Research On The Habitat Suitability Analysis And Artificial Breeding Of Hynobius Amjiensis In The Anhui Qingliangfeng National Nature Reserve

Hynobius amjiensis is the endemic endangered amphibian of China.A new distribution of such species was found by our research team in March2013,where it was located in the Anhui Qingliangfeng National Nature Reserve.The new locality will be helpful for the conservation and study of Hynobius amjiensis.A consecutive field monitoring was conducted during the breeding season in Anhui Qingliangfeng National Nature Reserve,on which it lasted four years from 2014 to 2017.In the study,a total of 83 quadrats(60 using samples,23 comparison samples)were established,and the size of each quadrat is 1m×1m.Furthermore,a total of 15 ecological factors associated with the wild survival of Hynobius amjiensis were selected and used to analyze the difference among every sample by the Mann-Whitney U method.In addition,by principal component analysis(PCA),the habitat characteristics were analyzed in this study.According to the above result of habitat quality and dynamic variation of population over the four years,we summarized the the current situation of Hynobius amjiensis in Qingliangfeng reserve.Meanwhile,some relevant suggestions on the conservation and management were also discussed in this study.A preliminary study of artificial breeding of Hynobius amjiensis was reported in this research.Some development data from 21 larvae were measured and recorded continuously,including weight(m),whole length(l1),tail length(l2),forelimblength(l3)and hind limb length(l4).The non-linear model was used to analyze and fit the growth curve of the larvae.Furthermore,on the basis of the allele-specific diagnostic PCR method,we also develop a new species-identification method for Hynobius amjiensis,which can resolve the identifying problem on the sampling of larvae(egg sacs,tadpoles)in the field.Main results of our research are described as follows:(1)By using the habitat factor distribution frequency,Mann-Whitney U method and principal component analysis,the results showed the air temperature(?6?)and water temperature(5?~7?)are the main factors for Hynobius amjiensis,and the second factors include the soil type and vegetation form,which indicates the native habitat of high mountain meadow plays a very important role in their survival during their breeding season.According to the quantity surveying from 2014 to 2017,101,91,85 and 82 egg sacs of Hynobius amjiensis were found in the Qingliangfeng reserve respectively.(2)We used 3 types of non-linear growth curve models,including Gompertz,Logistic and von Bertalanffy,to construct mathematical simulating of the larvae developmental changes.The results showed that the logistic curve model brought the best fitting effect of growth processes of the larvae weight(m),whole length(l1),tail length(l2),forelimb length(l3)and hind limb length(l4),and the fitting degrees were 0.9966,0.9973,0.9994,0.9854 and 0.9911 respectively.(3)A pair of Hynobius amjiensis' s species-specific primers,HA-idF/HA-idR,was designed based on the Cyt b gene sequences of Hynobius amjiensis and other related amphibians.By using the universal primer of 12 S rRNA gene(H1478/L1091)and the multiple PCR,the species of Hynobius amjiensis can be confirmed by the number of amplified bands quickly,which will be conductive to the identification of Hynobius amjiensislarva incluing egg sacs and tadpoles.