Evaluation Of Microsatellite Markers For Identification Of Cross-breeding Carp And Screening The Microarray Probe Of Immune-related Gene In The Cyprinus Carp

The application of microsatellite markers for parentage determination has beenapproved in aquaculture which not only can maintain the parentage information, butalso greatly reduce the cost in space and man-power and greatly improve breedingselection effect in communal environment, especially in the process of selectivebreeding. In this study, two selected cross-breeding families (the49full-sib families:7of C. carpio var. wuyuanensis♀×7of C. carpio L. mirror♂and the24full-sibfamilies:3of C. carpio L. mirror♀×8of C. carpio var. wuyuanensis♂) wereevaluated for genetic diversity and inheritance characteristics by21microsatellitesloci, and examined several properties of microsatellites which could be considered toaffect the accuracy of loci in parentage assignment, such as polymorphism, null allelefrequency and linkage equilibrium. Finally,13suitability loci were obtained forparentage assignment, and performance of them showed that:(1) These13screenedloci were high polymorphic. Mean number of alleles (K), mean expectedheterozygosity (He), mean observed heterozygosity (Ho) and mean polymorphicinformation content (PIC) were respectively0.505,0.648,0.791and0.5889in the49full-sib families and0.548,0.670,0.819and0.6138in the24full-sib families.(2)13screened loci were of high discriminatory power. The combined exclusion probability(CPE) was0.999997978in the49full-sib families and0.999999583in the24full-sibfamilies.(3) High efficiency of the loci subset was obtained. The CPE of4coremicrosatellite loci (MFW29, HLJ392, HLJ044, HLJ855) could be accurate to morethan99%in the two cross-breeding groups.(4) Combined non-exclusion probabilitieswhen the first parent was available (NE-1P), the second parent was available (NE-2P)and parent pairs were available (NE-PP) were respectively1.34E-02,3.91E-04and2.02E-06in the49full-sib families, and were respectively1.34E-02,3.91E-04and2.02E-06in the24full-sib families.(5) The simulations with99%confidenceindicated that100%offspring of49full-sib families could be assigned to a parent pairusing the11most polymorphic loci or more loci,99%offspring of24full-sib familiescould be assigned to a parent pair using the best9or more loci.(6) Thedistance-based method of cluster analysis showed that: the set of13loci could clearlydistinguish between the offspring of cross-breeding and the unrelated family in thepolyculture. Based on these results, a suitable microsatellite-based tool for parentageassignment in cross-breeding carp was obtained, and displayed a reliability and highefficacy alternative to physical tagging in subsequently selecting program. In the aquaculture, the resistance of varieties is a hot pints. One of the focus ishow to improve the immunity of individual for being cope with massive outbreaks ofdiseases. The problem how to reduce the mortality of varieties need be solved inbreeding carp. In this study, through the principle of bioinformatics and comparativegenomicsr, we have screened and obtained493candidate genes of immune relatedfrom the37number of known microarray of human immune and the15number ofpathway of human immune related. And six kinds of internal control genes（18SrRNA、beta-actin、RPL-7、RPL-13α、EF1-α、B2M）are suitable for the carp in thegene expression study. This study for the construction microarray of the carp immunerelated provide the clear purpose probe, and which for the subsequent immunemechanism study of carp provide the theory basis.