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Abstract: The purple-leaf line XY-PL,an advanced recombinant inbred line,was screened out from the progenies of interspecific hybridization between Brassica napus and Brassica juncea,had performance of stable purple leaf phenotype,high anthocyanin content and good agronomic traits and could be potentially used as intermediates for high anthocyanin content breeding programs.To explore the relationship between the purple-leaf trait of XY-PL and its APR2,a key enzyme for sulfur assimilation and metabolism,applying homologous cloning method and XY-PL and green leaf Zhongshuang No.11 (ZS11) as materials three APR2 homologs were successfully isolated,designated as PL-APR2,GL-APR 2.1 and GL-APR2.2 respectively.Results of sequence analyses showed that they encompassed four exons and three introns,the encoded proteins each had a typical chloroplast transport peptide domain and shared 98.2%—99.1% identities among them,and had the closest phylogenetic relationship with APR2 from cruciferous plants.Compared with GL-APR2.1 and GL-APR2.2,PL-APR2 had three amino acid residues (51SLK53) deletion in the chloroplast transport peptide transporter domain,which would affect the transmembrane transport of its precursor proteins and the efficiency of sulfur assimilation and utilization.Overexpression of PL-APR2 in Arabidopsis thaliana demonstrated that all transgenic lines did not exhibit purple-leaf phenotype from generation T0 to T2 perhaps because of the presence of wild-type AtAPR2 in transformation receptor material.Next,the Arabidopsis apr2 mutant will be used as genetic transformation receptor for further function verification and the transport function of chloroplast transport peptide of PL-APR2 will be analyzed deeply.

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摘要： 紫叶油菜品系XY-PL是通过甘芥杂交获得的一个高世代重组自交系，其紫叶表型稳定、花青素含量高、主要农艺性状良好，是一个潜在的高花青素育种的中间材料。为探讨XY-PL紫叶表型与硫同化代谢关键酶APR2的关系，采用同源克隆法分别克隆了XY-PL和绿叶油菜品种中双11号（ZS11）的APR2基因，共获得3条差异序列，分别命名为PL-APR2、GL-APR2.1和GL-APR2.2。序列分析表明，它们均由4个外显子和3个内含子组成，其编码产物均具有典型的叶绿体转运肽序列，序列之间的一致性为98.2%～99.1%，在系统进化关系上与十字花科植物来源的APR2亲缘关系最近。相对于GL-APR2.1和GL-APR2.2，PL-APR2在叶绿体转运肽转运功能区存在3个氨基酸残基（51SLK53）缺失，可能影响其前体蛋白的跨膜转运，进而影响硫的同化与利用效率。在拟南芥中过表达结果表明，PL-APR2转基因株系在T0—T2代均未表现出紫叶表型，可能与转化受体材料内存在野生型AtAPR2有关。下一步将采用拟南芥apr2突变体作为遗传转化受体进行继续验证，并将对其叶绿体转运肽转运功能进行深入分析。

ZHANG Hong, LI Gang, HAN Fengying, ZHAO Fuyong. Cloning and Genetic Transformation in Arabidopsis thaliana of Sulfate Assimilation Reductase Gene APR2 in a Purple-leaf Rapeseed Line[J]. Journal of Henan Agricultural Sciences, 2020, 49(2): 17-26.

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