Digestive Tract Morphology and Gut Microbiota Jointly Determine an Efficient Digestive Strategy in Subterranean Rodents: Plateau Zokor.

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.

Main Factors Influencing the Gut Microbiota of Datong Yaks in Mixed Group.

The Datong yak (Bos grunniens) is the first artificial breed of yaks in the world and has played an important role in the improvement of domestic yak quality on the Qinghai-Tibet Plateau. The Datong yak breeding farm in the Qinghai province of China is the main place for the breeding and feeding of Datong yaks. It hosts domestic Datong yaks and wild male yaks, mainly in mixed groups. Different managements have different effects on livestock. The gut microbiota is closely related to the health and immunity of Datong yaks, and mixed grouping can affect the composition and diversity of the gut microbiota of Datong yaks. To reveal the effects of mixed grouping on the gut microbiota of Datong yaks and wild yaks and identify the main dominant factors, we compared the gut microbial diversities of domestic males and females and wild males based on 16S rRNA V3-V4 regions using fresh fecal samples. The data showed significant differences in the gut microbial diversity of these three groups, and the α-diversity was the highest in wild males. Different factors influence the gut microbiota, and the main influencing factors were different in different groups, including sex differences, host genetics, and physical interactions. We also compared ecological assembly processes in the three groups. The results showed that mixed grouping contributed to the improvement of gut microbial diversity in domestic females. Our study provides effective and feasible suggestions for the feeding and management of the Datong yaks.

The complete chloroplast genome and phylogenetic analysis of Syringa reticulata subsp. amurensis (Rupr.) P.S.Green & M.C.Chang from Qinghai Province, China.

Syringa reticulata subsp. amurensis (Rupr.) P. S. Green & M. C. Chang (Oleaceae) is a shrub or tree with high medicinal value as well as great ecological significance as an urban garden plant. To better understand the molecular genetics and evolutionary of S. reticulata subsp. amurensis, its complete chloroplast genome was sequenced and annotated. The assembled chloroplast genome is a circular 156,141 bp sequence, consisting of 87,108 bp large single copy (LSC) region and 17,239 bp small single copy (SSC) region, which were flanked by a pair of 25,897 bp inverted repeats (IRs). The GC content of the chloroplast genome is 36.14%. Moreover, a total of 132 functional genes were annotated, including 88 protein-coding, 36 tRNA, and eight rRNA genes. Phylogenetic analysis showed that S. reticulata subsp. amurensis was most closely related to S. reticulata subsp. Pekinensis and the genus Syringa is paraphyletic group. This study provides important information for further phylogenetic studies on S. reticulata subsp. amurensis and its allies.

Gut Microbiome Changes in Captive Plateau Zokors (Eospalax baileyi).

Wild-caught animals must cope with drastic lifestyle and dietary changes after being induced to captivity. How the gut microbiome structure of these animals will change in response receives increasing attention. The plateau zokor (Eospalax baileyi), a typic subterranean rodent endemic to the Qinghai-Tibet plateau, spends almost the whole life underground and is well adapted to the environmental pressures of both plateau and underground. However, how the gut microbiome of the plateau zokor will change in response to captivity has not been reported to date. This study compared the microbial community structure and functions of 22 plateau zokors before (the WS group) and after being kept in captivity for 15 days (the LS group, fed on carrots) using the 16S rRNA gene via high-throughput sequencing technology. The results showed that the LS group retained 973 of the 977 operational taxonomic units (OTUs) in the WS group, and no new OTUs were found in the LS group. The dominant bacterial phyla were Bacteroides and Firmicutes in both groups. In alpha diversity analysis, the Shannon, Sobs, and ACE indexes of the LS group were significantly lower than those of the WS group. A remarkable difference (P < 0.01) between groups was also detected in beta diversity analysis. The UPGMA clustering, NMDS, PCoA, and Anosim results all showed that the intergroup difference was significantly greater than the intragroup difference. And compared with the WS group, the intragroup difference of the gut microbiota in the LS group was much larger, which failed to support the assumption that similar diets should drive convergence of gut microbial communities. PICRUSt revealed that although some functional categories displayed significant differences between groups, the relative abundances of these categories were very close in both groups. Based on all the results, we conclude that as plateau zokors enter captivity for a short time, although the relative abundances of different gut microbiota categories shifted significantly, they can maintain almost all the OTUs and the functions of the gut microbiota in the wild. So, the use of wild-caught plateau zokors in gut microbial studies is acceptable if the time in captivity is short.

Characteristics of complete chloroplast genome of a high-quality forage on Qinghai-Tibet Plateau, Medicago archiducis-nicolai Sirj. (Fabaceae：Trifolieae).

Medicago archiducis-nicolai Sirj. is a well-known high-quality forage as its good palatability and strong tolerance to drought, cold and saline-alkali stress. Here, the complete chloroplast genome sequence of M. archiducis-nicolai was reported. The size of the complete chloroplast genome is 127,072 bp in length. The chloroplast genome has no inverted repeat (IR) regions, which is very common in the family Fabaceae. The M. archiducis-nicolai chloroplast genome encodes 106 genes: 72 protein-coding genes, 30 tRNAs, and 4 rRNAs. The phylogenetic analysis result strongly suggested that M. archiducis-nicolai is a distinct lineage in Medicago, being sister to highly supported clade composed of three species (M. hybrida, M. papillosa and M. sativa).

Complete chloroplast genome of a high-quality forage in north China, Medicago ruthenica (Fabaceae：Trifolieae).

Medicago ruthenica is a well-known high-quality forage due to its good palatability and strong tolerance to drought, cold and saline-alkali stress. Here, the complete chloroplast genome sequence of M. ruthenica was reported. The chloroplast genome is 126,939 bp in length. This chloroplast genome has no inverted repeat (IR) regions, which is very common in the family Fabaceae. The M. ruthenica chloroplast genome encodes 107 genes, including 73 protein-coding genes, 30 tRNA genes, and 4 rRNA genes. Phylogenetic analysis result strongly suggested that M. ruthenica is a distinct lineage in Medicago, being sister to highly supported clade composed of three species (M. hybrida, M. papillosa and M. sativa).

The complete chloroplast genome sequence of Stellera chamaejasme f. chrysantha (Thymelaeaceae).

Stellera chamaejasme L. f. chrysantha S. C. Huang is a toxic perennial herb of Thymelaeaceae and has the potential for medicine as Stellera chamaejasme L. Here, we present the complete chloroplast genome sequence of S. chamaejasme f. chrysantha based on Illumina sequencing data. The complete chloroplast genome sequence is 173,364 bp in size and contains four subregions: a pair of inverted repeats (IRs, each for 41,978 bp), a large single-copy region (LSC, 86,558 bp), and a small single-copy region (SSC, 2,850 bp). 141 genes were recognized in the assembled sequence: 93 protein-coding genes (PCGs), 38 tRNAs, 8 rRNAs, and 2 pseudo genes. The phylogenetic analysis result strongly supported that Stellera chamaejasme f. chrysantha was closely related to S. chamaejasme L.

Quantitative Analyses of Nine Phenolic Compounds and Their Antioxidant Activities from Thirty-Seven Varieties of Raspberry Grown in the Qinghai-Tibetan Plateau Region.

In this work, an efficient method for the rapid extraction and separation of antioxidant phenols was developed and optimized. The method was then applied to extract and separate nine phenols from 37 varieties of raspberry, in which their antioxidant activities were further investigated. First, the extraction was conducted using ultra-sonication, which was then further separated using reversed-phase high-performance liquid chromatography/ultraviolet (RP-HPLC/UV) analysis. In this step, several key parameters (volume of the extraction reagent, time of extraction, and the temperature of extraction) affecting its efficiency were investigated and optimized using the response surface methodology (RSM) combined with the Box-Behnken design (BBD) so that the optimal conditions were obtained. According to the overall results of the optimization study, the optimal conditions were chosen as follows: volume of extraction reagent = 2.0 mL, time of extraction = 50.0 min, and temperature of extraction = 50 °C. The optimal conditions were then applied to extract nine phenols, including gallic acid, catechin, chlorogenic acid, vanillic acid, syringic acid, cumaric acid, ferulic acid, rosemary acid, and quercetin from 37 raspberry varieties. The extracted phenols were characterized and their antioxidant activities, including DPPH- and ABTS- free radical scavenging and intracellular reactive oxygen species (ROS) activity, using HepG2 cells as the model, were subsequently studied. The findings suggested that although their contents varied among most raspberry varieties, these phenols significantly contributed toward their antioxidant capacity and scavenging intracellular ROS activities. This study provides a scientific and theoretical basis for the selection of raspberry varieties and product development in Qinghai province.

Complete plastome sequence of Eustoma grandiflorum (Gentianaceae), a popular cut flower.

Eustoma grandiflorum (Raf.) Shinners is a popular cut flower due to its beautiful morphological characteristics and extended vase life. Here, the complete plastome sequence of E. grandiflorum was reported based on the Illumina HiSeq Platform. The plastome sequence is 154,230 bp in length with a typical quadripartite structure, containing a pair of inverted repeated (IR) regions (25,755 bp) that are separated by a large single copy (LSC) region of 84,297 bp, and a small single copy (SSC) region of 18,423 bp. A total of 130 functional genes were annotated, including 84 protein-coding genes, 38 tRNA genes, and 8 rRNA genes. The complete plastome sequence of E. grandiflorum will provide a valuable resource for its garden utilization and the phylogenetic studies of Gentianaceae.

Genetic variation in bitter taste receptor genes influences the foraging behavior of plateau zokor (Eospalax baileyi).

The ability to detect bitter tastes is important for animals; it can help them to avoid ingesting harmful substances. Bitter taste perception is mainly mediated by bitter taste receptor proteins, which are encoded by members of the Tas2r gene family and vary with the dietary preference of a specific species. Although individuals with different genotypes differ in bitterness recognition capability, little is known about the relationship between genetic variation and food selection tendencies at the intraspecific level. In this study, we examined the relationship between genotypes and diet in plateau zokor (Eospalax baileyi), a subterranean rodent endemic to the Qinghai-Tibet Plateau that caches food for the winter. We assayed the composition and taste profile of each plant contained in temporary caches and vicinity quadrats, which were representative of selected and available food, respectively. Bitter plant selection indices (E bitter) were estimated. We also sequenced 26 candidate Tas2r genes from zokors and determined their relationships with the E bitter of their caches. We identified four key results: (1) zokors varied considerably in both bitter food preference and Tas2r sequences; (2) five genes (zTas2r115,zTas2r119,zTas2r126,zTas2r134, and zTas2r136) exhibited allelic variation that was significantly associated with E bitter; (3) synonymous SNPs, nonsynonymous SNPs, and pseudogenization are involved in the genotype-phenotype relationship; (4) the minor genotypes of zTas2r115,zTas2r134, and zTas2r136 and the major genotypes of zTas2r119 and zTas2r126 cached more bitter plants. Our results link Tas2r variation with food selection behavior at the population level for the first time.