| Cheilinus undulatus belongs to Cheilinus genus (Osteichthyes:Actinopterygii: Perciformes:Labroidei:Labridae). C. undulatus is an important large coral reef fish distributed widely throughout the tropical Indo-Pacific, it had been listed as vulnerable in the IUCN, WWF, CITES for four times since1996to2005. Therefore, the resources investigation and conservation, the enhancement of artificial propagation are needed. Until now, the study of Cheilinus undulatus has mainly focused on biology, rarely know about germplasm resources. In this study, the sequences of D-loop, CO I, Cytb and ND1of mtDNA were used to analyzed four different geographical populations of Lingshui, Malaysia, Xisha, Nansha by the method of amplification, clone and sequencing, to estimate the genetic diversity and divergence of C. undulatus within and among populations comprehensively,15fluorescentlylabeled primers of microsatellite were also used in this study to measure the genetic diversity. The results were as follows:1. The analysis of the genetic diversity and divergence of the four C. undulatus populations revealed by D-loopThe software Dnasp was used to compute the parameters of genetic diversity of four different geographic populations which collected from Lingshui, Malaysia, Xisha and Nansha. The number of variable sites is63,35,14,53respectively, and110in totals. The number of haplotype(h) and haploytpe diversity(Hd) are29/0.996,17/0.979,9/0.934,29/0.984respectively, while the nucleotide diversity(Pi)and average number of nucleotide differences(K)are0.00660,0.00569,0.00661,0.00509and5.957,5.126,5.978,4.590respectively. The average K2-P distance within a geographic population range from0.00502to0.00670, and0.00525to0.00698between groups; The software Areliquin3.5is used to do the Tajima's selective and Fu's neutrality test, the result are-2.32975(P=0.001),-1.85574(P=0.019),-0.21586(P=0.426),-2.28163(P=0.001) and-23.79762(P=0.000),-9.77664(P=0.000),-0.74532(P=0.353),-22.33161(P=0.000) respectively. Genetic divergence also computed by the software, they are range from-7.88843to5.31207. From the analysis of AMOVA, we know that3.19%variation contributed by between-group and96.81%within-group. The base content, number of haplotype and their distribution and frequencies of the four groups were also calculated by the software. Then, the NJ tree of haplotype was done by the software MEGA5.0. 2. The analysis of the genetic diversity and divergence of the four C. undulatus populations revealed by COIThe software Dnasp was used to compute the parameters of genetic diversity of four different geographic populations which collected from Lingshui, Malaysia, Xisha and Nansha. The number of variable sites is10,0,2,15respectively, and23in totals. The number of haplotype(h) and haploytpe diversity(Hd) are9/0.798,1/0.000,3/0.275,14/0.768respectively, while the nucleotide diversity(Pi)and average number of nucleotide differences(K)are0.00092,0.00000,0.00018,0.00081and1.428.0.000,0.286,1.254respectively. The average K2-P distance within a geographic population range from0.00000to0.00086, and0.00042to0.00101between groups; The software Areliquin3.5is used to do the Tajima's selective and Fu's neutrality test, the result are-1.67131(P=0.025),0.00000(P=1.000),-1.47534(P=0.059),-2.12342(P=0.003) and-3.75881(P=0.010),-0.00000(P=N.A.)5-1.47534(P=0.018),-11.41372(P=0.000) respectively. Genetic divergence also computed by the software, they are range from-1.37161to0.05932. From the analysis of AMOVA, we know that13.20%variation contributed by between-group and86.80%within-group. The base content, number of haplotype and their distribution and frequencies of the four groups were also calculated by the software. Then, the NJ tree of haplotype was done by the software MEGA5.0.3. The analysis of the genetic diversity and divergence of the four C. undulatus populations revealed by CytbThe software Dnasp was used to compute the parameters of genetic diversity of four different geographic populations which collected from Lingshui, Malaysia, Xisha and Nansha. The number of variable sites is16,5,5,15respectively, and31in totals. The number of haplotype(h) and haploytpe diversity(Hd) are14/0.815,5/0.663,6/0.681,14/0.816respectively, while the nucleotide diversity(Pi)and average number of nucleotide differences(K)are0.00437,0.00081,0.00089,0.00142and1.916,0.921,1.011,1.614respectively. The average K2-P distance within a geographic population range from0.00082to0.00127, and0.00089to0.00128between groups; The software Areliquin3.5is used to do the Tajima's selective and Fu's neutrality test, the result are-2.47517(P=0.000),-0.49917(P=0.351),-1.16440(P=0.123),-1.56910(P=0.054) and-0.96945(P=0.381),-0.20857(P=0.451),-2.22298(P=0.040),-7.04791(P=0.000) respectively. Genetic divergence also computed by the software, they are range from-1.37161to0.05932. From the analysis of AMOVA, we know that1.11%variation contributed by between-group and98.89%within-group. The base content, number of haplotype and their distribution and frequencies of the four groups were also calculated by the software. Then, the NJ tree of haplotype was done by the software MEGA5.0.4. The analysis of the genetic diversity and divergence of the four C. undulatus populations revealed by ND1The software Dnasp was used to compute the parameters of genetic diversity of four different geographic populations which collected from Lingshui, Malaysia, Xisha and Nansha. The number of variable sites is7,2,2,5respectively, and13in totals. The number of haplotype(h) and haploytpe diversity(Hd) are9/0.456,3/0.195,2/0.143,6/0.262respectively, while the nucleotide diversity(Pi)and average number of nucleotide differences(K) are0.00053,0.00021,0.00029,0.00029and0.514,0.200,0.286,0.278respectively. The average K2-P distance within a geographic population range from0.00021to0.00053, and0.00025to0.00041between groups; The software Areliquin3.5is used to do the Tajima's selective and Fu's neutrality test, the result are-2.07171(P=0.002),-1.51284(P=0.051),-1.48074(P=0.067),-2.00709(P=0.002) and-9.16602(P=0.000),-1.86305(P=0.013),0.29648(P=0.305),-5.93882(P=0.000) respectively. Genetic divergence also computed by the software, they are range from--7.88843to5.31207. From the analysis of AMOVA, we know that-0.58%variation contributed by between-group and100.58%within-group. The base content, number of haplotype and their distribution and frequencies of the four groups were also calculated by the software. Then, the NJ tree of haplotype was done by the software MEGA5.0.5. The analysis of the genetic diversity and divergence of the four C. undulatus populations revealed by microsatellite15fluorescently labeled primers of microsatellite were used to measure the genetic diversity of the four groups. The software POPGEN32compute the Nei's genetic identity, genetic distance and construct UPGMA tree, the Nei's genetic identity range from0.7667to0.8644, and genetic distance0.1457to0.2656. The UPGMA tree demonstrated that the Lingshui group and Malaysia grouped together and then grouped together with Xisha; Nansha group comprised the other clade. The Ne, Ho, HE computed by the software are1.3431-7.3080/0.2258-0.7742/0.2597-0.8773,1.0000-8.6022/0.0000-0.6000/0.0000-0.9064,1.4152-6.1250/0.2143-0.8571/0.3042-0.8677,1.3388-7.9754/0.1389-0.9722/0.2567-0.8807respectively, meanwhile, likelihood ratio test for Hardy-Weinberg equilibrium of the15sites in the four groups, the value of the P are range from0.00512to0.81543,0.02252-0.95771,0.02244-0.99893,0.01823-0.40838respectively; The software was used to calculate the PIC of each group, the result are0.23535-0.85203,0.45531-0.87353(A3is monomorphic),0.53962-0.81300,0.24523-0.86359; The Genetic divergence computed by software Areliquin3.5range from0.17001(P=0.34545)-0.39153(P=0.40000), and the analysis of AMOVA show that35.75%variation contributed by between-group,35.75%among individuals within populations and60.90%within individuals.6. Even though there are certain differences between the result of the analysis of genetic diversity and genetic divergence by using four mitochondrial sequences and microsatellite DNA at the population level in this study, they come to a conclusion together, unconspicuous genetic divergence between populations and low diversity of Cheilinus undulatus. |
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