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玉米ZmPox3过氧化物酶基因的核苷酸多样性:外显子2中微型反向重复转座元件(MITE)插入与饲用玉米消化率变异之间的关系

Nucleotide diversity of the ZmPox3 maize peroxidase gene: relationships between a MITE insertion in exon 2 and variation in forage maize digestibility.

作者信息

Guillet-Claude Carine, Birolleau-Touchard Christelle, Manicacci Domenica, Rogowsky Peter M, Rigau Joan, Murigneux Alain, Martinant Jean-Pierre, Barrière Yves

机构信息

Centre de recherche en Biologie Forestière, Université Laval, Sainte-Foy, Québec, G1K 7P4, Canada.

出版信息

BMC Genet. 2004 Jul 16;5:19. doi: 10.1186/1471-2156-5-19.

Abstract

BACKGROUND

Polymorphisms were investigated within the ZmPox3 maize peroxidase gene, possibly involved in lignin biosynthesis because of its colocalization with a cluster of QTL related to lignin content and cell wall digestibility. The purpose of this study was to identify, on the basis of 37 maize lines chosen for their varying degrees of cell wall digestibility and representative of temperate regions germplasm, ZmPox3 haplotypes or individual polymorphisms possibly associated with digestibility.

RESULTS

Numerous haplotypes with high diversity were identified. Frequency of nucleotide changes was high with on average one SNP every 57 bp. Nucleotide diversity was not equally distributed among site categories: the estimated pi was on average eight times higher for silent sites than for non-synonymous sites. Numerous sites were in linkage disequilibrium that decayed with increasing physical distance. A zmPox3 mutant allele, carrying an insertion of a transposable element in the second exon, was found in lines derived from the early flint inbred line, F7. This element possesses many structural features of miniature inverted-repeat transposable elements (MITE). The mutant allele encodes a truncated protein lacking important functional sites. An ANOVA performed with a subset of 31 maize lines indicated that the transposable element was significantly associated with cell wall digestibility. This association was confirmed using an additional set of 25 flint lines related to F7. Moreover, RT-PCR experiments revealed a decreased amount of corresponding mRNA in plants with the MITE insertion.

CONCLUSION

These results showed that ZmPox3 could possibly be involved in monolignol polymerisation, and that a deficiency in ZmPox3 peroxidase activity seemingly has a negative effect on cell wall digestibility. Also, genetic diversity analyses of ZmPox3 indicated that this peroxidase could be a relevant target for grass digestibility improvement using specific allele introgressions.

摘要

背景

对ZmPox3玉米过氧化物酶基因内的多态性进行了研究,该基因可能参与木质素生物合成,因为它与一组与木质素含量和细胞壁消化率相关的QTL共定位。本研究的目的是基于37个因细胞壁消化率不同而选择的、代表温带地区种质的玉米品系,鉴定可能与消化率相关的ZmPox3单倍型或个体多态性。

结果

鉴定出了众多具有高度多样性的单倍型。核苷酸变化频率很高,平均每57 bp有一个SNP。核苷酸多样性在不同位点类别中分布不均:沉默位点的估计π值平均是非同义位点的八倍。许多位点处于连锁不平衡状态,且随着物理距离的增加而衰减。在源自早期硬粒自交系F7的品系中发现了一个zmPox3突变等位基因,该基因在第二个外显子中有一个转座元件插入。该元件具有许多微型反向重复转座元件(MITE)的结构特征。突变等位基因编码一种缺少重要功能位点的截短蛋白。对31个玉米品系的子集进行的方差分析表明,转座元件与细胞壁消化率显著相关。使用另外一组与F7相关的25个硬粒品系证实了这种关联。此外,RT-PCR实验显示,有MITE插入的植物中相应mRNA的量减少。

结论

这些结果表明,ZmPox3可能参与单木质醇聚合,并且ZmPox3过氧化物酶活性的缺乏似乎对细胞壁消化率有负面影响。此外,ZmPox3的遗传多样性分析表明,这种过氧化物酶可能是通过特定等位基因渗入来提高禾本科植物消化率的一个相关靶点。

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