage(Brassica campestris L.ssp.pekinensis)

Genetically stable population of recombination inbred line (RIL) was derived from a cross between a heat tolerant line 177 and a heat sensitive line 276 of Chinese cabbage (Brassica campestris L. ssp. pekinensis) by single seed descent. The RILs were analyzed using isozyme, RAPD and AFLP techniques in order to find molecular markers that are linked to heat tolerance quantitative trait loci (QTL). The results of variance analysis of single factor indicated that there were 9 molecular markers closely linked with heat tolerance QTL, including 5 AFLP markers, 3 RAPD markers and 1 PGM isozyme marker. Total genetic contribution of these makers to heat tolerance was 46.7%. Five of the nine markers distributed in one linkage group, the remaining 4 markers were located in separate groups. Thus the 9 heat tolerance linked markers distributed in 5 independent locations in the genome of Chinese cabbage.

Examine the Correlation between Heat Shock Protein IbpA and Heat Tolerance in Cronobacter sakazakii

We used a proteomic approach to identify IbpA in Cronobacter sakazokii （C. sakazaki）, which is related to heat tolerance in this strain. The abundance of IbpA in C. sakazakii strains strongly increased after heat shock. C sakazakii CMCC 45402 ibpA deletion mutants were successfully constructed. The C. sakazakii CMCC 45402 AibpA and wild-type strains could not be distinguished based on colony morphology on LB agar plates or biochemical assays. The growth of the C. sakazakii CMCC 45402 AibpA mutant in heat shock conditions was indistinguishable from that of the isogenic wild-type, but showed greater heat resistance than E. coil O157：H7 strain CMCC 44828. This study suggests that the absence of a single ibpA gene has no obvious effect on the phenotype or heat resistance of the strain C. sakazakii CMCC 45402.

Wheat genotypes tolerant to heat at seedling stage tend to be also tolerant at adult stage:The possibility of early selection for heat tolerance breeding

Wheat production is seriously influenced by extreme hot weather,which has attracted increasing attention.It is important to compare wheat responses to heat at seedling and reproductive stages,to explore the potential relationship between the performances at different growing stages and the possibility of early selection to accelerate heat tolerance breeding.In this study,forty wheat genotypes were screened under heat stress at both seedling and adult stages.It was found that root lengths at seedling stage were severely reduced by heat stress with significant variations among wheat genotypes.Heat-tolerant genotypes at seedling stage showed less root length decrease than susceptible ones.Wheat genotypes tolerant at seedling stage showed higher yield at adult stage after heat treatment.The performances of wheat genotypes screened under heat stress at seedling and adult stages were ranked by seedling damage index and adult damage index.A significant positive relationship was found between heat tolerance at seedling stage and at adult stage(r=0.6930),suggesting a similar tolerant/susceptible mechanism at different plant growth stages and the possibility of early selection at seedling stage for breeding heat tolerance.Extremely tolerant and susceptible genotypes with consistent performances at seedling and adult stages were genetically compared and associated SNP markers and linked candidate genes were identified.

Evaluation of a new method for quantification of heat tolerance in different wheat cultivars

Heat stress seriously affects wheat production in many regions of the world. At present, heat tolerance research remains one of the least understood fields in wheat genetics and breeding and there is a lack of effective methods to quantify heat stress and heat tolerance in different wheat cultivars. The objective of this study was to use various wheat cultivars to evaluate stress intensity(δ) and a new method for quantification of heat tolerance and compare this technique with three other currently utilized methods. This new parameter for heat tolerance quantification is referred to as the heat tolerance index(HTI) and is an indicator of both yield potential and yield stability. Heat treatments were applied in a controlled setting when anthesis had been reached for 80% of the wheat. The stress intensity evaluation indicated heat shock was the main factor associated with kernel weight reduction while grain yield reduction was mainly associated with chronic high temperature. The methods evaluation showed that a temperature difference of 5°C from natural temperatures was a suitable heat treatment to compare to the untreated controls. HTI was positively correlated with yield under heat stress(r=0.8657, δ=0.15, in 2009–2010; r=0.8418, δ=0.20, in 2010–2011; P<0.01), and negatively correlated with yield reduction rate(r=–0.8344, in 2009–2010; r=–0.7158, in 2010–2011; P<0.01). The results of this study validated the use of HTI and temperature difference control for quantifying wheat heat tolerance that included the yield potential and the stability of different wheat cultivars under heat stress. Additionally, 10 wheat cultivars showed high HTI and should be further tested for their heat confirming characteristics for use in wheat heat tolerance breeding.

Effect of the C.–1388 A>G polymorphism in chicken heat shock transcription factor 3 gene on heat tolerance

Heat stress is one of the main factors that inlfuence poultry production. Heat shock proteins (HSPs) are known to affect heat tolerance. The formation of HSPs is regulated by heat shock transcription factor 3 (HSF3) in chicken. A DNA pool was established for identifying single nucleotide polymorphisms (SNPs) of the chicken HSF3, and 13 SNPs were detected. The bioinformatic analysis showed that 8 SNPs had the capacity to alter the transcription activity of HSF3. The dual luciferase report gene assay showed that there was a signiifcant difference (P<0.01) in the Firelfy luciferase/Renil a luciferase ratio (F/R) of C.–1 703 A>G (S1) and C.–1 388 A>G (S4) sites at the 5′-untranslated region (UTR) of chicken HSF3. The elec-trophoretic mobility shift assay showed that the S4 site was a transcription binding factor. The analysis of the association of the S1 and S4 sites with heat tolerance index revealed that the S4 site was signiifcantly correlated with the CD3+T cel , corticosterone, and T3 levels in Lingshan chickens and with the heterophil/lymphocyte value in White Recessive Rock. These results showed that the S4 site at the 5′ UTR of chicken HSF3 might have an impact on heat tolerance in summer and could be used as a potential marker for the selection of chicken with heat tolerance in the future.

Male Parent Plays More Important Role in Heat Tolerance in Three-Line Hybrid Rice

Ten F1 combinations with their male and female parents were employed to evaluate their heat tolerance during the flowering and early grain filling stages. The rice plants were subjected to heat stress（39 °C–43 °C） for 1–15 d during flowering. Based on the heat stress index, heat tolerance was only observed in the F1 combinations H2（K22A × R207）, H3（Bobai A × R207） and H4（Bobai A × Minghui 63）, whereas the others received above 0.5000 of heat stress index. Both parents of the tolerant combination（heat-tolerant × heat-tolerant） possessed heat tolerance, whereas the susceptible combinations were crossed by heat-tolerant（sterile lines） × heat-susceptible（restorer lines）, heat-susceptible × heat-tolerant, or heat-susceptible × heat-susceptible parents. This result indicated that heat tolerance in rice was controlled by recessive genes. Thus, both parents should possess high temperature tolerance to develop heat-tolerant F1 combinations. Furthermore, the heat stress index of F1 combinations was significantly correlated with the heat stress index of restorer lines but not with the heat stress index of maintainer lines. This result suggested that male parents play a more important role in heat-tolerant combinations than female parents. Therefore, the heat susceptibility of the hybrid rice in China is mainly due to the wide application of low-heat-tolerant restorer lines with high yield in three-line hybrid rice breeding.

Mapping QTLs for Heat Tolerance in Rice (Oryza sativa L. ) at Heading Stage Using Chromosome Segment Substitution Lines

In the present study, a japonica rice （ Oryza sativa L. ） variety Nipponbare, an indica variety 9311 and a set of chromosome segment substitution lines （CSSLs） which were generated using Nipponbare as the recipient parent and 9311 as the donor parent were used as the experimental materials. The CSSLs were grown in 2012 （normal temperature condition） and 2013 （high temperature condition） in Yangzhou, Jiangsu, and were used to map the quantitative trait loci （QTLs） for heat tolerance, based on the heat tolerance index [ （The seed setting rate under normal temperature condition -The seed setting rate under high temper- ature condition） / The seed setting rate under normal temperature condition]. As a result, three QTLs related to heat tolerance in rice were mapped on chromo- somes 2, 4 and 12, respectively. They had LOD （logarithm of rntds） scores of 2.56, 4.02 and 2.79, and contributian rates of 4.95%, 7.99% and 5.44%. Among them, qHT12.1 showed positive effect, while qHT2.1 and qHT4. t showed negative effect on heat tolerance. The results lay a foundation for the fine mapping and cloning of the QTLs and genes related to heat tolerance, and for the breeding of heat-tolerant rice varieties.

Physiological Response to High Temperature and Evaluation of Heat Tolerance of Different Grape Cultivars

[Objectives]This study was conducted to investigate the differences in the physiological responses of different grape cultivars to high temperature.[Methods]The 19 tested cultivars were selected from the grape germplasm resources pool of Turpan Research Institute of Xinjiang Academy of Agricultural Sciences.Twelve physiological indexes including gas exchange parameters,chlorophyll content,antioxidant enzyme activity and proline content were determined in grape leaves under field conditions during the middle period of local natural high temperature period(July,daily maximum air temperature>35℃).The heat tolerance of different cultivars was evaluated by fuzzy membership function analysis and optimum partitioning clustering of ordered samples.[Results](1)Under natural high temperature conditions in Turpan,the 19 tested grape cultivars responded differently to high temperature.‘Red Globe’,‘Fujiminori’,‘Beta’,‘Hetianhuang’had strong heat tolerance,while‘Thompson Seedless’,‘Hongqi Tezaomeigui’,‘Shuijing Wuhe’,‘Victoria’,‘Yatomi Rosa’and‘Crimson Seedless’had weak heat tolerance.(2)Among the 12 physiological indexes,malondialdehyde content and antioxidant enzyme activity were mostly different among various grape cultivars.The grape cultivars with strong heat tolerance,‘Red Globe’and‘Fujiminori’,had relatively lower malondialdehyde contents,while‘Beta’and‘Hetianhuang’had relatively higher malondialdehyde contents.But they had higher activity of antioxidant enzymes.(3)The results of fuzzy membership function analysis showed that the cumulative membership value(AR)of each physiological index was consistent with its apparent heat tolerance performance,suggesting that AR can be a potential index for the evaluation of heat tolerance of grape cultivars.Further cluster analysis classified the tested cultivars as strong,medium and weak.‘Red Globe’,‘Fujiminori’,‘Beta’and‘Hetianhuang’had strong heat tolerance.[Conclusions]This study provides a reference for grape cultivation under high temperature and stress and breeding of heat-tolerant varieties.

Research Progress of Heat-tolerance of Brassica campestris ssp. Chinensis

Brassica campestris ssp. chinensis, also known as non-heading Chinese cabbage, is an important vegetable widely distributed in southern China. High temperature is the most common adversity factor in vegetable production, because Brassica campestris ssp. chinensis is a thermophilic vegetable, which can＇t well grow at high temperature. In summer and autumn, high temperature stress would prevent Brassica campestris ssp. chinensis from growing and result in poor quality of its seedlings, seriously influencing yield and quality of the vegetable in later period. In this paper, present situation and latest advances in heat resistance research of Brassica campestris ssp. chinensis in recent years were reviewed at home and abroad. The research tendency in Bassica campestris ssp. chinensis was also described, providing reference for breeding of heat-resistant Brassica campestris ssp. chinensis.

Mapping QTL for Heat-Tolerance at Grain Filling Stage in Rice

A mapping population of 98 lines (backcross inbred lines, BILs) derived from a backcross of Nipponbare/Kasalath// Nipponbare was planted at two experimental sites, Nanjing and Nanchang, and treated with high and optimal temperature during grain filling, respectively. The grain weight heat susceptibility index [GWHSI= (grain weight at optimum temperature-grain weight at high temperature) / grain weight at optimum temperature × 100] was employed to evaluate the tolerance of rice to heat stress. A genetic linkage map with 245 RFLP markers and a mixed linear-model approach was used to detect quantitative trait loci (QTLs) and their main effects, epistatic interactions and QTL× environment interactions (Q×E). The threshold of LOD score=2.0 was used to detect the significance of association between marker and trait. A total of 3 QTLs controlling heat tolerance during grain filling were detected, on chromosomes 1, 4 and 7, with LOD scores of 8.16, 11.08 and 12.86, respectively, and they explained the phenotypic variance of 8.94, 17.25 and 13.50 %, correspondingly. The QTL located in the C1100-R1783 region of chromosome 4 showed no QTL× environment interaction and epistatic effect, suggesting that it could be stably expressed in different environments and genetic backgrounds, and thus it would be valuable in rice breeding for heat tolerance improvement. This QTL allele, derived from Kasalath reduced 3.31% of the grain weight loss under heat stress. One located between R1613-C970 on chromosome 1 and the other between C1226-R1440 on chromosome 7, with additive effect 2.38 and 2.92%, respectively. The tolerance alleles of both these QTLs were derived from Nipponbare. Both of these QTLs had significant QTL× environment interactions, and the latter was involved in epistatic interaction also. Eight pairs of epistatic effect QTLs were detected, one pair each on chromosomes 1,2,3, 5, 7, 8, 10 and 12. The results could be useful for elucidating the genetic mechanism of heat-tolerance and the development of new rice varieties with heat tolerance during grain filling phase.

Arabidopsis immune-associated nucleotide-binding genes repress heat tolerance at the reproductive stage by inhibiting the unfolded protein response and promoting cell death

Plants are vulnerable to heat stress,especially during reproductive development.The heat shock response(HSR)in the cytosol and nucleus,as well as the unfolded protein response(UPR)in the endoplasmic reticulum(ER),are two mechanisms that enable plants to survive heat stress.Excessive heat or ER stresses lead to cell death when the UPR cannot repair stress damage,but the means by which cell survival or death is determined remains unclear.In this study,we used a genome-wide association study(GWAS)to identify that a cluster of five Immune-associated nucleotide-binding protein(IAN)genes(IAN2 to IAN6)is responsible for variation in heat tolerance at the reproductive stage in Arabidopsis thaliana.These IAN genes have both unique and overlapping functions in the negative regulation of heat tolerance,and their loss of function singly or in combination confers increased heat tolerance,measured by a lower number of barren siliques and a higher seedling survival rate under heat.The loss of rice IAN1 gene function also leads to enhanced heat tolerance,suggesting a conserved function of plant IANs.Transcriptome analysis revealed enhanced expression of HSR and UPR genes,as well as reduced cell death,under heat and ER stress in the mutant of IAN6,a major effect member in Arabidopsis.Furthermore,the IAN proteins were found to promote cell death induced by heat stress,ER stress,and cell death-inducing molecules.Thus,the Arabidopsis IAN genes repress heat tolerance,probably through the HSR and UPR and by enhancing the cell death pathway.The IAN2 to IAN6 proteins are partially localized to the ER,suggesting a direct role in the UPR and UPR-mediated cell death.In addition,a natural IAN6 variant from more heat-tolerant Arabidopsis accessions confers greater heat tolerance and induces less cell death compared with the natural variant from less heat-tolerant accessions.The heat-tolerant IAN6 variant is associated with a higher maximum temperature of the warmest month at its collection sites compared with the heat-sensitive variant.Taken together,these results reveal an important role of Arabidopsis IAN2 to IAN6 genes in the regulation of the HSR,UPR,and cell death,and suggest that their natural variations have adaptive functions in heat tolerance.

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.

Determination of the High Temperature Stress Tolerances of Bread Wheat Genotypes

Thirty bread wheat genotypes were used as material during the 2014-2015 cropping season. The experimental layout was a randomized complete block design with 3 replications. The sowing rate was 500 seeds square meter. Sowing was done in plots of 6 rows （1.2 m × 5 m, spaced 20 cm apart） in Namlk Kemal University, Faculty of Agriculture, Field Crops Department experimental area. Two sowing times were performed. First sowing was made in November suggested usual （standard） and second one was made in January as delayed sown in order to push growing stages of plants into periods in which heat stress is expected will be effected. Sowing times were allotted to main-plots while genotypes were allotted to sub-plots. When the bread wheat varieties and lines used in the experiment are evaluated in terms of tolerance to high temperature, it was shown that Dropia and Nota varieties and CIMMYT-HTN 2014/15-2, CIMMYT-HTN 2014/15 -6, CIMMYT-HTN 2014/15 - 10 lines were better tolerance to high temperature. However, it was noticed that these genotypes were not included in the first groups in terms of grain yield. It is possible to utilize these genotypes as a genitor in cross-breeding programs for breeding studies for tolerance to high temperatures.

Effect of Heat Stress on Photosynthetic Characteristics of Different Green Organs of Winter Wheat During Grain-filling Stage

Four winter wheat (Triticum aestivum L.) varieties ('JD 8', 'Jing 411','Centurk' and 'Tam 202') were used to study the effect of heat stress on photosynthetic characteristics of flag leaf blade, nag leaf sheath, peduncle, glume, lemma and awn during grain-filling stage. The results showed that heat acclimation during grain-filling stage increased thermotolerance of wheat with significant differences among different green organs. During heat stress, the decreases of the efficiency of primary light energy conversion (F-v/F-m) of PS II and pigment (chlorophyll and carotenoid) content were much slower in peduncle, flag leaf sheath and glume than in nag leaf blade, lemma and ann; and the percentage of decrease in net photosynthetic rate (P-n) of ear was lower than that of the nag leaf blade. The measured photosynthetic parameters (F-v/F-m, P-n and pigment content) of 'JD 8', a relatively heat tolerant variety, declined more slowly than those of the other three varieties during the whole heat stress period.

Pig pangenome graph reveals functional features of non‑reference sequences

Background The reliance on a solitary linear reference genome has imposed a significant constraint on our compre-hensive understanding of genetic variation in animals.This constraint is particularly pronounced for non-reference sequences(NRSs),which have not been extensively studied.Results In this study,we constructed a pig pangenome graph using 21 pig assemblies and identified 23,831 NRSs with a total length of 105 Mb.Our findings revealed that NRSs were more prevalent in breeds exhibiting greater genetic divergence from the reference genome.Furthermore,we observed that NRSs were rarely found within coding sequences,while NRS insertions were enriched in immune-related Gene Ontology terms.Notably,our investigation also unveiled a close association between novel genes and the immune capacity of pigs.We observed substantial differences in terms of frequencies of NRSs between Eastern and Western pigs,and the heat-resistant pigs exhibited a substantial number of NRS insertions in an 11.6 Mb interval on chromosome X.Additionally,we discovered a 665 bp insertion in the fourth intron of the TNFRSF19 gene that may be associated with the ability of heat tolerance in South-ern Chinese pigs.Conclusions Our findings demonstrate the potential of a graph genome approach to reveal important functional features of NRSs in pig populations.

Effects of Heat Stress at Vegetative and Reproductive Stages on Spikelet Fertility

Seed-setting rate, yield components and grain quality traits of 169 accessions from an exotic rice germplasm were tested under high temperatures from 40 oC to 45 oC for 6 h during the daytime at the vegetative and reproductive stages, respectively. The results showed that heat stress significantly decreased the seed-setting rate of all the accessions, but the heat stress effects varied among accessions. Based on the decreases in seed-setting rate at high temperatures, N22 was the most tolerant, followed by AUS17, M9962, SONALEE and AUS16. Moreover, the reductions in seed-setting rate and yield under heat stress were more serious at the vegetative stage （45 d before heading） than at the booting stage （15 d before heading）. In addition, heat stress also affected grain quality, especially by conferring chalkiness to most of the accessions, but SONALEE did not change much. The heat-tolerant accessions identified here and the phenotype protocols developed could be used in future genetic studies and breeding programmes focused on heat tolerance.