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.     

Interpreting genotype × environment interactions for grain yield of rainfed durum wheat in Iran

Clustering genotype × environment(GE) interactions and understanding the causes of GE interactions are among the most important tasks in crop breeding programs. Pattern analysis(cluster and ordination techniques) was applied to analyze GE interactions for grain yield of 24 durum wheat(Triticum turgidum L. var. durum) genotypes(breeding lines and old and new cultivars) along with a popular bread wheat(Triticum aestivum) cultivar grown in 21 different rainfed environments during the 2010–2013 cropping seasons. To investigate the causes of GE interaction, several genotypic and environmental covariables were used. In a combined ANOVA, environment was the predominant source of variation,accounting for 81.2% of the total sum of squares(TSS), and the remaining TSS due to the GE interaction effect was almost seven times that of the genetic effect. Cluster analysis separated the environments into four groups with similar discriminating ability among genotypes, and genotypes into five groups with similar patterns in yield performance.Pattern analysis confirmed two major environmental clusters(cold and warm), and allowed the discrimination and characterization of genotype adaptation. Within the cold-environment cluster, several subclusters were identified. The breeding lines were most adapted to moderate and warm environments, whereas the old varieties were adapted to cold environments. The results indicated that winter rainfall and plant height were among the environmental and genotypic covariables, respectively, that contributed most to GE interaction for grain yield in rainfed durum wheat.     

Comparisons on Genetic Diversity among the Isonuclear-Alloplasmic Male Sterile Lines and Their Maintainer Lines in Rice

Four sets of rice isonuclear-alloplasmic lines including 16 male sterile lines and their maintainer lines were analyzed by using 91 pairs of SSR primers to study the genetic diversity of nuclear genome and their relative relationships. A total of 169 alleles were detected in the 16 lines, with a frequency of polymorphic loci of 53.85% and an average number of alleles per locus of 1.8, and the average gene diversity was 0.228. Four sets of the isonuclear-alloplasmic male sterile lines shared 146 identical alleles, corresponding to 86.39% of the total alleles; meanwhile, there are 23 different alleles among the tested materials, being 13.61% of the total alleles. On average, 78.70% identical alleles and 21.30% different alleles of the total alleles were detected between the isonuclear-alloplasmic male sterile lines and their maintainer lines. There were 53.85% identical alleles and 46.15% different alleles of the total alleles among the homozygous allonucleus male sterile lines. The fingerprints were established for some male sterile lines and maintainer lines. All the materials tested were divided into three groups at the 0.2 genetic distance based on the cluster analysis. Eight lines of Huanong A and Huayu A (including Huanong B and Huayu B) were in the first group, four lines of Kezhen A (including Kezhen B) in the second group, and four lines of Zhenshan 97A (including Zhenshan 97B) in the third group. For the isonuclear-alloplasmic male sterile lines, the similarity coefficient between Y (Yegong) type and WA (wild abortive) type or between CW (Raoping wild rice) and WA type reached 87-98%.     

Identification of a major quantitative trait locus and its candidate underlying genetic variation for rice stigma exsertion rate

Stigma exsertion in male sterile lines of hybrid rice is important for seed yield.In the present study, ZS616 [Oryza sativa subsp.Xian(indica)], a male sterile line with a stigma exsertion rate(SER) as high as 94.5%, was crossed to DS552, a japonica line with almost no exserted stigmas.F3 plants with extremely low and high SER were sequenced to identify SER-associated quantitative trait loci(QTL).A major QTL for SER, qSER-3.1, was identified along with other QTL on chromosome 3 in a 3.9 Mb region.A total of 307 nonsynonymous single-nucleotide polymorphisms(SNPs) and 27 frame-shift insertion/deletions(InDels)differentiating ZS616 and DS552 were identified in the region containing qSER-3.1.Most SNPs(294) and InDels(25) were excluded after further analysis because they were shared by ZS616 and low(<2.0%) SER accessions in the Huazhong Agricultural University(HAU) core rice collection.Association analysis using the full HAU collection identified a 17-bp InDel in OS03 G0689400 as the most likely causal genetic variant underlying qSER-3.1.ZS616-type accessions(n = 54, with the 17-bp insertion) in the HAU collection had minimum(16.5%)and mean(39.6%) SERs significantly greater than those(n = 424) without the insertion(with minimum and mean SERs of 0.2% and 20.6%, respectively).Thus, this study identified a major QTL for stigma exsertion and revealed the mutation in a candidate gene for the QTL.     

Evaluation of Rice Seedling Tolerance to Constant and Intermittent Low Temperature Stress

M202,IR50 and advanced backcross(BC 4 F 6) lines carrying the cold tolerance QTLs qCTS4 and qCTS12 were evaluated using three low temperature stress assays(constant,intermittent and constant with recovery period).Under constant stress,two BC 4 F 6 lines(MIb 4853-9 and MIb 6885-2) exhibited differences in seedling growth and accumulation of stress-related compounds although both carry the two QTLs in the IR50 background.Differences between these lines were also observed in the constant with recovery period assay,although both of them performed comparably under intermittent stress.MIb 6885-2 performed more similarly to M202 in the constant and constant with recovery period assays,suggesting that this line contains an introgressed region(s) not present in MIb 4853-9.The three assays were also applied to assess variations in cold tolerance in a set of diverse germplasms.Performance metrics and visual ratings were comparable for evaluating tolerance to constant stress.However,differences in response to intermittent stress were more evident from growth and total chlorophyll measurements than from visual ratings.With regard to the constant with recovery period assay,about 15% of the germplasm accessions did not recover.However,some lines which exhibited chilling injuries of comparable severity were able to overcome them.This recovery phenotype may improve rice performance in the field and warrant further investigation.     

Evaluation and Bulked Segregant Analysis of Major Yield QTL qtl12.1 Introgressed into Indigenous Elite Line for Low Water Availability under Water Stress

Near isogenic lines carrying large-effect QTL (qtl12.1), which has a consistent influence on grain yield under upland drought stress conditions in a wide range of environments, were evaluated under water stress in the fields. The line which gave higher yield under drought was crossed with a local elite line, PMK3, and forwarded to F2:3 generation. Significant variation was found among the F2:3 lines for agronomic traits under water stress in the fields. Low to high broad sense heritability (H) for investigated traits was also found. Water stress indicators such as leaf rolling and leaf drying were negatively correlated with plant height, biomass and grain yield under stress. Bulked segregant analysis (BSA) was performed with the markers in the vicinity of qtl12.1, and RM27933 was found to be segregated perfectly well in individual components of drought resistant and drought susceptible bulks which were bulked based on yield under water stress among F2:3 lines. Hence, this simple and breeder friendly marker, RM27933, may be useful as a potentially valuable candidate marker for the transfer of the QTL qtl12.1 in the regional breeding program. Bioinformatic analysis of the DNA sequence of the qtl12.1 region was also done to identify and analyze positional candidate genes associated with this QTL and to ascertain the putative molecular basis of qtl12.1.     

Genetic analysis of blast resistance On rice cultivars with wide—spectrum resistance

In recent years,indirect selection based ontightly linked genetic markers has been sug-gested to pyramid the resistance genes.How-ever,resistance parents used in the linkagestudies were usually near isogenic lines possessing a single resistance gene derived fromthe original resistance donor.Little was knownon the interaction among the genes at differentloci.In the present study,the genetic controlof blast resistance in two wide—spectrum re-vistance rice cultivars was analyzed.A susceptible indica varieties Xianfenglwas used as the female parent,and the blastresistance cultivars Tetep and Hongjiaozhanwere used as the male parents.The threeparental lines were inoculated with each of 20races of Pyricularia oryzae Cav at tilleringstage.The race ZC_(13) was selected to inoculatethe 161 F_3 lines of Xianfeng1/Tetep and 175 F,lines of Xianfengl/Hongjiaozhan.Eighteen in-dividuals of each line were inoculated.In both populations,all the individualsdemonstrated extreme reaction to the blast dis-ease.In the F_3 po     

Genetic Diversity of Tropical Hybrid Rice Germplasm Measured by Molecular Markers

Investigation of genetic diversity and relationships among breeding lines is of great importance to facilitate parent selection in hybrid rice breeding programs.In this study,we characterized 168 hybrid rice parents from International Rice Research Institute with 207 simple sequence repeat (SSR) and 353 single nucleotide polymorphism (SNP) markers.A total of 1 267 SSR and 706 SNP alleles were detected with the averages of 6.1 (SSR) and 2.0 (SNP) alleles per locus respectively across all lines.Based on the genetic distances estimated from the SSR and SNP markers separately and combined,the unrooted neighbor-joining cluster and STRUCTURE analyses consistently separated the 168 hybrid rice parents into two major groups:B-line and R-line,which is consistent with known parent pedigree information.The genetic distance matrices derived from the SSR and SNP genotyping were highly correlated (r=0.81,P < 0.001),indicating that both of the SSR and SNP markers have distinguishable power to detect polymorphism and are appropriate for genetic diversity analysis among tropical hybrid rice parents.A subset of 60 SSR markers were also chosen by the Core Hunter with 368 alleles,and the cluster analysis based on the total and subset of SSR markers highly corresponded at r=0.91 (P < 0.001),suggesting that fewer SSR markers can be used to classify and evaluate genetic diversity among parental lines.     

Differential Expression of Whitebacked Planthopper Resistance in the japonicaJindica Doubled Haploid Rice Population under Field Evaluation and Seedbox Screening Test

Whitebacked planthopper (WBPH) -resistance in a japonica / indica doubled haploid (DH) rice population established from a cross between WBPH-resistant japonica Chunjiang 06 and susceptible indica TN1, was comparatively evaluated through a field experiment based on the WBPH immigrant density and standardized seedbox screening test (SSST). All the susceptible DH lines in the field experiment behaved accordingly in SSST. However, 35 of resistant 66 lines (53%) in the field, were categorized to susceptible groups in SSST. Likewise, there were no significant differences in WBPH immigrant densities among 70 DH lines that were highly resistant to susceptible in SSST. The results revealed that SSST could not evaluate properly WBPH resistance in the DH lines. Four QTLs for WBPH-resistance phenotyped by the immigrant density were detected on chromosomes 2, 3, 4, and 11. Of them, the QTL on chromosome 4 was the most effective (LOD 21.8, variance 78%). Five QTLs associated with seedling mortality were mapped on chromosomes 2, 3, 4, 5 and 6. In addition to the QTL (LOD 10.5, variance 68%) on chromosome 4, there was another major QTL (LOD 12.7, variance 71%) located on chromosome 5, which was SSST-specific but might be irrespective of the WBPH resistance traits.     

Analysis of Introgressed Segments in Near-isogenic Lines for F1 Pollen Sterility in Rice （Oryza sativa）

One hundred and fifty-eight microsatellite markers showing polymorphism among parents were used to survey the introgressed segments in the 50 near-isogenic lines of F1 pollen sterility. Two hundred and sixty introgressed segments were detected in 50 near-isogenic lines, each carrying 5.2 introgressed segments on an average. Among the 260 segments, one hundred carrying F1 pollen sterility loci concentrated on the region of F1 pollen sterility genes, and the remaining one hundred and sixty without F1 pollen sterility loci distributed randomly over 12 chromosomes.Both the average number and length of the introgressed segments decreased along with the increase of backcross generations. The number of introgressed segments was less than four and the length was less than 20 cM in the near-isogenic lines after backcrossing for four or more times.     

Analysis of genetic relationships between grain milling quality traits and plant agronomic traits of rice

Since seed and maternal plant are in different generations, the grain milling traits and plant agronomic traits ot rice are based on different genetic models. Using the genetic models which could estimate covariance components between two traits with unequal design matrices, we analyzed the gentic relationships between grain milling traits and plant agronomic traits. Nine cytoplasmic male sterile (CMS) lines (Zhenshan 97 A and others) and five restoring lines (T 49 and others) of indica rice were used in an incomplete diallel cross. Seedlings of parents and F_1s were planted in the field at Zhejiang Agri Univ in 1994. There were 24 plants in each plot with three replications. Plant agronomic traits were determined from eight plants in the middle part of the plot. Seed samples of parents, F_1 and F_2s were used for analyzing milling quality traits of rice. It was indicated by the results (See table) that stronger genotypic relationships were existed according to genotypic covariances (C_(G/G)) between th     

Quantitative Trait Loci and Epistatic Analysis of Seed Anoxia Germinability in Rice (Oryza sativa)

Anoxia germinability (AG) of 35 rice varieties was evaluated under different temperature and water submergence conditions. The shoot (including coleoptile) length of seedlings germinating under 30℃, 0.2 m water submergence for 5 days could be used as an optimal criterion for the AG evaluation of all the varieties. Differences were observed among the AGs of 359 varieties from different regions and subspecies with the optimized method. Moreover, 81 recombinant inbred lines derived from a cross of Kinmaze (japonica)/DV85 (indica) were used to detect quantitative trait loci (QTLs) conferring AG. A total of five QTLs for AG in the recombinant inbred population were detected on chromosomes 1, 2, 5 and 7, respectively. Phenotypic variations explained by each QTL ranged from 10.5% to 19.6%. Based on the directions of the additive effects, the alleles at three loci qAG-1, qAG-2 and qAG-7 from Kinmaze increased AG, while alleles at loci qAG-5a and qAG-5b from DV85 increased AG. Meanwhile, three pairs of epistatic loci were found to be located on chromosomes 2, 3, 5 and 11 with significant effects ranging from 16.7% to 48.8%, and the highest one 48.78%, was detected between C563–X182 on chromosome 3 and R830–X208 on chromosome 5.     

Genomic prediction of the performance of hybrids and the combining abilities for line by tester trials in maize

The two most important activities in maize breeding are the development of inbred lines with high values of general combining ability(GCA)and specific combining ability(SCA),and the identification of hybrids with high yield potentials.Genomic selection(GS)is a promising genomic tool to perform selection on the untested breeding material based on the genomic estimated breeding values estimated from the genomic prediction(GP).In this study,GP analyses were carried out to estimate the performance of hybrids,GCA,and SCA for grain yield(GY)in three maize line-by-tester trials,where all the material was phenotyped in 10 to 11 multiple-location trials and genotyped with a mid-density molecular marker platform.Results showed that the prediction abilities for the performance of hybrids ranged from 0.59 to0.81 across all trials in the model including the additive effect of lines and testers.In the model including both additive and non-additive effects,the prediction abilities for the performance of hybrids were improved and ranged from 0.64 to 0.86 across all trials.The prediction abilities of the GCA for GY were low,ranging between-0.14 and 0.13 across all trials in the model including only inbred lines;the prediction abilities of the GCA for GY were improved and ranged from 0.49 to 0.55 across all trials in the model including both inbred lines and testers,while the prediction abilities of the SCA for GY were negative across all trials.The prediction abilities for GY between testers varied from-0.66 to 0.82;the performance of hybrids between testers is difficult to predict.GS offers the opportunity to predict the performance of new hybrids and the GCA of new inbred lines based on the molecular marker information,the total breeding cost could be reduced dramatically by phenotyping fewer multiple-location trials.     

Analysis of Introgressed Segments in Near isogenic Lines for F1 Pollen Sterility in Rice (Oryza sativa)

One hundred and fifty eight microsatellite markers showing polymorphism among parents were used to survey the introgressed segments in the 50 near isogenic lines of F1 pollen sterility. Two hundred and sixty introgressed segments were detected in 50 near isogenic lines, each carrying 5.2 introgressed segments on an average. Among the 260 segments, one hundred carrying F1 pollen sterility loci concentrated on the region of F1 pollen sterility genes, and the remaining one hundred and sixty without F1 pollen sterility loci distributed randomly over 12 chromosomes. Both the average number and length of the introgressed segments decreased along with the increase of backcross generations. The number of introgressed segments was less than four and the length was less than 20 cM in the near isogenic lines after backcrossing for four or more times.     

QTLs Analysis of Cold Tolerance During Early Growth Period for Rice

The quantitative trait loci (QTLs) for cold tolerance during early growth period were identified using a F2:3 population (including 200 individuals and lines derived from a cross between indica rice and japonica rice ‘Milyang 23/Jileng 1’) with microsatellite markers. The cold tolerance at the seedling and tillering stages, and the growth ability of seedling under low temperature conditions were evaluated. All of the traits associated with cold tolerance at early growth stages appeared a continuous distribution near to normal in F3 lines, these were inherited as quantitative traits controlled by polygenes. Three QTLs on chromosomes 1, 5 and 9, which associated with cold tolerance at the seedling stage were detected. Among them, qCTS1 accounted for 15.5% of observed phenotypic variation; Five QTLs on chromosomes 2, 3, 7, 9 and 11, associated with cold tolerance at the tillering stage were found, which explained lower percentage of observed phenotypic variation; Four QTLs on chromosomes 1, 2, 11 and 12, which associated with the growth ability of seedling under low temperature conditions were found, among them, qGAS2 and qGAS12 explained 26.6 and 42.9% of observed phenotypic variation, respectively, which were major genes.     

Genetic Analysis of Early Generation Stability in Rice

The mechanism of early generation stability （EGS） in rice was studied via genetic analysis. Three types of crosses were made, namely between EGS varieties, EGS and conventional rice variety, and conventional rice varieties. The genetic analysis was based on the stable lines in F2 population. The stable lines may appear from some combinations of EGS rice crossing with each other and EGS rice crossing with conventional varieties at different frequencies, but stable lines didn＇t appear in conventional varieties crossing with conventional varieties. Genetic analysis results indicated that the EGS phenomena should just exist in special rice materials, and the frequency of stable lines was closely related to the EGS traits of parents. The EGS traits were neither qualitative nor quantitative traits, and they were controlled by neither dominant genes nor recessive genes. The EGS traits might be inherited by F1 single plant, and the traits of F3 and F4 were corresponded to those of F2 population, i.e. F3 and F4 lines derived from non-segregating F2 showed uniform agronomic traits, and those from segregating F2.did not. The agronomic traits of EGS lines were consistent with those of F1 single plant. On the other hand, when EGS lines occurred, the segregating lines in Mendelian manner were also observed in all F2 population of the same combination. It was suggested that the reason why the stable strains occurred might be a special factor to control （open/close） gene at the beginning of cell division in zygote, resulting in closing mitosis and opening somatic reduction. The somatic reduction of zygote resulted in recombination and homozygosity forming in F1 single plant, and some lines with uniform agronomic traits were observed in some lines of F2 population.     

Determination of Heterotic Groups and Heterosis Analysis of Yield Performance in indica Rice

To compare the heterosis levels among various groups of parental lines used extensively in China, identify foundational heterotic groups in parental pools and understand the relationship between genetic distance and heterosis performance, 16 parental lines with extensive genetic variation were selected from various sub-groups, and 39 hybrid combinations were generated and evaluated in Fujian and Hainan Provinces of China. The main results were as follows:(1) The 16 parental lines can be grouped into 7 sub-groups consisting of 1 maintainer sub-group and 6 restorer sub-groups;(2) Mean grain yield of the restorer lines was higher than that of the maintainer lines, and mean yield of parental lines was higher than that of the hybrid combinations;(3) The two best heterotic patterns were II-32A × G5 and II-32A × G6, moreover, the order of restorer sub-groups according to grain yield, from the highest to lowest, was G7, G6, G5, G4, G3 and G2; High specific combining ability values were observed for combinations of II-32A × G5, II-32A × G6 and Tianfeng A × G7;(4) Hybrid combinations derived from II-32A crossed with 13 restorer lines had higher yield trait values(mid-parent heterosis, better-parent heterosis, standard heterosis over check and specific combining ability) than any other combinations;(5) Genetic distance was positively correlated with panicle number, grain length and length-to-width ratio(P 0.05) and negatively correlated with grain width, grain yield, seed-setting rate, as well as mid-parent heterosis, standard heterosis over check, and specific combining ability for grain yield(P 0.01). These heterotic groups and patterns and their argonomic traits will provide useful information for future hybrid rice breeding programs.     

贵州常用玉米自交系的SSR遗传分析

Genetic variation was analyzed of 35 maize inbred lines in Guizhou with SSR.Selected 10 core-primers which could test 75 allele sites in sample of 35 inbred lines.The average number of allele per SSR locus was 7.5 with arrange from 5 to 14.The average of polymorphic information content(PIC) was 0.739.Gene locus among the 35 inbred lines ranged from 0.154 to 0.838 with an average of 0.513.The clustering results showed that the inbred lines could be classified into 7 groups.The clustering results were consistent with the groups based on the available.The local germplasm lines could be divided into three heterotic groups,showed the local germplasm had wide genetic basis.