• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    The present invention provides an analysis method for identification of different geographical populations of Chinese mitten crab, belonging to the technical field of molecular design breeding of Chinese mitten crab. According to the analysis method provided by the present invention, fatty acid composition in the Chinese mitten crabs are used for successfully distinguishing the Chinese mitten crabs from different geographical origins, overcoming the problem that the geographical populations cannot be distinguished by reduced representation genome sequencing (RRGS), and providing reference for commercial value sources of the Chinese mitten crabs from different geographical populations of lakes. This analysis method can be used for detecting the attribution problem of Chinese mitten crabs related to market value, and can also be used for analyzing the genetic and environmental interactions of different geographical populations.
    公开号:NL2026401A
    申请号:NL2026401
    申请日:2020-09-03
    公开日:2021-08-11
    发明作者:Xu Pao；Su Shengyan；Li Jianlin；Wang Meiyao；Tang Yongkai
    申请人:Freshwater Fisheries Res Ct Cafs；
    IPC主号:
    专利说明:

    ANALYSIS METHOD FOR IDENTIFICATION OF DIFFERENTGEOGRAPHICAL POPULATIONS OF CHINESE MITTEN CRAB
    TECHNICAL FIELD The invention belongs to the technical field of molecular design breeding in Chinese mitten crabs, particularly to an analysis method for identification of different geographical populations of Chinese mitten crabs.
    BACKGROUND As an aquatic delicacy widely appreciated by people, Chinese mitten crab (Eriocheir sinensis) has tasty meat, rich protein, less fat and carbohydrate, high cholesterol content in crab roe, rich trace elements such as calcium, phosphorus, potassium, sodium, magnesium, and selenium, and high edible value, medicinal value and economic value. Especially Yangcheng Lake hairy crabs are well- known throughout the country. With the gradual improvement of its economic value, the fishing volume of wild population has exacerbated the disappearance of wild species, therefore strengthening the protection and rational utilization of germplasm resources from different geographical populations of lakes has become a key issue that needs to be urgently solved. The identification of germplasm resources from different lakes has become the primary issue, especially for Jiangsu Province, in which most Chinese mitten crabs are cultivated. Different geographical populations of lakes has a large price variance, so there is the saying that Chinese mitten crabs "take a bath" (note: crabs from outside are transported to Yangcheng Lake, soaked in Yangcheng Lake for a period of time, and finally sold as "Yangcheng Lake Hairy Crab"). Therefore, it is necessary to identify these germplasm from a scientific point of view.
    Currently, methods for germplasm identification include phenotypic identification, molecular identification (such as SNPs, microsatellites, RFLP, etc.), and genomic identification. When the genomic identification was applied by the applicant, it was found that SNPs based on the genome range could not distinguish different geographical populations of lakes, and it was even more difficult to distinguish the populations using phenotypic identification. However, the economic value from different geographical populations differs greatly, which cannot meet the market's recognition of germplasm identification. As a result, there is a need for a method that can accurately distinguish Chinese mitten crabs from different geographical populations of lakes.
    SUMMARY In view of this, the invention aims at providing an analysis method capable of identifying different geographical populations of Chinese mitten crabs, overcoming the problem that the geographical populations cannot be distinguished by reduced representation genome sequencing (RRGS), and providing reference for commercial value sources of the Chinese mitten crabs from different geographical populations of lakes.
    The present invention provides an analysis method for identification of different geographical populations of Chinese mitten crabs, including the following steps of: 1) acquiring Chinese mitten crabs from different geographical populations of lakes; 2) separating crab meat from the Chinese mitten crabs acquired in the step 1); 3) extracting and measuring a content of fatty acid in the crab meat to obtain fatty acid profiles data of the Chinese mitten crabs; where the fatty acid includes C12:0, C14:0, C15:0, C16:0, C16:1, C17:0, C18:0, C18:1, C18:2, C18:3, C20:0, C20:1, C20:2, C20:3, C20:4, C20:5 and C22:0; and 4) performing principal component analysis (PCA) and cluster analysis on the fatty acid profiles data of the Chinese mitten crabs, and determining a geographical origin of lake of each Chinese mitten crab according to comprehensive results of PCA and cluster analysis.
    Preferably, the geographical populations of lakes includes two or more of the following lakes: Yangcheng Lake, Taihu Lake, Hongze Lake, Gucheng Lake, Changdang Lake and Gaoyou Lake.
    Preferably, the crab meat is separated from the second pair of appendages of Chinese mitten crabs.
    Preferably, a method for extracting fatty acid from the crab meat includes the following steps of: A. crushing the crab meat into minced meat, performing oscillatory extraction with a chloroform:methanol solution, filtering, and collecting filtrate; B. removing the organic solvent from the filtrate, and sequentially performing water bath extraction by using sodium hydroxide in methanol and boron trifluoride in methanol to obtain an extracting solution; and C. performing oscillatory extraction on the extracting solution by using n- hexane and a saturated sodium chloride solution, and collecting supernate for the measurement of fatty acid content.
    Preferably, a volume ratio of the chloroform to the methanol in the chloroform:methanol solution is 2:1; and oscillatory extraction is conducted for 30-60 min.
    Preferably, a molar concentration of the sodium hydroxide in methanol 18 preferably 0.5 mol/L; and a volume percent of the boron trifluoride in methanol is preferably 25%. Preferably, the water bath extraction is conducted at 60-65°C for 30-35 min.
    Preferably, a volume ratio of the n-hexane to the saturated sodium chloride solution is 1:1. Preferably, the content of fatty acid in the crab meat is determined by gas chromatography; where the gas chromatography specifically includes: Method: GC/FID; Chromatographic column: DB-WAX 30 M L D. 0.32 mm; Injection port temperature: 250°C; Detector temperature: 250°C; Temperature programming: starting with 100°C with a hold time of 3 min; rising to 180°C at a ramp rate of 10°C/min with a hold time of 1 min; and then to 240°C at 3°C/min with a hold time of 15 min; Carrier gas Na flow rate: 3 mL/min; Fuel gas Hz flow rate: 40 mL/min; Oxidant gas flow rate: 400 mL/min; Split ratio: 1:10; and Injection volume: 0.6 pL.
    Preferably, the Chinese mitten crab in step 1) is the one whose lakes geographical population origins cannot be distinguished through PCA by a reduced representation genome method.
    According to the analysis method for identification of different geographical populations of Chinese mitten crabs provided by the present invention, fatty acid composition in the Chinese mitten crabs are used for successfully distinguishing the Chinese mitten crabs from different geographical origins, overcoming the problem that the geographical populations cannot be distinguished by RRGS, and providing reference for commercial value sources of the Chinese mitten crabs from different geographical populations of lakes. The analysis method is applied to the fields of regional identification and commercial value development of other aquatic organism germplasm resources, which also provides ideas for study of the impact of geographical lake environments on aquatic organisms.
    BRIEF DESCRIPTION OF THE DRAWING FIG. 1 is a geographical distribution map of 6 geographical populations of lakes; FIG. 2 is principal component analysis (PCA) and cluster analysis plots of 3 geographical populations of Chinese mitten crabs based on fatty acid profiles data from 18 samples in Example 1, where FIG. 2-A is a cluster analysis plot of 3 geographical populations of Chinese mitten crabs; FIG. 2-B is a PCA plot of 3 geographical populations of Chinese mitten crabs; FIG. 3 is PCA and cluster analysis plots of 6geographical populations of Chinese mitten crabs based on fatty acid profiles data from 48 samples in Example 2, where FIG. 3-A is a cluster analysis plot of 6 geographical populations of Chinese mitten crabs; FIG. 3-B is a PCA plot of 6 geographical populations of Chinese mitten crabs; FIG. 4 is PCA and cluster analysis plots of 6 geographical populations of Chinese mitten crabs based on fatty acid profiles data from 6 samples in Example 3, where FIG. 4-A is a cluster analysis plot of 6 geographical populations of Chinese mitten crabs; FIG. 4-B is a PCA plot of 6 geographical populations of Chinese mitten crabs; FIG. 5 is a PCA plot of 6 geographical populations in Jiangsu Province based on RRGS, where C: Changdang Lake, H: Hongze Lake, G: Gucheng Lake,
    T: Taihu Lake, Gy: Gaoyou Lake, Y: Yangcheng Lake.
    DETAILED DESCRIPTION The invention provides an analysis method for identification of different geographical populations of Chinese mitten crabs, which includes the following 5 steps of: 1) acquiring Chinese mitten crabs from different geographical populations of lakes; 2) separating crab meat from the Chinese mitten crabs acquired in the step by; 3) extracting and measuring a content of fatty acid in the crab meat to obtain fatty acid profiles data of the Chinese mitten crabs; the fatty acid includes C12:0, C14:0, C15:0, C16:0, C16:1, C17:0, C18:0, C18:1, C18:2, C18:3, C20:0, C20:1, C20:2, C20:3, C20:4, C20:5 and C22:0;: and 4) performing PCA and cluster analysis on the fatty acid profiles data of the Chinese mitten crabs, and determining the geographical origin of lake of each Chinese mitten crab according to comprehensive clustering results of PCA and cluster analysis. In the present invention, Chinese mitten crabs are acquired from different geographical populations of lakes. The geographical populations of lakes preferably include two or more of the following lakes: Yangcheng Lake, Taihu Lake, Hongze Lake, Gucheng Lake, Changdang Lake and Gaoyou Lake. The body weight of the Chinese mitten crabs is preferably 130 g/crab to 160 g/crab. The number of Chinese mitten crabs acquired from each lake geographical population is not less than 8, including half males and half females. Preferably, the Chinese mitten crab is the one whose lakes geographical population origins cannot be distinguished through PCA by a reduced representation genome method. After being acquired, the crab meat is separated from the acquired Chinese mitten crabs. In the present invention, the crab meat is preferably separated from the second pair of appendages of Chinese mitten crabs. After the crab meat is separated, fatty acid in the crab meat is extracted and a content is measured to obtain fatty acid protiles data of the Chinese mitten crabs; In the present invention, a method for extracting fatty acid from the crab meat preferably includes the following steps of: A. crushing the crab meat into minced meat, performing oscillatory extraction with a chloroform:methanol solution, filtering, and acquiring filtrate; B. removing the organic solvent from the filtrate, and sequentially performing water bath extraction by using sodium hydroxide in methanol and boron trifluoride in methanol to obtain an extracting solution; and C. performing oscillatory extraction on the extracting solution by using n- hexane and a saturated sodium chloride solution, and acquiring supernate for the measurement of fatty acid content.
    In the present invention, a volume ratio of chloroform to methanol in a chloroform:methanol solution is preferably 2:1. A ratio of the minced meat mass to the chloroform:methanol solution volume is 2 g : (10-15 mL); the number of oscillatory extractions is preferably 3. Each oscillatory extraction is conducted for preferably 30-60 min. Oscillatory extraction is conducted three times, followed by filtering respectively, and filtrates are combined for subsequent experiments.
    A method for removing organic solvent is not particularly limited in the present invention, and methods for removing organic solvent well known in the art may be used, such as natural volatilization or concentration under reduced pressure. A volume of the sodium hydroxide in methanol and the boron triflooride in methanol for extraction of 2 g minced meat is 2 mL independently. A molar concentration of the sodium hydroxide in methanol is preferably 0.5 mol/L; a volume percent of the boron trifluoride in methanol is preferably 25%. The water bath extraction is conducted preferably at 60-65°C for preferably 30- 35 min.
    In the present invention, a volume ratio of the n-hexane to the saturated sodium chloride solution 18 preferably 1:1. A volume of the n-hexane and the saturated sodium chloride solution for extraction of 2 g minced meat is 2 mL independently. The mixture is stood for layering after extraction.
    In the present invention, the content of fatty acid in the crab meat is determined by gas chromatography (GC); where the gas chromatography specifically includes: Method: GC/FID; Chromatographic column: DB-WAX 30 ML D. 0.32 mm; Injection port temperature: 250°C; Detector temperature: 250°C; Temperature programming: starting with 100°C with a hold time of 3 min; rising to 180°C at a ramp rate of 10°C/min with a hold time of 1 min; and then to 240°C at 3°C/min with a hold time of 15 min; Carrier gas No flow rate: 3 mL/min; Fuel gas Hz flow rate: 40 mL/min; Oxidant gas flow rate: 400 mL/min; Split ratio: 1:10; and Injection volume: 0.6 pL.
    After the fatty acid profiles data of the Chinese mitten crabs is obtained in the present invention, PCA and cluster analysis on the data are performed, and the geographical origin of lake of each Chinese mitten crab are determined according to comprehensive clustering results of PCA and cluster analysis.
    In the present invention, the clustering result of the fatty acid profiles is consistent with the geographical distribution, and Gucheng lake, Yangcheng lake and Hongze lake can be distinguished. PCA and cluster analysis are performed for the data of 6 populations including Gucheng Lake, Yangcheng Lake, Hongze Lake, Gaoyou Lake, Changdang Lake and Taihu Lake, and the results are as follows: Yangcheng Lake, Taihu Lake and Gucheng Lake are clustered into one group firstly, while Changdang Lake, Gaoyou Lake and Hongze Lake are clustered into one group. This clustering result is slightly different from the geographical distribution, but judging from the results of PCA, its clustering result can not only clearly distinguish each geographical population, but the clustering result is similar to the actual geographical distribution. Therefore, cluster analysis using fatty acid profiles is more reliable.
    The analysis method for identification of different geographical populations of Chinese mitten crabs according to the present invention will be described in detail with reference to the following examples, but is not to be construed as limiting the scope of the present invention.
    Example 1 Based on the meat quality differences of Chinese mitten crabs populations in 3 different geographical lakes of Hongze Lake, Taihu Lake and Gucheng Lake in Jiangsu Province, the fatty acid composition were studied as follows: 8 individuals (half males and half females) were acquired from each population, numbered taking Taihu Lake population as example, T1, T2, T3, T4, T1.1, T2.1, T3.1 and T4.1 (female crabs with decimal points). After Chinese mitten crabs acquired between August 16th, 2018 and August 20th, 2018 were distinguished by a selected method successfully, then samples of 3 other populations were acquired in the same way at the same time; the geographical location and abbreviation information of the 6 lakes were shown in Table 1, where Taihu Lake population is represented by "T", Yangcheng Lake population is represented by "Y", Gaoyou Lake population is represented by "Gy", Gucheng Lake population is represented by "G", Hongze Lake population is represented by "H", and Changdang Lake population is represented by "C". Table 1 Sampling point information of Chinese mitten crabs in 6 geographical lakes in Jiangsu Province Sampling Lakes Habitat Area(km ) Longitude and latitude Quantity location CHANG DANG N31°59"- E119°52"- C Lake 89 8 HU 31°62 118°60" N31°14' E118°53" G GUCHENGHU Lake 65 8 31°18 118957 N32°42 E119°06" Gy GAOYOUHU Lake 674.7 8 33°41’ 119725 N33°06'- E118°10" H HONGZEHU Lake 1576.9 8 3304() 118°52' N30°55"- E119°52" T TAI HU Lake 2425 8 31°32 120°36' Y YANGCHENG Lake 119.04 N30°55"- E119°51" 8
    HU 30°32 120°36' The left and right second pair of appendages of the acquired Chinese mitten crabs samples were cut, meat was taken out and placed into ziploc bags marked in advance, and stored at -80°C; the sample was then cut up and dried, and then about 1.9 g of the sample was weighed and added with 15 mL, 10 mL, 10 mL of mixed solvent (chloroform/methanol = 2/1, V/V) respectively for oscillating and extracting three times, followed by filtering, and filtrates were combined for evaporation drying; 2 mL of 0.5 mol/L sodium hydroxide in methanol was added, and 60°C water bath was conducted for 30 min; after cooling, 2 mL of 25% boron trifluoride in methanol was added and 60°C water bath was conducted for 20 min; after cooling, 2 ml of n-hexane and 2 ml of saturated sodium chloride solution were added, oscillatory extraction was conducted, followed by standing for layering. The upper layer was analyzed by gas chromatography. The used instrument and equipment and the set parameters were as follows: Instrument: Shimadzu GC-2030; Method: GC/FID; Chromatographic column: DB-WAX 30 M 1. D. 0.32 mm; Injection port temperature: 250°C; Detector temperature: 250°C; Temperature programming: starting with 100°C with a hold time of 3 min; rising to 180°C at a ramp rate of 10°C/min with a hold time of 1 min; and then to 240°C at 3°C/min with a hold time of 15 min; Carrier gas (N2) flow rate: 3 mL/min; Fuel gas (H>) flow rate: 40 mL/min; Oxidant gas (Air) flow rate: 400 mL/min; Split ratio: 1:10; and Injection volume: 0.6 pL.
    (4) The data obtained in step (3) was subjected to ANOVA (used to analyze whether there are significant differences in fatty acid composition of different geographical populations), PCA and cluster analysis to obtain fatty acid composition profiles through R3 software for identification of Chinese mitten crab species from different geographical populations.
    PCA and cluster analysis were performed on 3 geographical populations of Chinese mitten crabs using fatty acid profiles data (excluding MUFA, PUFA, UFA and SFA) (see part of the data in Table 2). The results were shown in FIG.
    2. FIG. 2-A showed a cluster plot of the 3 populations based on the fatty acid profiles, and FIG. 2-B showed a PCA plot of the 3 populations based on the fatty acid. It can be clearly seen from the combination of FIG. 1 and FIG. 2 that Gucheng Lake, Yangcheng Lake and Hongze Lake may be distinguished using fatty acid profiles clustering, which is consistent with the geographical location distribution. Example 2 To verify the results in Example 1, 3 geographical populations of Chinese mitten crabs from Gaoyou Lake, Changdang Lake and Taihu Lake were tested using the method in Example 1, and the fatty acid profiles data of 48 Chinese mitten crabs from 6 geographical populations (see Table 2) were subjected to PCA and cluster analysis together. The results were shown in FIG. 3. Combined the basically consistent clustering results of FIG. 3-A and FIG. 3-B, the geographical populations of 48 samples may be determined. Therefore, cluster analysis using fatty acid profiles could enable identification of different geographical populations. Example 3 The fatty acid composition data among the 6 populations (Table 2) obtained by liquid chromatography were analyzed by ANOVA to obtain fatty acid composition profiles for identification of Chinese mitten crab species from different geographical populations; the obtained fatty acid composition among the 6 populations of Chinese mitten crab as well as the contents of saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs), polyunsaturated fatty acids (PUFAs) and unsaturated fatty acids (UFAs) were shown in Table 3. Table 3 Fatty Acid Composition of Chinese mitten crab Populations in 6 Geographical Lakes in Jiangsu Province Cc G Gy H T Y C12:0 0.01820.002: 0.0170.0032 0.01620.003% 0.01720.005 0.01240.002% 0.0100.003° C140 0.418+0.035 0.48320.017 0.411£0.023 0.514+0.029 0.460+0.019 0.515£0.036
    CIS:0 0.207 20.011° 0.27520.009°: 0.235£0.014% 0.399+0.033* 0.248£0.009% 0.28520.013° CI6:0 12.15740.237 12.43420.146 12.870+0.199 12.07620.144 12.677+0.236 12.222+0.206 CI6:1 4.38120.465® 3.34820. 1145 5.59620.6922 5.14420.676:® 3.50940.1715 3.25020.120° CI7:0 0.617£0.032 0.64440.028) 0.51540.042° 0.87820.092 2 0.542+0.025° 0.61240.0225 CI8:0 7.70720.205 7.36820.207 5 6.98820.186° 6.08040.255 7.328+0.154° 7.328+0.170® CIS:1 2297920747> 22.610+0295% 25708£0.765% 22.70240.962° 25.215£0.375% 23.026¢0.291b CI8:2 8.090+0.771° 5.6740.343% 4.59020.297° 5.183x1.190 5.73640.426% 6.669£0.433 CI8:3 1.37940.085® 1.225+0.117° 2.24540.269° 2.463x0.221* 034730,052P 0.9564), 046° C20:0 0.142+0.014 0.17420.014 0.1280.010 0.11820.015 0.160+0.042 0.197£0.018 C20:1 0.768+0.081° 0.8710.048" 0.79020.06 1" 1.08920.107% 1.0650.046% 1.367+0.103: CM:2 1.27340.12963 1,73520,053® 10040000 15840, 1Q5 a 121540,091 ¢ £866.) 082 2 20:3 0.58220.058% 0.4070.028 0.5870.026% 0.48620.051° 0.29120.022 0.35320.020°° C20:4 6.3370.252% 5.99120.221% 6.882+0.338 8.72120.894° 4.74120.366° 5.73520.218% C20:5 1I893340.650% 20:28040.2422 17.495+0.860° [7.933£0.822 [8.30820.261% 1873110386: C22:0 0.109£0.004 0.124+0.003 0.11240.015 0.090+0.006 0.116+0.006 0.113+0.010 SFA 25.75440.458% 256710306 26.87141.258% 25.31440.462% 25,05140.2990 253240259 MUFA 23.95640.76453 236804) 2640 26.71020.725 24.02550.994% 26.485£0.4228 24694), 2720bed PUFA 50289x1.175" 314534 3870 46419£1.168> SO.660£1.010° 48.463£0.610° 50.809:0.444 UFA 74.245£0458% 7513230306" 73.129£0.532% 7468620463 7494920299 75.468+0.259 Note: different letters in the table represent significant differences in this indicator among populations (comparison between rows). As can be seen from Table 3, no significant difference was detected in the C14:0, C16:0, C22:0 and C22:0 contents in all Chinese mitten crab populations. A content of C17:0 in population H was significantly higher than that in populations C, G, Gy. T and Y, while a content of C18:0 was significantly lower than that in these populations. A content of C15:0 in Hongze Lake population was higher than that in other populations. A content of saturated fatty acids in population Gy was significantly higher than that in populations C, G, T and Y.
    Regarding the content of monounsaturated fatty acids, significant difference was detected only in populations T and G: a content of MUFAs in population T was significantly higher than that in population G. Regarding the content of polyunsaturated fatty acids, a content of PUFAs in populations C, G, Hand Y was significantly higher than that in population Gy. A content of unsaturated fatty acids in populations G, T and Y was significantly higher than that in population Gy.
    Results from PCA and cluster analysis were shown in FIG. 4. Yangcheng Lake, Taihu Lake and Gucheng Lake were clustered into one group firstly, while Changdang Lake, Gaoyou Lake and Hongze Lake were clustered into one group. The clustering results were different from the geographical distribution, but according to the results of PCA, the clustering results which are similar to the actual geographical distribution can clearly distinguish each geographical population. Therefore, cluster analysis using fatty acid profiles is more reliable.
    Comparative Example 1 The Chinese mitten crabs acquired from Yangcheng Lake, Taihu Lake, Hongze Lake, Gucheng Lake, Changdang Lake and Gaoyou Lake in Jiangsu Province in Example 2 were subjected to RRGS (Beijing Novogene Science and Technology Co., Ltd.) and PCA to confirm whether it could be used to identify geographical attributes of different geographical populations.
    The results were shown in FIG. 5. As can be seen from FIG. 5, the Chinese mitten crabs from 6 different geographical populations were subjected to PCA with RRGS data. Most of the 48 Chinese mitten crabs samples were clustered together, and it was impossible to distinguish the geographical populations from their respective origin. The results showed that the genetic characteristics of Chinese mitten crabs from these populations were relatively similar, and the Chinese mitten crabs from these lake geographical populations could not be distinguished by PCA through a reduced representation genome method.
    The above description is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, many modifications and modifications can be made without departing from the principle of the present invention, and these modifications and modifications should also be regarded as the protection scope of the present invention.
    Table 2 Fatty acid profiles data for 6 populations
    Tete Ga en Tae Sade ge ae See Sa ra Tee igen iat a ei ea te Te Dati Ted es TR Sh TRY Jo i ia Tet Teen See oe ee Soe re Bo Ba fie de 1 Porro ne TOW INE TOW I Caen AIR ee DRE IH B00 IRIE TRG TEE DN A WS RA JR IE ELE TARE Hel DEN ES a DARA NO A TERY Ge Led Wa Be Baie EN Dd Dee ig ag Seed Dang fed ed inal ee Me RY Ne EY DRS SRD DRY ND RY MR Ih IAN DOD NNN Nn DA ION RN DY A Ih IE Ty MONON WENT AY Ne a a opr TTT TITTY RAY NN PUSAN Ay Ted Daag aad bend Dad teed DG TI IGN IND ATO SN NO UN NR FR EE MYON EWEN NY Gena EER TEE I UREN HERS HE SRN ENE RIE REIR A 00 § 8 NGI NER DEE re Corr DEE UU rr PUTT whe dei fen le tle Ried Vr DIT niet eh eh EE Ig Rien ee Re en en NR A ee NO ene en A A en er NK OM Be Sebo bebe eee ee dede Pee mins! Len ine ix en minis] eee dene Boe ben Den Dane ben Dene Dede eve Dede do de dies Dene len ee] mua dw enim Mavi les Dens fede Ee Ee melds IN
    EEE ER EI LEE CE NE NE MEE RE IRIE Ry nt Mme EXAM 1 ANNI N IRD MINIT 0 OR IE KD EN OND LEN RI EY HE ND Ed INS NN NIN OND KIND ND IKN Ind NN MDN NaN: in re nel Dem ES ESES wa lie Dali me me dre Da EEE EN ase ind DEN IA Ta TR DE DOI OE es JE DRS GC TE a TID Mle 0 In EE RCE Tee OR ee ie 00 XS oh it The ee dee Ge nceey NY I a fe eel baaien BT Da DOS Dah heh Dome lei] die Ie Led Fo RTE ATE ERIN 1, ea! . iedee, mina SE EE EE EE =S EEE SE EEE INO Da aN WE Re We Wes len mal ea I Nr Na Re Ea Re elen Du ies DRED gh ST en Kn dhe REED ed Dae Dl Des OF IU DRG Wee Mi IGE MIE TNE Ka Yad ROE Rd TES Se a ea IO et Ld NN Br IO RTD ln DI DA I hE Dy DN EC Oe SY DT Ie De TT C0 TER GER DE NN Rah JONI Dea Dea Yad wh OD OH GaN aT oh TE Ta Fo) hd ee IK ee TT ey TEI Ded DE Ga ed led Nd NN Ins lad Kn KN Kien Rind ina inn lee da Ne Nae len en jen Gadi kn EREN], Koko lew jes [es Fd anak fae BIT LT TT IT LT TT TT DT eT EE EEE PETE NT ar TE ry 2 ETE ma EE ESS Te RE RE IEE TR RR RIE Ee EES ESES ELEN SEE SES Reels dived RD IRE Ea be Kaede TEED Vale BE OCR DE24 Dd Der EEEN hd WED Vad EEE EE ¥a IND Ta Dear Nel a Ede Ne els a ade De LR NT TB le SEN Dh epe ee ee ene ee ee Dn ee ee ee Den ep ns ne ee en : EN oy EE Ths RS ERT LR Ka Idee re Tr fe ps I ne ee BORO TRING (Ra TR IE SR IRE IR RO RE I ee Dee EE Bee ILC EE EEE EE Tn EN EEEN UN be TTT ET ey Tea ET a Tien een Dn Oe Ded DOT Tin EEE ger SES EE ad TA 1 Ti Nor BE X00 Tel Ded ESE ESE TTA EAN Teer Veet 00D TAC WA URIDINE Doo Kd RCN ROY Male tela Darl ede leq Dab DI Ted DCA 23 Bd aie I SE I a Tat ud a3 DEE HRD Bh Ila Tee DR NOR east NS Deve Dede Dead Gave [Te DON ae Dad ne dnl ed RAEI ead Data Dd Da De DG Dei BE We Tle foe Tes GER XT SON GEE DN ee pa Tn Md ange be ind eta ea ien isa 0A Ten Teal Tear Tae Tat pal RY Tat ea Wan IO SR Dn fen Pt Tos xa sal ita Lo BR lag ige fas Teal Frenne BNR we RAR LT PONE TOR Aenean bent NR Rh Nn Et dt nn baw tf wt cher hr 2 NN appt ge Wa nN dn» rN FG Trad fev Dn en Deol De Td Uh IRE TRY 0 Ne HE NOTH Se Tea Tare Thur han DT Hee ET tae ear vai ene EE Le ud fal No ee Ree pe TE rar Dal De Ea Dac Te DEN Ded Das Eee dal Se In Ted nf Tac Tdi TN apd ee et [GO KE CH TR GHG Td 0 We LE DEE Tae $07 N08 $0 MAAS) TN BE I ee ee J ee a Na Nd ST BEN RT eT EE DD orn orn FE EE DD DE
    SE SEE ID NE SEE LEL EE EEn PETRIE Teun Teds en Gum Dee Dede lw law Ig em ae VD Dede te lemme Dd Dad DT ST Lod le Ded Lede MP 30 Wee Woe (008 ar nar Tae ne Dh Dod Mga dN hedge id eo GEN Tee The TCH a I hehe TR i nen Ga DN NE Deal at Te Dae Tre TN UGE Wh BT ee Ten 0a Dane TOR EE ent DIN AUN IU Bay de Ih Dn Da Non Na Nan bed 20022 FB Te en le ee Ie ES EN Dn Be WF Eg HET Rr TT EOC IRN RO ROR | EEE ed TE nt De fe Dee TdT hr MO EE et el EEE TE ad RS ER Pde re he Te Red BOE en REE TT nd eye nT lr aes ne EE TE a Toad Ted TEE a Dae Da BBR INR EERE ES Dead Dua Ia Dard Die Te Dds 3a See Ina Tee De DE Made Nan EE hog 3 Xe Ie Tee lip dade LO Toh he Tele Shr feed Doo den i RN Tale Dev ate Bd Th Day N3O0eer Gnd Day ter aon Won DT Dg Whe BY Ee Bd Id Dae ad dev bn an fede Da Inn Dike dr ed I Ina Dad Novi ee Kee Xe fel IE Tear Da DS per Dae SR Cet RE Dae Sed OR ee SENDER De ee dee Send Tem Dave Tene ed ew nS STE SES ESSE Wh a EE In Ine Re Nu Mw Re ie Wie SEE SE id DIN ST ae Ta SE wT gy Mwy SUT ET ed Tha Ka APRN VER DASU Than eG el TRL DANG ed Eins Be It TOT TE IRD Gast IR Th DID NSU NSU POE IRY Di DRG EY BB BI 0 eo tee YT 3 RY ed Dive Be Be Ray Ra i
    BO I I NEED ENE EE EE ENE EEE ESE SRI SE ER SIE SE ARR Ter Ded Dade thr Deed Tr Dn hoa In Ia To RN RO en Dad Ned Rd ESS em te Dan tna ies Den fee nies lee les ka emis daw dul fee Dew lan fon few olen Won im len ien iow lo dew few dus on ies io Ro lew ien iow dul sedan id IT ET TT TT TT IT TT TT RT ey IT TT I A gry 2 NT NTT HT TT LT gg WT ggg tT RTT TAT A LT TTT WT EEE TE En Deen Then DDE Iban el Be Ta Ta fee ee ant Be ITI TO IN Tae IR TE DST Ren Ber den ley ied teed FO Ee Ne Net D0 Rel le Tied Ties Be NT Re nena LY dis ET Sen Tule OOD IKE Be RR ae Tee Gln be he Daly OIG ICT ACY SO ee Tue Dn Dw Bd RT IE IRD TD RE den den aD TRE ER ED TT R30 00 pe TRE NN de RN Ti ie NEE TRE Ies Da Dat SRL INE a TRG DE GS Bn eis ELE NOTED DT De Bad Dae DR DOs Dace Fel wa gh On Du J es boi RE SEE en DEE HR Don Bh ee Ine Tal MR ae ae rh] DD] rine lee er ine Ded ins pagani Gapaies Roeder ed Des ed Dea indi The Eo do BO ve ip es hehehe aia ing dsl Goes ga Radel kor The TEN Dn Fan Dade DR ee CEE TEN) DRA EE ee OE DI SR DD LI INO Tg TaN GRE Da NR TE TOT Ee MN wn GR ale Ces im MA Yn nen La per ENO ne Fg LTA Te TREN CW RTE NA nL de FT rin lw Mw a ne FF rb bl TENS wa TOE Net Leh Teer ep Ne le ed AN RNY el NCTE vel Bd TCE CIN elle GN NGM IE eR LY OE ETE YN MD IED Nelo ele Je] ZR AE ble sss ee GATED fed ah ed BL ee Eek xe di en LO Lo ZE SP dw DRY Sn in Ten LR pa edad Ra Ea DE Te DE DT DN ee Cae Ea DI TR a NY DL te TI Ted Td TaN Oe Dad ME NET EE Ton Bd wh ne Re in te XS I ey De TN! EEE EES tie tes Te ds Pan ada les
    SEE EE EE EE SE EE EE EEE SE HF EE RES DIe eN ED a ND a AD DRE ENA DEREESE ERE SEE SES SSN ES EEE ES EEE SEENEIS EN ES DS ENE RT
    ESSE EENES SE SES SS EEE ESES EESTI miele] fea tas Brad dea Dae der der frm es Bede dre Deen Dre bene Jee ie] NN NN Nisee es Yer Be wie xl ee Gene lene late Ja de lene [en lad Be Be we ME Ind EE EE ES ERT CNT nd de hem CT En EEE fen Dore Ten © Bee ee kain DoW Lew beheksen See im he hai ud Ted Xe Ia ns Tea 3 Nek aI ED tee X00 ID Le Da MR ow dean 10 [Ei TEE TE TDA ESSE TE ae le TR a a Rr TU De TR Dae Dee a SE a Te i Den Tae 10) TG ERY Hee 0 oie DE Tle XA Se Ka wn ee 0B in 0 ESTEN ENE DAH TH Tend hala end Tend Ws al Ta NN nn ied ome I Ha a A FEAL NR Sn ROI J NI § Xun: iel II ed
    EERSEL SE NSE SEE NE NS EN ELSE SR SELS EE EN SE ad te SH en en a ee ee ee a es SD a DERE Sn ee De ee ar lan [Sa tin ie TRE en ND ene ies Tod ld DG EN TN a es Tear le a Del Ded [EN Se HE IER DA TE We Ne 0 ale Nii Die dn Ted NT ns | SE EE SES SS SEE NEN EIS ha SEE ESES tei ha haha ES dies NEN HNE NE NE SEE NE NEEN SE] ver Ds DI DE a en A SO ea en a DI SRE ee TE EI ED Ee ee De EONS CN TE TRIE SRI Eh PELE LANE a DE A I EE EE EE EDE pe EE EE SE sea a ede Te fe ee To Tee pee eea a ee Teen To i Wee eee a a en en en te et tie gee pa i De EEN Ds en DION DD (ER Ee DA EE A A TE OG IE DO RE A ACE TR EN EON TI ee MN] Dotan to fea been ta edi 01 lead aims ein! ief ai Die IR Dei eN ne [ee eee Palen re] Ln GN ien inn Ie eds Rede! EE TE EE EE EE LE nT RTT a a nT ST Te ee Bae Dear te dae Dany hae TN Tan ta ain da Te Ia Thue Tee TEE TORY Flo Sn en ee Sead Xd eerie etl Da nol la fen tee ee na iN PG EE Ti fed Te TERR Ded Re ele REESE ESES es Ee et Reg
    ER RI ESS SE SEE EE Tee BE Te Ten ee em fees Bann Ten eije] eG de ei Ny ESE SEE SE EE SES NI ISL I Dy vy BLAIR THEN Ae JRF IT WI a JRE KTR Be TIE LR DURE RRO a ee BE CRG (I LB AIL ee 0 TR UR CTR SIRE SCT 5 Bo AEE iva ind es Nin Nay 6 Sr Dad Te Sede Dh Dd GEN II Dee ede Sa Tar See Tea Do Dale Dol 6 TRY DCH Gary ne Dali le X07 X00 90 Soa Ad Sted ioe Ne xed lea Di Re a 0 Rada Vi BE AE ee ZE IE a a RAE ID a SL HERS Td A DD orn orn DN ee norm rr FE pe Ra Deir zeiss dre igsipe ins oie den pe Bre ien ie ies Tee fen Xe Be de fee fs je ape EEE Ibe nnie Xe dels es pe ha gad ee tr TooToo ne Rn ERE EEE hed eb REEL CEE EATER SE OU SCT DEELT gd Tech Teer Dig tw Uden Tage Sed a a din Tei a mae dod Malt dy ee Dia ear Ten Dee So Wo dae 00 Teer DEE ND br ie en iter A har hee WN Xi Tee lun Felon minded Tele the Dev Good Dede Dela den NIN Den edn Uh aN de ol Mr fear DR Igy DON Den I Ded he mv a dua tee Nas Fu Sen Tn Ma i hes En WIG I ly Na Nate Ted FE Es le Ee er Dee Te DP a EE RR ET IR TSR OR SN Ta DEN Dan ee DEEN ine Nn Di Fron ed AND TO Dh DT WE EN DN IOs DN aN NN a fen fate ml » ERR ROD zein ien ied WONT IT my VTL TTT LT BATT LT See FUENTES Tu Dann TOT IN De In RTE aN ON ETE Rey een EN D0 a TEE er Dead Nat Fr Tew DAR Ded Te EYER EEN PURER waa lod DOU Ded RY EE Rie Lon EEE Ea DRE DET a ea DO el A ee ede Eel Gn SE ar RE TO Dd dai Red MN Sel Der Gly TRE a ihe le 0 IGE dl aE Da Sed EE Cd lie WN A Rad JERE DHE TLC EEE DOH TENE Te D3 ed Tae TR 00 EES IS I tah den dln Neha om Oe ee SE WR ee CE A SE AE WEA SE A a A Ho RE EE EV Ln We Tn PA TL ET RN NE EE ee EE: Doron on ohn DIE DEE Et EE EEE Ww iam lene em Dene Te eine in Den weed Dele Nolen Nm ele wine IN me iam kl lo INo ine ioe Wawa ma mad ESSEN FLT he TT TT TT LT ATT rn pa eT eT eT eT TL eN NS NS Ge gre Var DH PEAY DN ESE TRE DR EN Da REE a SE EEE FW Iki D0 GE TRE Xana BLL Toon NaI DD Tale Need DCR EERE DAO ce EE ey EA IND NO a ase le od Ede ved £000 wef ed lade NON TEIE Tek OO Ia } (ob Ga ive TA IES a Toe ee Ed a Dean Tae Ba GN a ad rn Taal ae IS De TO ee ai RT di 3 ete De DE ie ee BD Se TR 0 ri LN MN RE ete TY aT Be Mp aR EE INGE EE RSE Es Ge eee eg LE RS Fe A EE IE EE B EEN B Ge Dante TA Dn Dali (al ed as Tag 0 ta Kiten DN I Dn Te TET TBD OER Dah Ml Jule Xan CE RTE Deen MY fer Mh Noh IND a ED le Nor Del Des Dole ae ed EEE dl BT Tn dee pen ETE IT Tn hk Eee FF nen EEE] TU Deer Tagd fen Dl Rd Gen Eas Ten eS SEE San ira 1 Dal Dee 30 War X32 0a fee Dita B02 a0 0 iad Doo 0 Die an Hed Then a Ten den Ne mean ee tes TEAR Dt Date Tan VINE DA Date UTE NE edd DEAD Ee Re i TN DRY Da ea a Ta Le eed Wee Be INT ned TR Wi Bo NT NL ER TE Late ee COE LEE dh TE Nhe a a a ie ea Nee TF OB J a (Ca le ie ee RON RN pr EA EH En a BER HEE ita] Teh hai Tea eh ENTER EEEN TEEVEE STE IE EN EVEN EE EE STD HE TE ENEN ENEN ENEN EN SISD IEN IEI NN EN ESES © TT ne TT Rn em nT eT nT nn RT EEE EEE EE Ix fee ae Dox lpia ins Tee kain DA US RINT Gr Fen Tea Dad Ted Deer IR I Dad M3 Nd er bon Tad Wijde haine fos fer fee Xan de len Deen [en Ge ease ed TOE TAL be TO UE DRI EN EE DAE Ee WL I Ln IRE ALE TRY TEAR Date HE NTN EAE ETE DE UE EAI AGN DR MEE TRY Tid Lint B J RN Ne Tee DCH an Seed Du TG STE a aa Tele DOE NN Re a TE Gear Tee Tn Dee Jee MN Yel Il De MAO LN We fat 0 a Dien 00 Ba Ie Tap ed Re EN Waan Dale Dela [CN ale leds IND! ION Tele MND Dale MMe lev Dela ld IRD Deen (ND Uwe BEER IMD IEN NN ND NDOT ba Dede ER Re IMD IRD Ie MN ND NNWaNN 3 TAT RT RTL LE Eh he En le Iie EE EEE Ten TROD DCN TOR EN) EE me I DI DED De I Tl TET ee Bod Rad ETH a 0 OE EE MXN EO lin Tot a DO EE Dh DO AERO AEE pad aR EE We I Ie edn Deh ade Ted Di da 3 RE od tah Lah Tha dE Sag Gy Sdn om eae ee wn IRD ESE Dev TE Th EER SEE Tah Bt BE a Tl Tet 0 a RY MA Beer Dad fed DR Tel Dr Dads 3a NRE Ted TY TRY NAO RON Ne in Re Tole 40 Xa a TD Tie Jel le aa ei] Ted Edn Ten ted Ien leien ies! Tad led 5 a Tle WB Ine en lee tee Do Dee GN Tend lel Wo Me etn Neo © Din lide lee pe list Ro ev boter wy EE DT IT RS BI ET ET I i IT ET eG ri Led Dry Dd Daan Dg Ga be tLe ge ee in Le XR Da a a Tee TL Wn bd on Die NX be fp ue TE IO RT ee TeX a Re Ro i PX LED EN DY Dl et CECE ED TIN Gy He DC aN DY Gs en ID CE Nae MONET Tat DOI MN RTH Neh SHR te bd Xa eT Tey ed 3 Sa RE ee ane DA SEI Tee RIOR Ge a Ta Teen ah ah RE NOs IR Dn LE Gr TON Te Daly wen oh Ne RY Thr Ee 1 ha ee deen Eada wee Re GR Dees ee ad Dea len Daan fra fae ae dren © fa tas ian en Sev Deen dee Ine tra Swan d (or tee eer Net New hen eer ii Yer Do Ges ger Gren Sen Sew aes New dent fer leet Yes edn] ie ents leien es lea ien Beinn lei zen ee es I SE EE ADA IP IEN PN TI PEP a nd SES SSS Dee Dis 0 ws wenden, Bo Th Ea TR TRAY DRE Del ner BEIGE Id DAD BCG i Ny el kd ake AE TE hE ee deter Dal gg NVE Kn SE Ed LODGE Dey De MEX) hada TR DAE TR Il Nv tl I Rg te tool Reh ae en Ee ee tee tA TO Rg ra RR a en XEN a IR We TE ETN RT ae Ne NT TERE MY Sn aes Na! Tea se Ge IGE A LT a aN ee dre en aN Tn Whe Le Tae Dah TEST TENE Td Re NE XE EA ea) lean BA XN Ee doe en AN aN Lae MN Nd TaN He FES 1 vd HAIRY RES SI EE BEX] ie ESS SS ed en ey TION en Eee EEEN DN IRD FRU FO NA ACR Th ZO gh IT © IE Nea Ln sij WONT ge AN B EE SE EES ON A ee Dae De Ee DA NN ag EN ee TN CEE CERN NON EE en ee GN AEL NA SETA IEN ETAT Ww ela ie ge Ee asia Pe = EE ee eg ER ee BL [a es PTO ps Rt BER RRR Ea a PE RST TCE DEE a ea De Ae a DIE INCE SE ea 1G DAN RN RN TO Ne DSR [MD EO Ne] Ee EE EE RE EE EEEN ENEN EES BEE EEE EN ea EE ESES EE EN EN EES EE ENE EEE EEEN EN EES SS EE] TRI LHD Cn RE I Ta ae Da ren EUW ANY I Te Ta pa ERY De DR XW dt DAT en EN XE Ee Re lg tee nA ol I DEAE TOA Bd ENE EN Ws ly TRI NO I NINE ae af rl HE IR UY A DER IE IA EI HE RO PRN J ERE Si Re RAE PR pe I Ae ie EER VER DONE Che GER URN IGE EE a lel TEN Fen le DEN Se TENT TRY Lge DRT De TR TR De BK I Ee IR hd I NA NR KT TGR ed es a WE Les I ee Slaw Tend TE Day Da Te Da Da TES ERY Dale 00 GH EG ST IAN ID Der TD a TY TG ERY BUX MN Ia DCE Dane X02 XU uh XH CR Boe En ND M0 leh Dat Dees To X01 Kanade | en De IE ei Dn Ie B De Ee YB EB TH IRIE RAR a BA PE NA IE RR CRUE PIE SPR rat
    权利要求:
    Claims (10)
    [1]
    Conclusions i. Analytical method for identifying different geographic populations of Chinese mitten crabs, comprising the following steps: 1) obtaining Chinese mitten crabs from different geographic populations from lakes; 2} separating crab meat from the Chinese mitten crabs obtained in step 1); 3) extracting and measuring a fatty acid content in the crab meat to obtain fatty acid profile data from the Chinese mitten crabs; wherein the fatty acid is C12:0, C14:0, C15:0, C16:0, C16:1, C17:0, C18:0, C18:1, C18:2, C18:3, C20:0, C20:1 , C20:2, C20:3, C20:4, C20:5 and C22:0; and 4) performing principal component analysis (PCA) and cluster analysis on the fatty acid profile data of the Chinese mitten crabs, and determining a geographic origin of one lake of each Chinese mitten crab according to comprehensive results from PCA and cluster analysis.
    [2]
    The analytical method of claim 1, wherein the geographic lake populations comprise two or more of the following lakes: Lake Yangcheng, Lake Taihu, Lake Hongze, Lake Gucheng, Lake Changdang, and Lake Gaoyou.
    [3]
    The analytical method of claim 1, wherein the crab meat is separated from the second pair of appendages of Chinese mitten crabs.
    [4]
    The analytical method of claim 1, wherein a method for extracting fatty acid from the crab meat comprises the steps of: A. crushing the crab meat into mincemeat, performing vibration extraction with a solution of chloroform:methanol, filtering , and collecting filtrate;
    B. removing the organic solvent from the filtrate, and sequentially conducting water bath extraction using sodium hydroxide in methanol and boron trifluoride in methanol to obtain an extraction solution; and C. performing vibrational extraction on the extraction solution using n-hexane and a saturated sodium chloride solution, and collecting supernatant to measure fatty acid content.
    [5]
    The analytical method of claim 4, wherein a volume ratio of the chloroform to the methanol in the chloroform:methanol solution is 2:1; and vibration extraction is performed for 30-60 min.
    [6]
    The analytical method according to claim 4, wherein a molar concentration of the sodium hydroxide in methanol is preferably 0.5 mol/l, and a volume percentage of the boron trifluoride in methanol is preferably 25%.
    [7]
    The analytical method according to claim 4 or 6, wherein the water bath extraction is performed at 60-65°C for 30-35 min.
    [8]
    The analytical method of claim 4, wherein a volume ratio of the n-hexane to the saturated sodium chloride solution is 1:1.
    [9]
    The analytical method according to claim 1 or 4, wherein the content of fatty acid in the crab meat is determined by gas chromatography; wherein the gas chromatography specifically comprises: method: GC/FID; chromatographic column: DB-WAX 30 M 1. D. 0.32 mm; injection port temperature: 250°C; temperature detector: 250°C; temperature programming: start at 100°C with a waiting time of 3 min; increase to 180°C with a ramp rate of 10°C/min and a hold time of 1 min; and then to 240°C at 3°C/min and a 15 min hold time; carrier gas flow rate N2: 3 ml/min;
    fuel gas flow rate H2: 40 ml/min; oxidation gas flow rate: 400 ml/min; split ratio: 1:10; and injection volume: 0.6 µl.
    [10]
    The analytical method according to any one of claims 1-6 and 8, wherein the Chinese mitten crab in step 1) is the one whose geographic origin from a lake population cannot be determined by PCA by a genome method with reduce the representation.
    类似技术:
    公开号 | 公开日 | 专利标题
    Rojas et al.2016|Bats | challenge a recent origin of extant neotropical diversity
    Santos et al.2008|Age and autochthonous evolution of the Sunsás Orogen in West Amazon Craton based on mapping and U–Pb geochronology
    Braun et al.2010|Early hominin diet included diverse terrestrial and aquatic animals 1.95 Ma in East Turkana, Kenya
    Lane et al.2015|The Late Quaternary tephrostratigraphy of annually laminated sediments from Meerfelder Maar, Germany
    McCulloch et al.2010|Proliferation and demise of deep-sea corals in the Mediterranean during the Younger Dryas
    Friedman2015|The early evolution of ray‐finned fishes
    Basolo1998|Evolutionary change in a receiver bias: a comparison of female preference functions
    Hamilton et al.2008|Geochemical investigation of gametogenic calcite addition in the planktonic foraminifera Orbulina universa
    Baker et al.1986|Sexual response of female great tits to variation in size of males' song repertoires
    Di Vito et al.2008|The late Pleistocene pyroclastic deposits of the Campanian Plain: new insights into the explosive activity of Neapolitan volcanoes
    CN103914546B|2017-10-27|Data-updating method and its device
    Alström et al.2008|Species delimitation based on multiple criteria: the Spotted Bush Warbler Bradypterus thoracicus complex |
    Florisbal et al.2012|Space–time relation of post-collisional granitic magmatism in Santa Catarina, southern Brazil: U–Pb LA-MC-ICP-MS zircon geochronology of coeval mafic–felsic magmatism related to the Major Gercino Shear Zone
    Bengtson1986|Siliceous microfossils from the Upper Cambrian of Queensland
    Martin et al.2010|Recent advances in the comprehension of the biogeography of Cretaceous European eusuchians
    Herridge et al.2012|Extreme insular dwarfism evolved in a mammoth
    RUNGE et al.1996|Characterization of the pelagic ecosystem in surface waters of the northern Gulf of St, Lawrence in early summer: the larval redfish‐Calanus‐microplankton interaction
    Jiang et al.2013|Female‐only sex‐linked amplified fragment length polymorphism markers support ZW/ZZ sex determination in the giant freshwater prawn Macrobrachium rosenbergii
    NL2026401A|2021-08-11|Analysis method for identification of different geographical populations of chinese mitten crab
    Wynn et al.1996|Evolution of sex differences in spatial cognition
    Buffetaut et al.2009|Late Palaeozoic and Mesozoic continental ecosystems of SE Asia: an introduction
    Carruthers et al.2014|Estimating tag reporting rates for tropical tuna fleets of the Indian Ocean
    MacFadden et al.2013|Middle Pleistocene age of the fossiliferous sedimentary sequence from Tarija, Bolivia
    Wheeler et al.2016|Using circulating reproductive hormones for sex determination of Atlantic sturgeon | in the Saco River estuary, Maine
    Jukar et al.2021|Late Quaternary extinctions in the Indian subcontinent
    同族专利:
    公开号 | 公开日
    CN110879263A|2020-03-13|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    CN103675136A|2013-12-11|2014-03-26|南昌大学|Soft-shelled turtle fatty acid analysis method|
    CN108717497A|2018-05-23|2018-10-30|大连海事大学|Imitative stichopus japonicus place of production discrimination method based on PCA-SVM|
    CN109828056A|2019-03-22|2019-05-31|中国水产科学研究院淡水渔业研究中心|A kind of discrimination method of Eriocheir japonica sinensis feed mode|
    法律状态:
    优先权:
    申请号 | 申请日 | 专利标题
    CN201911240254.XA|CN110879263A|2019-12-06|2019-12-06|Analysis method for identifying different geographical groups of eriocheir sinensis|
    [返回顶部]