【摘要】 2014年8月,在江西武夷山国家级自然保护区采集到3号蛇类标本,基于形态学比较,并结合线粒体细胞色素b基因(Cytb)序列的分子系统关系重建,确定这3号标本为缅甸钝头蛇(Pareas hamptoni),为江西省蛇类分布新记录种。更多还原

【Abstract】 Three individuals of snake were collected from Wuyi Mountain National Nature Reserve, Jiangxi Province in August 2014. All specimens were preserved in 75% ethanol for permanent storage after liver tissues were taken and preserved in 85% ethanol. External morphology was examined and recorded on one of the three(YBU14573). Genomic DNA was extracted from liver tissue using OMEGA Kit and cytochrome b(Cyt b) was targeted and amplified using primers H16064(5′-CTT TGG TTT ACA AGA ACA ATG CTT TA-3′) and L14910(5′-GAC CTG TGA TMT GAA AAC CAY CGT TGT-3′). Polymerase chain reaction(PCR) were performed at 25 μl volume, and amplified DNA was produced after initial denaturing for 4 min at 95 ℃, 35 cycles of denaturation for 45 s at 94 ℃, annealing for 45 s at 58 ℃, extension for 1 min at 72 ℃, final extension for 7 min at 72 ℃, and ending at 10 ℃. The PCR products were purified and sequenced by a commercial company. Sequences editing and management were performed in Lasergene7.1 manually. Finally, all three sequences were obtained and deposited in GenBank(HM46857﹣HM46859; Fig. 3). Bayesian inference(BI) and Maximum Likelihood(ML) analysis were performed in MrBayes 3.2.2 and RAxML 7.0.4 respectively. The DNA substitution model was chosen by Partition Finder 2 using Akaike Information Criterion(AIC), which is a TVM + I + G model for the first codon position, a GTR + I + G model for the second codon position and a TIM + G model for the third position. In BI analysis, three independent runs of four Markov Chains for 5 000 000 generations were summarized, and sampled every 100 generations, and the first 25% were discarded as burnin. The ML analyses were performed in RAx ML 7.0.4 under the most complex substitution model(GTRGAMMA) based on the AIC model assessment results. Partitions were unlinked and bootstrap proportions(BSP) were investigated with 1 000 bootstrap replicates using the fast bootstrapping algorithm, and default parameters with others. Both morphological comparison and molecular data consistently indicated these specimens were Pareas hamptoni, which is a species new to Jiangxi Province, China. The new discovery extends the geographical distribution of P. hamptoni to more than 600 kilometers eastward.更多还原