【摘要】：紫石房蛤,是一種海洋雙殼貝類(lèi),分布在中國(guó),日本和韓國(guó)的周邊地區(qū)。由于它的高營(yíng)養(yǎng)和經(jīng)濟(jì)價(jià)值,使它成為一種重要的經(jīng)濟(jì)海產(chǎn)品。近年來(lái),由于過(guò)度捕撈和環(huán)境惡化,紫石房蛤的自然資源一直呈下降趨勢(shì),為了保護(hù)野生群體,開(kāi)發(fā)和利用分子標(biāo)記對(duì)紫石房蛤野生群體進(jìn)行遺傳分析是必要的。在本研究中,我們通過(guò)Illumina Hiseq 2500測(cè)序平臺(tái)對(duì)紫石房蛤的鰓進(jìn)行轉(zhuǎn)錄組文庫(kù)測(cè)序,旨在獲得轉(zhuǎn)錄組數(shù)據(jù)開(kāi)發(fā)分子標(biāo)記,為這個(gè)物種遺傳多樣性分析和分子標(biāo)記輔助育種奠定基礎(chǔ)。在測(cè)序報(bào)告中,我們獲得了100,480,000條原始讀段,去除低質(zhì)量序列、rRNA和接頭序列后,得到68,080,636條干凈讀段。利用這些讀段進(jìn)行組裝,共獲得到了5,115,494條重疊群,120,479條轉(zhuǎn)錄本和66,388條unigenes。通過(guò)與NCBI數(shù)據(jù)庫(kù)進(jìn)行比對(duì),一共有26,781條unigenes被注釋。利用生物信息學(xué)軟件,從轉(zhuǎn)錄組數(shù)據(jù)庫(kù)的66,388條unigene序列中進(jìn)行篩選獲得414個(gè)微衛(wèi)星序列,其中二堿基重復(fù)135個(gè),三堿基重復(fù)217個(gè),四堿基重復(fù)58個(gè),分別占微衛(wèi)星序列總數(shù)的32.6%,52.4%和14%。隨機(jī)選取35個(gè)微衛(wèi)星序列設(shè)計(jì)引物,26個(gè)引物擴(kuò)增出目的產(chǎn)物,利用30個(gè)紫石房蛤個(gè)體檢測(cè)微衛(wèi)星的多態(tài)性,其中13個(gè)證明有多態(tài)性位點(diǎn)。進(jìn)一步結(jié)果分析顯示等位基因從3到6不等,平均每個(gè)位點(diǎn)4.69個(gè)等位基因;觀測(cè)雜合度和期望雜合度為0.4667-0.6667和0.5847-0.8322,沒(méi)有觀察到連鎖不平衡位點(diǎn)(LD);2個(gè)位點(diǎn)顯著偏離哈迪溫伯格平衡(PHWE0.01)。轉(zhuǎn)錄組測(cè)序平臺(tái)展示了對(duì)紫石房蛤物種快速開(kāi)發(fā)分子資源的能力。這些新的多態(tài)性微衛(wèi)星標(biāo)記對(duì)進(jìn)一步的種群研究和保護(hù)遺傳學(xué)提供基礎(chǔ)資料。
[Abstract]:Purple clam, a kind of marine bivalve shellfish, is distributed in the surrounding areas of China, Japan and South Korea. Because of its high nutrition and economic value, it has become an important economic seafood. In recent years, due to overfishing and environmental deterioration, the natural resources of clam have been declining. In order to protect the wild population, it is necessary to develop and use molecular markers to carry out genetic analysis of the wild population of clam. In this study, we sequenced the Gill of clam by Illumina Hiseq 2500 sequencing platform in order to obtain the data of transcriptional group and develop molecular markers, which laid a foundation for genetic diversity analysis and molecular marker assisted breeding of this species. In the sequencing report, 100480000 original reading segments were obtained, and 68080636 clean reading segments were obtained after removing low quality sequences, rRNA and connector sequences. Using these reading segments to assemble, a total of 5115494 overlapping groups, 120479 transcripts and 66388 unigenes. were obtained. Compared with the NCBI database, a total of 26781 unigenes have been commented on. Using bioinformatics software, 414 microsatellite sequences were screened from 66388 unigene sequences in the transcriptional group database, including two base repeats, three base repeats and 58 four base repeats, accounting for 32.6%, 52.4% and 14% of the total microsatellite sequences, respectively. Thirty-five microsatellite sequences were randomly selected to design primers, and 26 primers were used to amplify the target products. The microsatellite polymorphism was detected by 30 individuals of the clam, 13 of which proved to have polymorphism sites. The further results showed that the alleles varied from 3 to 6, with an average of 4.69 alleles per locus, and the heterozygosity and expected heterozygosity were 0.467 鈮，

本文編號(hào)：2500742