Ueda Tadamasa, Sato Tadashi, Hidema Jun, Hirouchi Tokuhisa, Yamamoto Kazuo, Kumagai Tadashi, Yano Masahiro
National Institute of Agrobiological Sciences, Kannondai 2-1-2, Tsukuba, Ibaraki 305-8602, Japan.
Genetics. 2005 Dec;171(4):1941-50. doi: 10.1534/genetics.105.044735. Epub 2005 Jun 18.
Rice qUVR-10, a quantitative trait locus (QTL) for ultraviolet-B (UVB) resistance on chromosome 10, was cloned by map-based strategy. It was detected in backcross inbred lines (BILs) derived from a cross between the japonica variety Nipponbare (UV resistant) and the indica variety Kasalath (UV sensitive). Plants homozygous for the Nipponbare allele at the qUVR-10 locus were more resistant to UVB compared with the Kasalath allele. High-resolution mapping using 1850 F(2) plants enabled us to delimit qUVR-10 to a <27-kb genomic region. We identified a gene encoding the cyclobutane pyrimidine dimer (CPD) photolyase in this region. Activity of CPD photorepair in Nipponbare was higher than that of Kasalath and nearly isogenic with qUVR-10 [NIL(qUVR-10)], suggesting that the CPD photolyase of Kasalath was defective. We introduced a genomic fragment containing the CPD photolyase gene of Nipponbare to NIL(qUVR-10). Transgenic plants showed the same level of resistance as Nipponbare did, indicating that the qUVR-10 encoded the CPD photolyase. Comparison of the qUVR-10 sequence in the Nipponbare and Kasalath alleles revealed one probable candidate for the functional nucleotide polymorphism. It was indicated that single-base substitution in the CPD photolyase gene caused the alteration of activity of CPD photorepair and UVB resistance. Furthermore, we were able to develop a UV-hyperresistant plant by overexpression of the photolyase gene.
水稻qUVR-10是位于第10号染色体上的一个抗紫外线B(UVB)的数量性状基因座(QTL),通过图位克隆策略被克隆出来。它是在由粳稻品种日本晴(抗UV)和籼稻品种卡萨拉斯(对UV敏感)杂交产生的回交自交系(BILs)中检测到的。在qUVR-10基因座上纯合日本晴等位基因的植株比卡萨拉斯等位基因的植株对UVB更具抗性。利用1850株F₂植株进行的高分辨率定位使我们能够将qUVR-10定位到一个小于27kb的基因组区域。我们在该区域鉴定出一个编码环丁烷嘧啶二聚体(CPD)光解酶的基因。日本晴中CPD光修复的活性高于卡萨拉斯以及与qUVR-10近等基因系【NIL(qUVR-10)】,这表明卡萨拉斯的CPD光解酶存在缺陷。我们将包含日本晴CPD光解酶基因的基因组片段导入NIL(qUVR-10)。转基因植株表现出与日本晴相同水平的抗性,表明qUVR-10编码CPD光解酶。对日本晴和卡萨拉斯等位基因中qUVR-10序列的比较揭示了一个可能的功能核苷酸多态性候选位点。结果表明,CPD光解酶基因中的单碱基替换导致了CPD光修复活性和UVB抗性的改变。此外,我们通过光解酶基因的过表达培育出了一种对UV超抗性的植株。