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荧光显微观察表明,20℃水温下,栉孔扇贝(Chlamys farreri)的卵与海湾扇贝(Argopecten irradians)的精子可以正常受精和发育,具备人工诱导三倍体的可行性。亲贝充分促熟后,分开催产,以20:1的精卵比授精;在50%的受精卵排出第1极体时,以60mg/L6-二甲基氨基嘌呤(6-DMAP)处理受精卵10-25min,可诱导75.23%-92.14%的三倍体;6-DMAP处理15min综合诱导效果最好,三倍体诱导率可达88.56%,孵化率可达53.52%。得到的三倍体幼虫经基因组原位杂交(Genomic in situ hybridization,GISH)验证,为含有2套栉孔扇贝染色体组和1套海湾扇贝染色体组的异源三倍体。孵化后诱导组与杂交对照组(未经6-DMAP处理)幼虫生长越来越缓慢,受精后14d其幼虫存活率分别下降到0.00067%和0.00224%,没有幼虫度过附着变态期。GISH分析显示,栉孔扇贝与海湾扇贝不同倍性的杂种后代早在担轮幼虫阶段就出现母本偏向性染色体转变。
Fluorescence microscopic observation showed that the eggs of Chlamys farreri and the sperm of Argopecten irradians could fertilize normally at 20 ℃, and the feasibility of artificial induction of triploidy was proved. After the progeny were fully ripened, they were separated and made to fertilize at a sperm-to-egg ratio of 20: 1. When the first polar body was ejected from 50% of the fertilized eggs, they were fertilized with 60 mg / L 6-dimethylaminopurine Eggs 10-25min can induce 75.23% -92.14% triploids; 6-DMAP treatment 15min comprehensive induction of the best, triploid induction rate up to 88.56%, the hatching rate of up to 53.52%. The obtained triploid larvae were verified by Genomic in situ hybridization (GISH) and were allotetraploids containing 2 Chlamys farreri genomes and 1 set of Gulf scallop genomes. After hatching, the larvae of the induction group and the hybridization control group (without 6-DMAP treatment) grew slower and slower, and the larvae survival rate decreased to 0.00067% and 0.00224% respectively after 14 days of fertilization. No larvae passed the metamorphosis period. GISH analysis showed that the offspring of hybrids with different ploidy of the chlamys farreri and the Gulf scallop appeared the maternal biased chromosomal transition as early as the larval larval stage.