Increased sugar content impairs pollen fertility and reduces seed-setting in high-photosynthetic-efficiency rice

Crop yield depends on biomass,which is primarily associated with photosynthesis.We previously demonstrated that two photorespiratory bypasses,i.e.,GOC(glycolate oxidase+oxalate oxidase+catalase)and GCGT(glycolate oxidase+catalase+glyoxylate carboligase+tartronic semialdehyde reductase),significantly increased photosynthesis,biomass,and grain yield,but decreased seed-setting rates in rice.This study explored the underlying mechanism of how elevated photosynthetic efficiency impacted the seed-setting.First,pollen germination assessed in vivo and in vitro,revealed a reduced germination rate in GCGT rice.Subsequent analysis found that photosynthates highly accumulated in the leaves and stems;sucrose and soluble sugar levels were increased but the starch level was reduced in the anthers.Uridine diphosphate glucose(UDP-Glc)was increased but uridine diphosphate galactose(UDP-Gal)was unaltered,thus causing an imbalance in the UDP-Glc/UDP-Gal ratio in GCGT anthers.Most anthers in GCGT plants had two locules in contrast to four in the wild-type(WT).Pollen tapetum was developmentally abnormal,and genes related to sucrose synthesis,transport,and tapetal programmed cell death(PCD)were upregulated,whereas those involved in starch synthesis and conversion were downregulated in GCGT anthers.Taken together,our results demonstrated that an increase in sugar content was the primary factor causing reduced seed-setting rates in high photosynthetic efficiency rice,during which metabolic disorder of sugars and UDP sugar imbalance in anthers lead to impaired pollen fertility.

Quantitative Trait Loci Associated with Pollen Fertility under High Temperature Stress at Flowering Stage in Rice (Oryza sativa)

High temperature stress (HTS), an increasingly important problem in rice production, significantly reduces rice yield by reducing pollen fertility and seed setting rate. Breeding rice varieties with tolerance to HTS at the flowering stage is therefore essential for maintaining rice production as the climate continues to become warm. In this study, two quantitative trait loci (QTLs) underlying tolerance to HTS were identified using recombinant inbred lines derived from a cross between an HTS-tolerant rice cultivar 996 and a sensitive cultivar 4628. Pollen fertility was used as a heat-tolerance indicator for the lines subjected to HTS at the flowering stage in field experiments. Two QTLs that affected pollen fertility, qPF4 and qPF6, were detected between RM5687 and RM471 on chromosome 4, and between RM190 and RM225 on chromosome 6, by using the composite interval mapping (CIM) analysis. The two QTLs explained 15.1% and 9.31% of the total phenotypic variation in pollen fertility, and increased the pollen fertility of the plants subjected to HTS by 7.15% and 5.25%, respectively. The positive additive effects of the two QTLs were derived from the 996 alleles. The two major QTLs identified would be useful for further fine mapping and cloning of these genes and for molecular marker-assistant breeding of heat-tolerant rice varieties.

Effect of Temperature on Pollen Fertility in Inter-Subspecific Rice Hybrids

Seventeen rice varieties and hybrids of different types (indica, japonica, javanica, indica hybrid, japonica hybrid and inter-subspecific hybrid) were evaluated to determine the effect of temperature on pollen fertility in inter-subspecfic hybrids. The pollen fertility of inter-subspecific hybrids was greatly reduced when average daily temperature dropped to 22.0 - 23.0℃ at meiosis stage, and the extent of pollen fertility reduction varied greatly with respect to different hybrids. However, the pollen fertility reduction of indica and japonica hybrids and conventional varieties was not obvious under the same regime of temperature conditions. When the average daily temperature dropped to 20℃, the pollen development of conventional varieties and hybrids was also affected. Correlation analysis revealed that there existed a positive correlation between pollen fertility and average daily temperature. A significant difference (P<0.01) was also found between the two correlation coefficients i.e. inter-subspecific hybrids and conventional varieties. Temperature at meiosis stage of pollen mother cell was a key factor in pollen developing, and the pollen fertility of inter-subspecific hybrids was more sensitive to low temperature than that of traditional variety.

Effect of day length and temperature on the pollen fertility in photo(thermo)-sensitive genic male-sterile rice

The effect of day length and temperature on the pollen fertility of five photoperiod-sensitive genic male-sterile japonica rice lines (PGMSR) and three temperature-sensitive genic malesterile indica rice lines (TGMSR) were investigated in phytotron. The light source used for illumination was xenon lamp, and the light intensity which plant accepted on the leaf surface was 300—350μmol photons ms. The results indicated that pollens of PGMSR 7001S and E47S aborted completely whereas a little part of 31116S pollens appeared normal under long day photoperiod (LD,25℃,15h) (Table 1). High temperature (HT, 30℃, 12h) and lower temperature (LT,

Sporophytic control of tapetal development and pollen fertility by a mitogen-activated protein kinase cascade in rice

Tapetum,the innermost layer of the anther wall,provides essential nutrients and materials for pollen development.Timely degradation of anther tapetal cells is a prerequisite for normal pollen development in flowering plants.Tapetal cells facilitate male gametogenesis by providing cellular contents after highly coordinated programmed cell death(PCD).Tapetal development is regulated by a transcriptional network.However,the signaling pathway(s)involved in this process are poorly understood.In this study,we report that a mitogen-activated protein kinase(MAPK)cascade composed of OsYDA1/OsYDA2-OsMKK4-OsMPK6 plays an important role in tapetal development and male gametophyte fertility.Loss of function of this MAPK cascade leads to anther indehiscence,enlarged tapetum,and aborted pollen grains.Tapetal cells in osmkk4 and osmpk6 mutants exhibit an increased presence of lipid body-like structures within the cytoplasm,which is accompanied by a delayed occurrence of PCD.Expression of a constitutively active version of OsMPK6(CA-OsMPK6)can rescue the pollen defects in osmkk4 mutants,confirming that OsMPK6 functions downstream of OsMKK4 in this pathway.Genetic crosses also demonstrated that the MAPK cascade sporophyticly regulates pollen development.Our study reveals a novel function of rice MAPK cascade in plant male reproductive biology.

A Novel C2-Domain Phospholipid-Binding Protein, OsPBP1, Is Required for Pollen Fertility in Rice

Pollen fertility is a crucial factor for successful pollination and essential for seed formation. Recent studies have suggested that a diverse range of internal and external factors, signaling components and their related pathways are likely involved in pollen fertility. Here, we report a single C2-domain containing protein, OsPBP1, initially identified through cDNA microarray analysis. OsPBP1 is a single copy gene and preferentially expressed in pistil and pollen but downregulated by pollination. OsPBP1 had a calcium concentration-dependent phospholipid-binding activity and was localized mainly in cytoplasm and nucleus, but translocated onto the plasma membrane in response to an intracellular Ca^2＋ increase. Pollen grains of antisense OsPBP1 transgenic lines were largely nonviable, germinated poorly in vitro and of low fertility. OsPBP1 protein was localized in a region peripheral to pollen wall and vesicles of elongating pollen tube, and its repressed expression reduced substantially this association and led to alteration of microfilament polymerization during pollen germination. Taken together, these results indicate that OsPBP1 is a novel functional C2-domain phospholipids-binding protein that is required for pollen fertility likely by regulating Ca^2＋ and phospholipid signaling pathways.

Research on Flowering Habits and Fertility of Gaozhou Common Wild Rice (Oryza rufipogon) in Lingshui of Hainan Province

[Objective] The paper was to explore the flowering habits and fertility of Gaozhou common wild rice （Oryza rufipogon Griff.） in Lingshui of Hainan Province. [Method] The flowering habits, pollen fertility and seed setting rate of Gaozhou common wild rice in Lingshui of Hainan Province were studied, and compared with the flowering habits of common wild rice in Hainan Province. [Result] Gaozhou common wild rice was able to flower twice a year. The first flowering stage was from mid-April to mid-June for about 60 d, and the daily peak flowering dates were from May 13 to May 22. The daily flowering time was mainly between 11：20 and 13：00, and the peak flowering was at 11：30. The second flowering stage was from late August to late October for about 45 d, and the peak flowering dates lasted from September 8 to September 18. The daily flowering time was mainly from 11：15 to 12：50, and the daily peak flowering was at 11：30. In both flowering stages, the panicles had the same flowering characteristics. The stigma exsertion reached a high value of 95.37% with single （46.23%） and dual （49.14%） stigmas exsertion respectively. A spikelet remained open for 1.5-2.5 h, the angle of spikelet opening varied from 29.9° to 32.4°. At the two flowering stage the average pollen fertility rates were 89.67% and 85.62%, the average seed setting rates were 58.81% and 55.79%, respectively. Hainan population flowered only once （from mid-October to mid-December）, this had significant difference with the performance of Gaozhou common wild rice which flowered twice a year. [Conclusion] The results provided basis for the research on gene flow frequency from transgenic rice to wild rice and its relationship with flowering habits of common wild rice.

Less and shrunken pollen 1(LSP1) encodes a member of the ABC transporter family required for pollen wall development in rice(Oryza sativa L.)

Pollen fertility is an agronomic trait that strongly influences rice yield. Recent studies have revealed that the development of the pollen wall is required for pollen fertility and is regulated by several genes. However, the mechanisms underlying pollen and pollen wall development in rice remain largely unknown. In the present study, a point mutation in a gene on chromosome 1 was identified that resulted in the production of less and shrunken pollen(LSP) and led to defects in pollen wall formation. This gene was named LSP1 and was found to encode a member of the adenosine triphosphate-binding cassette(ABC)transporter G subfamily, OsABCG3. Two other loss-of-function mutants of LSP1/OsABCG3,generated using CRISPR/Cas9 technology, showed the same male sterile phenotype. The LSP1/OsABCG3 gene showed a spatio-temporal expression pattern in the developing anthers, and is an ortholog of the Arabidopsis genes At ABCG1 and At ABCG16, which play an important role in pollen wall development. Mutation of LSP1/OsABCG3 affected the expression of several genes involved in pollen and pollen wall formation. These results suggest that LSP1/OsABCG3 is critical for normal pollen fertility and shed light on the molecular mechanisms underlying rice pollen wall development.

Research on Pollen Grains of Tetraploid Lines and Diploid Control Line of Chrysanthemum cinerariifolium (Trev.) Vis.

[Objective] The aim was to study characters of pollen grains of tetraploid lines and diploid control line of Chrysanthemum cinerariifolium (Trev.) Vis.,morphological characters,fertility of pollen grain and germination percentage of seeds. [Method] Pollen grains were prepared by sulphuric acid-acetyl oxide decomposition method. The lengths of polar axis and equatorial axis of pollen grains were determined with general optical microscope. The morphology of pollen grains was observed with SEM (scanning electron microscope) and the typical visual fields of 2 500× (or 2 000×),7 000× were taken pictures. [Result] Comparing with the diploid control line,the pollen grains of five tetraploid lines which were tested were different from the diploid line in morphology,sculpture,etc.. 4 of the 5 tested samples were significant larger than the diploid line in size and one was similar to the diploid line. [Conclusion] This research provided references for breeding tetraploid improved varieties of Chrysanthemum cinerariifolium (Trev.) Vis. with good fertility and high germination percentage.

Test on Indica-compatibility of Candidate Indicacompatible Japonica Lines in Rice

[Objective] The aim was to breed the indica-compatible japonica lines（ICJLs）.[Method] The indica-compatibility of 5 candidate ICJLs were tested based on the pollen fertility and spikelet fertility of F1 hybrids with 6 indica testers,6 japonica testers and 6 middle testers.[Result] Candidate ICJL G2123,G2417,G2410 and G3005 were grouped into the exceptionally（Specific）indica-compatible lines based on their high indica-compatibility and low japonica-compatibility.Candidate ICJL G2615 was grouped into the non-compatible lines on their low indica-compatibility and low japonica-compatibility.[Conclusion] The testers,fertility identification standards,test methods of pollen fertility and spikelet fertility were discussed when the compatibility of CICJLs was tested.The reason of the indica-compatibility in G2417 was analyzed.

Perennial aneuploidy as a potential material for gene introgression between maize and Zea perennis

Hybridization,which allows for gene flow between crops,is difficult between maize and Zea perennis.In this study,we aim to initiate and study gene flow between maize and Z.perennis via a special aneuploid plant（MDT） derived from an interspecific hybrid of the two species.The chromosome constitution and morphological characters of MDT as well as certain backcross progenies were examined.Results from genomic in situ hybridization（GISH） indicate that aneuploid MDT consisted of nine maize chromosomes and 30 Z.perennis chromosomes.The backcross progenies of MDTxmaize displayed significant diversity of vegetative and ear morphology;several unusual plants with specific chromosome constitution were founded in its progenies.Some special perennial progeny with several maize chromosomes were obtained by backcrossing MDT with Z.perennis,and the first whole chromosome introgression from maize to Z.perennis was detected in this study.With this novel material and method,a number of maize-tetraploid teosinte addition or substitution lines can be generated for further study,which has great significance to maize and Z.perennis genetic research,especially for promoting introgression and transferring desirable traits.

GPS in pollen tubes: Found!

Sexual reproduction of flowering plants depends on the delivery of two immotile sperm cells to the female gametophyte（FG）,i.e.the embryo sac,through the growth of a pollen tube,a long cylindrical cellular extension from a pollen grain.The journey of pollen tubes toward the embryo sacs is led by female cues,which guide the path of the pollen tubes.

Characterization of a novel DUF1618 gene family in rice

Domain of unknown function （DUF） proteins represent a number of gene families that encode functionally uncharacterized proteins in eukaryotes. For example, DUF1618 family members in plants possess a 56-199-amino acid conserved domain and this family has not been described previously. Here, we report the characterization of 121 DUF1618 genes identified in the rice genome. Based on phylogenetic analysis, the rice DUF1618 family was divided into two major groups, each group consisting of two clades. Most DUF1618 genes with close phylogenetic relationships are located in gene clusters on the chromosomes, indicating that gene duplications increased the number of DUF1618 genes. A search for DUF1618 genes in genomic and/or expressed sequence tag databases for 35 other plant species showed that DUF1618 genes are only present in several monocot plants, suggesting that DUF1618 is a new gene family that originated after the dicot-monocot divergence. Based on public microarray databases, most rice DUF1618 genes are expressed at relatively low levels. Further experimental analysis showed that the transcriptional levels of some DUF1618 genes varied in different cultivars, and some responded to stress and hormone conditions, suggesting their important roles for development and fitness in rice （Oryza sativa L.）.

Long non-coding RNAs play an important role in regulating photoperiod-and temperature-sensitive male sterility in rice

Rice(Oryza sativa L.)is a major food crop for more than half of the world’s population.Therefore,increasing rice production is of great importance.The successful development of hybrid rice in the 1970s was an important achievement for increasing yield potential,since hybrid rice varieties produce higher yields than inbred varieties(Cheng et al.,2007).In plants,male sterility refers to the inability of producing dehiscent anthers and functional pollen(Chen and Liu,2014).