A full-length cDNA library from the testis of dark-spotted frogs （ Rana nigromaculata ） was constructed with the SMART （switching mechanism at 5＇ end of RNA transcript） technique. Total RNA was extracted from the...A full-length cDNA library from the testis of dark-spotted frogs （ Rana nigromaculata ） was constructed with the SMART （switching mechanism at 5＇ end of RNA transcript） technique. Total RNA was extracted from the testis and reverse transcripted into full-length cDNA using PowerScript reverse transcriptase. The first-strand cDNA was amplified using long-distance PCR （LD-PCR）. After Sfi Ⅰ digestion and fractionation, cDNA （ 〉 500 bp） was ligated to λ TriplEx2 vector and packaged with GigapackⅢ Gold Packaging Extract. The titers of optimal primary libraries were 2.0×10^6 pfu/mL and 2.4 × 10^6 pfu/mL and the tlters of the amplified libraries were 0.48 × 10^9 pfu/mL and 3.0 × 10^9 pfu/ mL, respectively. The percentages of recombinant clones of primary libraries and amplified libraries were all over 90%. The libraries were converted into pTriplEx2 plasmids in E. coli BM 25.8 strain. The insert sizes were measured by PCR which showed most fragments were over 500 bp and the average length was 1.0 kb approximately. A positive clone of 1 171 bp was sequenced and named RnUb based on sequence similarity with the known ubiquitin genes in Gen- Bank. This sequence was a full-length cDNA with complete coding sequences, which indicated that the library built a base for screening the full-length cDNA. These data showed that this library attained to the requirements of a standard cDNA library. This library provided a useful resource for the functional genomic research of Rana nigromaculata.展开更多
Changes in distribution density,morphology and secretory content of endocrine cells in the gastro-entero-pancreatic system of black-spotted frogs Rana nigromaculata before and after fasting were investigated using imm...Changes in distribution density,morphology and secretory content of endocrine cells in the gastro-entero-pancreatic system of black-spotted frogs Rana nigromaculata before and after fasting were investigated using immunohistochemistry and antisera to six gut hormones.Six types of endocrine cells were detected in the digestive tract of Rana nigromaculata,including 5-hydroxytryptamine(5-HT),gastrin(GAS),somatostatin(SOM),glucagon(GLU),pancreatic polypeptide(PP)and vasoactive intestinal polypeptide(VIP)cells.After fasting,the density of 5-HT cells in the esophagus,cardia and fundus,GAS cells in the fundus and pylorus,PP cells in the fundus decreased significantly(P<0.01),while SOM cells in the cardia,GLU cells in the rectum increased significantly(P<0.01).The cytoplasmic processes of 5-HT cells became shorter or not detectable.The secretory content of GAS cells reduced in the cardia.The positive immunostaining reaction in the perinuclear region of SOM cells in the cardia,fundus and pylorus became weaker,while the staining intensity in the periphery of these cells became stronger.VIP cells were not detectable in the whole digestive tract after fasting.Five types of endocrine cells were found in the pancreas of Rana nigromaculata,including 5-HT,GAS,SOM,GLU and PP cells.After fasting,the density of 5-HT cells decreased slightly(P>0.05),while SOM,GAS,GLU and PP cells increased significantly(P<0.01).Furthermore,the secretory content of GLU cells increased significantly.Considering their functionalities,our results indicate that the changes of GEP endocrine cells in Rana nigromaculata responded adaptively to starvation-induced stress.展开更多
A skeletochronological study was conducted for the age, longevity and growth of a Rana nigromaculata population in northeastern Sichuan, China. Of 139 specimens, 119(68 males and 51 females) exhibited distinct arreste...A skeletochronological study was conducted for the age, longevity and growth of a Rana nigromaculata population in northeastern Sichuan, China. Of 139 specimens, 119(68 males and 51 females) exhibited distinct arrested growth lines in phalanges. Age at first reproduction was found to be two years of age for males and three for females. Maximum longevity was estimated to be at least six years in males and seven years in females. Average age did not differ between males and females. Breeding females were significantly older than breeding males. Females were larger in body size and heavier in body mass than males. Age was not correlated with body size and body mass in males, but a significant correlation was found among age, body size and body mass in females. The growth curve appeared significantly different between the sexes, with a higher growth rate throughout life and a larger asymptotic size in females. Moreover, analysis of the growth model indicated that the population studied was relatively stable.展开更多
High density cell intensive culture pattern is an ecological, efficient, planting and breeding combined new breeding pattern of Rana nigromaculata. It protects racial continuation and population expansion of R. nigrom...High density cell intensive culture pattern is an ecological, efficient, planting and breeding combined new breeding pattern of Rana nigromaculata. It protects racial continuation and population expansion of R. nigromaculata, beneficial for environment construction. Meanwhile, it greatly increases the farmers＇ income, conducive to economic development. The economic, ecological and social benefit of this culture pattern is remarkable.展开更多
对黑斑蛙肌肉一般营养成分、氨基酸、脂肪酸和矿物元素进行测定,旨在比较分析野生与养殖黑斑蛙肌肉的营养品质。试验结果表明,养殖黑斑蛙肌肉粗蛋白与粗脂肪含量高于野生黑斑蛙,水分含量则低于野生黑斑蛙,但两者间差异不显著( P >0.0...对黑斑蛙肌肉一般营养成分、氨基酸、脂肪酸和矿物元素进行测定,旨在比较分析野生与养殖黑斑蛙肌肉的营养品质。试验结果表明,养殖黑斑蛙肌肉粗蛋白与粗脂肪含量高于野生黑斑蛙,水分含量则低于野生黑斑蛙,但两者间差异不显著( P >0.05);野生与养殖黑斑蛙肌肉除谷氨酸和酪氨酸外的氨基酸含量均无显著性差异( P >0.05);野生黑斑蛙肌肉的总氨基酸、非必需氨基酸和鲜味氨基酸含量高于养殖黑斑蛙。根据氨基酸评分和化学评分结果,野生和养殖黑斑蛙肌肉中赖氨酸含量相对较高,第一限制性氨基酸均为蛋氨酸+胱氨酸,必需氨基酸指数分别为70.25和72.42。野生黑斑蛙饱和脂肪酸含量显著高于养殖黑斑蛙( P <0.05),而养殖黑斑蛙肌肉的多不饱和脂肪酸、必需脂肪酸、二十碳五烯酸、二十二碳六烯酸、n-3和n-6多不饱和脂肪酸含量均高于野生黑斑蛙( P >0.05)。野生与养殖黑斑蛙肌肉14种矿物元素中,锌、铝和硒含量有显著差异( P <0.05),重金属元素(砷、镉、铅)含量均在限量范围以内。由此可知,养殖黑斑蛙营养组成价值接近于野生黑斑蛙。展开更多
基金The National Natural Science Foundation of China(30640048)the Natural Science Foundation in Anhui Province(01043202)
文摘A full-length cDNA library from the testis of dark-spotted frogs （ Rana nigromaculata ） was constructed with the SMART （switching mechanism at 5＇ end of RNA transcript） technique. Total RNA was extracted from the testis and reverse transcripted into full-length cDNA using PowerScript reverse transcriptase. The first-strand cDNA was amplified using long-distance PCR （LD-PCR）. After Sfi Ⅰ digestion and fractionation, cDNA （ 〉 500 bp） was ligated to λ TriplEx2 vector and packaged with GigapackⅢ Gold Packaging Extract. The titers of optimal primary libraries were 2.0×10^6 pfu/mL and 2.4 × 10^6 pfu/mL and the tlters of the amplified libraries were 0.48 × 10^9 pfu/mL and 3.0 × 10^9 pfu/ mL, respectively. The percentages of recombinant clones of primary libraries and amplified libraries were all over 90%. The libraries were converted into pTriplEx2 plasmids in E. coli BM 25.8 strain. The insert sizes were measured by PCR which showed most fragments were over 500 bp and the average length was 1.0 kb approximately. A positive clone of 1 171 bp was sequenced and named RnUb based on sequence similarity with the known ubiquitin genes in Gen- Bank. This sequence was a full-length cDNA with complete coding sequences, which indicated that the library built a base for screening the full-length cDNA. These data showed that this library attained to the requirements of a standard cDNA library. This library provided a useful resource for the functional genomic research of Rana nigromaculata.
基金supported by Open Fund of Key Laboratory of Bio-Macromolecular Evolution(No.2006B001)Foundation for outstanding academic team of Zoological Biology in Anhui Normal UniversitySpecial Foundation for Provincial Key Laboratories of Biotic Environment and Ecological Safety and Key Laboratory for Conservation and Use of Important Biological Resources of Anhui Province
文摘Changes in distribution density,morphology and secretory content of endocrine cells in the gastro-entero-pancreatic system of black-spotted frogs Rana nigromaculata before and after fasting were investigated using immunohistochemistry and antisera to six gut hormones.Six types of endocrine cells were detected in the digestive tract of Rana nigromaculata,including 5-hydroxytryptamine(5-HT),gastrin(GAS),somatostatin(SOM),glucagon(GLU),pancreatic polypeptide(PP)and vasoactive intestinal polypeptide(VIP)cells.After fasting,the density of 5-HT cells in the esophagus,cardia and fundus,GAS cells in the fundus and pylorus,PP cells in the fundus decreased significantly(P<0.01),while SOM cells in the cardia,GLU cells in the rectum increased significantly(P<0.01).The cytoplasmic processes of 5-HT cells became shorter or not detectable.The secretory content of GAS cells reduced in the cardia.The positive immunostaining reaction in the perinuclear region of SOM cells in the cardia,fundus and pylorus became weaker,while the staining intensity in the periphery of these cells became stronger.VIP cells were not detectable in the whole digestive tract after fasting.Five types of endocrine cells were found in the pancreas of Rana nigromaculata,including 5-HT,GAS,SOM,GLU and PP cells.After fasting,the density of 5-HT cells decreased slightly(P>0.05),while SOM,GAS,GLU and PP cells increased significantly(P<0.01).Furthermore,the secretory content of GLU cells increased significantly.Considering their functionalities,our results indicate that the changes of GEP endocrine cells in Rana nigromaculata responded adaptively to starvation-induced stress.
基金Financial support was provided by the Foundation of Key Laboratory of Southwest China Wildlife Resources Conservation (Ministry of Education)
文摘A skeletochronological study was conducted for the age, longevity and growth of a Rana nigromaculata population in northeastern Sichuan, China. Of 139 specimens, 119(68 males and 51 females) exhibited distinct arrested growth lines in phalanges. Age at first reproduction was found to be two years of age for males and three for females. Maximum longevity was estimated to be at least six years in males and seven years in females. Average age did not differ between males and females. Breeding females were significantly older than breeding males. Females were larger in body size and heavier in body mass than males. Age was not correlated with body size and body mass in males, but a significant correlation was found among age, body size and body mass in females. The growth curve appeared significantly different between the sexes, with a higher growth rate throughout life and a larger asymptotic size in females. Moreover, analysis of the growth model indicated that the population studied was relatively stable.
基金Supported by Innovative Entrepreneurial Training Project of National College StudentsFund Project of Zhujiang College of South China Agricultural University(201512623001)
文摘High density cell intensive culture pattern is an ecological, efficient, planting and breeding combined new breeding pattern of Rana nigromaculata. It protects racial continuation and population expansion of R. nigromaculata, beneficial for environment construction. Meanwhile, it greatly increases the farmers＇ income, conducive to economic development. The economic, ecological and social benefit of this culture pattern is remarkable.
文摘对黑斑蛙肌肉一般营养成分、氨基酸、脂肪酸和矿物元素进行测定,旨在比较分析野生与养殖黑斑蛙肌肉的营养品质。试验结果表明,养殖黑斑蛙肌肉粗蛋白与粗脂肪含量高于野生黑斑蛙,水分含量则低于野生黑斑蛙,但两者间差异不显著( P >0.05);野生与养殖黑斑蛙肌肉除谷氨酸和酪氨酸外的氨基酸含量均无显著性差异( P >0.05);野生黑斑蛙肌肉的总氨基酸、非必需氨基酸和鲜味氨基酸含量高于养殖黑斑蛙。根据氨基酸评分和化学评分结果,野生和养殖黑斑蛙肌肉中赖氨酸含量相对较高,第一限制性氨基酸均为蛋氨酸+胱氨酸,必需氨基酸指数分别为70.25和72.42。野生黑斑蛙饱和脂肪酸含量显著高于养殖黑斑蛙( P <0.05),而养殖黑斑蛙肌肉的多不饱和脂肪酸、必需脂肪酸、二十碳五烯酸、二十二碳六烯酸、n-3和n-6多不饱和脂肪酸含量均高于野生黑斑蛙( P >0.05)。野生与养殖黑斑蛙肌肉14种矿物元素中,锌、铝和硒含量有显著差异( P <0.05),重金属元素(砷、镉、铅)含量均在限量范围以内。由此可知,养殖黑斑蛙营养组成价值接近于野生黑斑蛙。
