萝卜ALMT基因家族的鉴定与表达

doi:10.3969/j.issn.1004-1524.2022.04.11

浙江农业学报 ›› 2022, Vol. 34 ›› Issue (4): 746-755.DOI: 10.3969/j.issn.1004-1524.2022.04.11

萝卜ALMT基因家族的鉴定与表达

刘同金¹(), 徐铭婕¹, 汪精磊², 刘良峰³, 崔群香¹, 包崇来²^,^*(), 王长义¹^,^*()

1.金陵科技学院园艺园林学院,江苏南京 210038
2.浙江省农业科学院蔬菜研究所,浙江杭州 310021
3.江宁区农业农村局,江苏南京 211100

收稿日期:2021-07-30 出版日期:2022-04-25 发布日期:2022-04-28
作者简介:包崇来,E-mail: baocl@mail.zaas.ac.cn
*王长义,E-mail: cywang@jit.edu.cn;
刘同金(1985—),男,山东德州人,博士,讲师,研究方向为蔬菜种质资源与分子生物学。E-mail: tongjinliu@163.com
通讯作者: 包崇来,王长义
基金资助:
国家自然科学基金(31801858);国家级大学生创新训练计划(202113573014Z);金陵科技学院高层次人才科研启动项目(jit-b-202009);国家现代农业产业技术体系(CARS-23-A-14)

Genome-wide identification and expression analysis of ALMT gene family in radish

LIU Tongjin¹(), XU Mingjie¹, WANG Jinglei², LIU Liangfeng³, CUI Qunxiang¹, BAO Chonglai²^,^*(), WANG Changyi¹^,^*()

1. College of Horticulture, Jinling Institute of Technology, Nanjing 210038, China
2. Institute of Vegetable, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
3. Agricultural and Rural Affairs Bureau of Jiangning District, Nanjing 211100, China

Received:2021-07-30 Online:2022-04-25 Published:2022-04-28
Contact: BAO Chonglai,WANG Changyi

摘要/Abstract

摘要：

为探究萝卜铝激活苹果酸转运蛋白(ALMT)家族成员对生物和非生物胁迫的响应,本研究通过生物信息学方法对萝卜ALMT家族成员进行鉴定,并利用转录组数据对其进行表达分析。结果显示,萝卜基因组中包含17个ALMT基因,分布于8条染色体。该家族成员外显子数量为5~7个,预测N-端含有5~6个跨膜结构,在染色体上的分布不均匀。萝卜ALMT基因家族成员启动子上有多种激素响应、胁迫响应、组织和器官生长发育与环境响应的顺式作用元件,表明萝卜ALMT基因家族成员的表达受多种条件的调控。对它们在不同组织和发育时期,以及对Agrobacterium tumefaciens的侵染和重金属(铅、镉和铬)胁迫的响应研究发现,RsALMT1和RsALMT13可能参与抵抗A. tumefaciens的侵染;RsALMT3可能调控叶片气孔的开放,而RsALMT7、RsALMT11和RsALMT14可能调控叶片气孔的关闭;RsALMT15可能参与绿肉萝卜根肉中叶绿素的积累;RsALMT16在抵御铅胁迫过程中发挥重要作用。以上结果将为进一步研究萝卜ALMT家族成员在生物和非生物胁迫响应中的功能与分子机制奠定基础。

关键词: 萝卜, ALMT基因家族, 系统进化, 基因表达

Abstract:

In order to explore the response of aluminium activated malate transporter (ALMT) genes to biotic and abiotic stress in radish,ALMT genes of radish were genome-wide identified by bioinformatics method and expression patterns were analyzed based on RNA-seq data. The results showed that there were 17 ALMT genes in radish genome, and distributed on eight chromosomes. Exon number of these genes ranged from 5 to 7, and 5-6 transmembrane domains were predicted in their N-terminal region. Number of RsALMT genes on chromosomes was different. Expression analysis of RsALMT genes in different tissues and developments stages,treatment with Agrobacterium tumefaciens and heavy metal (cadmium, chromium and lead) showed that RsALMT1 and RsALMT13 might be involved in resistance to the infection of A. tumefaciens, RsALMT3 might control stomatal open, while RsALMT7, RsALMT11 and RsALMT14 mightbe involved in stomatal closure.RsALMT15 might regulate the accumulation of chlorophyll in flesh of radish and RsALMT16 might be involved in resistance to leadstress.Therefore, the above results would provide a theoretical basis for further study on the functions and molecular mechanisms of radish ALMT family members in response to biotic and abiotic stress.

Key words: radish, ALMT gene family, phylogenetic evolution, gene expression

中图分类号:

S631.1

刘同金, 徐铭婕, 汪精磊, 刘良峰, 崔群香, 包崇来, 王长义. 萝卜ALMT基因家族的鉴定与表达[J]. 浙江农业学报, 2022, 34(4): 746-755.

LIU Tongjin, XU Mingjie, WANG Jinglei, LIU Liangfeng, CUI Qunxiang, BAO Chonglai, WANG Changyi. Genome-wide identification and expression analysis of ALMT gene family in radish[J]. Acta Agriculturae Zhejiangensis, 2022, 34(4): 746-755.

图/表 9

表1 萝卜ALMT家族成员基本信息

Table 1 Information of ALMT gene family members in radish

基因名称 Gene name	登录号 Accession No.	氨基酸数量 Number of amino acids/aa	分子量 Molecular weight/u	等电点 Isoeletric point	不稳定指数 Instability index	亲水性 GRAVY	脂肪指数 Aliphatic index	跨膜结构 Transmembrane domain	拟南芥同源基因 Homologue of AtALMT
RsALMT1	Rsa10013723	525	58 320.75	6.40	36.07	0.010	91.22	5	AT2G17470
RsALMT2	Rsa10016259	542	60 681.27	8.99	42.72	-0.072	96.88	5	AT5G46600
RsALMT3	Rsa10035068	621	70 102.62	6.18	36.65	-0.043	89.98	5	AT3G18440
RsALMT4	Rsa10034640	491	54 643.38	8.77	32.83	-0.027	99.61	5	AT3G11680
RsALMT5	Rsa10021670	479	53 569.17	8.30	36.44	0.014	89.37	5	AT4G00910
RsALMT6	Rsa10022591	481	53 733.47	9.04	32.19	-0.020	90.58	6	AT4G00910
RsALMT7	Rsa10025402	543	60 482.70	8.84	40.46	-0.071	95.91	6	AT4G17970
RsALMT8	Rsa10010851	618	68 136.96	7.59	38.48	-0.174	90.95	5	AT5G46610
RsALMT9	Rsa10020182	404	45 923.80	8.41	30.84	-0.125	94.03	6	AT2G27240
RsALMT10	Rsa10011190	489	54 469.32	9.10	32.90	0.032	104.03	5	AT3G11680
RsALMT11	Rsa10039953	580	65 364.87	7.21	40.72	-0.098	86.60	6	AT1G25480
RsALMT12	Rsa10038217	603	67 587.76	6.32	46.06	-0.036	90.10	6	AT1G18420
RsALMT13	Rsa10037498	505	56 082.17	6.53	40.90	-0.049	97.84	5	AT1G08440
RsALMT14	Rsa10040558	515	58 157.37	6.11	41.97	-0.104	88.62	5	AT1G25480
RsALMT15	Rsa10041290	510	56 863.94	6.69	38.69	-0.123	94.43	5	AT1G08440
RsALMT16	Rsa10041291	471	52 544.32	6.11	42.05	-0.020	98.09	5	AT1G08430
RsALMT17	Rsa10003573	484	54 188.99	8.59	31.96	0.067	93.22	6	AT4G00910

图1 萝卜ALMT基因家族成员的染色体定位

Fig.1 Chromosomal position of the ALMT gene family members in radish

图2 萝卜ALMT基因家族成员进化树与基因结构

Fig.2 Phylogenetic tree and exon distribution of ALMT family members in radish

图3 萝卜(Rs)、拟南芥(AT)与甘蓝型油菜(Bna)ALMT基因家族成员编码的蛋白质进化树

Fig.3 Phylogenetic tree of proteins coded by ALMT gene family members in Raphanus sativus(Rs), Arabidopsis(AT) and Brassica napus(Bna)

图4 萝卜ALMT基因家族成员启动子顺式作用元件预测

Fig.4 Predicted cis-elements in RsALMT promoters

图5 萝卜ALMT基因家族成员的组织表达模式 ESS、SS、EES、RES和MS分别为芽期、破肚期、膨大前期、膨大盛期和成熟期。

Fig.5 Expression profiles of RsALMT genes in various tissues ESS, seedling stage; SS, splitting stage; EES, early expanding stage; RES, rapid expanding stage; MS, mature stage.

图6 绿肉萝卜与白肉萝卜不同发育时期肉质根ALMT基因家族成员的表达 S1,9月25日,S2,10月2日,S3,10月9日,S4,10月16日,S5,10月23日。

Fig.6 Expression profiles of RsALMT genes in different growing stage of taproot of white and green flesh S1, September 25th; S2, October 2nd; S3, October 9th; S4, October 16th; S5, October 23th.

图7 抗和感根癌病萝卜接种A. tumefaciens 7 d后下胚轴ALMT基因家族成员的表达 Line_18和Line_19分别为抗和感根癌病萝卜株系;CK为接种LB培养基7 d,T为接种A. tumefaciens处理7 d。

Fig.7 Expression profiles of ALMT gene family members in 7 d after incubated with A. tumefaciens of hypocotyls Line_18 and Line_19 was resistance and susceptible radish inbred lines to A. tumefaciens, respectively. CK, 7 d after incubated with LB medium; T, 7 d after incubated with A. tumefaciens.

图8 重金属(铅、镉和铬)胁迫萝卜根中ALMT基因家族成员的表达

Fig.8 Expression profiles of ALMT gene family members in radish root with heavy metal (lead, cadmium and chromium) stress treatment

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萝卜ALMT基因家族的鉴定与表达

Genome-wide identification and expression analysis of ALMT gene family in radish

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