The Hooded Crane (Grus monacha) is a waterbird wintering in the wetlands of the middle and lower reaches of the Yangtze River, China. The gradual habitat loss resulting from wetland degradation may have posed negative...The Hooded Crane (Grus monacha) is a waterbird wintering in the wetlands of the middle and lower reaches of the Yangtze River, China. The gradual habitat loss resulting from wetland degradation may have posed negative effects on the structure of our wintering populations. For its effective protection, it is important to conduct an intensive study on the genetic structure of this population. A total of 221 faecal samples, nine feather samples and four muscle samples of Hooded Cranes from four wintering populations, i.e., from Caizi Lake and Shengjin Lake in Anhui, Poyang Lake in Jiangxi and Chongming Dongtan in Shanghai, were collected for this study. Full-length 1103–1104 bp mtDNA D-loop sequences from 72 samples were amplified using PCR. Based on our amplified D-loop sequences and the sequences of two individual birds obtained from GenBank (AB017625 and AB023813), we analyzed the genetic structure of these four wintering Hooded Crane populations. Twenty six variable sites were found among 72 target sequences in the four wintering populations and 23 haplotypes were defined. Genetic diversity analyses showed that the haplotype diversity of Hooded Cranes was 0.823 ± 0.042 with a nucleotide diversity of 0.00157 ± 0.00021. The FST values of the four populations show that there is no significant genetic differentiation among the populations of Hooded Cranes wintering in the middle and lower reaches of the Yangtze River. Tajima’s D and Fu’s tests suggest that the Hooded Crane populations may have experienced population expansion in their evolutionary history.展开更多
Extracting DNA from faecal sample is a newly developed technology of recent years. Although several methods to extract DNA from faeces have been developed home and abroad, they addressed fresh samples and it is still ...Extracting DNA from faecal sample is a newly developed technology of recent years. Although several methods to extract DNA from faeces have been developed home and abroad, they addressed fresh samples and it is still blank in extracting DNA from old faeces yet. Extracting old faeces’ DNA can make the full use of transfers out or the death animal’s genetic information and expanding the application scope of Molecular Scatology. In this experiment two groups of faeces were launched into DNA extracting. One group had been preserved by absolute ethanol for 3 years and the other group had been preserved by absolute ethanol for half past 4 years. Aiming at the old faecal’s characteristic of small number of cells and high degrading of DNA, several repetitions(4 tubes) were carried out at the same time in this experiment. Then 4 different DNA extracting methods (Preprocess-phenol/chloroform extraction method, Guanidine thiocyanate method, CTAB method, QIAamp DNA Stool Mini Kit, Qiagen) were followed. From these parallel methods and their corresponding results, we could get the best method while the possibility of extracting DNA from old faecal was studied. After DNA was dissolved in TE we got the original DNA. Finally we collected the original DNA from the 4 repetitions then purified and concentrated it by using DNA purification kit. In the end, the results of PCR amplification shows that only Preprocess-phenol/chloroform extraction method can extract enough DNA for PCR from two groups of faeces. In our study, we extracted DNA from old faeces successfully and got some genetic information from mtDNA. This result confirms the possibility of putting old faecal into molecular scatology study.展开更多
[Objective] To introduce an improved method for DNA extraction from the faeces of red deer. [Method] Based on the traditional method of CTAB lysis, we proposed an improved DNA extraction method according to the charac...[Objective] To introduce an improved method for DNA extraction from the faeces of red deer. [Method] Based on the traditional method of CTAB lysis, we proposed an improved DNA extraction method according to the characteristics of red deer faeces. [Result] This improved method extracted high-quality fecal DNA from Tianshan red deer and amplified the mitochondrial cytochrome b gene. With the muscle and fur DNA of red deer as the control, the sequencing results further con- firmed the reliability of the method. [Conclusion] The method requires no proteinase K in the process of extraction, and the extracted DNA can be used for PCR ampli- fication directly without the purification of DNA purification kit, thus, it is cost-saving.展开更多
基金supported by the National Natural Science Foundation of China (Grant No 31172117)the Anhui Academic and Technical Leaders Fund
文摘The Hooded Crane (Grus monacha) is a waterbird wintering in the wetlands of the middle and lower reaches of the Yangtze River, China. The gradual habitat loss resulting from wetland degradation may have posed negative effects on the structure of our wintering populations. For its effective protection, it is important to conduct an intensive study on the genetic structure of this population. A total of 221 faecal samples, nine feather samples and four muscle samples of Hooded Cranes from four wintering populations, i.e., from Caizi Lake and Shengjin Lake in Anhui, Poyang Lake in Jiangxi and Chongming Dongtan in Shanghai, were collected for this study. Full-length 1103–1104 bp mtDNA D-loop sequences from 72 samples were amplified using PCR. Based on our amplified D-loop sequences and the sequences of two individual birds obtained from GenBank (AB017625 and AB023813), we analyzed the genetic structure of these four wintering Hooded Crane populations. Twenty six variable sites were found among 72 target sequences in the four wintering populations and 23 haplotypes were defined. Genetic diversity analyses showed that the haplotype diversity of Hooded Cranes was 0.823 ± 0.042 with a nucleotide diversity of 0.00157 ± 0.00021. The FST values of the four populations show that there is no significant genetic differentiation among the populations of Hooded Cranes wintering in the middle and lower reaches of the Yangtze River. Tajima’s D and Fu’s tests suggest that the Hooded Crane populations may have experienced population expansion in their evolutionary history.
文摘Extracting DNA from faecal sample is a newly developed technology of recent years. Although several methods to extract DNA from faeces have been developed home and abroad, they addressed fresh samples and it is still blank in extracting DNA from old faeces yet. Extracting old faeces’ DNA can make the full use of transfers out or the death animal’s genetic information and expanding the application scope of Molecular Scatology. In this experiment two groups of faeces were launched into DNA extracting. One group had been preserved by absolute ethanol for 3 years and the other group had been preserved by absolute ethanol for half past 4 years. Aiming at the old faecal’s characteristic of small number of cells and high degrading of DNA, several repetitions(4 tubes) were carried out at the same time in this experiment. Then 4 different DNA extracting methods (Preprocess-phenol/chloroform extraction method, Guanidine thiocyanate method, CTAB method, QIAamp DNA Stool Mini Kit, Qiagen) were followed. From these parallel methods and their corresponding results, we could get the best method while the possibility of extracting DNA from old faecal was studied. After DNA was dissolved in TE we got the original DNA. Finally we collected the original DNA from the 4 repetitions then purified and concentrated it by using DNA purification kit. In the end, the results of PCR amplification shows that only Preprocess-phenol/chloroform extraction method can extract enough DNA for PCR from two groups of faeces. In our study, we extracted DNA from old faeces successfully and got some genetic information from mtDNA. This result confirms the possibility of putting old faecal into molecular scatology study.
基金Supported by the National Natural Science Foundation of China(31060152)the Natural Science Foundation of the Xinjiang Uygur Autonomous Region,China(2010211A02)the Key Program for Animal Sciences of Xinjiang University,China~~
文摘[Objective] To introduce an improved method for DNA extraction from the faeces of red deer. [Method] Based on the traditional method of CTAB lysis, we proposed an improved DNA extraction method according to the characteristics of red deer faeces. [Result] This improved method extracted high-quality fecal DNA from Tianshan red deer and amplified the mitochondrial cytochrome b gene. With the muscle and fur DNA of red deer as the control, the sequencing results further con- firmed the reliability of the method. [Conclusion] The method requires no proteinase K in the process of extraction, and the extracted DNA can be used for PCR ampli- fication directly without the purification of DNA purification kit, thus, it is cost-saving.
