Spiders of the genus Sinopoda Jäger, 1999 from Honghe Hani and Yi Autonomous Prefecture, Yunnan Province, China are studied. A total of four species are reported and illustrated of which two, Sinopoda honghe Yu & Zhong, sp. nov . and Sinopoda kuan Yu & Zhong, sp. nov ., are described as new to science. The other two previously described species from this region: S. tengchongensis Fu & Zhu, 2008 and S. tumefacta Zhong, Jäger, Chen & Liu, 2019 are also illustrated. Detailed descriptions, diagnoses, illustrations and DNA barcodes of the two new species are given. A distribution map of these four species in Honghe is provided.

A further study of the spider genus Pseudopoda Jäger, 2000, from Honghe Hani and Yi Autonomous Prefecture, Yunnan, southwest China, is presented. A total of six species are here addressed based on morphology and five methods of molecular species delimitation, increasing the number of known species of the genus in this area from 10 to 13. They comprise three new species, namely P. xiaozhua J. Zhang, H. Yu & Y. Zhong, sp. nov. (♂♀), P. yangae J. Zhang, H. Zhang & Y. Zhong, sp. nov. (♂♀), and P. ying J. Zhang, H. Zhang & Y. Zhong, sp. nov. (♂), and three known species: P. huanglianensis Zhang, Jäger & Liu, 2023, P. mamillaris Zhang, Jäger & Liu, 2023, and P. oliviformis Zhang, Jäger & Liu, 2023. The males of the three known species are described for the first time. Detailed descriptions, diagnoses, photographs of the six species, and a distribution map for all 13 Pseudopoda species in Honghe Prefecture are provided. The DNA barcodes of the six species were obtained for species delimitation, sex matching, and future use.

Background Sparassidae Bertkau, 1872 is the tenth largest family of Araneae, with 11 genera and 288 species known in China.

Background:Euryeidonconsideratum Dankittipakul & Jocqué, 2004 was described, based on a single female from Doi Luang National Park, Thailand and no additional specimens have been recorded since. New information:Euryeidonconsideratum is re-described and illustrated, based on new material from Xishuangbanna, China. The male is described and illustrated for the first time. In addition, this paper further illustrates the female and provides a supplementary description, confirming that the median plate of the epigyne is absent in this species, rather than broken off as suggested in the original publication. The distribution map of this species is given.

Background:Sinopoda Jäger, 1999 is a relatively large spider genus that currently comprises 141 species distributed worldwide. However, the genus remains inadequately studied because nearly half of the species are known from a single sex or juvenile specimens. Sinopodacurva Zhong, Jäger, Chen & Liu, 2019 was described, based on two male specimens from Damingshan National Nature Reserve, Guangxi Zhuang Autonomous Region, China and no additional specimens have been recorded since. New information:Recently, new materials of huntsman spiders have been collected from Mt. Wuyishan, including specimens of both sexes. Several males were identified as S.curva, based on morphological comparison with the holotype. Based on morphological characters and DNA barcodes, we confidently matched the females and males as S.curva. Herein, S.curva is re-described, based on these new materials and the female is described and illustrated for the first time.

Spiders of the family Philodromidae Thorell, 1869 from Guizhou and Hubei provinces, China are studied. A total of three genera and seven species are reported and illustrated, comprising Sinodromuslanyue sp. nov. (newly recorded genus for Hubei), and all known species from both provinces: Philodromusauricomus L. Koch, 1878, P.guiyang Long & Yu, 2022, P.subaureolus Bösenberg & Strand, 1906, P.paiki Jang, Lee, Yoo & Kim, 2024 (the previously records of P.spinitarsis Simon, 1895 from Guizhou and Hubei are presumed to be misidentifications, and should belong to P.paiki), P.rufus Walckenaer, 1826 (new record for Hubei) and Tibellusjaponicus Efimik, 1999 (new record for Hubei). Detailed descriptions, diagnoses, and illustrations of S.lanyue sp. nov. and P.guiyang are given, and the male of P.guiyang is diagnosed and described in English for the first time. The other five species are also re-illustrated. Their DNA barcodes were obtained for species delimitation, matching of sexes and future use.

Background:Selenopsab Logunov & Jäger, 2015 was originally described, based on a male specimen collected from Vietnam, with the female remaining unknown. New information:Based on specimens collected from Shenzhen City, Guangdong Province, the female of Selenopsab Logunov & Jäger, 2015 is described for the first time. This species is newly recorded from China.

Southeast China is one of the core areas of the Sino-Japanese floristic region and a hotspot of biodiversity in East Asia. This region has been considered as both a museum and a cradle of woody genera in China. Why a region with highly stable topography and climate could be a cradle of species diversity remains unclear. In this study, the phylogeographic pattern and genetic structure of Wikstroemia monnula, an endemic species to southeast China, were analyzed by sequencing four chloroplast DNA fragments (4679 bp in total) of 836 individuals from 39 populations. Extremely high diversity and endemism of chlorotypes were found. Out of 54 chlorotypes, 51 (94.44%) were private, with genetic diversity index (H-T) near 1 (H-T = 0.992), and 96.51% of the genetic variation occurring among populations, indicating that this species has undergone strong intraspecific differentiation and very limited migration. The correlational analysis showed that the population differentiation of W. monnula was driven not only via isolation by distance (IBD), but more importantly via isolation by environment (IBE). A significant phylogeographic structure was not found (N-ST = 0.979, G(ST) = 0.833, P > 0.05); however, the spatial pattern of the sublineages of chlorotypes was largely consistent with the biogeographic boundaries on a smaller scale in the region. These results suggest that habitat heterogeneity in southeast China not only promoted speciation on a long time-scale, but still continues to impact population differentiation in recent times.

One new species of the genus Pseudopoda Jäger, 2000, Pseudopodadeformis Gong & Zhong, sp. nov. (♂, ♀), is described and documented with digital images from Shennongjia Forestry District, Hubei Province, China, based on morphology and DNA barcodes. This new species is separated from other Pseudopoda species by the unique type of internal ducts of the female vulva that are curved longitudinally, forming a narrow triangle or trapezoidal shape. In addition, DNA barcodes for this species are provided.

Three species of spider genus Sinopoda Jäger, 1999 are reported from southern China. Two of them are described as new to science: S.guiyang Zhang, Yu & Zhong, sp. nov. and S.xishui Zhang, Yu & Zhong, sp. nov., both from Guizhou Province. The male of S.horizontalis Zhong, Cao & Liu, 2017 is described for the first time based on new material from the type locality, Wuyishan National Nature Reserve, Fujian Province, China. Detailed descriptions, diagnoses, photographs and a distribution map of the three species are provided.

Membrane-localized transporters constitute important components for specialized metabolism in plants. However, due to the vast array of specialized metabolites produced by plants, and the large families of transporter genes, knowledge about the intracellular and intercellular transport of plant metabolites is still in its infancy. Cucurbitacins are bitter and defensive triterpenoids produced mainly in the cucurbits. Using a comparative genomics and multi-omics approach, a MATE gene (CsMATE1), physically clustered with cucurbitacin C (CuC) biosynthetic genes, was identified and functionally shown to sequester CuC in cucumber leaf mesophyll cells. Notably, the CuC transport process is strictly co-regulated with CuC biosynthesis. CsMATE1 clustering with bitterness biosynthesis genes may provide benefits and a basis for this feedback regulation on CuC sequestration and biosynthesis. Identification of transport systems for plant-specialized metabolites can accelerate the metabolic engineering of high-value-added compounds by simplifying their purification process.

Four new species of the huntsmen spider genus Pseudopoda are described: P. hongqi sp. n. (male), P. shuyue sp. n. (male, female), P. wuxi sp. n. (male, female) and P. yintiaoling sp. n. (male, female). Morphological descriptions, photos of body and copulatory organs, as well as a locality map are provided.

Underground microbial ecosystems have profound impacts on plant health 1 – 5 . Recently, essential roles have been shown for plant specialized metabolites in shaping the rhizosphere microbiome 6 – 9 . However, the potential mechanisms underlying the root-to-soil delivery of these metabolites remain to be elucidated 10 . Cucurbitacins, the characteristic bitter triterpenoids in cucurbit plants (such as melon and watermelon), are synthesized by operon-like gene clusters 11 . Here we report two Multidrug and Toxic Compound Extrusion (MATE) proteins involved in the transport of their respective cucurbitacins, a process co-regulated with cucurbitacin biosynthesis. We further show that the transport of cucurbitacin B from the roots of melon into the soil modulates the rhizosphere microbiome by selectively enriching for two bacterial genera, Enterobacter and Bacillus , and we demonstrate that this, in turn, leads to robust resistance against the soil-borne wilt fungal pathogen, Fusarium oxysporum . Our study offers insights into how transporters for specialized metabolites manipulate the rhizosphere microbiota and thereby affect crop fitness.

The basic helix-loop-helix (bHLH) family of transcription factors (TFs) participate in a variety of biological regulatory processes in plants, and have undergone significant expansion during land plant evolution by gene duplications. In cucurbit crops, several bHLH genes have been found to be responsible for agronomic traits such as bitterness. However, the characterization of bHLH genes across the genomes of cucurbit species has not been reported, and how they have evolved and diverged remains largely unanswered. Here we identified 1,160 bHLH genes in seven cucurbit crops and performed a comprehensive comparative genomics analysis. We determined orthologous and paralogous bHLH genes across cucurbit crops by syntenic analysis between or within species. Orthology and phylogenetic analysis of the tandem-duplicated bHLH genes in the Bt cluster, which regulate the biosynthesis of cucurbitacins, suggest that this cluster is derived from three ancestral genes after the cucurbit-common tetraploidization event. Interestingly, we identified a new conserved cluster paralogous to the Bt cluster that includes two tandem bHLH genes, and the evolutionary history and expression profiles of these two genes in the new cluster suggest the involvement of one gene (Brp) in the regulation of cucurbitacin biosynthesis in roots. Further biochemical and transgenic assays in melon hairy roots supported the function of Brp. This study provides useful information for further investigating the functions of bHLH TFs and novel insights into the regulation of cucurbitacin biosynthesis in cucurbit crops and other plants.

Population genetic structure can provide valuable insights for conserving genetic resources and understanding population evolution, but it is often underestimated when using the most popular method and software, STRUCTURE and delta K, to assess. Although the hierarchical STRUCTURE analysis has been proposed early to overcome the above potential problems, this method was just utilized in a few studies and its reliability needs to be further tested. In this study, the genetic structure of populations of Wikstroemia monnula was evaluated by sequencing 12 nuclear microsatellite loci of 905 individuals from 38 populations. The STRUCTURE analysis suggested the most likely number of clusters was two, but using multi-hierarchical structure analysis, almost every population was determined with an endemic genetic component. The latter result is consistent with the extremely low gene flow among populations and a large number of unique cpDNA haplotypes in this species, indicating one level of structure analysis would extremely underestimate its genetic component. The simulation analysis shows the number of populations and the genetic dispersion among populations are two key factors to affect the estimation of K value using the above tools. When the number of populations is more than a certain amount, K always is equal to 2, and when a simulation only includes few populations, the underestimation of K value also may occur only if these populations consist of two main types of significantly differentiated genetic components. Our results strongly support that the hierarchical STRUCTURE analysis is necessary and practicable for the species with lots of subdivisions.

Programmed cell death (PCD) in tapetum provides nutrients for pollen development. Once this process becomes abnormal, the pollen will be aborted, and sterile lines will be formed. Lycium barbarum L. is a well-known medicinal and edible plant, and male sterile lines play an important role in the cultivation of its new varieties by utilizing hybrid vigor. However, the male sterility mechanism of L. barbarum is poorly understood. In this study, the main characteristic changes during the pollen development of L. barbarum sterile line (Ningqi No. 5) and fertile line (Ningqi No. 1) were compared through anatomical observation, physiological detection and gene expression analysis. Anatomical observation showed that compared with that of the fertile line, the tapetum of the sterile line persisted during pollen development, the organelle morphology and number of tapetal cells changed remarkably, and the PCD was remarkably delayed. Membranous peroxidation caused by reactive oxygen species (ROS) in the sterile line occurred from the late tetrad to the pollen grain stage, and that in Ningqi No. 1 transpired in the tetrad stage. This difference in the dynamic changes of ROS affected the redox balance of microspore development. qPCR detection of DYT1 and MS1 genes regulating tapetum development showed that compared with those in the fertile line, the expression levels of both genes in the sterile line changed significantly from pollen mother cell stage to pollen grain stage. This finding may be associated with the start-up delay of tapetal PCD. All these results suggested that abnormal tapetal PCD is an important mechanism leading to male sterility in L. barbarum.

The corticalis group is one of most diverse species-group in genus Clubiona Latreille, 1804. Currently, a total of 81 corticalis group species are known worldwide, amongst them 67 were recorded from China. However, the diversity of this group in China is still insufficiently known.Clubiona xianning sp. nov. is described as a new species of the C. corticalis species-group collected from Hubei Province, China.

Due to its special ways of hiding and lifestyle, Thelcticopis is a type of spider that is very difficult to collect. In 2018, we collected two huntsman spiders in Guangxi. After comparison with other Thelcticopis species, such as anterior median eye larger than other eyes, they were found to belong to the genus of Thelcticopis.Currently, four Thelcticopis species are reported from China, T. severa (L. Koch, 1875), T. zhengi Liu, Li & Jäger, 2010, T. dahanensis Zhu & Zhong, 2020 and T. unciformis Zhu & Zhong, 2020. They are mainly distributed in the tropical or subtropical areas of China (Fujian, Guangdong, Guangxi, Hainan, Hongkong, Hunan, Taiwan, Yunnan and Zhejiang). In this paper, we diagnose and describe a new species, Thelcticopis pinmini sp. nov., which was collected from Guangxi Province, China.

The diversity of axon guidance (AG) receptors reflects gains in complexity of the animal nervous system during evolution. Members of the Roundabout (Robo) family of receptors interact with Slit proteins and play important roles in many developmental processes, including AG and neural crest cell migration. There are four members of the Robo gene family. However, the evolutionary history of Robo family genes remain obscure. We analyzed the distribution of Robo family members in metazoan species ranging in complexity from hydras to humans. We undertook a phylogenetic analysis in metazoans, synteny analysis, and ancestral chromosome mapping in vertebrates, and detected selection pressure and functional divergence among four mammalian Robo paralogs. Based on our analysis, we proposed that the ancestral Robo gene could have undergone a tandem duplication in the vertebrate ancestor; then one round of whole genome duplication events occurred before the divergence of ancestral lamprey and gnathostome, generating four paralogs in early vertebrates. Robo4 paralog underwent segmental loss in the following evolutionary process. Our results showed that Robo3 paralog is under more powerful purifying selection pressure compared with other three paralogs, which could correlate with its unique expression pattern and function. Furthermore, we found four sites under positive selection pressure on the Ig1-2 domains of Robo4 that might interfere with its binding to Slits ligand. Diverge analysis at the amino acid level showed that Robo4 paralog have relatively greater functional diversifications than other Robo paralogs. This coincides with the fact that Robo4 predominantly functions in vascular endothelial cells but not the nervous system.

Sinopoda spiders are a diverse group with limited dispersal ability. They are remarkably sympatric among related species, which often results in misidentification and incorrect matching of sexes. In order to understand the evolutionary relationships and revise the taxonomy problems in this genus, we offer the first molecular phylogeny of Sinopoda. Our results strongly support the monophyly of Sinopoda and its sister relationship with Spariolenus and reject the monophyly of the S. okinawana species group. We establish three new species groups based on both molecular and morphological data. Our phylogeny also illuminates some taxonomic issues and clarifies some species limits: (1) Supporting the newly revised matching of sexes in S. longiducta and S. yaanensis by Zhong et al. (2019). (2) The original description of S. campanacea was based on mismatched sexes. S. changde is proposed as a junior synonymy of S. campanacea, while the original female 'S. campanacea' is here described as a new species: S. papilionaceous Liu sp. nov. (3) The type series of S. serpentembolus contains mismatched sexes. The female is considered as S. campanacea, while we here report the correctly matched females of S. serpentembolus. (4) We describe one additional new species: S. wuyiensis Liu sp. nov. Our first molecular phylogeny of Sinopoda provides a tool for comparative analyses and a solid base for the future biodiversity and taxonomic work on the genus.