Isolation of mutant lines with decreased numbers of chloroplasts per cell from a tagged mutant library of the moss Physcomitrella patens
Isolation of mutant lines with decreased numbers of chloroplasts per cell from a tagged mutant library of the moss Physcomitrella patens
复制标题
DOI：
10.1055/s-2005-837691
复制
发表时间：
2005-05-01
期刊：
影响因子：
3.9
通讯作者：
Takano, H
中科院分区：
文献类型：
--
作者：
Hayashida, A;Takechi, K;Takano, H
通过显微镜观察，从8800个标记的小立碗藓突变株系中分离出11个叶绿体数目减少的突变株系。在野生型植物中的叶绿体线亚顶端细胞有48个叶绿体的平均值，而在突变株系215和222的亚顶端细胞中的叶绿体数目减少到野生型的75%。七个突变系--473、122、221、129、492、207和138的叶绿体数量约为野生型的一半。突变体11号有几个明显增大的叶绿体，突变体347号有各种大小的叶绿体。在突变株系222、473、221、129、492和207中，细胞体积与野生型相同，而其它突变株的细胞体积增加。当能够形成雄配子体时，各突变系叶细胞的叶绿体数量与绿丝细胞的叶绿体数量相同。氨苄青霉素处理降低了所有突变株系中叶绿体的数量。以标签中的DNA为探针进行Southern杂交，结果表明突变体473和221中只有一个插入。为了确定标记的DNA是否插入到已知的质体分裂基因中，我们分离了PpMinD 1、PpMinD 2和PPMinE 1基因。基因组聚合酶链反应分析表明，PpFtsZ和PpMinD/E基因没有被破坏的标签插入突变株系11和347，分别。
Eleven mutant lines exhibiting decreased numbers of chloroplasts per cell were isolated from 8800 tagged mutant lines of Physcomitrella patens by microscopic observations. Chloronema subapical cells in wild-type plants had a mean of 48 chloroplasts, whereas chloroplast numbers in subapical cells in mutant lines 215 and 222 decreased to 75% of that in the wild type. Seven mutant lines - 473, 122, 221, 129, 492, 207, and 138 had about half as many chloroplasts as the wild type. Mutant line 11 had a few remarkably enlarged chloroplasts, and mutant line 347 had chloroplasts of various sizes. Whereas the cell volume was the same as in the wild type in mutant lines 222, 473, 221, 129, 492, and 207, the cell volume of the other mutants increased. The chloroplast number of leaf cells was the same as that of chloronema cells in each mutant line when gametophores could be formed. Treatment with ampicillin decreased the number of chloroplasts in all mutant lines. Southern hybridization using DNA in tags as probes showed that only one insertion occurred in mutant lines 473 and 221. To determine whether the tagged DNA inserted into the known genes for plastid division, we isolated the PpMinD1, PpMinD2, and PPMinE1 genes. Genomic polymerase chain reaction analysis showed that the PpFtsZ and PpMinD/E genes were not disrupted by the insertion of the tags in mutant lines 11 and 347, respectively.