Nishimura Hideki, Ahmed Nisar, Tsugane Kazuo, Iida Shigeru, Maekawa Masahiko
Research Institute for Bioresources, Okayama University, Kurashiki, 710-0046, Japan.
Theor Appl Genet. 2008 Feb;116(3):395-405. doi: 10.1007/s00122-007-0677-z. Epub 2007 Dec 4.
An endogenous 0.6-kb rice DNA transposon, nDart1, has been identified as a causative element of a spontaneous mutable virescent allele pyl-v conferring pale-yellow leaves with dark-green sectors in the seedlings, due to somatic excision of nDart1 integrated into the OsClpP5 gene encoding the nuclear-coded chloroplast protease. As the transposition of nDart1 depends on the presence of an active autonomous aDart element in the genome, the plants exhibiting the leaf variegation carry the active aDart element. As several mutable alleles caused by nDart1 insertions have subsequently been identified, nDart1-promoted gene tagging has been proven to be an effective system. At present, the nDart/aDart system appears to be the only endogenous rice DNA transposon system whose transposition activity can be controlled under natural growth conditions without any artificial treatments, including tissue cultures. To apply the nDart/aDart tagging system in various cultivated rice varieties, we explored the presence and distribution of an active autonomous aDart element in 19 temperate japonica, 30 tropical japonica, and 51 indica varieties. Only eight temperate japonica varieties were found to bear a single copy of an active aDart element, and no aDart activity could be detected in the indica varieties examined. Six of seven japonica varieties appear to carry the active aDart element at the identical site on chromosome 6, whereas the remaining one contains aDart on chromosome 5. Leaf variegations in the plants with the mutable pyl-v allele and the excision frequencies of endogenous nDart1 elements indicated that the aDart element on chromosome 6 is more active than that on chromosome 5. The findings described here are an important step in the development of a new and efficient nDart1-promoted gene-tagging system in various rice cultivars.
一个内源性的0.6kb水稻DNA转座子nDart1,已被鉴定为一个自发突变的淡绿等位基因pyl-v的致病元件,该等位基因导致幼苗出现淡黄色叶片并带有深绿色斑块,这是由于整合到编码核编码叶绿体蛋白酶的OsClpP5基因中的nDart1发生体细胞切除所致。由于nDart1的转座依赖于基因组中活跃的自主aDart元件的存在,表现出叶片斑驳的植株携带活跃的aDart元件。随后又鉴定出了几个由nDart1插入引起的突变等位基因,nDart1促进的基因标签已被证明是一个有效的系统。目前,nDart/aDart系统似乎是唯一一种内源性水稻DNA转座子系统,其转座活性在自然生长条件下无需任何人工处理(包括组织培养)即可得到控制。为了在各种栽培水稻品种中应用nDart/aDart标签系统,我们研究了19个温带粳稻品种、30个热带粳稻品种和51个籼稻品种中活跃的自主aDart元件的存在和分布情况。仅发现8个温带粳稻品种携带单个活跃的aDart元件拷贝,在所检测的籼稻品种中未检测到aDart活性。7个粳稻品种中有6个似乎在6号染色体的相同位点携带活跃的aDart元件,而其余一个在5号染色体上含有aDart元件。具有突变pyl-v等位基因的植株的叶片斑驳以及内源性nDart1元件的切除频率表明,6号染色体上的aDart元件比5号染色体上的更活跃。本文所述的研究结果是在各种水稻品种中开发新的高效nDart1促进基因标签系统的重要一步。