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通过显性负抑制错配修复蛋白 OsPMS1 快速生成水稻突变体。

Rapid generation of rice mutants via the dominant negative suppression of the mismatch repair protein OsPMS1.

机构信息

Guangdong Provincial Key Lab of Biotechnology for Plant Development, College of Life Sciences, South China Normal University, Guangzhou 510631, People's Republic of China.

出版信息

Theor Appl Genet. 2012 Sep;125(5):975-86. doi: 10.1007/s00122-012-1888-5. Epub 2012 Jun 12.

Abstract

Mismatch repair (MMR) is a conservative pathway for maintaining the genome integrity of different organisms. Although suppression of MMR has resulted in various mutation phenotypes in Arabidopsis, the use of this strategy for mutation breeding in major crops has not been reported. Here, we overexpressed a truncated version of the OsPMS1 protein in rice; this approach is expected to suppress the rice MMR system through a dominant negative mechanism. We observed a wide spectrum of mutation phenotypes in the progeny of the transgenic plants during seed germination and the plant growth stages. Genomic variations were detected with inter-simple sequence repeat (ISSR), and sequencing of the differential ISSR bands revealed that the mutation occurred as a point mutation or as microsatellite instability at high frequencies. Plant lines with agronomically important traits, such as salt and drought tolerance, various tiller number, and early flowering, were obtained. Furthermore, we obtained mutants with important traits that are free of the transgene. Together, these results demonstrate that MMR suppression can be used as an efficient strategy for mutation breeding in rice.

摘要

错配修复(MMR)是维持不同生物基因组完整性的保守途径。尽管抑制 MMR 在拟南芥中导致了各种突变表型,但在主要作物的突变育种中尚未报道这种策略的应用。在这里,我们在水稻中过表达了一个截短的 OsPMS1 蛋白,预计通过显性负作用机制抑制水稻 MMR 系统。我们在转基因植物的后代在种子萌发和植物生长阶段观察到广泛的突变表型。利用简单重复间序列(ISSR)检测到基因组变异,对差异 ISSR 带的测序表明,突变以高频发生点突变或微卫星不稳定性。获得了具有农艺重要性状的植物系,如耐盐性和耐旱性、不同分蘖数和早开花。此外,我们还获得了无转基因的具有重要性状的突变体。总之,这些结果表明,抑制 MMR 可作为水稻突变育种的有效策略。

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