芥菜型油菜AHK家族成员的全基因组鉴定及调控粒质量性状的初步探索

doi:10.7668/hbnxb.20194481

摘要/Abstract

摘要：

为探究芥菜型油菜AHK家族蛋白的结构特征及其与粒质量性状相关的调控功能,采用生物信息学方法对芥菜型油菜AHK家族成员进行了全基因组鉴定、理化性质、系统发育树、蛋白结构、启动子顺式元件和不同组织表达谱等分析。结果表明,在芥菜型油菜基因组中共鉴定到19条BjAHK蛋白序列,系统发育树分析将BjAHK家族成员分为了4个分支,即G1(BjAHK2~BjAHK4)、G2(BjAHK5)、G3(BjAHK1)和G4(BjCKI1)。功能结构域研究发现,BjAHK家族成员具有6个核心基序(Motif1~Motif6),与HisKA、HATPase_c和REC保守结构域相对应,其中BjAHK1~BjAHK4和BjCKI1蛋白的N端含有较为保守的跨膜结构域,可能与其跨膜作用功能有关。不同组织表达谱研究发现,细胞分裂素受体蛋白基因(BjAHK2~BjAHK4)为泛表达基因,且在根组织中表达量最高。结合已有的芥菜型油菜粒质量性状转录组数据,成功挖掘出4个可能参与粒质量性状调控的基因BjA07.AHK3、BjB03.AHK3、BjA10.AHK5和BjB05.AHK5。其中,BjA07.AHK3和BjB03.AHK3可能正向调控种子发育,而BjA10.AHK5和BjB05.AHK5则负向调控。结合BjAHK基因家族分析和转录组测序结果,推测BjAHK3和BjAHK5基因在调控芥菜型油菜种子发育方面发挥重要功能。

关键词: 芥菜型油菜, AHK家族, 粒质量性状, 表达分析

Abstract:

In order to investigate the structural characteristics of Brassica juncea AHK family proteins and their regulatory functions related to seed weight traits,it employed bioinformatics methods to perform whole-genome identification,physicochemical properties analysis,phylogenetic tree construction,protein structure analysis,promoter cis-elements analysis,and differential tissue expression profiling of the BjAHK family members.The main findings were as follows:a total of 19 BjAHK protein sequences were identified from the B.juncea genome,and phylogenetic analysis classified the BjAHK family members into four branches:G1 (BjAHK2—BjAHK4),G2 (BjAHK5),G3 (BjAHK1),and G4 (BjCKI1).Analysis of functional domains revealed the presence of six conserved motifs (Motif1—Motif6) in BjAHK family members,corresponding to the HisKA,HATPase_c,and REC conserved domains.The N-terminal regions of BjAHK1—BjAHK4 and BjCKI1 proteins exhibited relatively conserved transmembrane domains,which might be associated with their transmembrane function.Differential tissue expression profiling revealed that the genes encoding cytokinin receptor proteins (BjAHK2—BjAHK4) were constitutively expressed genes,with the highest expression levels observed in root tissues.Through integration with existing transcriptomic data on seed weight traits in B.juncea,four candidate genes potentially involved in the regulation of seed weight traits were identified:BjA07.AHK3,BjB03.AHK3,BjA10.AHK5,and BjB05.AHK5.BjA07.AHK3 and BjB03.AHK3 might positively regulate seed development,while BjA10.AHK5 and BjB05.AHK5 might have the opposite effect.Based on the integration of gene family analysis and transcriptome result,it was inferred that BjAHK3 and BjAHK5 genes play an important role in the regulation of seed development in B.juncea.

Key words: Brassica juncea, AHK family, Seed weight traits, Expression analysis

游亮, 陈浩, 肖华贵, 梁峰豪, 张超, 王璐璐, 唐容, 杨斌. 芥菜型油菜AHK家族成员的全基因组鉴定及调控粒质量性状的初步探索[J]. 华北农学报, 2024, 39(1): 8-17. doi: 10.7668/hbnxb.20194481.

YOU Liang, CHEN Hao, XIAO Huagui, LIANG Fenghao, ZHANG Chao, WANG Lulu, TANG Rong, YANG Bin. Genome-wide Identification of AHK Family Members in Brassica juncea and Preliminary Exploration of Regulating Seed Weight Traits[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(1): 8-17. doi: 10.7668/hbnxb.20194481.

http://www.hbnxb.net/CN/Y2024/V39/I1/8

图/表 9

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基因 Genes	引物序列 (5'—3') Sequence of primers (5'—3')
β-actin	Forward: AGCTCCGTGTTGCCCCTGAAGA Reverse: TGGATAGCAACATACATGGCAGGG
BjA10.AHK5	Forward: CATGCTCGCAAATACGGTGG Reverse: TCCTTCGGTTGTGTCGTCTG
BjB03.AHK3	Forward: TCTATTGATGTAGAGACATCATCATCT Reverse: GAAAGGTGTAAAAATACGGGACTGC

基因 Genes	引物序列 (5'—3') Sequence of primers (5'—3')
β-actin	Forward: AGCTCCGTGTTGCCCCTGAAGA Reverse: TGGATAGCAACATACATGGCAGGG
BjA10.AHK5	Forward: CATGCTCGCAAATACGGTGG Reverse: TCCTTCGGTTGTGTCGTCTG
BjB03.AHK3	Forward: TCTATTGATGTAGAGACATCATCATCT Reverse: GAAAGGTGTAAAAATACGGGACTGC

基因名称 Gene name	基因编号 Gene ID	氨基酸 Amino acid	等电点 pI	分子量/ku Molecular weight	亚细胞定位 Sub-celluluar localization	同系物 Homolog	一致性/% Identity
BjA07.AHK1	BjuA07g02470S	1 194	6.17	133.89	细胞质膜	AHK1	87.92
BjB03.AHK1	BjuB03g03770S	1 200	6.50	134.46	细胞质膜		86.83
BjA03.AHK2	BjuA03g20320S	1 123	5.66	125.32	细胞质膜	AHK2	81.50
BjA04.AHK2	BjuA04g08390S	1 843	6.36	205.56	细胞质膜		81.02
BjB01.AHK2	BjuB01g23760S	1 182	6.44	131.53	细胞质膜		71.22
BjB02.AHK2	BjuB02g08880S	1 066	5.45	119.46	细胞质膜		68.27
BjA07.AHK3	BjuA07g10690S	1 017	6.69	114.20	细胞质膜	AHK3	85.04
BjB03.AHK3	BjuB03g32520S	1 033	6.05	115.76	细胞质膜		83.89
BjB02.AHK4	BjuB02g52930S	1 059	6.20	117.43	内质网和细胞核	AHK4	88.39
BjA02.AHK5	BjuA02g39830S	908	5.12	102.41	细胞核和细胞质	AHK5	86.70
BjA03.AHK5	BjuA03g53640S	887	5.16	100.01	细胞核和细胞质		85.24
BjA10.AHK5	BjuA10g23440S	898	4.99	101.18	细胞核和细胞质		86.04
BjB02.AHK5	BjuB02g41860S	898	4.97	101.12	细胞核和细胞质		80.37
BjB05.AHK5	BjuB05g40600S	966	5.28	108.60	叶绿体和细胞核		82.73
BjB08.AHK5	BjuB08g57290S	941	5.14	105.78	细胞核和细胞质		82.82
BjA04.CKI1	BjuA04g06800S	1 056	8.81	116.94	细胞质膜	CKI	70.44
BjA05.CKI1	BjuA05g00190S	1 077	6.67	118.76	细胞质膜		68.01
BjB06.CKI1a	BjuB06g57680S	1 111	7.82	123.04	细胞质膜		71.20
BjB06.CKI1b	BjuB06g01950S	999	6.73	110.89	细胞质膜		67.08

基因名称 Gene name	基因编号 Gene ID	氨基酸 Amino acid	等电点 pI	分子量/ku Molecular weight	亚细胞定位 Sub-celluluar localization	同系物 Homolog	一致性/% Identity
BjA07.AHK1	BjuA07g02470S	1 194	6.17	133.89	细胞质膜	AHK1	87.92
BjB03.AHK1	BjuB03g03770S	1 200	6.50	134.46	细胞质膜		86.83
BjA03.AHK2	BjuA03g20320S	1 123	5.66	125.32	细胞质膜	AHK2	81.50
BjA04.AHK2	BjuA04g08390S	1 843	6.36	205.56	细胞质膜		81.02
BjB01.AHK2	BjuB01g23760S	1 182	6.44	131.53	细胞质膜		71.22
BjB02.AHK2	BjuB02g08880S	1 066	5.45	119.46	细胞质膜		68.27
BjA07.AHK3	BjuA07g10690S	1 017	6.69	114.20	细胞质膜	AHK3	85.04
BjB03.AHK3	BjuB03g32520S	1 033	6.05	115.76	细胞质膜		83.89
BjB02.AHK4	BjuB02g52930S	1 059	6.20	117.43	内质网和细胞核	AHK4	88.39
BjA02.AHK5	BjuA02g39830S	908	5.12	102.41	细胞核和细胞质	AHK5	86.70
BjA03.AHK5	BjuA03g53640S	887	5.16	100.01	细胞核和细胞质		85.24
BjA10.AHK5	BjuA10g23440S	898	4.99	101.18	细胞核和细胞质		86.04
BjB02.AHK5	BjuB02g41860S	898	4.97	101.12	细胞核和细胞质		80.37
BjB05.AHK5	BjuB05g40600S	966	5.28	108.60	叶绿体和细胞核		82.73
BjB08.AHK5	BjuB08g57290S	941	5.14	105.78	细胞核和细胞质		82.82
BjA04.CKI1	BjuA04g06800S	1 056	8.81	116.94	细胞质膜	CKI	70.44
BjA05.CKI1	BjuA05g00190S	1 077	6.67	118.76	细胞质膜		68.01
BjB06.CKI1a	BjuB06g57680S	1 111	7.82	123.04	细胞质膜		71.20
BjB06.CKI1b	BjuB06g01950S	999	6.73	110.89	细胞质膜		67.08

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