To help resolve the systematic position of Myospalacine (zokors) and to clarify the phylogenetic relationships among zokor species, complete mitochondrial 12S rRNA and cytochrome b gene sequences were determined for seven zokor species from China. Phylogenetic relationships among all extant zokors, except the Siberian zokor (Myospalax myospalax) and Smith's zokor (Eospalax smithi), were reconstructed with the Chinese bamboo rat (Rhizomys sinensis) as an outgroup. Our results support the generic status of Eospalax and also support the validity of the specific status of Eospalax cansus, Eospalax baileyi and Eospalax rufescens.
The application of animal medicinal materials has a centuries-old history in China. Zokors (Myospalacinae) were recorded as a type of animal crude drug in Bencaogangmu, a famous work written by Li Shizhen about 400 years ago during the Ming Dynasty, which identified 1892 kinds of medicines. The flesh and bone of the zokors were reported to cure many kinds of inflammation.1) The dried bone of plateau zokor (Myospalax baileyi) is called Sailonggu, and has been used medicinally by local Tibetans in the Qinghai Province in China.2) In the middle of the 1970s, it was found that the Tibetan drug Sailonggu could cure rheumatic diseases and is an ideal substitute for the Chinese crude drug "tiger bone." 3,4) Sailong Rheumatic Liquor, made from the bone of plateau zokor, has been produced since 1995 by the Beijing Tongrentang Medicinal Liquor Factory, Beijing Tongrentang Medicine Industry Limited Group.Plateau zokor (M. baileyi) is found in plateau habitats in Qinghai, Gansu and Sichuan Provinces. 5) However, it shares habitats with the Gansu zokor (M. cansus), the bamboo rat (Rhizomys sinensis), and several other rodents such as the black lipped pika (Ochotona curzoniae). Consequently, Sailonggu is mingled with adulterants in the course of crude drug collection. The identification of animal crude drugs has always been problematical because their substitutes or adulterants are very similar to the quality goods, both in morphological characteristics and in organic or inorganic components, especially when cut into pieces. The mtDNA sequences of 12S rRNA and cytochrome b (cyt b) genes have been used successfully by Wang and Zhou 6) as molecular markers in the authentication of a variety of animal crude drugs. These were investigated in the present study in order to develop molecular genetic markers for the identification of Sailonggu. MATERIALS AND METHODSSample Collection Twenty-one individuals, representing seven species of zokors of the genus Myospalax, were used in this study (Table 1). One Chinese bamboo rat (Rhizomys sinensis) and one black lipped pika (Ochotona curzoniae) were also included. The original animals of the samples were identified according to morphological characteristics. The crude drug Sailonggu samples were provided by Beijing Tongrentang Medicinal Liquor Factory, Beijing Tongrentang Medicine Industry Limited Group. All specimens are deposited in the collection of Nanjing Normal University (NNU). Tissue samples derived from muscle are preserved in 95% ethanol.DNA Extraction Total genomic DNA was extracted from the ethanol preserved muscle samples as described in Sambrook et al. 7) For isolation of total DNA from the crude drug Sailonggu, about 1.0 g bone pieces taken from each sample were washed with water and ddH 2 O twice, respectively, and dried at 80°C. The bone pieces were exposed under a 20 W ultraviolet lamp for 40 min in order to remove exo-DNA contaminations. The samples were ground to a fine powder using liquid nitrogen. One milliliter of 0.5 M EDTA (pH 8.0) was added to 0.5 g bone powder in...
MicroRNAs (miRNAs) are now recognized as key post-transcriptional regulators in regulation of phenotypic diversity. Qinlingacris elaeodes is a species of the alpine grasshopper, which is endemic to China. Adult individuals have three wing forms: wingless, unilateral-winged and short-winged. This is an ideal species to investigate the phenotypic plasticity, development and evolution of insect wings because of its case of unilateral wing form in both the sexes. We sequenced a small RNA library prepared from mesothoraxes of the adult grasshoppers using the Illumina deep sequencing technology. Approximately 12,792,458 raw reads were generated, of which the 854,580 high-quality reads were used only for miRNA identification. In this study, we identified 49 conserved miRNAs belonging to 41 families and 69 species-specific miRNAs. Moreover, seven miRNA*s were detected both for conserved miRNAs and species-specific miRNAs, which were supported by hairpin forming precursors based on polymerase chain reaction. This is the first description of miRNAs in alpine grasshoppers. The results provide a useful resource for further studies on molecular regulation and evolution of miRNAs in grasshoppers. These findings not only enrich the miRNAs for insects but also lay the groundwork for the study of post-transcriptional regulation of wing forms.
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