桃GRF基因家族的序列及其组织特异性表达分析
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中国农业科学院郑州果树研究所,河南农业大学,中国农业科学院郑州果树研究所,中国农业科学院郑州果树研究所,中国农业科学院郑州果树研究所,中国农业科学院郑州果树研究所,中国农业科学院郑州果树研究所,中国农业科学院郑州果树研究所

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中央级科研院所基本科研业务费专项(1610192016201);中国农业科学院科技创新工程项目[CAAS-ASTIP-2017-ZFRI-(01)]


The sequence character of GRF gene and its tissue-specific expression pattern in peach
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Zhengzhou Fruit Research Institute,Chinese Academy of Agricultural Sciences,Henan Agricultural University,Zhengzhou Fruit Research Institute,Chinese Academy of Agricultural Sciences,Zhengzhou Fruit Research Institute,Chinese Academy of Agricultural Sciences,Zhengzhou Fruit Research Institute,Chinese Academy of Agricultural Sciences,Zhengzhou Fruit Research Institute,Chinese Academy of Agricultural Sciences,Zhengzhou Fruit Research Institute,Chinese Academy of Agricultural Sciences,Zhengzhou Fruit Research Institute,Chinese Academy of Agricultural Sciences

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    摘要:

    转录因子家族基因GRF(growth-regulating factor)是植物中特有的一类生长调控因子基因,在水稻和拟南芥中,GRF家族基因编码的蛋白质具有正向调控茎和叶生长发育的功能。为了探究在桃中GRF基因是否参与了狭叶桃“金蜜狭叶”的叶片发育,本试验首先分析桃基因组中鉴定出的GRF基因的序列信息;然后,以正常叶片的“上海水蜜”桃为对照,通过荧光定量PCR检测了这些GRF基因在“金蜜狭叶”桃的组织特异性表达情况;最后,利用126对简单序列重复标记对1个F2杂交群体进行连锁分析,定位狭叶性状,辅助筛选狭叶性状的候选基因。结果表明:桃基因组GRF基因包括10个成员,分布在除4和8外的其他染色体上,编码蛋白的氨基酸长度在191至612之间。蛋白序列分析表明,除ppa011917m的QLQ结构域中的Leu残基被Phe代替外,这10个GRF的N端均含保守的QLQ和WRC结构域。系统进化树将桃上的10个GRF基因分为3组。从10个GRF基因的表达结果可以看出,6个基因在茎尖的表达量高于幼叶,也高于其他成熟组织和器官;其中2个基因在狭叶桃中的表达显著高于正常叶对照。通过连锁分析将“金蜜狭叶”的狭叶性状定位在第6染色体的15.7至21.1 Mb之间,该区间仅含有1个GRF基因ppa003017m,是狭叶性状的候选基因。本研究将为“金蜜狭叶”桃狭叶性状基因的鉴定和高光效种质筛选奠定研究基础。

    Abstract:

    The growth-regulating factor (GRF) transcription factor is a plant-specific gene family that regulates plant growth. In Arabidopsis and rice, the GRF gene family-encoding proteins have a positive function in regulating stem and leaf growth and development. In the present study, we speculated that the GRF genes might be more important and responsible for the formation of narrow leaves. To ask if the GRF genes are the causal mechanism in the development of a narrow-leaf peach variety (Prunus persica ‘Jin Mi Xia Ye’), GRF genes in the peach genome were identified by BLAST search according to the sequence homology. The expression pattern of these GRF genes was determined in the shoot apical meristem, young leaves, mature leaves, seeds, flesh of fruit, and phloem in ‘Jin Mi Xia Ye’ and the normal-leaf variety ‘Chinese Cling’. Meanwhile, the genetic mapping of narrow-leave was conducted in an F2 population by using 126 simple sequence repeats (SSR) markers. Ten members of the GRF genes were found in the peach genome. The deduced proteins of GRFs arranged from 191 and 612 amino acids. Sequence analysis showed that the N terminal of these proteins maintained two conserved motifs QLQ (Gln-Leu-Gln) and WRC (Trp、Arg、Cys), except that the Leu residue of QLQ in ppa011917m was replaced by Phe. While the C terminal of those sequences varied, for instance, a TQL and a GGPL motifs. A phylogenetic tree using a neighbor-joining method placed the GRF genes into three groups. According the expression result of 10 GRF genes in peach, six of them exhibited higher expression levels in the shoot apical meristem than in young leaves and other mature tissues and organs. Only a few genes showed slight expression levels in the flesh of fruit, mature leaves, and phloem, such as ppa022199m in flesh and ppa019752m in phloem. Only two genes, ppa019623m and ppa003017m, expressed higher levels in peach varieties with narrow leaves than in those with normal leaves. Meanwhile, we located the narrow-leaf trait in the region between two markers (UDP98-407 and BPPCT025 of linkage group 6) using linkage analysis based on SSR markers. Based on the linkage analysis, we then chose the ppa003017m as one of the key candidates in regulating leaf development in the ‘Jin Mi Xia Ye’ variety. However, the sequence amplification found that, among 30 seedings with different leaf phenotypes, no variation existed in the promoter of the key candidate gene, ppa003017m. The results indicate that there is no relationship between the expression of ppa003017m and sequence variation in its promoter regions. This study highlights the importance of identifying genes that regulate the narrow-leaf trait in peach, as well as the importance of selecting the excellent peach germplasm resources with high photosynthesis efficiency using molecular marker assisted selection in the future.

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曹 珂,薛灵姿,王 蛟,等.桃GRF基因家族的序列及其组织特异性表达分析[J].植物遗传资源学报,2018,19(3):578-586.

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  • 收稿日期:2017-09-28
  • 最后修改日期:2018-03-15
  • 录用日期:2018-01-05
  • 在线发布日期: 2018-05-08
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