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三裂叶薯SPL基因家族鉴定、表达及miR156的调控分析
王婷1, 唐锐敏1, 王瑞晋2, 贺立恒3, 刘霞宇1, 刘世芳1, 李鹏飞1, 李润植3, 贾小云1
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(1山西农业大学生命科学学院,太谷 030801;2山西省太谷中学,太谷 030801;3 山西农业大学农学院,太谷 030801)
摘要:
SQUAMOSA promoter binding protein-like(SPL)是一类广泛存在于植物中的转录因子,均含有一个高度保守的SBP结构域(SQUAMOSA-PROMOTER BINDING PROTEIN)。本研究通过SBP结构域的隐马尔可夫模型、Blastp、CDD 和 SMART等程序从栽培甘薯的二倍体近缘野生种三裂叶薯(Ipomoea triloba L.)全基因组中鉴定出26个SPL基因家族成员。它们不均匀分布于三裂叶薯的12条染色体上。利用系统进化分析将新鉴定的26个三裂叶薯ItbSPL基因和来源于苔藓植物、单子叶植物、双子叶植物的116个SPL基因构建进化树,并根据拟南芥AtSPL基因家族的分类标准将26个ItbSPL基因家族分为7组。GSDS 2.0基因结构和MEME 4.12.0保守基序分析表明,不同进化分支中的ItbSPL基因的外显子/内含子数目及其蛋白基序组成差异明显。利用拟南芥中已知功能的SPL对ItbSPL基因的功能进行预测,发现三裂叶薯ItbSPL家族基因成员可能与植物开花、逆境胁迫、次生代谢产物等生物学过程相关。通过预测microRNA156(miR156)的作用位点发现,在26个ItbSPL基因中14个含有miR156的作用位点,其中13个ItbSPL基因具有PCR扩增产物。利用qRT-PCR检测13个候选靶基因及miR156在三裂叶薯叶、茎、根中的表达量,发现13个ItbSPL均在茎中的表达量最低,显著低于叶与根中的表达,而miR156则在茎中的表达量最高,在根中的表达量最低,初步推测这13个ItbSPL是miR156的靶基因。以上研究结果为六倍体栽培甘薯SPL基因家族成员的鉴定、进化分析及功能研究提供了方法和基础。
关键词:  三裂叶薯  SPL基因家族  全基因组鉴定  miR156  表达分析
DOI:10.13430/j.cnki.jpgr.20181204002
投稿时间:2018-12-04修订日期:2019-03-16
基金项目:国家重点研发专项(2018YFD1000705, 2018YFD1000700);山西省自然基金项目(201801D121238);山西省重点研发项目(201803D221008-6);山西省优秀博士来晋工作奖励资金科研项目(SXYBKY2018034);山西农谷建设科研专项(SXNGJSKYZX201701-03);山西省农业科技成果转化和推广示范项目(SXNKTG201812);山西农业大学科技创新基金(2018yz001)
Genome-wide Identification and Expression Analysis of SPL Gene Family Regulated by miR156 in Ipomoea triloba
WANG Ting1, TANG Rui-min1, WANG Rui-jin2, HE Li-heng3, LIU Xia-yu1, LIU Shi-fang1, LI Peng-fei1, LI Run-zhi3, JIA Xiao-yun1
(1College of Life Sciences, Shanxi Agricultural University, Taigu 030801;2Taigu high school, Taigu 030801;3College of Agriculture, Shanxi Agricultural University,Taigu 030801)
Abstract:
SQUAMOSA promoter binding protein-like (SPL) is one of the most important transcription factors in plant. The members of this family contain a highly conserved SBP (Squamosa-promoter Binding Protein) domain, and involve in regulating plant growth, secondary metabolism, hormonal signal transduction and abiotic stress. In this study, twenty-six SPL family genes (ItbSPL) were identified using the SBP Hidden Markov Model (HMM), Blastp, CDD and SMART from Ipomoea triloba (I.triloba) genome, which is a wild diploid progenitor of the sweetpotato. The ItbSPL members were unevenly distributed on 12 of the 15 chromosomes of I.triloba. Phylogenetic analysis with the newly identified 26 ItbSPL genes and 116 SPL genes from bryophytes, dicotyledons and monocotyledons showed that the 26 ItbSPL genes were classified into 7 sub-groups based on the similarity of conserved SBP domain with the orthologs in Arabidopsis thaliana. By analyzing gene structure and motif composition, the number of exons and motif of ItbSPL were different among different evolutionary clades. SPL genes from I.triloba and Arabidopsis were grouped together, suggesting a similar function of SPL genes. PsRNA Target prediction suggested 14 of the 26 ItbSPL genes contained complementary sequencesof miR156t, and 13 ItbSPL genes of them had PCR production. Furthermore, qRT-PCR showed the 13 ItbSPL genes had lowest expression in stem, where the expression of miR156 was highly accumulated, suggesting that the 13 ItbSPL genes might be the targets of miR156. Taken together, these results will be helpful for future studies on the identification, evolutionary and function analysis of SPL genes in hexaploid sweetpotato.
Key words:  Ipomoea triloba  SPL transcription factor  miR156  expression analysis  genome-wide analysis

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