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Zokors (Myospalacinae) continue to be the center of systematics discussions. Phylogenetic schemes based on molecular data do not always agree with each other, nor can phylogenetic schemes based on paleontological material be complete due to the only-partial description of West Siberian zokors. This paper tries to fill this gap and presents a description of the West Siberian lineage from the late early Pleistocene to the present, together with an analysis of molar development in other zokor lineages. We describe two new species and one subspecies (Myospalax myospalax krukoveri n. ssp., Myospalax convexus n. sp., Prosiphneus razdoleanensis n. sp.) ancestral to the extant Myospalax myospalax Laxmann, 1769. We also reveal differences in the ontogeny of molars of modern species that were not previously detailed. These differences, together with paleontological data, indicate that in the West Siberian lineage, peramorphosis occurred in the structure of the chewing surface (with the exception of the lower m1), whereas in all other zokor lineages, there was pedomorphosis. On the basis of these results, we suggest a new view on the systematics of Myospalacinae. UUID: http://zoobank.org/b06d6c99-1648-454b-9b95-4d869bfe8bdc

Phylogenetic relationships between zokors living in different territories of Russia: Altai zokors Myospalax myospalax from “Altai” (Altai Republic and Altaiskii Krai) and “Priobie” from the River Ob zone (Tomsk oblast and Novosibirsk oblast) and subspecies M. m. tarbagataicus from Kazakhstan (ridge Tarbagatai) and M. aspalax and M. psilurus from Zabaikalskii Krai were determined on the basis of craniometrical and molecular analysis of the mitochondrial 12S rRNA gene. The comparison of the craniometrical and molecular data revealed significant differences between the Altai zokors of the “Priobie” (the River Ob zone) and “Altai” populations. The importance of geographic isolation to the formation of morphometric and genetic differentiation of distinct geographic forms of the investigated zokors is shown. Specific ecological and morphological adaptations and distinct genetic features of two forms of zokors indicate the existence of separate subspecies of the species M. myospalax.

Rodents’ lifestyles vary in different environments, and to adapt to various lifestyles specific digestion strategies have been developed. Among these strategies, the morphology of the digestive tracts and the gut microbiota are considered to play the most important roles in such adaptations. However, how subterranean rodents adapt to extreme environments through regulating gut microbial diversity and morphology of the digestive tract has yet to be fully studied. Here, we conducted the comparisons of the gastrointestinal morphology, food intake, food assimilation, food digestibility and gut microbiota of plateau zokor Eospalax baileyi in Qinghai-Tibet Plateau and laboratory rats Rattus norvegicus to further understand the survival strategy in a typical subterranean rodent species endemic to the Qinghai-Tibet Plateau. Our results revealed that plateau zokor evolved an efficient foraging strategy with low food intake, high food digestibility, and ultimately achieved a similar amount of food assimilation to laboratory rats. The length and weight of the digestive tract of the plateau zokor was significantly higher than the laboratory rat. Particularly, the weight and length of the large intestine and cecum in plateau zokor is three times greater than that of the laboratory rat. Microbiome analysis showed that genus (i.e., Prevotella, Oscillospira, CF231, Ruminococcus and Bacteroides), which are usually associated with cellulose degradation, were significantly enriched in laboratory rats, compared to plateau zokor. However, prediction of metagenomic function revealed that both plateau zokor and laboratory rats shared the same functions in carbohydrate metabolism and energy metabolism. The higher digestibility of crude fiber in plateau zokor was mainly driven by the sizes of cecum and cecum tract, as well as those gut microbiota which associated with cellulose degradation. Altogether, our results highlight that both gut microbiota and the morphology of the digestive tract are vital to the digestion in wild rodents.

Zokors are one of the subterranean rodents in the subfamily Myospalacinae. Extant zokors include two major genera, Eospalax and Myospalax, which can be distinguished by skull shape. however, due to few available studies on skull morphology of zokors, the interspecific differentiation of their skulls is still unclear. To differentiate among species and to describe the sexual dimorphism within each species, we measured morphological variation using one-way analysis of variance and cluster analysis in four species of zokor including Eospalax cansus, Eospalax rothschildi, Eospalax baileyi, and Myospalax aspalax. We also used principal component analysis and dichotomy to explore key measurements which could reflect skull differences to the maximum. In addition, we tested the correlations between environmental factors and skull measurements for each species. The results of variance analysis indicated that three zokors showed male-biased sexual size dimorphism except for E. baileyi. We also found that there were significant differences of linear measurements among four species. The results of cluster analysis showed that the four species of zokors divided into two branches, one including E. cansus and E. rothschildi, the other E. baileyi and M. aspalax. The results of principal component analysis showed that the variables that contribute to the first principal component were related to skull size, and the variables which contribute to the second principal component were different between sexes. Thus, linear measurements that reflect the skull size such as cranial length, basal length and cranial base could be key measurements for cluster and classification. Zygomatic breadth could be considered as quantifiable differences between the two genera Eospalax and Myospalax. We suggest that the clustering patterns could be related to climate and geographical factors. finally, it was found that most of the skull measurements were positively correlated with latitude but negatively correlated with annual mean temperature and precipitation, which follows Bergmann's rule. This work provides a significant reference value to scholars who apply themselves to pest control and to the classification and identification of zokors using morphology. Keywords: Cluster, Morphology, Skull, Zokor.

Background Mammals exhibit diverse adaptations to varied habitats, yet the genomic mechanisms underlying these transitions remain incompletely understood. Here, we present a comparative genomic, epigenomic, and three-dimensional chromatin analysis of two closely related zokor species, Myospalax aspalax and Myospalax psilurus , which inhabit arid and humid subterranean environments, respectively. Results We identified subterranean lineage-specific positively selected and rapidly evolving genes enriched for DNA repair, hypoxia response, blood vessel development, and fructose metabolism. M. psilurus showed a notable loss of olfactory receptor genes, expansion of pheromone-related gene families, and widespread activation of a specific ERVK transposable element family, accompanied by elevated DNA methylation, suggesting enhanced epigenetic defenses and ecological specialization. In contrast, M. aspalax showed selection in kidney and lipid metabolism genes. Large chromosomal inversions (> 1 Mb) overlapped islands of high genetic divergence (high- F ST or high D xy ) and were enriched for sensory, reproductive, and DNA repair genes. Although chromatin compartments remained largely stable, large inversions disrupted topologically associating domains and preferentially occurred in regions characterized by longer chromatin loops. Inversion breakpoints were flanked by inverted repeats and segmental duplications—features characteristic of the formation mechanism of non-allelic homologous recombination, suggesting that both the spatial proximity and sequence architecture of these regions may predispose them to large inversions. Conclusions Overall, our results illuminate the structural, regulatory, and evolutionary mechanisms underlying ecological divergence and highlight how genome architecture contributes to adaptive evolution in subterranean mammals.

Background/Objectives: Seasonal reproduction in mammals is primarily regulated by the hypothalamic–pituitary–ovarian (HPO) axis, yet its molecular mechanisms in subterranean rodents living in light-restricted environments remain poorly understood. This study aimed to characterize the transcriptional regulation of the HPO axis during seasonal estrus in the Manchurian zokor (Myospalax psilurus, M. psilurus), a fossorial rodent exhibiting distinct breeding cycles despite perpetual darkness. Methods: Hypothalamic, pituitary, and ovarian tissues were collected from female zokors during estrus and anestrus (n = 5 per group). RNA sequencing was performed, followed by de novo transcriptome assembly and bioinformatic analyses. Differentially expressed genes (DEGs) were identified using edgeR, and functional enrichment was assessed via GO and KEGG analyses. Key DEGs were validated by RT-qPCR. Results: A total of 513, 292, and 138 DEGs were identified in the hypothalamus, pituitary, and ovary, respectively. GO analysis highlighted enrichment in G-protein-coupled receptor signaling, oxidation–reduction processes, and calcium ion binding. KEGG pathway analysis revealed significant enrichment of the neuroactive ligand–receptor interaction pathway across all three tissues. Key candidate genes included Trh and Mc3r in the hypothalamus, Pitx2 and NR4A2 in the pituitary, and PTGER2 and Sphk1 in the ovary. Conclusions: This study provides the first comprehensive transcriptomic profile of the HPO axis in Manchurian zokors during seasonal estrus. The neuroactive ligand–receptor interaction pathway appears central to reproductive regulation, and several tissue-specific genes were identified as potential regulators of seasonal breeding. These findings enhance our understanding of reproductive adaptation in subterranean mammals and offer a foundation for further functional studies.

In order to manage Eospalax (Myospalax) baileyi (Plateau zokors) pests with physical method, the efficacy of four different arrow traps was tested to control the plateau zokor in Sichuan Northwest Prairie, China. Population densities of 39.7-42.1 ha-1 were found in the test plots. Such densities require control measures since pasture damages and economic losses are huge. Traps differed in trigger rates, from 83.8% (three-arrow type), 93.8% (infrared type), 93.9% (T-type), to 94.3% (one-arrow type). Capture rates ranged from 84.1 (three-arrow type), 91.8% (infrared type), 93.7% (T-type) to 93.8% (one-arrow type). The single-arrow trap has the highest efficacy and is also the cheapest, which is important knowledge for zokor control in western China.

Animals living at high altitudes have evolved distinct phenotypic and genotypic adaptations against stressful environments. We studied the adaptive patterns of altitudinal stresses on transcriptome turnover in subterranean plateau zokors ( Myospalax baileyi ) in the high-altitude Qinghai-Tibetan Plateau. Transcriptomes of zokors from three populations with distinct altitudes and ecologies ( L ow: 2846 m, M iddle: 3282 m, H igh: 3,714 m) were sequenced and compared. Phylogenetic and principal component analyses classified them into three divergent altitudinal population clusters. Genetic polymorphisms showed that the population at H , approaching the uppermost species boundary, harbors the highest genetic polymorphism. Moreover, 1056 highly up-regulated UniGenes were identified from M to H . Gene ontologies reveal genes like EPAS1 and COX1 were overexpressed under hypoxia conditions. EPAS1 , EGLN1 , and COX1 were convergent in high-altitude adaptation against stresses in other species. The fixation indices ( F ST and G ST )-based outlier analysis identified 191 and 211 genes, highly differentiated among L , M , and H . We observed adaptive transcriptome changes in Myospalax baileyi , across a few hundred meters, near the uppermost species boundary, regardless of their relatively stable underground burrows’ microclimate. The highly variant genes identified in Myospalax were involved in hypoxia tolerance, hypercapnia tolerance, ATP-pathway energetics, and temperature changes.

As the most abundant group of mammals, rodents possess a very rich ecotype, which makes them ideal for studying the relationship between diet and host gut microecology. Zokors are specialized herbivorous rodents adapted to living underground. Unlike more generalized herbivorous rodents, they feed on the underground parts of grassland plants. There are two species of the genus Myospalax in the Eurasian steppes in China: one is Myospalax psilurus, which inhabits meadow grasslands and forest edge areas, and the other is M. aspalax, which inhabits typical grassland areas. How are the dietary choices of the two species adapted to long-term subterranean life, and what is the relationship of this diet with gut microbes? Are there unique indicator genera for their gut microbial communities? Relevant factors, such as the ability of both species to degrade cellulose, are not yet clear. In this study, we analyzed the gut bacterial communities and diet compositions of two species of zokors using 16S amplicon technology combined with macro-barcoding technology. We found that the diversity of gut microbial bacterial communities in M. psilurus was significantly higher than that in M. aspalax, and that the two species of zokors possessed different gut bacterial indicator genera. Differences in the feeding habits of the two species of zokors stem from food composition rather than diversity. Based on the results of Mantel analyses, the gut bacterial community of M. aspalax showed a significant positive correlation with the creeping-rooted type food, and there was a complementary relationship between the axis root-type-food- and the rhizome-type-food-dominated (containing bulb types and tuberous root types) food groups. Functional prediction based on KEGG found that M. psilurus possessed a stronger degradation ability in the same cellulose degradation pathway. Neutral modeling results show that the gut flora of the M. psilurus has a wider ecological niche compared to that of the M. aspalax. This provides a new perspective for understanding how rodents living underground in grassland areas respond to changes in food conditions.

Zokor is a group of subterranean rodents that are adapted to underground life and feed on plant roots. Here, we investigated the intestinal microbes of five zokor species (Eospalax cansus, Eospalax rothschildi, Eospalax smithi, Myospalax aspalax, and Myospalax psilurus) using 16S amplicon technology combined with bioinformatics. Microbial composition analysis showed similar intestinal microbes but different proportions among five zokor species, and their dominant bacteria corresponded to those of herbivores. To visualize the relationships among samples, PCoA and PERMANOVA tests showed that the intestinal microbes of zokors are largely clustered by host species, but less so by genetics and geographical location. To find microbes that differ among species, LefSe analysis identified Lactobacillus, Muribaculaceae, Lachnospiraceae_NK4A136_group, unclassified_f_Christensenellaceae, and Desulfovibrio as biomarkers for E. cansus, E. rothschildi, E. smithi, M. aspalax, and M. psilurus, respectively. PICRUSt metagenome predictions revealed enriched microbial genes for carbohydrate and amino acid metabolism in E. cansus and E. smithi, and for cofactor and vitamin metabolism as well as glycan biosynthesis and metabolism in E. rothschildi, M. aspalax, and M. psilurus. Our results demonstrated differences in the microbial composition and functions among five zokor species, potentially related to host genetics, and host ecology including dietary habits and habitat environment. These works would provide new insight into understanding how subterranean zokors adapt to their habitats by regulating intestinal microbes.