坛紫菜品系间杂交子代杂种优势的ISSR分析
ISSR analysis on the heterosis in hybrids of Porphyra haitanensis
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摘要: 以野生型坛紫菜(♀)和经过人工诱变选育获得的红色型(♂)坛紫菜进行杂交获得丝状体和叶状体,从中挑选5株具有一定生长优势和其它经济性状的藻体分别进行体细胞克隆和丝状体培育,获得5个性状稳定的子代品系(品系A,B,C,D,E)。然后应用ISSR标记技术对杂交亲本及这5个子代品系的杂种优势进行研究,从67个引物中筛选出了21条能扩增出清晰、稳定条带的引物,共得到366个扩增位点,其中347个位点具有多态性,多态位点百分率达94.8%,扩增的条带大小在400~4 000 bp之间。通过对扩增带谱的统计分析,发现5个坛紫菜杂种子代品系同父本和母本的遗传相似性平均值分别为0.519 1和0.555 1,都明显低于两亲本间的遗传相似性0.6120;并且5个杂种子代的有效等位基因数1.550 1,期望杂合度0.320 0,Shannon多样性指数0.472 0都要明显高于两亲本的有效等位基因数1.390 7,期望杂合度0.195 4和Shannon多样性指数0.270 8。由此表明杂交使得坛紫菜的遗传多样性水平明显提高,产生了显著的杂种优势。从扩增带谱的统计结果,还可看出5个杂种子代同两亲本的遗传距离不是对等的,全部都稍偏向母本,聚类分析结果也同样证明了这一点,说明细胞质基因在坛紫菜遗传性状的表达中也发挥了重要作用。文章最后对ISSR技术在杂种优势预测中的应用也进行了讨论。Abstract: The wild strain (♀) of Porphyrahaitanensis was crossbred with the red strain (♂) one which selected through induction of mutation to get the hybrid conchocelies and thallus.Of all the hybrids, 5 healthy thallus which have predominance of growth and other economic characteristics were selected to digest with snail enzymes, and through somacloning and conchocelies culturing, 5 s rains (A、B、C、D、E) which have steady characters were obtained.And then, ISSR method was used to study the heterosis of the P.haitanensis parents and these 5 hybrid strains, 21 ISSR primers(were screened from 67 primers) gave rise to 366 discernible DNA bands of which 347 (94.8%) were polymorphic, the size of ISSR markers were highly reproducible ranged from 400 to 4000 bp.By statistic and analyzing these amplified bands, it can find that the average genetic similarity between the 5 hybrids and female parent and male parent are 0.519 1 and 0.555 1, which less the genetic similarity between the parents (0.612 0).The effective number of alleles, expected heterozygosity and shannon's information index among 5 hybrids were 1.550 1, 0.320 0 and 0.472 0 respectively, which also greater than the corresponding values (1.390 7, 0.195 4 and 0.270 8 respectively) between the parents evidently.From these datas, it can conclude that the hybrid can enhance the genetic diversity of P.haitanensis and produce distinct heterosis.From these datas, it also can find that the genetic distances between the 5 hybrids and female parent and male parent are no equality, all secund to the female parent, the reason maybe is the cytoplasmic gene also participate in the expressing and regulation of P.haitanensis genetic characters.At last, the application of ISSR method in fo recasting heterosis was discussed.
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Key words:
- Porphyrahaitanensis /
- hybrid /
- heterosis /
- ISSR
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