毛竹SQUAMOSA启动子结合蛋白SBP家族基因的鉴定及表达模式分析

doi:10.11707/j.1001-7488.20171206

摘要/Abstract

摘要： [目的]SBP家族基因编码的一类转录因子可识别并结合MADS-box基因SQUAMOSA（SQUA）的启动子，参与植物营养生长和生殖生长的调控。系统地鉴定和分析毛竹SBP家族基因，有助于理解SBP家族基因在毛竹营养生长和生殖生长过程中的调控作用，为毛竹SBP家族基因功能分析奠定基础。[方法]利用已公布的毛竹基因组数据，通过生物信息学方法鉴定毛竹SBP家族基因。利用MEGA 6.0、DNAMAN、psRNATarget等生物信息学软件获得毛竹SBP家族基因基本生物学信息。通过实时荧光定量PCR分析SBP家族基因在毛竹根茎叶、花序发育不同时期和实生苗受到高盐胁迫时的表达模式。[结果]在毛竹基因组中共鉴定到18条SBP家族基因，这些基因都有单一且高度保守的SBP结构域，其中有10条SBP家族基因具有miR156靶位点。毛竹SBP结构域有74个氨基酸残基，包括2个锌指结构域和1个NLS结构域。毛竹SBP转录因子进化关系、基序分布和DNA结构结果表明，进化关系相近的毛竹SBP转录因子的Motif和基因结构均具相似性。对不同物种来源的SBP家族基因进行进化分析发现，毛竹SBP家族基因与水稻SBP家族基因进化关系较近。对SBP基因在毛竹根茎叶中的相对表达量进行研究发现，PeSBP11和PeSBP16表达量分别在茎和根中最高。在花序发育P1-P4期（P1：花序形成初期；P2：小穗的分化；P3：颖花的分化；P4：小穗的生长），有6个基因（PeSBP1，PeSBP2，PeSBP3，PeSBP7，PeSBP15和PeSBP17）表达量较低，且无明显的表达量变化；7个基因（PeSBP5，PeSBP9，PeSBP10，PeSBP12，PeSBP14，PeSBP16和PeSBP18）在P1-P2期表达量上升，其中PeSBP10与PeSBP14的高表达延续到P3期；7个基因（PeSBP6，PeSBP8，PeSBP9，PeSBP10，PeSBP14，PeSBP16和PeSBP18）在P3期表达值较高；5个基因（PeSBP4，PeSBP5，PeSBP9，PeSBP13和PeSBP16）在P4期表达值较高。在高盐胁迫环境下，随着高盐胁迫处理时间的延长，PeSBP2，PeSBP3，PeSBP4，PeSBP8，PeSBP10，PeSBP11，PeSBP12与PeSBP14相对表达量逐渐上升。[结论]毛竹SBP家族基因在进化上非常保守，在进化过程中毛竹基因组可能并未出现特异的SBP基因。实时荧光定量PCR分析显示毛竹SBP家族基因参与了毛竹的营养器官生长，而且在毛竹开花过程中也发挥了重要的作用。在逆境盐胁迫条件下，有4个毛竹SBP基因的表达量发生了明显上调，表明毛竹SBP家族基因可能参与了毛竹对高盐的响应。

关键词: 毛竹, SBP家族基因, 基因表达, 花序发育, 盐胁迫

Abstract: [Objective] The SBP family genes encode a class of transcription factors which recognize and bind to the promoter of the MADS-box gene SQUAMOSA (SQUA), and regulate plant growth and reproductive growth. Systematic identification and analysis of Phyllostachys edulis SBP family genes can help to understand the role of them in vegetative growth and reproductive growth of P. edulis, and lay a foundation for gene functional analysis of SBP family genes in P. edulis.[Method] The SBP family genes were identified by bioinformatics methods using published P. edulis genomic data. The basic biological information of P. edulis SBP family genes was analyzed by using MEGA 6.0,DNAMAN, psRNATarget and other bioinformatics software. The expression patterns of P. edulis SBP family genes were studied by using the quantitative real-time fluorescence PCR (qPCR) in different tissues including leaf, root and stem, during floral development and under high salt stress.[Result] A total of 18 SBP family genes were identified in the genome of P. edulis, and these genes had a single and highly conserved SBP domain, of which 10 SBP family genes had miR156 target sites. There are 74 amino acid residues in the SBP domain of P. edulis, including two zinc finger domains and one NLS domain. The SBP transcription factors with close evolutionary relationship have similar motif distribution pattern and gene structure. The phylogenetic analysis of the SBP family gene of different species showed that the SBP family genes of P. edulis were closely related to the SBP family in rice. The qPCR result showed that, the relative expression levels of PeSBP11 and PeSBP16 were the highest in stems and roots, respectively. There were six genes (PeSBP1, PeSBP2, PeSBP3, PeSBP7, PeSBP15 and PeSBP17) with relatively low expression levels and no obvious change of expression abundances at P1-P4 stages (early in the formation of inflorescence (P1), the differentiation of spikelet (P2), the differentiation of floret (P3), the growth of spikelet (P4)) during the development of inflorescences. And seven genes (PeSBP5, PeSBP9, PeSBP10, PeSBP12, PeSBP14, PeSBP16 and PeSBP18) expression level increased in the P1-P2, of which high expression level of PeSBP10 and PeSBP14 continued to P3 phase. Seven genes (PeSBP6, PeSBP8, PeSBP9, PeSBP10, PeSBP14, PeSBP16 and PeSBP18) had a higher expression level than other genes in P3. Five genes (PeSBP4, PeSBP5, PeSBP9, PeSBP13 and PeSBP16) had the higher expression value than other genes at stage P4. The relative expression levels of PeSBP2, PeSBP3, PeSBP4, PeSBP8, PeSBP10, PeSBP11, PeSBP12 and PeSBP14 increased gradually with the prolonging of treatment time under high salt stress.[Conclusion] P. edulis SBP family genes were relatively conserved in evolution, and there is not novel SBP genes found in P. edulis genome. qPCR analysis showed that the SBP family genes of P. edulis were involved in the growth of vegetative organs of P. edulis, and played an important role in the flowering process of P. edulis. Under the high salt stress, the expression of four SBP family genes were significantly increased, which were involved in the response to high salt stress.

Key words: Phyllostachys edulis, SBP family genes, gene expression pattern, development of inflorescences, salt stress

中图分类号:

S718.46

王凯利, 傅鹰, 周明兵. 毛竹SQUAMOSA启动子结合蛋白SBP家族基因的鉴定及表达模式分析[J]. 林业科学, 2017, 53(12): 50-61.

Wang Kaili, Fu Ying, Zhou Mingbing. Identification and Expression Pattern of SQUAMOSA Promoter Binding Protein (SBP) Family Genes in Phyllostachys edulis[J]. Scientia Silvae Sinicae, 2017, 53(12): 50-61.

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