Chromosome-Scale Genome Assembly for Clubrush (Bolboschoenus planiculmis) Indicates a Karyotype with High Chromosome Number and Heterogeneous Centromere Distribution.

Bolboschoenus planiculmis (F.Schmidt) T.V.Egorova is a typical wetland plant in the species-rich Cyperaceae family. This species contributes prominently to carbon dynamics and trophic integration in wetland ecosystems. Previous studies have reported that the chromosomes of B. planiculmis are holocentric; i.e. they have kinetic activity along their entire length and carry multiple centromeres. This feature was suggested to lead to a rapid genome evolution through chromosomal fissions and fusions and participate to the diversification and ecological success of the Bolboschoenus genus. However, the specific mechanism remains uncertain, partly due to the scarcity of genetic information on Bolboschoenus. We present here the first chromosome-level genome assembly for B. planiculmis. Through the integration of high-quality long-read and short-read data, together with chromatin conformation using Hi-C technology, the ultimate genome assembly was 238.01 Mb with a contig N50 value of 3.61 Mb. Repetitive elements constituted 37.04% of the genome, and 18,760 protein-coding genes were predicted. The low proportion of long terminal repeat retrotransposons (∼9.62%) was similar to that reported for other Cyperaceae species. The Ks (synonymous substitutions per synonymous site) distribution suggested no recent large-scale genome duplication in this genome. The haploid assembly contained a large number of 54 pseudochromosomes with a small mean size of 4.10 Mb, covering most of the karyotype. The results of centromere detection support that not all the chromosomes in B. planiculmis have multiple centromeres, indicating more efforts are needed to fully reveal the specific style of holocentricity in cyperids and its evolutionary significance.

The chromosome-scale genome of Kobresia myosuroides sheds light on karyotype evolution and recent diversification of a dominant herb group on the Qinghai-Tibet Plateau.

Kobresia species are common in meadows on the Qinghai-Tibet Plateau. They are important food resources for local livestock, and serve a critical foundation for ecosystem integration. Genetic resources of Kobresia species are scarce. Here, we generated a chromosome-level genome assembly for K. myosuroides (Cyperaceae), using PacBio long-reads, Illumina short-reads, and Hi-C technology. The final assembly had a total size of 399.9 Mb with a contig N50 value of 11.9 Mb. The Hi-C result supported a 29 pseudomolecules model which was in consistent with cytological results. A total of 185.5 Mb (44.89% of the genome) transposable elements were detected, and 26,748 protein-coding genes were predicted. Comparative analysis revealed that Kobresia plants have experienced recent diversification events during the late Miocene to Pliocene. Karyotypes analysis indicated that the fission and fusion of chromosomes have been a major driver of speciation, which complied with the lack of whole-genome duplication (WGD) in K. myosuroides genome. Generally, this high-quality reference genome provides insights into the evolution of alpine sedges, and may be helpful to endemic forage improvement and alpine ecosystem preservation.

Genomic Data Reveals Profound Genetic Structure and Multiple Glacial Refugia in Lonicera oblata (Caprifoliaceae), a Threatened Montane Shrub Endemic to North China.

Characterizing genetic diversity and structure and identifying conservation units are both crucial for the conservation and management of threatened species. The development of high-throughput sequencing technology provides exciting opportunities for conservation genetics. Here, we employed the powerful SuperGBS method to identify 33, 758 high-quality single-nucleotide polymorphisms (SNP) from 134 individuals of a critically endangered montane shrub endemic to North China, Lonicera oblata. A low level of genetic diversity and a high degree of genetic differentiation among populations were observed based on the SNP data. Both principal component and phylogenetic analyses detected seven clusters, which correspond exactly to the seven geographic populations. Under the optimal K = 7, Admixture suggested the combination of the two small and geographically neighboring populations in the Taihang Mountains, Dongling Mountains, and Lijiazhuang, while the division of the big population of Jiankou Great Wall in the Yan Mountains into two clusters. High population genetic diversity and a large number of private alleles were detected in the four large populations, while low diversity and non-private alleles were observed for the remaining three small populations, implying the importance of these large populations as conservation units in priority. Demographic history inference suggested two drastic contractions of population size events that occurred after the Middle Pleistocene Transition and the Last Glacial Maximum, respectively. Combining our previous ecological niche modeling results with the present genomic data, there was a possible presence of glacial refugia in the Taihang and Yan Mountains, North China. This study provides valuable data for the conservation and management of L. oblata and broadens the understanding of the high biodiversity in the Taihang and Yan Mountains.

Characterization of the complete chloroplast genome of Carex myosuroides Villars, 1779 (Cyperaceae).

Carex myosuroides Villars, 1779 is a typical alpine sedge with both ecological and agricultural value. The work reported here is the first complete chloroplast genome of this species. The chloroplast genome, with a total size of 185,609 bp, consists of two inverted repeats (IRs, 38,374 bp) separated by a large single-copy (LSC, 99,911 bp) region, and a small single-copy (SSC, 8950 bp) region. The overall genome GC content is 34.12%. The genome contains 125 genes, consisting of 82 protein-coding genes, 35 tRNA genes, and eight rRNA genes. Phylogenetic analysis supports the taxonomic treatment of incorporating genus Kobresia to a broader circumscription of Carex. Our work could be helpful to future research on Cyperaceae.

Reproductive biology of an endangered lithophytic shrub and implications for its conservation.

BACKGROUND: Plants in cliff habitats may evolve specific reproductive strategies to cope with harsh environments, and unraveling these reproductive characteristics can improve our understanding of survival strategies and lithophyte evolution. This understanding is especially important for efforts to protect rare and endemic plants. Here, we investigated the reproductive biology of Lonicera oblata, an endangered lithophytic shrub that is scattered in highly fragmented and isolated cliff habitats of the Taihang and Yan mountains in North China. RESULTS: Flowers of L. oblata are herkogamous and protandrous, characteristics that can prevent autogamy at the single-flower level, and insects are necessary for pollination. The outcrossing index, pollen/ovule ratio, and the results of hand pollination were measured and all revealed a mixed mating system for L. oblata, that combines cross-fertilization and partial self-fertilization. The floral traits of L. oblata of zygomorphic and brightly yellowish corolla, heavy fragrance, and rich nectar, suggest an entomophilous pollination system. Sweat bees were observed as the most effective pollinators but their visiting frequencies were not high. Pollen limitation may limit the reproductive success of L. oblata. CONCLUSIONS: We determined the reproductive characteristics of L. oblata, a critically endangered species endemic to cliffs in North China, providing insight into its endangerment and suggesting conservation strategies. L. oblata has highly pollinator-dependent self-fertilization as part of a mixed mating system. Floral features such as low-flowering synchrony, asynchronous anthers dehiscence, and high duration of stigma receptivity, improve pollination efficiency in the case of low pollinator service. Our work provides reference information to understand the survival strategies and conservation of L. oblata and other lithophytes.

The complete chloroplast genome of Lonicera oblata, a critically endangered species endemic to North China.

Lonicera oblata, a critically endangered species endemic to North China with about 30 wild individuals, has long been ignored for conservation since its publication because of little attention on its living situation. In this study, we characterized the complete chloroplast (cp) genome of L. oblata. The cp genome was 155,481 bp in length, included a large single-copy (LSC) region of 89,139 bp, a small single-copy (SSC) region of 18,676 bp, and two inverted repeat (IR) regions of 23,833 bp each. The genome contains 130 genes, including 85 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. Phylogenetic position of L. oblata was also investigated based on cp genome phylogeny of Lonicera representatives. This study is valuable for molecular phylogenetic study and conservation of Lonicera and related taxa.

Cloning and functional analysis of glutathione peroxidase gene in red swamp crayfish Procambarus clarkii.

Glutathione peroxidases (GPxs) are key enzymes in the antioxidant defense systems of living organisms, including crustaceans. The red swamp crayfish Procambarus clarkii is the most commonly farmed freshwater crayfish in Chinese inland nowadays due to its commercial value. However, high stocking density has resulted in adverse effects in growth performance and health. To investigate the function of GPxs in immune defense of the crayfish, we cloned and characterized a full length GPx (PcGPx) from P. clarkii by a reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). The 931 bp PcGPx cDNA contains a 38 bp 5'-untranslated region (UTR), a 519 bp coding sequence (CDS) and a 375 bp 3'-UTR with a selenocysteine insertion sequence (SECIS). The PcGPx was predicted to encode 172 amino acids, and its putative molecular mass was 20.9 kDa with a pI of 4.37. A selenocysteine (Sec) encoded by the unusual stop codon, TGA, was in the protein coding region. Phylogenetic analysis showed that PcGPx clustered with the GPxs from the penaeid shrimp Metapenaeus ensis and Caenorhabditis elegans, sharing much higher similarity with vertebrate GPx1 and GPx2 than with GPx3 and GPx5. Quantitative RT-PCR revealed that PcGPx was extremely highly expressed in ovary and early embryos. In addition, the levels of PcGPx mRNA and reactive oxygen species (ROS) significantly increased after challenge with gram-negative Vibrio harveyi, gram-positive Staphyloccocus aureus or white spot syndrome virus (WSSV). These results suggest that PcGPx may play important roles not only in immune defense, but also in oogenesis in the crayfish.