Fu-Min Zhang

Resequencing 50 accessions of cultivated and wild rice yields markers for identifying agronomically important genes.

Rice is a staple crop that has undergone substantial phenotypic and physiological changes during domestication. Here we resequenced the genomes of 40 cultivated accessions selected from the major groups of rice and 10 accessions of their wild progenitors (Oryza rufipogon and Oryza nivara) to >15 × raw data coverage. We investigated genome-wide variation patterns in rice and obtained 6.5 million high-quality single nucleotide polymorphisms (SNPs) after excluding sites with missing data in any accession. Using these population SNP data, we identified thousands of genes with significantly lower diversity in cultivated but not wild rice, which represent candidate regions selected during domestication. Some of these variants are associated with important biological features, whereas others have yet to be functionally characterized. The molecular markers we have identified should be valuable for breeding and for identifying agronomically important genes in rice.

Analysis of 142 genes resolves the rapid diversification of the rice genus

BackgroundThe completion of rice genome sequencing has made rice and its wild relatives an attractive system for biological studies. Despite great efforts, phylogenetic relationships among genome types and species in the rice genus have not been fully resolved. To take full advantage of rice genome resources for biological research and rice breeding, we will benefit from the availability of a robust phylogeny of the rice genus.ResultsThrough screening rice genome sequences, we sampled and sequenced 142 single-copy genes to clarify the relationships among all diploid genome types of the rice genus. The analysis identified two short internal branches around which most previous phylogenetic inconsistency emerged. These represent two episodes of rapid speciation that occurred approximately 5 and 10 million years ago (Mya) and gave rise to almost the entire diversity of the genus. The known chromosomal distribution of the sampled genes allowed the documentation of whole-genome sorting of ancestral alleles during the rapid speciation, which was responsible primarily for extensive incongruence between gene phylogenies and persisting phylogenetic ambiguity in the genus. Random sample analysis showed that 120 genes with an average length of 874 bp were needed to resolve both short branches with 95% confidence.ConclusionOur phylogenomic analysis successfully resolved the phylogeny of rice genome types, which lays a solid foundation for comparative and functional genomic studies of rice and its relatives. This study also highlights that organismal genomes might be mosaics of conflicting genealogies because of rapid speciation and demonstrates the power of phylogenomics in the reconstruction of rapid diversification.

Evolutionary rate patterns of the Gibberellin pathway genes

BackgroundAnalysis of molecular evolutionary patterns of different genes within metabolic pathways allows us to determine whether these genes are subject to equivalent evolutionary forces and how natural selection shapes the evolution of proteins in an interacting system. Although previous studies found that upstream genes in the pathway evolved more slowly than downstream genes, the correlation between evolutionary rate and position of the genes in metabolic pathways as well as its implications in molecular evolution are still less understood.ResultsWe sequenced and characterized 7 core structural genes of the gibberellin biosynthetic pathway from 8 representative species of the rice tribe (Oryzeae) to address alternative hypotheses regarding evolutionary rates and patterns of metabolic pathway genes. We have detected significant rate heterogeneity among 7 GA pathway genes for both synonymous and nonsynonymous sites. Such rate variation is mostly likely attributed to differences of selection intensity rather than differential mutation pressures on the genes. Unlike previous argument that downstream genes in metabolic pathways would evolve more slowly than upstream genes, the downstream genes in the GA pathway did not exhibited the elevated substitution rate and instead, the genes that encode either the enzyme at the branch point (GA20ox) or enzymes catalyzing multiple steps (KO, KAO and GA3ox) in the pathway had the lowest evolutionary rates due to strong purifying selection. Our branch and codon models failed to detect signature of positive selection for any lineage and codon of the GA pathway genes.ConclusionThis study suggests that significant heterogeneity of evolutionary rate of the GA pathway genes is mainly ascribed to differential constraint relaxation rather than the positive selection and supports the pathway flux theory that predicts that natural selection primarily targets enzymes that have the greatest control on fluxes.

Draft genome of the living fossil Ginkgo biloba

BackgroundGinkgo biloba L. (Ginkgoaceae) is one of the most distinctive plants. It possesses a suite of fascinating characteristics including a large genome, outstanding resistance/tolerance to abiotic and biotic stresses, and dioecious reproduction, making it an ideal model species for biological studies. However, the lack of a high-quality genome sequence has been an impediment to our understanding of its biology and evolution.FindingsThe 10.61 Gb genome sequence containing 41,840 annotated genes was assembled in the present study. Repetitive sequences account for 76.58% of the assembled sequence, and long terminal repeat retrotransposons (LTR-RTs) are particularly prevalent. The diversity and abundance of LTR-RTs is due to their gradual accumulation and a remarkable amplification between 16 and 24 million years ago, and they contribute to the long introns and large genome. Whole genome duplication (WGD) may have occurred twice, with an ancient WGD consistent with that shown to occur in other seed plants, and a more recent event specific to ginkgo. Abundant gene clusters from tandem duplication were also evident, and enrichment of expanded gene families indicates a remarkable array of chemical and antibacterial defense pathways.ConclusionsThe ginkgo genome consists mainly of LTR-RTs resulting from ancient gradual accumulation and two WGD events. The multiple defense mechanisms underlying the characteristic resilience of ginkgo are fostered by a remarkable enrichment in ancient duplicated and ginkgo-specific gene clusters. The present study sheds light on sequencing large genomes, and opens an avenue for further genetic and evolutionary research.

Reproductive success of non-rewarding Cypripedium japonicum benefits from low spatial dispersion pattern and asynchronous flowering

BACKGROUND AND AIMS Outcrossing animal-pollinated plants, particularly non-rewarding species, often experience pollinator limitation to reproduction. Pollinator visitation is affected by various factors, and it is hypothesized that reproduction in non-rewarding plants would benefit from low spatial flower abundance and asynchronous flowering. In order to test this hypothesis, the influence of spatial pattern and flowering phenology on male and female reproductive success (RS) was investigated in a non-rewarding orchid, Cypripedium japonicum, in central China over two flowering seasons. METHODS The probabilities of intrafloral self-pollination and geitonogamy caused by pollinator behaviours were estimated from field observations. Pollinator limitation was evaluated by hand-pollination experiments. RS was surveyed in different spatial flower dispersal patterns and local flower densities. The effects of flowering phenological traits on RS were assessed by univariate and multivariate regression analyses. KEY RESULTS Hand-pollination experiments revealed that fruit production was strongly pollen limited throughout the entire reproductive season - over two seasons, 74.3 % of individuals set fruit following hand pollination, but only 5.2-7.7 % did so under natural conditions. Intrafloral self-pollination and geitonogamy within the potential clones might be rare. Both male and female fitness were substantially lower in clustered plants than in those growing singly. An increase in local conspecific flower density significantly and negatively influenced male RS, but had no effect on female RS. Phenotypic selection analysis indicated that individuals flowering earlier have the greatest probability of RS. Over 85 % of sampled flowering individuals had a flowering synchrony value >0.7; however, highly synchronous flowering was not advantageous for RS, as indicated by the negative directional selection differentials and gradients, and by the positive quadratic selection gradients. CONCLUSIONS These results support the hypothesis that, as a consequence of density-dependent selection, low spatio-temporal flower abundance is advantageous for attracting pollinators and for reproduction in natural populations of non-rewarding C. japonicum.

Mountains of Southern China as “Plant Museums” and “Plant Cradles”: Evolutionary and Conservation Insights

Abstract The occurrence of areas or centers of endemism is commonly attributed to the existence of suitable refugia in which plant lineages survived while others evolved during the late Neogene and Quaternary global cooling. In China, several studies performed since the 1980s have identified the mountains of central and southern China as the main centers of endemism in the country. A recent work studied the patterns of endemism separately for palaeoendemics and neoendemics and found that these tend to be located in different mountain ranges. Whereas, young endemics are preferably located in the mountain ranges of the eastern fringe of the Tibetan Plateau (“plant cradles”), old endemics tend to occur in the mountains of central, south central, and southeastern China (“plant museums”), although there are some exceptions. This pattern seems to be related to the different geological history of the mountain ranges. The eastern fringe of the Tibetan Plateau clearly constitutes the “evolutionary front” of China, probably due to the uninterrupted uplift of the plateau from the late Neogene. In contrast, the relative tectonic stability in central and southern China during most of the Tertiary may have maximized the persistence of relict plant lineages. These results have significant implications in the conservation of the endemic flora, which are briefly discussed.

Phylogeny and species delimitation of the C-genome diploid species in Oryza

Abstract  The diploid Oryza species with C‐genome type possesses abundant genes useful for rice improvement and provides parental donors of many tetraploid species with the C‐genome (BBCC, CCDD). Despite extensive studies, the phylogenetic relationship among the C‐genome species and the taxonomic status of some taxa remain controversial. In this study, we reconstructed the phylogeny of three diploid species with C‐genome (Oryza officinalis, O. rhizomatis, and O. eichingeri) based on sequences of 68 nuclear single‐copy genes. We obtained a fully resolved phylogenetic tree, clearly indicating the sister relationship of O. officinalis and O. rhizomatis, with O. eichingeri being the more divergent lineage. Incongruent phylogenies of the C‐genome species found in previous studies might result from lineage sorting, introgression/hybridization and limited number of genetic markers used. We further applied a recently developed Bayesian species delimitation method to investigate the species status of the Sri Lankan and African O. eichingeri. Analyses of two datasets (68 genes with a single sample, and 10 genes with multiple samples) support the distinct species status of the Sri Lankan and African O. eichingeri. In addition, we evaluated the impact of the number of sampled individuals and loci on species delimitation. Our simulation suggests that sampling multiple individuals is critically important for species delimitation, particularly for closely related species.

No Correlation Between Heterozygosity and Vegetative Fitness in the Narrow Endemic and Critically Endangered Clematis acerifolia (Ranunculaceae)

The relationship between heretozygosity and vegetative fitness was explored in the narrow endemic and threatened Clematis acerifolia (Ranunculaceae), both at individual and population levels. The relationships between fitness, habitat factors, and population size were also analyzed. Allozyme electrophoresis was used to quantify the levels of heterozygosity of nearly 200 surveyed individuals belonging to the nine extant populations of this species. Six parameters of vegetative fitness were measured: plant height, shrub diameter, length of the largest leaf, width of the largest leaf, mean number of leaves/stem, and total number of stems. The percentage of tree cover (light availability) was measured as an indicator of habitat quality. A principal component analysis reduced the original fitness variables to two uncorrelated principal components. None of these correlated significantly with both heterozygosity and population size, in contrast to the expected result. Nevertheless, one of the principal components showed a positive relationship with light availability, which may indicate that habitat quality may have significant effects on the performance of this species. Thus, to ensure the viability of this endangered species, maintenance of adequate habitat quality (by avoiding further fragmentation) may be more important than maximizing genetic diversity within populations.

Synthesis of porosity-free nanocrystalline materials with ultrafine grain size by annealing amorphous alloy under high pressure

A porosity-free nanocrystalline Fe735CulNb3Si135B9 alloy with an ultrafine grain size of 4-6 nm has been synthesized. Experiments were carried out by annealing amorphous Fe735CulNb3Si135B9 alloy at a temperature of 823 K under a pressure of 5 GPa in a belt-type pressure apparatus. The microstructure of the samples annealed respectively under high pressure and normal pressure was investigated using X-ray diffraction (XRD), electron diffraction (ED) and transmission electron microscopy (TEM). Analysis results show that the high pressure makes the size of crystalline phase become much smaller,from 10-12 nm for the normal pressure to 4-6 nm for 5 GPa, without changing the lattice type of the crystalline phase, which is attributed to the effects of pressure on the crystallization process of amorphous alloy