苹果PLATZ基因家族鉴定及MdPLATZ9基因功能研究

doi:10.13560/j.cnki.biotech.bull.1985.2024-0903

摘要/Abstract

摘要：

目的从苹果全基因组中鉴定MdPLATZs转录因子家族，分析其表达特点，为后续深入研究MdPLATZs转录因子的生物学功能奠定基础。方法通过生物信息学方法从苹果全基因组鉴定出17个PLATZ基因家族成员，并依据其序列特征将其划分为4个亚家族，并对其理化特征、系统进化关系、基因结构、染色体定位和启动子顺式作用元件等进行分析。本研究测定了苹果不同组织中PLATZ家族成员的表达水平，并通过异源表达及干旱胁迫处理来鉴定MdPLATZ9的基因功能。结果生物信息学分析结果表明，MdPLATZs分子量范围为17.23-29.39 kD，等电点介于8.24-9.51之间，且多数成员定位在细胞核内。这些基因分布于苹果的13条染色体上，同一亚家族的成员具有相似的基因结构并具有较高的序列同源性，且各组之间存在明显的共线性关系。PLATZ结构域在MdPLATZs家族中高度保守。基因表达模式分析显示，MdPLATZs家族成员存在组织表达特异性，MdPLATZs在花中均有表达，MdPLATZ7在花和茎中表达量最高。进一步分析显示，过表达MdPLATZ9的拟南芥植株过氧化物酶和过氧化氢酶活性显著增强，同时丙二醛和过氧化氢含量降低，抗坏血酸水平升高，并且超氧阴离子的生成速率减缓，这些结果表明，过表达MdPLATZ9提高了拟南芥的抗旱性。结论从苹果全基因组中鉴定出17个MdPLATZs转录因子家族成员，该家族成员在基因结构、蛋白基序表现出高度的相似性和同源性，并且存在显著的片段重复。过表达MdPLATZ9的拟南芥植株表现出明显的抗旱性。

关键词: 苹果, MdPLATZ9, 转录因子, 生物信息学, 表达模式分析

Abstract:

Objective To identify MdPLATZs transcription factor families from the whole apple (Malus domestica) genome, to analyze their expression characteristics, and to lay the foundation for the subsequent in-depth study of the biological functions ofMdPLATZstranscription factors. Method Seventeen members of the PLATZ gene family were identified from the whole apple genome by bioinformatics methods and divided into four subfamilies based on their sequence features, and their physicochemical characteristics, phylogenetic relationships, gene structures, chromosomal localisation and promoter cis-acting elements were analyzed. The expressions of PLATZ family members in different tissues of apple were determined, and the gene functions of MdPLATZ9 by heterologous expression and drought stress treatment were identified. Result Bioinformatics analyses showed that the molecular weight of MdPLATZs ranged from 17.23-29.39 kD, the isoelectric point ranged from 8.24-9.51, and most of the members were localised in the nucleus. These genes were distributed on 13 chromosomes of apple, and members of the same subfamily had similar gene structures and high sequence homology, with obvious covariance between groups. The PLATZ structural domains were highly conserved in the MdPLATZs family. Gene expression pattern analysis showed that there was tissue expression specificity among MdPLATZs family members, with MdPLATZs being expressed in flowers, and MdPLATZ7 being the most highly expressed in flowers and stems. Further analysis showed that peroxidase and catalase activities were significantly enhanced in Arabidopsis plants overexpressing MdPLATZ9, while malondialdehyde and hydrogen peroxide levels reduced, ascorbic acid levels increased, and the rate of superoxide anion production was slowed down, which indicated that the overexpression of MdPLATZ9 improved drought tolerance in Arabidopsis. Conclusion Seventeen members of the MdPLATZs transcription factor family are identified from the whole apple genome, and the members of this family show high similarity and homology in gene structure, protein motifs, and significant segmental duplications. Arabidopsis plants overexpressing MdPLATZ9 demonstrates significant drought resistance.

Key words: apple, MdPLATZ9, transcription factor, bioinformatics, expression pattern analysis

王天禧, 杨炳松, 潘荣君, 盖文贤, 梁美霞. 苹果PLATZ基因家族鉴定及MdPLATZ9基因功能研究[J]. 生物技术通报, 2025, 41(4): 176-187.

WANG Tian-xi, YANG Bing-song, PAN Rong-jun, GAI Wen-xian, LIANG Mei-xia. Identification of the Apple PLATZ Gene Family and Functional Study of the MdPLATZ9 Gene[J]. Biotechnology Bulletin, 2025, 41(4): 176-187.

图/表 10

表1 苹果MdPLATZ基因家族成员信息

Table 1 Information of MdPLATZ gene family member in apple

基因ID Gene ID	基因名称 Gene name	等电点 pI	分子量 Molecular weight/Da	亚细胞定位预测 Localization prediction
MD00G1059100	MdPLATZ1	8.55	28 544.59	Nucleus
MD02G1017000	MdPLATZ2	8.56	29 394.72	Nucleus
MD02G1208800	MdPLATZ3	8.82	24 671.44	Nucleus
MD03G1129200	MdPLATZ4	9.51	21 971.14	Nucleus
MD05G1248500	MdPLATZ5	8.39	28 487.59	Chloroplast
MD06G1001300	MdPLATZ6	8.78	28 441.46	Chloroplast, nucleus
MD06G1035500	MdPLATZ7	9.25	24 878.17	Nucleus
MD07G1117000	MdPLATZ8	8.81	27 385.73	Nucleus
MD10G1229000	MdPLATZ9	8.39	28 548.56	Chloroplast, nucleus
MD11G1151300	MdPLATZ10	9.30	24 312.98	Nucleus
MD12G1103000	MdPLATZ11	8.88	22 043.09	Nucleus
MD13G1017800	MdPLATZ12	9.50	24 708.19	Nucleus
MD14G1097400	MdPLATZ13	8.52	21 996.09	Nucleus
MD15G1161500	MdPLATZ14	8.75	29 101.53	Nucleus
MD16G1015800	MdPLATZ15	9.02	25 758.81	Nucleus
MD16G1273400	MdPLATZ16	9.33	25 264.61	Nucleus
MD17G1254800	MdPLATZ17	8.24	17 230.53	Nucleus

图1 MdPLATZs家族成员的基因染色体定位（A）、基因结构和保守基序分析（B）

Fig. 1 Chromosomal location of genes (A), gene structure and conserved motif analysis (B) of MdPLATZs family members

图2 苹果中PLATZ基因的同源性分析灰色曲线表示苹果基因组中的共线区域，红色曲线表示发生片段重复的基因对

Fig. 2 Homology analysis of PLATZ genes in appleGrey curves indicate regions of colinearity in the apple genome, and red curves indicate pairs of genes where segmental duplication occurs

图3 苹果、拟南芥和玉米的PLATZ系统发育树▲表示苹果PLATZ蛋白；★表示玉米PLATZ蛋白；●表示拟南芥PLATZ蛋白；数字代表亲缘关系的远近

Fig. 3 PLATZ phylogenetic tree of apple, Arabidopsis and maize▲ indicates apple's PLATZ protein. ★ indicates maize's PLATZ protein. ● indicates Arabidopsis's PLATZprotein. Numbers indicate the closeness of kinship

图4 苹果MdPLATZs基因在不同组织表达水平WS：整株幼苗；FR：果实；L：叶；SA：茎尖；FL：花；S：茎；EDB：早期休眠芽；FDB：盛开芽；YL：幼叶

Fig. 4 Expressions of MdPLATZs gene in different tissues of appleWS: Whole seedling; FR: fruit; L: leaf; SA: stem tip; FL: flower; S: stem; EDB: early dormant bud; FDB: bloom bud; YL: young

图5 MdPLATZs基因启动子中顺式作用元件分析MdPLATZs启动子种类及数量可视化

Fig. 5 Analysis of cis-acting elements in the promoter of the MdPLATZs geneVisualized types and number of MdPLATZs promoters

图6 MdPLATZs与ORESARA15进化关系图中数字分别代表亲缘关系的远近以及分支长度

Fig. 6 Evolutionary relationship between MdPLATZs and ORESARA15The numbers in the figure indicate the proximity of the relatives and the branch lengths, respectively

图7 转基因拟南芥与野生型拟南芥的比较A：野生型拟南芥与过表达拟南芥生长14 d表型；B：野生型拟南芥与过表达拟南芥MdPLATZ9表达量；C：野生型拟南芥与过表达拟南芥生长14 d后根长数据统计；WT：野生型拟南芥；OE1：过表达拟南芥株系；OE2：过表达株系2；OE3：过表达株系3；下同；数据为平均值±标准差，n=3；* P<0.05，** P<0.01，*** P<0.001

Fig. 7 Comparison of transgenic A. thaliana with wild-type A. thalianaA: Phenotypes of wild-type Arabidopsisthaliana and overexpressed A. thaliana grown for 14 d. B: Expression of MdPLATZ9 in wild-type A. thaliana and overexpressed A. thaliana. C: Statistics of root length data of wild-type A. thaliana and overexpressed A. thaliana grown for 14 d. WT: Wild-type A. thaliana; OE1: overexpressed A. thaliana line; OE2: overexpressed line 2; OE3: overexpressed line 3; the same below. The data are means±standard deviation (SD), n=3. * P<0.05; ** P<0.01, *** P<0.001

图8 干旱胁迫下野生型与过表达株系拟南芥的表型及生理指标测定A：野生型拟南芥与过表达拟南芥在干旱胁迫下的表型； B‒G：野生型拟南芥与过表达拟南芥在干旱胁迫下O2-生成速率、H2O2含量、CAT活性、POD活性、Vc活性、MDA含量。数据为3个重复样本的平均值±标准偏差，不同字母表示在P<0.05水平差异显著。下同

Fig. 8 Determination of phenotypic and physiological parameters of wild-type and overexpressed strains of A. thaliana under drought stressA: Phenotypes of wild-type A. thaliana and overexpressed A. thaliana under drought stress. B‒G: Rate of O2- production, H2O2 content, CAT activity, POD activity, Vc activity, and MDA content of wild-type A. thaliana and overexpressed A. thaliana under drought stress. Data are the mean±SD of three replicates. Different letters indicate significant differences at P<0.05 level. The same below

图9 四个不同基因在拟南芥野生型和过表达植株中的表达量

Fig. 9 Expressions of four different genes in Arabidopsis wild-type and overexpressed plants

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