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Enhanced neovasculogenesis, especially vasculogenic mimicry (VM), contributes to the development of triple-negative breast cancer (TNBC). Breast tumor-initiating cells (BTICs) are involved in forming VM; however, the specific VM-forming BTIC population and the regulatory mechanisms remain undefined. We find that tumor endothelial marker 8 (TEM8) is abundantly expressed in TNBC and serves as a marker for VM-forming BTICs. Mechanistically, TEM8 increases active RhoC level and induces ROCK1-mediated phosphorylation of SMAD5, in a cascade essential for promoting stemness and VM capacity of breast cancer cells. ASB10, an estrogen receptor ERα trans-activated E3 ligase, ubiquitylates TEM8 for degradation, and its deficiency in TNBC resulted in a high homeostatic level of TEM8. In this work, we identify TEM8 as a functional marker for VM-forming BTICs in TNBC, providing a target for the development of effective therapies against TNBC targeting both BTIC self-renewal and neovasculogenesis simultaneously., Vasculogenic mimicry (VM) contributes to the development of triple-negative breast cancer. In this study, the authors show that TEM8 is expressed in VM-forming breast cancer stem cells and it promotes stemness and VM differentiation capacity through a RhoC/ROCK1/SMAD5 axis

Stents made of nearly equiatomic NiTi alloy are used to treat malignant obstruction caused by cancer, but prevention of re-obstruction after surgery is still a challenge because the bare stents possess poor anticancer and antibacterial properties to inhibit cancer/bacteria invasion. The present work aims at endowing the NiTi alloy with anticancer and antibacterial abilities by surface modification. Ni–Ti–O nanoporous layers with different thicknesses were prepared on NiTi by anodization, and biological experiments were conducted to evaluate the effects on gram-positive Staphylococcus aureus, human lung epithelial cancer cells (A549), as well as human endothelial cells (EA. hy926). The nanoporous layer with a thickness of 10.1 μm inhibits growth of cancer cells and kill bacteria but shows little adverse effects on normal cells. Such selectivity is related to the larger amount of Ni ions leached from the sample in the acidic microenvironment of cancer cells in comparison with normal cells. The Ni–Ti–O nanoporous layers are promising as coatings on NiTi stents to prevent re-obstruction after surgery.

Stauntonia obovatifoliola Hayata subsp. urophylla is perennial, evergreen vine producing large edible fruits commonly known as “Jiuyuehuang” (September yellow) in China. In fact, rich nutritional value and favorite taste of S. obovatifoliola Hayata subsp. urophylla give it new potential as a new edible fruit crop. Investigation on genetic diversity is the basis for the conservation and utilization of genetic resources in agriculture. In the current study, we compared 16 morphological traits including single fruit weight (SFW), fruit length (FL), fruit width (FW), peel thickness (PT), peel weight (PW), soluble solids content (SSC), seed weight, edible ratio (ER), hundred-seed weight, seed length, seed width, seed number, petiole length of compound leaf, petiolule length, leaflet length and leaflet width of S. obovatifoliola Hayata subsp. urophylla populations collected from four regions in Hunan Province, China. The results showed that the fruit weight in the four populations ranged from 70.38 to 263.27 g with an average of 155.99 g, FL and FW ranged from 69.85 to 154.16 mm and 40.54 to 63.38 mm, respectively. The SSC and ER varied from 10.10 to 20.60% and 24.84 to 68.14%, respectively. ANOVA indicated that the variances of SFW, FL, FW, PT, and PW that related to fruit traits between populations were greater than within populations while the fruit traits SSC, ER has no significant differences between populations. The fruit weight showed highly positive correlations with FL (r = 0.89, p

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus is highly contagious and causes lymphocytopenia, but the underlying mechanisms are poorly understood. We demonstrate here that heterotypic cell-in-cell structures with lymphocytes inside multinucleate syncytia are prevalent in the lung tissues of coronavirus disease 2019 (COVID-19) patients. These unique cellular structures are a direct result of SARS-CoV-2 infection, as the expression of the SARS-CoV-2 spike glycoprotein is sufficient to induce a rapid (approximately 45.1 nm/sec) membrane fusion to produce syncytium, which could readily internalize multiple lines of lymphocytes to form typical cell-in-cell structures, remarkably leading to the death of internalized cells. This membrane fusion is dictated by a bi-arginine motif within the polybasic S1/S2 cleavage site, which is frequently present in the surface glycoprotein of most highly contagious viruses. Moreover, candidate anti-viral drugs could efficiently inhibit spike glycoprotein processing, membrane fusion, and cell-in-cell formation. Together, we delineate a molecular and cellular rationale for SARS-CoV-2 pathogenesis and identify novel targets for COVID-19 therapy.

The interspecies relationships within Actinidia remain controversial due to lack of sufficient taxon sampling and molecular markers. Intergenic spacers (IGS) in the chloroplast genome have been shown to yield substantial variability and were successfully used in phylogenetic study in diverse plants. Hence, in this study, we develop IGS markers by comparing the available chloroplast genomes of Actinidia to explore the interspecific relationships within Actinidia. A total of 46 species were sampled, and three phylogenetic analyses methods (MP, ML and Bayesian) were used. Compared to the plastomes of other congeneric species, twenty-four highly variable intergenic spacers are identified, and four intergenic spacers with high polymorphism [ndhF-rps32, ndhC-trnV (UAC), rps16-trnQ (UUG) and trnE (UUC)-trnT (GGU)] were used to reconstruct phylogenetic relationships within Actinidia. The phylogeny reveals two main groups within Actinidia, and the four traditionally recognized sections are not monophyletic. Two sections (Leiocarpae and Maculatae) can be proposed, among which Leiocarpae is the earliest diverged lineage in the genus. As compared to previous work, our study provides a robust phylogenetic hypothesis that aids in our understanding of the interspecific relationships of Actinidia. Several closely related species are placed in disparate parts of the phylogeny. Moreover, the same species sampled at least two varieties are polyphyletic in our cpDNA trees, which suggests frequent hybridization/introgression events occurred in the genus of Actinidia.

Angiosperm mitochondrial genomes (mitogenomes) are notable for their extreme diversity in both size and structure. However, our current understanding of this diversity is limited, and the underlying mechanism contributing to this diversity remains unclear. Here, we completely assembled and compared the mitogenomes of three kiwifruit (Actinidia) species, which represent an early divergent lineage in asterids. We found conserved gene content and fewer genomic repeats, particularly large repeats (>1 kb), in the three mitogenomes. However, sequence transfers such as intracellular events are variable and dynamic, in which both ancestral shared and recently species-specific events as well as complicated transfers of two plastid-derived sequences into the nucleus through the mitogenomic bridge were detected. We identified extensive whole-genome rearrangements among kiwifruit mitogenomes and found a highly variable V region in which fragmentation and frequent mosaic loss of intergenic sequences occurred, resulting in greatly interspecific variations. One example is the fragmentation of the V region into two regions, V1 and V2, giving rise to the two mitochondrial chromosomes of Actinidia chinensis. Finally, we compared the kiwifruit mitogenomes with those of other asterids to characterize their overall mitogenomic diversity, which identified frequent gain/loss of genes/introns across lineages. In addition to repeat-mediated recombination and import-driven hypothesis of genome size expansion reported in previous studies, our results highlight a pattern of dynamic structural variation in plant mitogenomes through global genomic rearrangements and species-specific fragmentation and mosaic loss of intergenic sequences in highly variable regions on the basis of a relatively large ancestral mitogenome.

Akebia trifoliata is perennial and deciduous vines, its fruit, seed, stem and root have been used in Chinese herbalism for at least 2000 years. As a high-value medicinal, the A. trifoliata should be exploited and domestication is underway as edible fruit crop. In the present study, a wild seedlings population and a three-generation selected population have been evaluated and characterized for a set of morphological traits in order to evaluate the effectiveness of recurrent selection method in achieving selection progress, maintaining genetic variability, and increasing the potential for selection of superior genotypes in A. trifoliata. The result showed that the fruit weight increased 33.16% which ranged from 237.67 to 598.54 g with an average of 310.86 g in the selected population. The leaf of A. trifoliata in the selected population became much larger range from 91.24 to 170.43 mm with an average of 117.33 mm, the leaf length, leaf width and petiole length increased 15.72%, 7.91% and 9.33%, respectively. Correlation analysis showed most of the measured characters are highly correlated each other. The principal component analysis (PCA) indicated that the first five components can explain 83.90% of the total variance. Cluster analysis revealed two distinct clusters among the selected population. The present study showed the effectiveness of the recurrent selection method when applied in A. trifoliata breeding, although some refinements in the selection program could further improve its efficiency.

The species of Akebia Decne are well-known medicinal plants and have been used in Chinese herbalism for at least 2000 years. The Akebia fruits have a delicious flavor and high nutritional value and are especially rich in antineoplastic, diuretic and antiphlogistic properties, making them worthy of being exploited as a new high-value crop. However, the nature habitats of the Akebia species have been fragmented due to rapid and intensive expansion of agricultural land. In this study, population genetics based on simple sequence repeat markers were conducted on the three most economically important Akebia species in order to reveal the genetic effects of habitat fragmentation to gain useful information for conservation and management of genetic resources of Akebia. Three Akebia species maintained high within-population genetic variation and low level of genetic differentiation. Mantel’s test indicated that there was a significantly positive correlation between genetic distance and geographic distance. Our results reveal the resilience of genetic diversity within extant populations of three Akebia species to chronic forest fragmentation. For breeding purpose, germplasm collection for sampling as many individuals as possible from many populations to maximize genetic diversity of the species is recommended. In addition, natural introgression events have been revealed by Cluster and Structure analysis where admixed ancestry occurred across three marginal populations of A. trifoliata ssp. australis, suggesting these populations should be given priority for germplasm collection.

Background Plant phylogeographic studies of species in subtropical China have mainly focused on rare and endangered species, whereas few studies have been conducted on taxa with relatively wide distribution, especially polyploid species. We investigated the cytotype and haplotype distribution pattern of the Actinidia chinensis complex, a widespread geographically woody liana with variable ploidy in subtropical China comprising two varieties, with three chloroplast fragments DNA (ndhF-rpl132, rps16-trnQ and trnE-trnT). Macroevolutionary, microevolutionary and niche modeling tools were also combined to disentangle the origin and the demographic history of the species or cytotypes. Results The ploidy levels of 3338 individuals from 128 populations sampled throughout the species distribution range were estimated with flow cytometry. The widespread cytotypes were diploids followed by tetraploids and hexaploids, whereas triploids and octoploids occurred in a few populations. Thirty-one chloroplast haplotypes were detected. The genetic diversity and genetic structure were found to be high between varieties (or ploidy races) chinensis and deliciosa. Our results revealed that these two varieties inhabit significantly different climatic niche spaces. Ecological niche models (ENMs) indicate that all varieties’ ranges contracted during the Last Inter Glacial (LIG), and expanded eastward or northward during the Last Glacial Maximum (LGM). Conclusions Pliocene and Plio-Pleistocene climatic fluctuations and vicariance appear to have played key roles in shaping current population structure and historical demography in the A. chinensis complex. The polyploidization process also appears to have played an important role in the historical demography of the complex through improving their adaptability to environmental changes.

The whole complete chloroplast (cp) genome sequence of Actinidia kolomikta (formerly A. maloide) was sequenced and assembled from Illumina paired-end sequencing. The cp genome of A. arguta was 157,425 bp long, containing a large single copy region (LSC) of 88,498 bp and a small single copy region (SSC) of 20,475 bp and a pair of inverted repeat regions (IR) of 242,266 bp. It contained 113 different genes, including 79 protein-coding genes, 30 tRNA genes, 4 ribosomal RNA genes. Phylogenetic analysis using concatenated alignments of whole cp genome sequences revealed that A. kolomikta was the sister group to all other groups of Actinidia

Plastome sequences are rich sources of information for resolving difficult phylogenetic relationships and provide genomic data for conservation studies. Here, the complete plastome sequence of Alniphyllum eberhardtii Guillaumin is reported, representing the first plastome of the basal asterid family Styracaceae (Ericales). The plastome is 155,384 bp in length and contains 79 protein-coding genes, 30 tRNA genes and 4 rRNA genes, totaling 113 unique genes with 19 genes in the inverted repeat region. Unusual features of the plastome include the presence a large 20-kb inversion in the Large Single-Copy region, the pseudogenization of the accD gene, and the loss of the second intron from clpP. The 20-kb inversion includes 14 genes and has not been previously reported in other Ericales plastomes. Thirty-nine plastid simple sequence repeats (SSRs) that may provide genetic resources for the conservation of this economically import timber plant are characterized. Phylogenetic results inferred from ML and MP analyses of 66 plastid genes and 26 taxa reveal that the Styracaceae are sister to a clade including Actinidiaceae and Ericaceae and suggest that complete plastomes are likely to be very helpful in resolving the basal relationships among Ericales families, which have resisted resolution in smaller phylogenetic data sets.

Torreya grandis Fort. ex Lindl. cv. Merrillii (Taxaceae) is an important economic tree species grown for dried nuts in China. However, it was listed as national second-grade key protected wild plants due to overexploitation of wild resources. To enhance knowledge about genetic characteristics useful for the conservation and sustainable utilization of genetic resource of this species, microsatellite loci were developed by analyzing a reference transcriptome de novo assembled from RNA-seq data of T. grandis. A total of 1713 SSRs containing repeats from di-to pentanucleotides were detected in 46,000 unigenes. Trinucleotide (46.9%) was the most common repeat unit followed by the di (19.7%), pentanucleotides (15.1%), hexa (8.9%), and tetra (6.5%). From these, 108 primer pairs were designed, among which nineteen polymorphic microsatellites were developed and characterized. The number of alleles per locus ranged from 2 to 4, with an average of 2.6 alleles per locus. The observed and expected heterozygosity ranged from 0.10 to 1.00 and 0.13 to 0.70, respectively. Six loci showed significant departure from HWE after Bonferroni correction. The microsatellite markers described here will be useful for evaluating the genetic structure of T. grandis as well as to estimate fine-scale gene flow rates.

Akebia trifoliata belongs to the Lardizabalaceae family of flowering plants, has tremendous potential as a new fruit crop for further domestication and commercialization in China. However, the selection of A. trifoliata is extremely limited due to lack of information about basic inheritance of fruit traits. In this study, 11 key fruit characters, including single fruit weight (SFW), fruit length (FL), fruit width (FW), peel thickness (PT), peel weight (PW), soluble solids content (SSC), seed weight (SW), pulp weight (PuW), edible ratio (ER), hundred-grain weight (HGW) and seed number (SN) were evaluated, and genetic parameters were analyzed. The results showed that the 11 tested traits have high repeatability range from 0.87 to 0.95. Correlation analysis showed that SFW was positively correlated with FL, FW, PT, PW and PuW, but negatively correlated with ER. The Smith–Hazel-index based multitrait selection was used to select superior lines. The SH selection index identified superior lines with high SFW and ER, and low SN. Taken together, our results provide important information for domestication and breeding program of Akebia trifoliate.

Coating materials applied for intraosseous implants must be optimized to stimulate osseointegration. Osseointegration is a temporal and spatial physiological process that not only requires interactions between osteogenesis and angiogenesis but also necessitates a favorable immune microenvironment. It is now well-documented that hierarchical nano-micro surface structures promote the long-term stability of implants, the interactions between nano-micro structure and the immune response are largely unknown. Here, we report the effects of microporous titanium (Ti) surfaces coated with nano-hydroxyapatite (HA) produced by micro-arc oxidation and steam-hydrothermal treatment (SHT) on multiple cell behavior and osseointegration. By altering the processing time of SHT it was possible to shift HA structures from nano-particles to nano-rods on the microporous Ti surfaces. Ti surfaces coated with HA nano-particles were found to modulate the inflammatory response resulting in an osteoimmune microenvironment more favorable for osteo-/angio-genesis, most likely via the activation of certain key signaling pathways (TGF-β, OPG/RANKL, and VEGF). By contrast, Ti surfaces coated with nano-rod shaped HA particles had a negative impact on osteo-/angio-genesis and osteoimmunomodulation. In vivo results further demonstrated that Ti implant surfaces decorated with HA nano-particles can stimulate new bone formation and osseointegration with enhanced interaction between osteocytes and implant surfaces. This study demonstrated that Ti implants with micro-surfaces coated with nano-particle shaped HA have a positive impact on osseointegration. Statement of Significance Osteo-/angio-genesis are of importance during osteointegration of the implants. Recent advances unravel that immune response of macrophages and its manipulated osteoimmunomodulation also exerts a pivotal role to determine the fate of the implant. Surface nano-micro modification has evidenced to be efficient to influence osteogenesis, however, little is known links nano-microstructured surface to immune response, as well the osteoimmunomodulation. This study demonstrates that the nano-particles decorated micro-surface, compared with the nano-rods decorated micro-surface enables osteogenesis and angiogenesis concurrently that has not been investigated previously. This study also unravels that the immune response of macrophages can be manipulated by the nano-micro surface, especially the nano-dimension matters, leading to a differential effect on osteointegration. The additional knowledge obtained from this study may provide foundation and reference for future design of the coating materials for implantable materials.

The complete chloroplast (cp) genome sequence of Actinidia eriantha was sequenced and assembled using Illumina paired-end data. The cp genome from A. eriantha is 156,964 bp in length, compo...

Kiwifruit is a perennial horticultural crop species of the Actinidiaceae family and has high nutritional value. For a species with a long generation time, traditional breeding and genetic improvement is predicted to take more than 20 years to obtain superior cultivars. Thus, marker-assisted selection (MAS) should be used to accelerate the breeding process. Development of a genetic linkage map and molecular markers are pre-requisites for MAS of crop species. Here, we report a genome-wide SNP-based genetic map of kiwifruit by analysing next-generation restriction-site-associated DNA sequencing (RADseq) reads. To construct a genetic linkage map, a 102 F1 line mapping population of Actinidia chinensis (2n = 58) was derived by combining parents that had contrasting phenotypic traits. The maternal map contained 4112 SNP loci and spanned a distance of 3821 cM, with an average adjacent-marker interval length of 0.929 cM. The map length of the 29 linkage groups ranged from 78.3 to 169.9 cM, with an average length of 131.8 cM. High levels of collinearity between the 29 genetic maps with the kiwifruit reference genome were found. The genetic map developed in this study can serve as an important platform to improve kiwifruit research, including anchoring unmapped scaffolds of the kiwifruit genome sequence and mapping QTLs (quantitative trait loci) that control economically important traits.

Kiwifruit (Actinidia chinensis Planchon) is an important specialty fruit crop that suffers from narrow genetic diversity stemming from recent global commercialization and limited cultivar improvement. Here, we present high-density RAD-seq-based genetic maps using an interspecific F1 cross between Actinidia rufa 'MT570001' and A. chinensis 'Guihai No4'. The A. rufa (maternal) map consists of 2,426 single-nucleotide polymorphism (SNP) markers with a total length of 2,651 cM in 29 linkage groups (LGs) corresponding to the 29 chromosomes. The A. chinensis (paternal) map consists of 4,214 SNP markers over 3,142 cM in 29 LGs. Using these maps, we were able to anchor an additional 440 scaffolds from the kiwifruit draft genome assembly. Kiwifruit is functionally dioecious, which presents unique challenges for breeding and production. Three sex-specific simple sequence repeats (SSR) markers can be used to accurately sex type male and female kiwifruit in breeding programmes. The sex-determination region (SDR) in kiwifruit was narrowed to a 1-Mb subtelomeric region on chromosome 25. Localizing the SDR will expedite the discovery of genes controlling carpel abortion in males and pollen sterility in females.

Interspecific hybridization is widespread among plants. Understanding the phylogenetic relationships among species is necessary for revealing the potential hybridization events. Actinidia, best known as kiwifruit genus found throughout a wide range in eastern Asian from Indonesia to Siberia. In this study, phylogenetic relationships of Actinidia species with sympatric distributions were investigated using three chloroplast introns (trnL-F, atpB-rbcL and rpl32-trnL) and three Exon primed intron-crossing (EPIC) markers. Chloroplast phylogeny supports non-monophyly of the five species studied excluding Actinidia fulvicoma var. lanata. The non-monophyly was also revealed by EPIC markers. Our results showed EPIC markers are more variable and informative for phylogenetic inference than that of chloroplast markers. The incongruences between loci from the plastid and nuclear DNA phylogenic trees may stem from incomplete lineage sorting or historical introgression hybridization. Incomplete lineage sorting may explain the non-monophyly between Actinidia chrysantha (section Maculatae) and other four species (section Stellatae), and introgression hybridization and high level of interspecific gene flow may explain the non-monophyly among the species of sect. Stellatae. Thus, natural hybridization and introgression may be common in Actinidia with sympatric distribution.

Actinidia eriantha Benth. is a diploid perennial woody vine native to China and is recognized as a valuable species for commercial kiwifruit improvement with high levels of ascorbic acid as well as having been used in traditional Chinese medicine. Due to the lack of genomic resources for the species, microsatellite markers for population genetics studies are scarce. In this study, RNASeq was conducted on fruit tissue of A. eriantha, yielding 5,678,129 reads with a total output of 3.41 Gb. De novo assembly yielded 69,783 non-redundant unigenes (41.3 Mb), of which 21,730 were annotated using protein databases. A total of 8,658 EST-SSR loci were identified in 7,495 unigene sequences, for which primer pairs were successfully designed for 3,842 loci (44.4 %). Among these, 183 primer pairs were assayed for PCR amplification, yielding 69 with detectable polymorphism in A. eriantha. Additionally, 61 of the 69 polymorphic loci could be successfully amplified in at least one other Actinidia species. Of these, 14 polymorphic loci (mean NA = 6.07 ± 2.30) were randomly selected for assessing levels of genetic diversity and population structure within A. eriantha. Finally, a neighbor-joining tree and Bayesian clustering analysis showed distinct clustering into two groups (K = 2), agreeing with the geographical distributions of these populations. Overall, our results will facilitate further studies of genetic diversity within A. eriantha and will aid in discriminating outlier loci involved in local adaptation.

Relationships among the genera of the small, woody family Styracaceae and among families of the large, diverse order Ericales have resisted complete resolution with sequences from one or a few genes. We used plastome sequencing to attempt to resolve the backbone relationships of Styracaceae and Ericales and to explore plastome structural evolution. Complete plastomes for 23 species are newly reported here, including 18 taxa of Styracaceae and five of Ericales (including species of Sapotaceae, Clethraceae, Symplocaceae, and Diapensiaceae). Combined with publicly available complete plastome data, this resulted in a data set of 60 plastomes, including 11 of the 12 genera of Styracaceae and 12 of 22 families of Ericales. Styracaceae plastomes were found to possess the quadripartite structure typical of angiosperms, with sizes ranging from 155 to 159 kb. Most of the plastomes were found to possess the full complement of typical angiosperm plastome genes. Unusual structural features were detected in plastomes of Alniphyllum and Bruinsmia, including the presence of a large 20-kb inversion (14 genes) in the Large Single-Copy region, the loss or pseudogenization of the clpP and accD genes in Bruinsmia, and the loss of the first exon of rps16 in B. styracoides. Likewise, the second intron from clpP was found to be lost in Alniphyllum and Huodendron. Phylogenomic analyses including all 79 plastid protein-coding genes provided improved resolution for relationships among the genera of Styracaceae and families of Ericales. Styracaceae was strongly supported as monophyletic, with Styrax, Huodendron, and a clade of Alniphyllum + Bruinsmia successively sister to the remainder of the family, all with strong support. All genera of Styracaceae were recovered as monophyletic, except for Halesia and Pterostyrax, which were each recovered as polyphyletic with strong support. Within Ericales, all families were recovered as monophyletic with strong support, with Balsaminaceae sister to remaining Ericales. Most relationships recovered in plastome analyses are congruent with previous analyses based on smaller data sets. Our results demonstrate the power of plastid phylogenomics to improve phylogenetic hypotheses among genera and families, and provide new insight into plastome evolution across Ericales.

Background Kiwifruit is a perennial, deciduous and functionally dioecious plant. However, very little is known about the whole-genome molecular mechanisms contributing to distinct sexual phenotypes. To gain a global view of genes differentially expressed between male and female flowers, we analyzed genome-wide gene expression profiles in the flowers of male and female plants using high-throughput RNA sequencing. Results A total of 53.5 million reads were generated. Based on the alignments of unigenes to kiwifruit genome predicted genes, a total of 39,040 unique genes with a mean length of 970 bp were identified. There were 2,503 UniGenes differentially expressed between female and male flowers, with 1,793 up-regulated and 710 down-regulated in the female flowers. Moreover, the gene expression pattern of 17 out of 19 unigenes differentially expressed between male and female flowers revealed by RNA-Seq was confirmed by real-time quantitative PCR (qRT-PCR). Conclusions Here, we obtained a large number of EST sequences from female and male flowers of kiwifruit. This comparative transcriptome analysis provides an invaluable resource for gene expression, genomics, and functional genomic studies in A. chinensis and its related species. This study also represents a first step toward the investigation of genes involved in kiwifruit sex determination.

Psilopeganum sinense is a perennial herb endemic to Three-Gorges Reservoir Area (TGRA) in China. Genetic diversity of this endangered species was assessed by using 11 nuclear microsatellites and three chloroplast microsatellite (cpSSR) markers. A total of 8 haplotypes were identified in a survey of 212 individuals sampled from nine populations encompassing most of the natural range of the species. A low level of genetic diversity was detected: HE = 0.301 for SSR and HE = 0.28 for cpSSR. Populations were highly differentiated from one another: an AMOVA analysis that showed that 56.3% and 68.2% genetic variation resided between populations based on SSR and cpSSR analysis, respectively, and FST and FSTc (0.467 for SSR and 0.644 for cpSSR, respectively) were high. Significant differences were found between estimates of haplotypic differentiation calculated by using unordered alleles (GSTc = 0.857) and ordered alleles (NSTc = 0.728), which indicated the existence of phylogeographical structure in P. sinense. The indirect ratio of pollen flow/seed flow derived from estimates of haplotypic and nuclear DNA differentiation indicated that gene flow via pollen is less efficient than via seed. Two distinct evolutionary lineages (evolutionary significant units, ESUs) were recognized for P. sinense on the basis of both the PCoA and NCA analysis. Sampling strategies for conserving this endangered plant were discussed.

Understanding the extent and partitioning of crop genetic diversity is necessary for conserving and utilizing their genetic potentials for breeding. In the present study, fluorescence-labeled amplified fragment length polymorphism markers were used to characterize the genetic diversity and relationships of 79 cultivars and also of 122 F1 hybrids which resulted from six kiwifruit interploid crosses. A high level of mean genetic diversity was detected (Hj > 0.22) for all cultivars investigated, without significant differences among diploids (2x), tetraploids (4x), and hexaploids (6x). This suggested that no significant genetic erosion occurred in these cultivars, which were directly selected from natural resources or created from crosses. The Unweighted Pair Group Method with Arithmetic Mean analysis of the genetic dissimilarity between cultivars showed three main groups mostly based on their three ploidy levels. Among these, the red-fleshed cultivars which were originally derived from ‘Hongyang’ clustered into one subgroup of group I, suggesting their unique genetic background despite they were marked as different cultivars used in the current kiwifruit industry. By analyzing the genetic variation of hybrids with variable ploidy levels, our genetic analyses further revealed that interploid crosses can increase the genetic diversity of F1 offsprings, especially from the parental combinations of 6x–2x and 6x–4x, in which both parents showed significant differences in morphology and genetic backgrounds. Based on these findings, strategies were proposed for the conservation and utilization of the current kiwifruit genetic resources for future breeding programs.

Reticulate speciation caused by interspecific hybridization is now recognized as an important mechanism in the creation of biological diversity. However, depicting the patterns of phylogenetic networks for lineages that have undergone interspecific gene flow is challenging. Here we sequenced 25 taxa representing natural diversity in the genus Actinidia with an average mapping depth of 26× on the reference genome to reconstruct their reticulate history. We found evidence, including significant gene tree discordance, cytonuclear conflicts, and changes in genome-wide heterozygosity across taxa, collectively supporting extensive reticulation in the genus. Furthermore, at least two separate parental species pairs were involved in the repeated origin of the hybrid lineages, in some of which a further phase of syngameon was triggered. On the basis of the elucidated hybridization relationships, we obtained a highly resolved backbone phylogeny consisting of taxa exhibiting no evidence of hybrid origin. The backbone taxa have distinct demographic histories and are the product of recent rounds of rapid radiations via sorting of ancestral variation under variable climatic and ecological conditions. Our results suggest a mode for consecutive plant diversification through two layers of radiations, consisting of the rapid evolution of backbone lineages and the formation of hybrid swarms derived from these lineages.

Eucommia ulmoides, a traditional Chinese medicinal plant, is endangered as a consequence of long-term and widespread harvest in the late 20th century. It has been widely cultivated as a source of herbal medicine and for use in the organic chemical industry in China. In this study, eight microsatellite markers were applied to investigate genetic diversity in E. ulmoides. Three hundred individuals from one semi-wild population and nine cultivated populations across its main production area were collected. A high level of genetic diversity at population levels (HE = 0.716) was observed. The highly outcrossed mating system, high longevity of E. ulmoides and seed admixture may be responsible for high genetic variation within populations. A genetic bottleneck was observed in one population. Populations were only slightly differentiated from one another (FST = 0.063); this was also supported by AMOVA, which revealed that 94.05% of the total variation resided within populations. This is probably attributable to long-distance gene flow mediated by the exchange of seeds by local farmers. Implications of these results for the conservation of genetic resources of E. ulmoides are discussed. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173, 775–785.

a b s t r a c t Myricaria laxiflora was once considered to be restricted to the Three Gorges Reservoir Area (TGRA), China, and was thought to be the only species to become extinct due to the construction of the Three Gorges Dam (TGD). Recently, three natural populations of this species were found along the Yangtze River valley downstream of the TGD. We studied the genetic diversity and genetic differentiation of three remnant populations and nine extirpated populations, the material of which were collected from natural popula- tions before the impoundment of the Three Gorges Reservoir, using inter-simple sequence repeat (ISSR). High levels of genetic diversity were revealed within populations, both at the species (percentage of poly- morphic bands (PPB) = 95.9%) and population level (PPB varied from 58.5% to 80.3%). For the three remnant populations, the Yidu (MYD) and Yichang (MYC) populations showed comparatively higher genetic diver- sity among the 12 populations, whereas the Zhijiang (MZJ) population showed lower genetic variation. 97.96% of total genetic variation of the species was contained in the three remnant populations. The MZJ population appeared to be distinct from the other 11 populations, whereas the MYD and MYC popula- tions had a close genetic relationship with the nine extirpated populations. A moderate level of genetic differentiation was detected among populations by different estimation approaches. The high genetic diversity of the three natural remnant populations indicates that in situ conservation of this species may enable it to successfully maintain its evolutionary potential, especially in the present case that the habitat of transplanted populations are not suitable for individuals' survival.

Species ranges are influenced by past climate oscillations, geographical constraints and adaptive potential to colonize novel habitats at range limits. This study used Liriodendron chinense, an important temperate Asian tree species, as a model system to evaluate the roles of biogeographic history and marginal population genetics in determining range limits. We examined the demographic history and genetic diversity of 29 L. chinense populations using both chloroplast and nuclear microsatellite loci. Significant phylogeographic structure was recovered with haplotype clusters coinciding with major mountain regions. Long-term demographical stability was suggested by mismatch distribution analyses, neutrality tests and ecological niche models (ENM) and suggested the existence of LGM refuges within mountain regions. Differences in genetic diversity between central and marginal populations were not significant for either genomic region. However, asymmetrical gene flow was inferred from central populations to marginal populations, which could potentially limit range adaptation and expansion of L. chinense.

Understanding adaptive genetic variation and its relation to environmental factors are important for understanding how plants adapt to climate change and for managing genetic resources. Genome scans for the loci exhibiting either notably high or low levels of population differentiation (outlier loci) provide one means of identifying genomic regions possibly associated with convergent or divergent selection. In this study, we combined Amplified Fragment Length Polymorphism (AFLP) genome scan and environmental association analysis to test for signals of natural selection in natural populations of Liriodendron chinense (Chinese Tulip Tree; Magnoliaceae) along a latitudinal transect. We genotyped 276 individuals from 11 populations of L. chinense using 987 AFLP markers. Both frequency-based (Dfdist and BayeScan) and correlation-based (MLM) methods were applied to detect outlier loci. Our analyses recovered both neutral and potentially adaptive genetic differentiation among populations of L. chinense. We found moderate genetic diversity within populations and high genetic differentiation among populations with reduced genetic diversity toward the periphery of the species ranges. Nine AFLP marker loci showed evidence of being outliers for population differentiation for both detection methods. Of these, six were strongly associated with at least one climate factor. Temperature, precipitation, and radiation were found to be three important factors influencing local adaptation of L. chinense. The outlier AFLP loci are likely not the target of natural selection, but the neighboring genes of these loci might be involved in local adaptation. Hence, these candidates should be validated by further studies.

Simple sequence repeats (SSRs) have been widely used in the construction of linkage maps, quantitative trait loci (QTLs) mapping, and marker-assisted selection (MAS). The availability of the sequenced Actinidia chinensis (kiwifruit) genome allows for the inexpensive and efficient development of microsatellite markers. In this study, a total of 49,067 SSRs were identified and characterized in the genome sequences of kiwifruit. Dinucleotide repeats are the most abundant SSRs, with the AG/TC motif accounting for 44.2 % of all SSRs in the genome. Fifty-five newly derived SSRs, together with 46 previously available SSRs, were integrated into linkage maps of an interspecific kiwifruit population. In addition, eight sex-linked SSR markers (including one previously published SSR) were mapped in the sex-related region on the LG25, suggesting that recombination is partially suppressed to maintain dioecy in kiwifruit. The SSRs developed from this study are a valuable resource for kiwifruit genetics and will contribute to the use of MAS in early sex determination of dioecious plant breeding.

Plants have three genomes in their cells: the nuclear genome, the mitochondrial genome (mtDNA), and the chloroplast genome (cpDNA). They replicate independently on cell division and independently of the DNA located in the nucleus. There are few mitochondrion genome sequences until now owing to high structure variation and abundant repeat sequences found in the mitochondrion genome when compared to chloroplast genome. In this chapter, we outline some general features of the complete chloroplast genome sequences of Actinidia chinensis var. chinensis and Actinidia chinensis var. deliciosa obtained through de novo assembly of Illumina paired-end reads and produced by total DNA sequencing. The cp genomes share the typical quadripartite cp structure, with a pair of inverted repeats each separated by a small-single-copy region and a large-single-copy region. Gene content is typical for the plastomes of dicotyledonous angiosperms. In Actinidia, only mitochondria are inherited from the mother, while chloroplasts are inherited from the father, with very rare cases of maternal or biparental inheritance. These patterns of inheritance allow interspecies relationships within the genus Actinidia to be investigated using mt and cp DNA sequences.

Sinojackia, a member of the family Styracaceae, is an endangered genus endemic to China. The number of populations and population size of Sinojackia have decreased sharply because of habitat fragmentation and destruction. We studied the genetic diversity of extant populations in two different cohorts (adult and seedling) using eight microsatellite markers to investigate the genetic footprints of habitat fragmentation in four recognized Sinojackia spp. and to develop appropriate conservation measures. Data on intrapopulational genetic diversity suggest that Sinojackia populations have maintained relatively high levels of genetic diversity and low levels of genetic differentiation despite severe fragmentation. The high genetic diversity may be explained by the outcrossing mating system and high longevity of Sinojackia spp. The amount of genetic variation is not associated with population size, which was also supported by bottleneck analysis. In the species studied, there was no significant difference in the genetic diversity between the two cohorts analysed. However, inbreeding increased from adult trees to seedling populations, suggesting that the higher proportion of biparental inbreeding in the recent generations of seedlings is the result of restricted current genetic flow caused by habitat fragmentation. Average seed set per population was not significantly correlated with either population size or genetic diversity. Conservation management should aim to monitor inbreeding and outbreeding depression carefully to ensure the in situ and ex situ conservation of Sinojackia spp. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, ••, ••–••.

Eucommia ulmoides, the only species of Eucommiaceae, has been used as Chinese medicinal plant for more than 2,000 years, and is endangered as a consequence of long-term overexploitation. In this study, genetic diversity within and among the semi-wild and cultivated populations of E. ulmoides collected from its main production area was investigated using two cpSSR and 227 AFLP loci. A moderate level of within-population diversity was observed (hS = 0.549 for cpSSR and H eB = 0.183 for AFLPs), which could be explained by the dioecious breeding system, longevity, and human mediated seed exchange. Compared to the semi-wild population, the cultivated population possessed relatively low genetic diversity, which suggests cultivation practice may have reduced the genetic diversity of E. ulmoides. Populations were low to moderately differentiated from one another (G ST = 0.281 for cpSSR, and θ B = 0.100 for AFLPs), which was also supported by the AMOVA analysis that 62.52 and 83.48 % total variation resided within populations based on cpSSR and AFLP analysis, respectively. An isolation by distance analysis revealed that geographical distances was not significantly correlated with genetic distance. Human activities such as seed selection and seed exchange could account for the genetic structure of cultivated populations. In addition, two genetic clusters were detected by the Structure analysis. For conservation purpose, an ex situ conservation measure for conserving the genetically distant populations to maximize genetic diversity of the species is recommended.

Objective— Inflammation is closely linked to angiogenesis, and Toll-like receptors (TLRs) are the key mediators of inflammatory responses. However, the impact of TLRs on angiogenesis is incompletely understood. In this study, we determined the involvement of TLRs in angiogenesis. Methods and Results— In a mouse model of alkali-induced corneal neovascularization (CNV), we found that CNV was attenuated in TLR4 −/− but not TLR2 −/− mice. Further study revealed that the absence of TLR4 led to decreased production of proangiogenic factors in association with reduced accumulation of macrophages at the site of wounds, which was associated with reduced expression of high-mobility group box-1 (HMGB1) protein, an endogenous ligand for TLR4. Topical application of HMGB1 to the injured cornea promoted CNV with increased macrophage accumulation in wild-type mice but not in TLR4 −/− mice. HMGB1 treatment in vitro also promoted the production of proangiogenic factors by mouse macrophages in a TLR4-dependent manner. Furthermore, antagonists of HMGB1 and TLR4 reduced CNV and macrophage recruitment in the injured cornea of wild-type mice. Conclusion— Our results suggest that the release of HMGB1 in the wounds initiates TLR4-dependent responses that contribute to neovascularization. Thus, targeting HMGB1-TLR4 signaling cascade may constitute a novel therapeutic approach to angiogenesis-related diseases.

Populations of Sinojackia rehderiana are highly threatened and have small and scattered distribution due to habitat fragmentation and human activities. Understanding changes in genetic diversity, the fine-scale spatial genetic structure (SGS) at different life stages and gene flow of S. rehderiana is critical for developing successful conservation strategies for fragmented populations of this endangered species. In this study, 208 adults, 114 juveniles and 136 seedlings in a 50 × 100-m transect within an old-growth forest were mapped and genotyped using eight microsatellite makers to investigate the genetic diversity and SGS of this species. No significant differences in genetic diversity among different life-history stages were found. However, a significant heterozygote deficiency in adults and seedlings may result from substantial biparental inbreeding. Significant fine-scale spatial structure was found in different life-history stages within 19 m, suggesting that seed dispersal mainly occurred near a mother tree. Both historical and contemporary estimates of gene flow (13.06 and 16.77 m) indicated short-distance gene dispersal in isolated populations of S. rehderiana. The consistent spatial structure revealed in different life stages is most likely the result of limited gene flow. Our results have important implications for conservation of extant populations of S. rehderiana. Measures for promoting pollen flow should be taken for in situ conservation. The presence of a SGS in fragmented populations implies that seeds for ex situ conservation should be collected from trees at least 19-m apart to reduce genetic similarity between neighbouring individuals.

Sinojackia xylocarpa is a Chinese endemic species that is extinct in the wild but extant in botanical gardens. Microsatellites were used to investigate the genetic diversity and mating system of this species for future use in a reintroduction program. Ex situ conserved populations of S. xylocarpa maintain intermediate levels of genetic diversity ( H E = 0.570–0.640). However, a general and significant heterozygote excess was found, with a mean F IS of −0.103. S. xylocarpa was determined to be predominantly outcrossing ( t m = 0.992; t s = 1.092). Population size and genetic diversity were found to be positively correlated ( r = 0.991; P = 0.084). Principal coordinate analysis (PCA) suggests that all extant individuals are derived from two source populations. Reintroduction strategies of S . xylocarpa were proposed on the basis of these results.

The Akebia Decne belongs to the Lardizabalaceae family of flowering plants, which contains approximately nine genera and 50 species (Delectis Florae Reipublicae Popularis Sinicae Agendae Academiae Sinicae Edita, 2001). The family is widespread in East Asia, whereas there are two monotypic genera occurring in Chile. The plants of the genus Akebia are perennial, deciduous vines producing large edible fruits commonly known as ‘‘wild bananas’’ in China. The fruit resembles a small bunch of thick bananas that crack open longitudinally when ripe in Chinese lunar August. It is therefore called ‘‘Bayuezha’’ (August crack) by the local people and has been prized for its delicious sweet taste for centuries in China. Species of Akebia are commonly referred to as ‘‘chocolate vine’’ in United States, are grown by gardeners as ornamental climbing vines, are well-known medicinal plants, and have been used in Chinese herbalism for at least 2000 years. Three species (subspecies) (A. quinata, A. trifoliata ssp. trifoliata, and A. trifoliata ssp. australis) are listed in Chinese Pharmacopoeia (Pharmacopoeia Commission of PRC, 2005). The value of the Akebia fruit and its natural compounds with antineoplastic, diuretic, and antiphlogistic properties (Jiangsu New Medical College, 1985) is of great potential for development of an alternative high-value crop. However, few studies on Akebia breeding and cultivation have been conducted and limited information is available apart from a few reports on phytochemical analyses (Gao and Wang, 2006; Kawasaki and Higuchi, 1976a, 1976b). Synthesis of knowledge on these species is needed to provide insights for developing management options that enhance their conservation and contribution to fruit production. Recent investigations suggest that Akebia is worthy of being exploited as a new high-value fruit crop in China for its health benefits and other newly found fruit properties. This article provides a broad review on the most economically important Akebia species and discusses the value of exploring these species for domestication and commercial development.

Systemic anaplastic large cell lymphoma (ALCL) can be divided into two subgroups, anaplastic lymphoma kinase (ALK)-positive and ALK-negative, based on the expression of ALK protein. Expression of this protein is due to genetic alterations of ALK at 2p23. Overall, observations on ALK protein, ALK mRNA, ALK-associated genetic alterations and their relationships, to one another are not often reported in the literature. In this study, we investigated the expression of ALK protein, mRNA and fusion transcripts involving ALK and their relationships in ALCL and analyzed formalin-fixed, paraffin-embedded tissues. Forty-five human cases were analyzed with immunohistochemistry for the ALK protein and RT-PCR for ALK mRNA and seven kinds of ALK involved fusion transcripts. Our results showed that the expression of ALK protein, ALK mRNA and ALK fusion transcripts were significantly related to one another (P0.01). Consistent with the expression of ALK protein, patients presenting with ALK mRNA or ALK involved fusion transcripts were significantly younger than those lacking ALK gene alteration (P0.01). This study demonstrates expression of both ALK protein and ALK mRNA are positively correlated with expression of ALK-associated fusion transcripts. Combined detection of ALK protein, ALK mRNA and ALK fusion transcripts can complement each other to aid in the diagnosis of ALCL.

Eleven polymorphic microsatellite loci were isolated and characterized from an AC-enriched genomic library of Akebia trifoliate ssp. australis. The number of allele per locus ranged from 3 to 14. The observed heterozygosity and expected heterozygosity at population level were 0.196–1.000 and 0.522–0.902, respectively. In addition, this set of microsatellites produced robust cross-species amplification in other two related taxa, suggesting these microsatellite markers should provide a useful tool for genetic and conservation studies of the Akebia species.

Polymorphic simple sequence repeat (SSR) markers were used to investigate the impact of habitat fragmentation on the population structure and gene flow of Changiostyrax dolichocarpa, a critically endangered tree in central China. Intrapopulation genetic diversity, population structure and gene flow in the five extant populations of this species were analysed by eight SSR markers. Intrapopulation genetic diversity results suggest that C. dolichocarpa remnants maintained a relatively high degree of genetic diversity despite severe fragmentation. Low genetic differentiation among populations was found based on Wright's F(ST) and amova analysis. Both the F(ST)-based estimate and private allele method revealed high historical gene flow among the remnant populations. Recent immigrants, detected by assignment tests, tend to decrease from the grandparent generation to the current generation. The potentially highly restricted current gene flow among fragments may render the fragmented populations of C. dolichocarpa at a higher risk of local extinction several generations after fragmentation. Both in situ and ex situ conservation management for the remnant populations of C. dolichocarpa are therefore urgently needed to rescue remaining genetic diversity.

Background and Aims The phylogeography of plant species in sub-tropical China remains largely unclear. This study used Tapiscia sinensis, an endemic and endangered tree species widely but disjunctly distributed in sub-tropical China, as a model to reveal the patterns of genetic diversity and phylogeographical history of Tertiary relict plant species in this region. The implications of the results are discussed in relation to its conservation management. Methods Samples were taken from 24 populations covering the natural geographical distribution of T. sinensis. Genetic structure was investigated by analysis of molecular variance (AMOVA) and spatial analysis of molecular variance (SAMOVA). Phylogenetic relationships among haplotypes were constructed with maximum parsimony and haplotype network methods. Historical population expansion events were tested with pairwise mismatch distribution analysis and neutrality tests. Species potential range was deduced by ecological niche modelling (ENM). Key Results A low level of genetic diversity was detected at the population level. A high level of genetic differentiation and a significant phylogeographical structure were revealed. The mean divergence time of the haplotypes was approx. 1·33 million years ago. Recent range expansion in this species is suggested by a star-like haplotype network and by the results from the mismatch distribution analysis and neutrality tests. Conclusions Climatic oscillations during the Pleistocene have had pronounced effects on the extant distribution of Tapiscia relative to the Last Glacial Maximum (LGM). Spatial patterns of molecular variation and ENM suggest that T. sinensis may have retreated in south-western and central China and colonized eastern China prior to the LGM. Multiple montane refugia for T. sinense existing during the LGM are inferred in central and western China. The populations adjacent to or within these refugia of T. sinense should be given high priority in the development of conservation policies and management strategies for this endangered species.

Actinidia chinensis is an important economic plant belonging to the basal lineage of the asterids. Availability of a complete Actinidia chloroplast genome sequence is crucial to understanding phylogenetic relationships among major lineages of angiosperms and facilitates kiwifruit genetic improvement. We report here the complete nucleotide sequences of the chloroplast genomes for Actinidia chinensis and A. chinensis var deliciosa obtained through de novo assembly of Illumina paired-end reads produced by total DNA sequencing. The total genome size ranges from 155,446 to 157,557 bp, with an inverted repeat (IR) of 24,013 to 24,391 bp, a large single copy region (LSC) of 87,984 to 88,337 bp and a small single copy region (SSC) of 20,332 to 20,336 bp. The genome encodes 113 different genes, including 79 unique protein-coding genes, 30 tRNA genes and 4 ribosomal RNA genes, with 16 duplicated in the inverted repeats, and a tRNA gene (trnfM-CAU) duplicated once in the LSC region. Comparisons of IR boundaries among four asterid species showed that IR/LSC borders were extended into the 5’ portion of the psbA gene and IR contraction occurred in Actinidia. The clap gene has been lost from the chloroplast genome in Actinidia, and may have been transferred to the nucleus during chloroplast evolution. Twenty-seven polymorphic simple sequence repeat (SSR) loci were identified in the Actinidia chloroplast genome. Maximum parsimony analyses of a 72-gene, 16 taxa angiosperm dataset strongly support the placement of Actinidiaceae in Ericales within the basal asterids.

Synucleins are emerging as central players in the fundamental neural processes and in the formation of pathologically insoluble deposits characteristic of neurodegenerative diseases. However, synuclein gamma (SNCG), previously identified as a breast cancer specific gene (BCSG1), is also highly expressed in breast carcinomas, but not expressed in normal or benign breast tissues. We analyzed SNCG gene expression in 93 clinical breast specimens and associated it with clinical outcome. Overall SNCG mRNA expression was detectable in 36% breast cancers. However, 81% of stage III/IV breast cancers were positive for SNCG expression, while only 15% of stage I/II breast cancers were positive for SNCG expression. In contrast, SNCG was undetectable in benign breast lesions. Expression of SNCG in the primary tumor also significantly associated with lymph node involvement and metastasis. There was no significant correlation between SNCG gene expression and age, menstruation, and status of ER, PR, PCNA, and HER-2. Patients whose tumors expressed SNCG had a significantly shorter DFS and a high probability of death when compared with those whose tumors did not express SNCG. The hazard ratio of metastasis or recurrence according to the SNCG status was 4.515 (95% CI, 1,188-17.154; P = 0.027). Cox multivariate analysis showed that SNCG had independent prognostic significance above and beyond conventional variables. This study suggests that the expression of SNCG is an independent predictive marker for recurrence and metastasis in breast cancer progression. SNCG is expected to be a useful marker for breast cancer progression and a potential target for breast cancer treatment.

Spontaneous hybridization in ex situ facilities can undermine the genetic integrity of ex situ collections and potentially contaminate open-pollinated seeds or seedlings destined for the reintroduction of endangered plant species into the wild. In the present study, the potential risk of hybridization between two endangered Chinese endemic species, namely Sinojackia xylocarpa Hu and S. rehderiana Hu, which are naturally allopatric species but were conserved ex situ in Wuhan Botanical Garden (WBG), Wuhan, China, were investigated over three consecutive years from 2003 to 2005. The entire overlapping flowering period of the two species was 14–20 d and the two species shared the same pollinator insects during the entire flowering season in WBG. The floral isolation between the two species was not an issue in the ex situ collection at WBG. The results suggest an opportunity for pollen transfer between species and a potential risk of genetic introgression and loss of genetic identity of open-pollinated seeds produced in the ex situ collection of these two endangered species. An artificial reciprocal cross between S. xylocarpa and S. rehderiana confirmed that the two congener species could readily set seeds, indicating no post-pollination barriers to hybridization and the importance of spatial isolation as a barrier to inter-specific crossing. Therefore, to manage these crossable species with overlapping flowering times and shared pollination vectors in ex situ facilities, spatial isolation should be carefully considered to minimize the possibility of spontaneous hybridization. (Managing editor: Wei Wang)

Twenty-four polymorphic microsatellite loci were isolated and characterized from an AAG-enriched genomic library of Sinojackia xylocarpa. The average allele number of these microsatellites was 3.3 per locus, ranging from two to seven. The observed and expected heterozygosities at population level were 0.10–0.83 and 0.10–1.00, respectively. In addition, successful cross-species amplification of this set of microsatellites in three other species of Sinojackia and a closely related taxon, Changiostyrax dolichocarpa, suggested that this set of microsatellite markers should provide a useful tool for genetic and conservation studies of Sinojackia species and other closely related taxa in the Styracaceae.

Fourteen polymorphic microsatellite loci in the endangered fern Adiantum reniforme var. sinense were developed and characterized using the Fast Isolation by AFLP of Sequences Containing repeats (FIASCO) protocol. Polymorphism of each locus was assessed in a bulked sample of 30 individuals from 8 natural populations. The number of alleles per locus varied from 2 to 11, with an average value of 6.2. The ranges of observed and expected heterozygosity were 0.000–0.895 and 0.226–0.868, respectively. These microsatellite markers provide useful tools for the ongoing conservation genetic studies of Adiantum reniforme var. sinense.

This article reviewed the advances in research of modern biotechnology on kiwifruit, which mostly summarized the leaf culture, stem segment culture, shoot tip culture, endosperm culture, embryo culture, protoplast culture and fusion, gene engineering and so on. The application of DNA molecular mark on genetic diversity and gender identification were also involved. The isolated and cloned target genes in transgenic kiwifruit were mostly storage-resistant and related to fruit development and fruit ripening. Agrobacterium mediated transformation system was widely used, and PEG mediated transformation system or particle bombardment system is also involved for obtaining transgenic kiwifruit. Modern biotechnology is believed to be a major means of kiwifruit breeding, and bring the kiwifruit industry with an exciting future. This paper also put forward some existing problems and prospects for modern biotechnology study of kiwifruit.

A generalized logistic model of individual tree mortality was developed for trembling aspen (Populus tremuloides Michx.), white spruce (Picea glauca (Moench) Voss), and lodgepole pine (Pinus contorta Dougl. ex Loud. var. latifolia Engelm.) in Alberta boreal mixedwood forests based on an empirical data base of permanent sample plots. The model is suitable for observations from unequal remeasurement intervals. The maximum likelihood estimation was used to fit the model, the likelihood ratio test was combined with our understanding of mortality process to select the important variables, and the Hosmer-Lemeshow goodness-of-fit test was conducted to evaluate the fit. The fitted model predicts the survival probability of an individual tree based on the tree diameter at breast height, annual diameter increment, stand basal area, species composition, and site productivity.

† Background and Aims Natural selection and genetic drift are important evolutionary forces in determining genetic and phenotypic differentiation in plant populations. The extent to which these two distinct evolutionary forces affect locally adaptive quantitative traits has been well studied in common plant and animal species. However, we know less about how quantitative traits respond to selection pressures and drift in endangered species that have small population sizes and fragmented distributions. To address this question, this study assessed the relative strengths of selection and genetic drift in shaping population differentiation of phenotypic traits in Psilopeganum sinense, a naturally rare and recently endangered plant species. † Methods Population differentiation at five quantitative traits (QST) obtained from a common garden experiment was compared with differentiation at putatively neutral microsatellite markers (FST) in seven populations of P. sinense. QST estimates were derived using a Bayesian hierarchical variance component method. † Key Results Trait-specific QST values were equal to or lower than FST. Neutral genetic diversity was not correlated with quantitative genetic variation within the populations of P. sinense. † Conclusions Despite the prevalent empirical evidence for QST . FST, the results instead suggest a definitive role of stabilizing selection and drift leading to phenotypic differentiation among small populations. Three traits exhibited a significantly lower QST relative to FST, suggesting that populations of P. sinense might have experienced stabilizing selection for the same optimal phenotypes despite large geographical distances between populations and habitat fragmentation. For the other two traits, QST estimates were of the same magnitude as FST, indicating that divergence in these traits could have been achieved by genetic drift alone. The lack of correlation between molecular marker and quantitative genetic variation suggests that sophisticated considerations are required for the inference of conservation measures of P. sinense from neutral genetic markers.

Premise of the study: The genus Ficus (fig trees) comprises ca. 750 species of trees, vines, and stranglers found in tropical forests throughout the world. Fig trees are keystone species in many tropical forests, and their relationship with host-specific wasp pollinators has received much attention, although many questions remain unresolved regarding the levels of host specificity, cospeciation, and the role of hybridization in fig and wasp speciation. We developed exon-primed intron-crossing (EPIC) markers to obtain phylogenetic resolution needed to address these questions. Methods and Results: Expressed sequence tags (ESTs) from F. elastica were compared to Arabidopsis and Populus genomes to locate introns and to design primers in flanking exons. Primer pairs for 80 EPIC markers were tested in samples from divergent clades within Ficus and the outgroup Poulsenia (Moraceae). Conclusions: Thirty-one EPIC markers were successfully sequenced across Ficus, and 29 of the markers also amplified in Poulsenia, indicating broad transferability within Moraceae. All of the EPIC markers were polymorphic and showed levels of polymorphism similar to that of the widely used internal transcribed spacer (ITS).

Nine polymorphic microsatellite loci were isolated and characterized from an AC-enriched genomic library of Disanthus cercidifolius var. longipes. Microsatellite polymorphism was investigated using 24 individuals from one natural population. The observed number of alleles per locus ranged from two to four. Observed and expected heterozygosities ranged from 0.17 to 0.92 and from 0.16 to 0.72, respectively. These polymorphic microsatellite loci provide useful tools for the ongoing population genetic studies of D. cercidifolius var. longipes.