丹参HD-ZIP基因家族的鉴定与表达分析

Identification and Expression Analysis of HD⁃ZIP Family in Salvia miltiorrhiza

为筛选丹参(Salvia miltiorrhiza)中响应高温胁迫的HD-ZIP基因,利用生物信息学研究方法,对丹参全基因组的HD-ZIP基因进行了鉴定和分析,并以天丹1号为材料,通过qPCR技术检测了丹参HDZIP基因在高温胁迫下的表达模式。结果表明,丹参全基因组中包含44个HD-ZIP基因(SmHD-ZIP),不均匀地分布于8条染色体上。SmHD-ZIP可分为HD-ZIPⅠ、HD-ZIPⅡ、HD-ZIPⅢ和HD-ZIPⅣ4个亚家族,序列分析结果表明,这些蛋白质的氨基酸残基数为180~982个,相对分子质量为20.947~109.620 ku。SmHD-ZIP均为亲水蛋白,无跨膜结构域,绝大多数不含信号肽,等电点为4.48~10.91,且几乎都在细胞核表达,蛋白质稳定性较差。44个SmHD-ZIP基因中有10个基因复制事件,均为纯化选择。结构和模体分析结果表明,同一亚家族成员的外显子数目基本一致,HD-ZIPⅢ亚家族成员的外显子数目最多,模体数也最多。模体1、6是SmHD-ZIP的保守模体,模体10、11、12、13、15为HD-ZIPⅢ亚家族特有,模体5为HD-ZIPⅣ亚家族特有。通过分析高温(37℃)胁迫下SmHD-ZIP的表达模式发现,表达量升高和下降的基因数基本一致,在上调表达的基因中,SmHD-ZIP1.11、SmHD-ZIP1.13、SmHD-ZIP2.2、SmHD-ZIP2.5、SmHD-ZIP3.1、SmHD-ZIP3.4、SmHD-ZIP4.9、SmHD-ZIP4.10和SmHD-ZIP4.12的表达量提高了10倍以上,可作为提高丹参耐热性的候选基因资源。

To screen HD⁃ZIP genes(SmHD‑ZIPs)in Salvia miltiorrhiza in response to high temperature stress,SmHD‑ZIPs were identified and analyzed by bioinformatics,and the expression pattern of SmHD‑ZIPs under high temperature stress was detected by qPCR.The results showed that there were 44 HD⁃ZIP genes in S.miltiorrhiza,most of which were unevenly distributed on 8 chromosomes.SmHD⁃ZIPs could be divided into four subfamilies:HD⁃ZIPⅠ,HD⁃ZIPⅡ,HD⁃ZIPⅢ,HD⁃ZIPⅣ.The sequence analysis results showed that the amino acid residues of these proteins ranged from 180 to 982,and the relative molecular weight of these proteins ranged from 20.947 ku to 109.620 ku.SmHD⁃ZIPs were hydrophilic proteins without transmembrane domain,and most of them did not contain signal peptides.The isoelectric points of SmHD⁃ZIPs were between 4.48 and 10.91,and they were almost expressed in the nucleus with the poor protein stability.Among the 44 SmHD‑ZIP genes,there were 10 gene duplication events,all of which were purified selection.The results of structure and motif analysis showed that the exon number of members of the same subfamily was basically the same,and the members of HD⁃ZIPⅢsubfamily had the largest exon number and the largest motif number.Motifs 1 and 6 were conserved motifs of SmHD⁃ZIP,motifs 10,11,12,13 and 15 was unique to the HD⁃ZIPⅢsubfamily,while motif 5 were unique to the HD⁃ZIP IV subfamily.By analyzing the expression patterns of SmHD‑ZIPs under high temperature(37℃)stress,it was found that the number of genes with increased and decreased expression was basically the same.Among the genes with up⁃regulated expression,the expression levels of SmHD‑ZIP1.11,SmHD‑ZIP1.13,SmHD‑ZIP2.2,SmHD‑ZIP2.5,SmHD‑ZIP3.1,SmHD‑ZIP3.4,SmHD‑ZIP4.9,SmHD‑ZIP4.10 and SmHD‑ZIP4.12 were increased by more than 10 times,which could be used as candidate gene resources to improve the heat resistance of S.miltiorrhiza.