Stigma Exsertion Research Articles

Dissection of closely linked QTLs controlling stigma exsertion rate in rice by substitution mapping

Key messageThrough substitution mapping strategy, two pairs of closely linked QTLs controlling stigma exsertion rate were dissected from chromosomes 2 and 3 and the four QTLs were fine mapped.Stigma exsertion rate (SER) is an important trait affecting the outcrossing ability of male sterility lines in hybrid rice. This complex trait was controlled by multiple QTLs and affected by environment condition. Here, we dissected, respectively, two pairs of tightly linked QTLs for SER on chromosomes 2 and 3 by substitution mapping. On chromosome 2, two linkage QTLs, qSER-2a and qSER-2b, were located in the region of 1288.0 kb, and were, respectively, delimited to the intervals of 234.9 kb and 214.3 kb. On chromosome 3, two QTLs, qSER-3a and qSER-3b, were detected in the region of 3575.5 kb and were narrowed down to 319.1 kb and 637.3 kb, respectively. The additive effects of four QTLs ranged from 7.9 to 9.0%. The epistatic effect produced by the interaction of qSER-2a and qSER-2b was much greater than that of qSER-3a and qSER-3b. The open reading frames were identified within the maximum intervals of qSER-2a, qSER-2b and qSER-3a, respectively. These results revealed that there are potential QTL clusters for SER in the two regions of chromosome 2 and chromosome 3. Fine mapping of the QTLs laid a foundation for cloning of the genes of SER.

Molecular mapping of quantitative trait loci for stigma exsertion trait in rice (Oryza sativa L.)

Stigma exsertion is the most important floral trait of female parent/cytoplasmic male sterile (CMS) line for out crossing which increases hybrid seed production at farmer’s field. The present study was aimed to map the genomic regions controlling the stigma exsertion in F2 mapping population derived from IR68897B, a recipient maintainer line of IR68897A with low stigma exsertion (36.78 %) and BF-16B, a donor parent for high stigma exsertion (80.25%). Phenotyping and genotyping of stigma exsertion trait was done for 188 individuals of F2 population developed from IR68897B X BF16B. Frequency distribution of F2 population showed continuous variation which indicated polygenic nature of inheritance. Phenotypic evaluation revealed that significant variation for stigma exsertion trait and transgressive segregation was observed. A total of 15 QTLs were identified for total stigma exsertion rate (TSE) which includes single and dual stigma exsertion distributed on chrm 2, 3, 4, 5, 6, 8, 11 and 12 through QTL mapping. The phenotypic variance and LOD value explained by each QTL ranged from 1.19 to 5.83% and 2.54 to 9.29 respectively. In the present study, total 15 QTLs were minor effect QTLs s

Morphological and anatomical characteristics of exserted stigma sterility and the location and function of SlLst (Solanum lycopersicum Long styles) gene in tomato.

Anatomical changes in and hormone roles of the exserted stigma were investigated, and localization and functional analysis of SlLst for the exserted stigma were performed using SLAF-BSA-seq, parental resequencing and overexpression of SlLst in tomato. Tomato accession T431 produces stigmas under relatively high temperatures (> 27°C, the average temperature in Harbin, China, in June-August), so pollen can rarely reach the stigma properly. This allows the percentage of male sterility exceed 95%, making the use of this accession practical for hybrid seed production. To investigate the mechanism underlying the exserted stigma male sterility, the morphological changes of, anatomical changes of, and comparative endogenous hormone (IAA, ABA, GA3, ZT, SA) changes in flowers during flower development of tomato accessions DL5 and T431 were measured. The location and function of genes controlling exserted stigma sterility were analyzed using super SLAF-BSA-seq, parental resequencing, comparative genomics and the overexpression of SlLst in tomato. The results showed that an increase in cell number mainly caused stigma exsertion. IAA played a major role, while ABA had an opposite effect on stigma exertion. Moreover, 26 candidate genes related to the exserted stigma were found, located on chromosome 12. The Solyc12g027610.1 (SlLst) gene was identified as the key candidate gene by functional analysis. A subcellular localization assay revealed that SlLst is targeted to the nucleus and cell membrane. Phenotypic analysis of SlLst-overexpressing tomato showed that SlLst plays a crucial role during stigma exsertion.

QTLs detection and pyramiding for stigma exsertion rate in wild rice species by using the single-segment substitution lines

QTLs detection and pyramiding for stigma exsertion rate in wild rice species by using the single-segment substitution lines

ATP-Binding Cassette G Transporters SGE1 and MtABCG13 Control Stigma Exsertion.

Stigma exsertion is an important agricultural trait that facilitates the application of heterosis in crop breeding. Although several quantitative trait loci associated with stigma exsertion have been fine-mapped or cloned, the underlying genetic basis, particularly in legumes, remains unclear. In this study, we identified and characterized the exserted stigma mutant stigma exsertion1 (sge1) in the model legume Medicago truncatula The exserted stigma phenotype of sge1 is mainly caused by physical interaction between floral organs, in which normal petal and stamen elongation are inhibited due to flower cuticle defects. SGE1 encodes an ATP-binding cassette G (ABCG) transporter that plays a critical role in regulating floral cutin and wax secretion in M. truncatula SGE1 physically interacts with another half-size transporter, MtABCG13, to form a functional heterodimer. Mutation of MtABCG13 results in flower cuticle defects similar to those in sge1 as well as stigma exsertion, indicating that SGE1 and MtABCG13 are indispensable for flower cuticle secretion and collaboratively control stigma exsertion in M. truncatula Our findings reveal novel functions for ABCG transporters in determining stigma exsertion by affecting the physical interactions of floral organs, providing insight into the molecular mechanism underlying stigma exsertion in leguminous plants with complex zygomorphic flowers.

Genetic Control of Reproductive Traits in Tomatoes Under High Temperature.

Global climate change is increasing the range of temperatures that crop plants must face during their life cycle, giving negative effects to yields. In this changing scenario, understanding the genetic control of plant responses to a range of increasing temperature conditions is a prerequisite to developing cultivars with increased resilience. The current work reports the identification of Quantitative Trait Loci (QTL) involved in reproductive traits affected by temperature, such as the flower number (FLN) and fruit number (FRN) per truss and percentage of fruit set (FRS), stigma exsertion (SE), pollen viability (PV) and the incidence of the physiological disorder tipburn (TB). These traits were investigated in 168 Recombinant Inbred Lines (RIL) and 52 Introgression Lines (IL) derived from the cross between Solanum lycopersicum var. “MoneyMaker” and S. pimpinellifolium accession TO-937. Mapping populations were cultivated under increased temperature regimen conditions: T1 (25°C day/21°C night), T2 (30°C day/25°C night) and T3 (35°C day/30°C night). The increase in temperature drastically affected several reproductive traits, for example, FRS in Moneymaker was reduced between 75 and 87% at T2 and T3 when compared to T1, while several RILs showed a reduction of less than 50%. QTL analysis allowed the identification of genomic regions affecting these traits at different temperatures regimens. A total of 22 QTLs involved in reproductive traits at different temperatures were identified by multi-environmental QTL analysis and eight involved in pollen viability traits. Most QTLs were temperature specific, except QTLs on chromosomes 1, 2, 4, 6, and 12. Moreover, a QTL located in chromosome 7 was identified for low incidence of TP in the RIL population, which was confirmed in ILs with introgressions on chromosome 7. Furthermore, ILs with introgressions in chromosomes 1 and 12 had good FRN and FRS in T3 in replicated trials. These results represent a catalog of QTLs and pre-breeding materials that could be used as the starting point for deciphering the genetic control of the genetic response of reproductive traits at different temperatures and paving the road for developing new cultivars adapted to climate change.

Identification and fine mapping of a stigma exsertion mutant gene (Bolsem) in ornamental kale (Brassica oleracea var. acephala)

Stigma exsertion is a plant trait wherein stigmas emerge from flower buds before anthesis. It is a key determinant of the ornamental kale mating system, greatly increases the efficiency of hybrid seed production, and represents a technique to promote crossing pollination. The present study investigated a stigma exsertion mutant (‘Y009-10-5’) in ornamental kale and determined that the trait was determined by a recessive loss-of-function allele at the Bolsem (B. oleracea stigma exsertion mutant) locus. Then, using linkage analysis, with 350 F2 progeny and 98 SSR markers, the Bolsem locus was mapped to a 65.64-kb region of Chromosome 7, between the markers SSRCWS5 and SSRLS3. Sequence analysis and Brassica Database analysis identified two genes in this region: Bol005242, a gene of unknown function, and Bol005241, an ortholog of the Arabidopsis gene ABAP1, which controls cell proliferation and was, therefore, considered a potential candidate for Bolsem. Further sequence analysis identified two single-nucleotide polymorphisms (SNPs) in Bol005241. One SNP was found in the third exon, with the ‘Y009-10-5’ sequence yielding a TAA stop codon; therefore, it is predicted as the significant effects on stigma exsertion. Bol005241 also fully co-segregated with the exon SNP, and quantitative real-time PCR indicated that Bol005241 exhibited pistil-specific expression. Pollination experiments were performed to determine the effect of pollination timing on hybrid seed production. The results of this study provide a basis for elucidating the molecular mechanism underlying stigma exsertion, which could significantly improve hybrid seed production in ornamental kale.

Fine mapping a quantitative trait locus, qSER-7, that controls stigma exsertion rate in rice (Oryza sativa L.)

BackgroundStigma exsertion rate (SER) is a key determinant of outcrossing in hybrid rice seed production. A quantitative trait locus (QTL) for stigma exsertion rate in rice, qSER-7, has previously been detected on chromosome 7 by using a F2 population derived from two indica cytoplasmic male sterility (CMS) maintainers, Huhan 1B and II-32B.ResultsThe chromosomal location of qSER-7 was precisely delimited by fine-scale mapping. Near-isogenic lines (NILs) were established, one of which isolated the locus in the qSER-7II-32B line, which contains an introgressed segment of II-32B in the Huhan 1B genetic background, and exhibits a significantly higher stigma exsertion rate than that of the recurrent parent. Using 3192 individuals from the BC4F2 segregation population, the QTL qSER-7 was narrowed down to a 28.4-kb region between the markers RM3859 and Indel4373 on chromosome 7. According to the rice genome annotation database, three genes were predicted within the target region. Real-time PCR analysis showed significantly higher expression levels of LOC_Os07g15370 and LOC_Os07g15390 in II-32B than in Huhan 1B. LOC_Os07g15370(OsNRAMP5) was a previously reported gene for Mn and Cd transporter. The stigma exertion rates of OsNRAMP5-overexpressing plants were significantly higher than that of wild type plants, in contrast, a T-DNA insertion mutant osnramp5 showed a lower stigma exertion rate.ConclusionsIn the present study, the QTL qSER-7 was isolated to a region between the markers RM3859 and Indel4373. Two candidate genes were selected based on the expression difference between the two parents, which can facilitate the further cloning of the gene underlying the quantitative trait associated with qSER-7 as well as the marker-assisted transfer of desirable genes for stigma exsertion rate improvement in rice.

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 heat-sensitive 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.

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 OS03G0689400 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.

Detection of dominant QTLs for stigma exsertion ratio in rice derived from Oryza rufipogon accession 'W0120'.

Stigma exsertion can enhance the outcrossing fertility in rice. Dominant genes for this trait are expected to be essential for the effective application of a novel outcrossing-based breeding system that uses male sterility in rice, which is normally autogamous. Because reduction of stigma exsertion is a domestication trait, we screened wild rice species as possible donors of genes or QTLs for stigma exsertion. We used in silico image-based screening and selected the Oryza rufipogon accession ‘W0120’. A single F1 individual derived from a cross between the japonica rice cultivar ‘Akidawara’ and ‘W0120’ was used to generate F2 and BC1F1 populations. QTL analysis performed using 114 F2 individuals detected QTLs on chromosomes 2, 3, 4, 8, and 11. Only two major QTLs on chromosomes 3 and 8 showed higher degrees of dominance. On the other hand, there were no QTLs near GS3, which is well known as a gene for stigma exsertion. Validation of these QTLs using 188 BC1F1 individuals provided clear evidence for their dominance. Genotypes of the markers nearest to the two QTLs were also related to grain length. We expect the genes responsible for these QTLs to be promising tools for improving outcrossing-based breeding in rice.

QSE7 is a major quantitative trait locus (QTL) influencing stigma exsertion rate in rice (Oryza sativa L.)

Stigma exsertion is a key determinant to increase the efficiency of commercial hybrid rice seed production. The major quantitative trait locus (QTL) qSE7 for stigma exsertion rate was previously detected on the chromosome 7 using 75 Chromosome Segment Substitution Lines (CSSLs) derived from a cross between the high stigma exsertion indica maintainer XieqingzaoB (XQZB) and low stigma exsertion indica restorer Zhonghui9308 (ZH9308). The C51 line, a CSSL population with an introgression from XQZB, was backcrossed with ZH9308 to produce the secondary F2 (BC5F2) and F2:3 (BC5F2:3) populations. As a result, the Near Isogenic Line (NIL qSE7XB) was developed. Analysis indicated qSE7 acted as a single Mendelian factor and decreased the stigma exsertion. We hypothesized qSE7 regulates single, dual, and total stigma exsertion rate, provided experimental support. qSE7 was mapped and localized between RM5436 and RM5499 markers, within a physical distance of 1000-kb. With use of new insertion-deletion (InDel) markers and analysis of the heterozygous and phenotypic data, it was ultimately dissected to a 322.9-kb region between InDel SER4-1 and RM5436. The results are useful for additional identification and isolation of this candidate gene controlling stigma exsertion rate, and provide a basis for further fine mapping, gene cloning, and Marker Assisted Selection (MAS) breeding later.

Tomato stigma exsertion induced by high temperature is associated with the jasmonate signalling pathway.

High temperature (HT) is becoming an increasingly serious factor in limiting crop production with global climate change. During hot seasons, owing to prevailing HT, cultivated tomatoes are prone to exhibiting stigma exsertion, which hampers pollination and causes fruit set failure. However, the underlying regulatory mechanisms of the HT-induced stigma exsertion remain largely unknown. Here, we demonstrate that stigma exsertion induced by HT in cultivated tomato is caused by more seriously shortened stamens than pistils, which is different from the stigma exsertion observed in wild tomato species. Under the HT condition, the different responses of pectin, sugar, expansin, and cyclin cause cell wall remodelling and differentially localized cell division and selective cell enlargement, which further determine the lengths of stamens and pistils. In addition, auxin and jasmonate (JA) are implicated in regulating cell division and cell expansion in stamens and pistils, and exogenous JA instead of auxin treatment can effectively rescue tomato stigma exsertion through regulating the JA/COI1 signalling pathway. Our findings provide a better understanding of stigma exsertions under the HT condition in tomato and uncover a new function of JA in improving plant abiotic stress tolerance.

Breeding strategies for hybrid rice parental line improvement

In our country, significant progress has been made in the development of hybrid rice parental lines both female and male parents with good floral traits. An array of hybrids with good grain quality (both unique Basmati type and medium slender (MS) grain type) is developed. Efforts are focussed on the development of restorer lines through exploitation of inter sub-specific hybridization, recurrent selection and population improvement that will help in the development of highly heterotic rice hybrids. The female parental lines are being improved for traits like stigma exsertion and high outcrossing that helps in higher seed yields, thereby reducing the hybrid seed cost. Efforts are on to develop appropriate parental lines that can help in the development of suitable hybrids in late duration group and also for unfavourable ecologies. Many newly developed parental lines are being fortified with resistance genes for major diseases such as BB, blast and insect pests such as BPH that will help in the development of hybrids having resistance to major pests and diseases.

Identification and expression profiling of microRNAs involved in the stigma exsertion under high-temperature stress in tomato

BackgroundAutogamy in cultivated tomato varieties is a derived trait from wild type tomato plants, which are mostly allogamous. However, environmental stresses can cause morphological defects in tomato flowers and hinder autogamy. Under elevated temperatures, tomato plants usually exhibit the phenotype of stigma exsertion, with severely hindered self-pollination and fruit setting, whereas the inherent mechanism of stigma exsertion have been hitherto unknown. Numerous small RNAs (sRNAs) have been shown to play significant roles in plant development and stress responses, however, none of them have been studied with respect to stamen and pistil development under high-temperature conditions. We investigated the associations between stigma exsertion and small RNAs using high-throughput sequencing technology and molecular biology approaches.ResultsSixteen sRNA libraries of Micro-Tom were constructed from plants stamen and pistil samples and sequenced after 2 d and 12 d of exposure to heat stress, respectively, from which a total of 110 known and 84 novel miRNAs were identified. Under heat stress conditions, 34 known and 35 novel miRNAs were differentially expressed in stamens, and 20 known and 10 novel miRNAs were differentially expressed in pistils. GO and KEGG pathway analysis showed that the predicted target genes of differentially expressed miRNAs were significantly enriched in metabolic pathways in both stamen and pistil libraries. Potential miRNA-target cleavage cascades that correlated with the regulation of stigma exsertion under heat stress conditions were found and validated through qRT-PCR and RLM-5′ RACE.ConclusionOverall, a global spectrum of known and novel miRNAs involved in tomato stigma exsertion and induced by high temperatures were identified using high-throughput sequencing and molecular biology approaches, laying a foundation for revealing the miRNA-mediated regulatory network involved in the development of tomato stamens and pistils under high-temperature conditions.

Genetic Dissection of the Major Quantitative Trait Locus (qSE11), and Its Validation As the Major Influence on the Rate of Stigma Exsertion in Rice (Oryza sativa L.).

The rate of stigma exsertion (SE) is an important trait in rice breeding because the efficiency of hybrid rice seed production can be improved by increasing the percentage of stigmas that exsert. In this study, we developed a near isogenic line (NIL) from two parents, XieqingzaoB (XQZB) and Zhonghoi9308 (ZH9308), which have high and low SE rates in that order. In our previous study, we employed 75 chromosome segment substitution lines (CSSLs) and analyzed quantitative trait loci (QTLs) for their influence on SE rate. The single gene QTL (qSE11), which is located on chromosome 11, was responsible for this trait. In this study, we focused on one of the CSSLs (C65), namely, the NIL (qSE11XB). It contains an introgression segment of XQZB in the genetic background of ZH9308, and exhibits a significantly higher SE rate than that of the parents. We demonstrated that qSE11 regulated both the single and the dual SE rates. They both contribute to the total SE rate. Genetic analysis revealed that qSE11 acted as a single Mendelian factor and that the allele from XQZB increased the SE rate. The validity of our conclusions was established when C65 was used to develop secondary F2 (BC5F2) and F2:3 (BC5F2:3) populations by backcrossing to ZH9308, with subsequent selfing. We entered 3600 plants from the F2 population and 3200 from the F2:3 populations into a genetic dissection program and dissected the major QTL qSE11 to a 350.7-kb region located on chromosome 11. This study will contribute to the future isolation of candidate genes of SE and will play a vital role in future hybrid rice seed production programs.

Genetic mapping of quantitative trait loci for the stigma exsertion rate in rice (Oryza sativa L.)

The efficiency of hybrid rice seed production can be improved by increasing the percentage of exserted stigmas. To identify quantitative trait loci (QTLs) for this trait, we conducted QTL mapping using 75 chromosome segment substitution lines (CSSLs) developed from a cross between the donor parent, Xieqingzao B (XQZB), a maintainer line which has high stigma exsertion and the recurrent parent, Zhonghui 9308 (ZH9308), a restorer line which has low stigma exsertion. A total of nine QTLs (qSSE5, qSSE10, qSSE11, qDSE10, qDSE11, qTSE5, qTSE6, qTSE10, and qTSE11) for single stigma exsertion (SSE), dual stigma exsertion (DSE) and total stigma exsertion (TSE) were assessed in two environments (Hainan and Zhejiang). Six of these QTLs (qSSE10, qSSE11, qDSE10, qDSE11, qTSE10, and qTSE11) were found in both environments, while one QTL (qTSE6) was found in only Hainan, and two QTLs (qSSE5 and qTSE5) were found in only Zhejiang. The qSSE10, qSSE11, qDSE10, qDSE11, qTSE6, qTSE10, and qTSE11 alleles, which are derived from the parent XQZB, exhibited a positive additive effect. In contrast, the qSSE5 and qTSE5 alleles, which are derived from the parent ZH9308, exhibited a negative additive effect. The SSE, DSE and TSE traits were significantly correlated with each other in an environmentally dependent manner. These results indicated that the lines showing higher values for SSE were more likely to exhibit increased values for DSE, which would ultimately increase TSE. To evaluate the advantage of exserted stigmas for cross-pollination, single, dual and total stigma exsertion should be considered separately in future attempts at genetic improvement to achieve increased production of rice hybrid seeds. This study also provides information for fine mapping, gene cloning and particularly marker-assisted selection (MAS), on the latter and with an emphasis the phenotypic effects and implications of the QTLs for practical use in hybrid rice breeding.

Genome-wide association study of outcrossing in cytoplasmic male sterile lines of rice

Stigma exsertion and panicle enclosure of male sterile lines are two key determinants of outcrossing in hybrid rice seed production. Based on 43,394 single nucleotide polymorphism markers, 217 cytoplasmic male sterile lines were assigned into two subpopulations and a mixed-group where the linkage disequilibrium decay distances varied from 975 to 2,690 kb. Genome-wide association studies (GWAS) were performed for stigma exsertion rate (SE), panicle enclosure rate (PE) and seed-setting rate (SSR). A total of 154 significant association signals (P < 0.001) were identified. They were situated in 27 quantitative trait loci (QTLs), including 11 for SE, 6 for PE, and 10 for SSR. It was shown that six of the ten QTLs for SSR were tightly linked to QTLs for SE or/and PE with the expected allelic direction. These QTL clusters could be targeted to improve the outcrossing of female parents in hybrid rice breeding. Our study also indicates that GWAS-base QTL mapping can complement and enhance previous QTL information for understanding the genetic relationship between outcrossing and its related traits.

Genome-wide Association Analyses Reveal the Genetic Basis of Stigma Exsertion in Rice

Stigma exsertion, a key determinant of the rice mating system, greatly contributes to the application of heterosis in rice. Although a few quantitative trait loci associated with stigma exsertion have been fine mapped or cloned, the underlying genetic architecture remains unclear. We performed a genome-wide association study on stigma exsertion and related floral traits using 6.5 million SNPs characterized in 533 diverse accessions of Oryza sativa. We identified 23 genomic loci that are significantly associated with stigma exsertion and related traits, three of which are co-localized with three major grain size genes GS3, GW5, and GW2. Further analyses indicated that these three genes affected the stigma exsertion by controlling the size and shape of the spikelet and stigma. Combinations of GS3 and GW5 largely defined the levels of stigma exsertion and related traits. Selections of these two genes resulted in specific distributions of floral traits among subpopulations of O. sativa. The low stigma exsertion combination gw5GS3 existed in half of the cultivated rice varieties; therefore, introducing the GW5gs3 combination into male sterile lines is of high potential for improving the seed production of hybrid rice.

Mapping and application of the twin-grain1 gene in rice.

The map-based cloning and application of a flower organ number gene twin - grain1 provide great potential for improving seed production in hybrid rice. A new germplasm for high-yield rice breeding, the twin-grain1 (tg1) mutant with more than one grain in a glume, was obtained from the Zhejing 22 rice variety via physical mutagenesis. The mapping results showed that TG1 is allelic to FLORAL ORGAN NUMBER2 (FON2)/FLORAL ORGAN NUMBER4 (FON4), a flower organ number gene located at 88.7cM on chromosome 11. The novel tg1 gene allele was introgressed into the cytoplasmic male sterility (CMS) line Zhejing 22A, giving rise to a new CMS line Zhejing 22-tg1A. The Zhejing 22-tg1A line showed enhanced glume opening and stigma exsertion, which increased the outcrossing rate in hybrid rice. A small-scale hybrid rice seed production test demonstrated that the grain yield of the Zhejing 22-tg1A/Zhejinghui 5 line was significantly increased compared to that of the Zhejing 22A/Zhejinghui 5 line. The plot yield evaluation of the F1 hybrid lines showed a higher yield for the Zhejing 22-tg1A/Zhejinghui 5 line than that of the Zhejing 22A/Zhejinghui 5 line. The results implied great potentials for the tg1 gene inhybridrice breeding.