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园艺学报 ›› 2018, Vol. 45 ›› Issue (4): 627-640.doi: 10.16420/j.issn.0513-353x.2017-0443

• 研究论文 • 上一篇    下一篇

苹果全基因组CRF家族成员鉴定及在不定根发育过程中的表达分析

  珂,刘  桢,雷  超,左超然,董  凤,孟  媛,毛江萍,韩明玉,张  *   

  1. 西北农林科技大学园艺学院,陕西杨凌 712100
  • 出版日期:2018-04-25 发布日期:2018-04-25
  • 基金资助:

    陕西省重点研发项目(2017NY0055);西北农林科技大学“仲英青年学者”项目;国家现代农业产业技术体系建设专项资金项目(CARS-27);国家苹果改良中心杨凌分中心项目;陕西省果业发展协同中心和陕西果业发展项目

Genome-wide Identification and Expression Analysis of CRF Family Gene During Adventitious Root Development in Apple

LI Ke,LIU Zhen,LEI Chao,ZUO Chaoran,DONG Feng,MENG Yuan,MAO Jiangping,HAN Mingyu,and ZHANG Dong*   

  1. College of Horticulture,Northwest A & F University,Yangling,Shaanxi 712100,China
  • Online:2018-04-25 Published:2018-04-25

摘要:

通过生物信息学分析,从‘金冠’苹果基因组鉴定得到了11个细胞分裂素响应因子(CRF)基因。根据基因在染色体上的位置,将其依次命名为MdCRF1 ~ MdCRF11,并对其进行理化特性、基因结构、系统进化和启动子元件分析。MdCRF编码的蛋白在137 ~ 435 aa之间,等电点在4.26 ~ 9.68之间。基因结构分析发现,MdCRF1有2个外显子1个内含子,MdCRF11有3个外显子2个内含子,其余基因都只含有外显子,没有内含子;保守基序列分析发现,MdCRF蛋白的保守基序数量在5 ~ 9。通过系统进化分析将家族成员分为G1、G2和G3共3个亚组,每个亚组内成员数量相对均匀。组织特异性表达分析发现11个基因在不同杂交种苹果和不同器官中的表达量存在明显差异。以苹果砧木‘T337’为材料,qPCR验证了MdCRF家族基因大部分在根、茎、愈伤组织中高表达,同时也响应生根处理表达量发生显著下调。对11个MdCRF蛋白之间的网络调控关系进行了研究,分析预测了相关基因间的潜在联系。结合分析可知,其中MdCRF8、MdCRF10和MdCRF11可能参与调控苹果砧木不定根发育。

关键词: 苹果, CRF, 不定根发育, 生物信息学

Abstract:

In the current study,a total of 11 cytokinin response factor genes(CRF)were identified from the apple genome by bioinformatics. Genes were named MdCRF1–MdCRF11 by the position of the gene in the chromosome. Additionally,this study systematically analyzed its physical and chemical characteristics,gene structures,evolutionary relationships and promoter elements. MdCRF proteins contained 137–435 amino acids and the isoelectric points is from 4.26 to 9.68. Results of exon-intron structure analysis showed that the MdCRF1 contained 2 exons and 1 intron;MdCRF11 contained 3 exons and 2 introns;other genes only have exons and without introns. Conservative base sequence analysis showed that 5–9 conserved motifs existed in the MdCRF proteins. Phylogenetic analysis showed that MdCRF family genes could be classified into three subgroups(G1,G2 and G3)according to its homology. In addition,the number of the members of each subgroup were approximately equal. Tissue-specific expressions indicated that 11 MdCRF had significantly different expression patterns in different genotypes,as well as different tissues. The materials were‘T337’apple rootstock,qPCR confirmed most of genes were highly expressed in root,stem and callus. At the same time the expression of these genes were down-regulated by rooting treatment. Additionally,we systematically analyzed the connection network of MdCRF proteins and predicted the potential link between genes. The results showed that MdCRF8,MdCRF10 and MdCRF11 might be involved in regulating adventitious root development in apple rootstock.

Key words: apple, CRF, adventitious root development, bioinformatic

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