梨Dof家族基因鉴定及其在宿存与脱落萼片中的表达分析

doi:10.16420/j.issn.0513-353x.2021-0642

摘要/Abstract

摘要：

根据苹果、拟南芥等植物中的Dof转录因子基因序列,利用BLAST软件鉴定到梨基因组中51个PbDof基因,分为9个亚家族。基因结构分析表明,PbDof的内含子数为 1 ~ 3个;启动子区域含有5种胁迫响应和9种植物激素的顺式调控元件;基因复制结果显示,PbDof在物种进化过程中主要受纯化选择的作用。以蒸馏水为对照,于‘库尔勒香梨’盛花期喷施外源GA₃和PP₃₃₃,并采qRT-PCR技术检测各处理条件下宿萼组和脱萼组子房和萼片中PbDof的表达情况。结果表明,处理9 d后,PbDof34和PbDof51的表达量在GA₃处理宿萼组萼片中均显著高于PP₃₃₃处理脱萼组,且PbDof34显著高于对照;而PbDof16、PbDof19和PbDof44的表达量在GA₃处理宿萼组萼片中均低于PP₃₃₃处理脱萼组;此外,PbDof16和PbDof44的表达量在PP₃₃₃处理脱萼组萼片中均显著高于对照。因此,PbDof34和PbDof51可能正向调控萼片宿存,PbDof16、PbDof19和PbDof44可能正向调控萼片脱落。

关键词: 梨, Dof基因家族, 萼片宿存, 萼片脱落

Abstract:

Dof genes in pear genome were identified by BLAST software based on Dof genes from domestic apple and Arabidopsis thaliana. TBtools,MEME and MEGA-X software were used for bioinformatics analysis of Dof protein and gene sequences. The results showed that a total of 51 PbDof genes were identified,which belong to nine subfamilies. Structural analysis showed that the number of introns in PbDof gene varies from one to three. The promoter region contains five cis-regulatory elements related to stress response and nine cis-regulatory elements related to plant hormones. Synteny analysis of PbDof genes showed that PbDof was mainly affected by purification selection in the process of species evolution. To provide theoretical basis for understanding what roles the Dof genes play in calyx persistence and abscission of pear,the qRT-PCR method was used to detect the relative expression of Dof genes in ovary and sepals after sprayed with distilled water,PP₃₃₃ and GA₃ at full bloom stage. qRT-PCR analysis revealed that the expression levels of PbDof34 and PbDof51 after GA₃treatment were significantly higher than those after PP₃₃₃ treatment,and the expression levels of PbDof34 were significantly higher than those after distilled water treatment（control） in calyxes on 9 d. The expression levels of PbDof16,PbDof19 and PbDof44 after GA₃treatment were all lower than those after PP₃₃₃ treatment in calyxes on 9 d. The expression levels of PbDof16 and PbDof44 after PP₃₃₃ treatment was significantly lower than those of control in calyxes on 9 d. We speculated that PbDof34 and PbDof51 may promote the calyxes persistence whereas PbDof16,PbDof19 and PbDof44 may promote the calyxes abscission in‘Korla Fragrant Pear’.

Key words: pear, Dof gene family, calyx persistence, calyx abscission

中图分类号:

S661.2

刘金明, 郭彩华, 袁星, 亢超, 全绍文, 牛建新. 梨Dof家族基因鉴定及其在宿存与脱落萼片中的表达分析[J]. 园艺学报, 2022, 49(8): 1637-1649.

LIU Jinming, GUO Caihua, YUAN Xing, KANG Chao, QUAN Shaowen, NIU Jianxin. Genome-wide Identification of Dof Family Genes and Expression Analysis Sepal Persistent and Abscission in Pear[J]. Acta Horticulturae Sinica, 2022, 49(8): 1637-1649.

图/表 7

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基因	上游引物（5′-3′）	下游引物（5′-3′）
Gene	Forward primer	Reverse primer
PbDof11	AACTCATGCCCTAAGCCTGC	TTGAACCACCTCCAACTGGG
PbDof16	CCGGTGTTCTGTGGCCTTAT	CCCTCGAATGCTTCCCCAAT
PbDof19	CTCCTTGGCCTTACCCTTGG	TCGAATGTTTCCCCAGGGTG
PbDof34	AGGTCCAAAACGGCCAAGAT	CAGCCCTTGCAGAAGTGTCT
PbDof44	CTCCTTGGCCTTACCCTTGG	CGGTGGAGCTTCTTCCTCAG
PbDof51	CGGCACTTCTGCAAATCCTG	ACTCAACTGTAAGGCCGCTC
Actin	CCATCCAGGCTGTTCTCTC	GCAAGGTCCAGACGAAGG

基因	上游引物（5′-3′）	下游引物（5′-3′）
Gene	Forward primer	Reverse primer
PbDof11	AACTCATGCCCTAAGCCTGC	TTGAACCACCTCCAACTGGG
PbDof16	CCGGTGTTCTGTGGCCTTAT	CCCTCGAATGCTTCCCCAAT
PbDof19	CTCCTTGGCCTTACCCTTGG	TCGAATGTTTCCCCAGGGTG
PbDof34	AGGTCCAAAACGGCCAAGAT	CAGCCCTTGCAGAAGTGTCT
PbDof44	CTCCTTGGCCTTACCCTTGG	CGGTGGAGCTTCTTCCTCAG
PbDof51	CGGCACTTCTGCAAATCCTG	ACTCAACTGTAAGGCCGCTC
Actin	CCATCCAGGCTGTTCTCTC	GCAAGGTCCAGACGAAGG

基因 Gene	ID	染色体位置 Chromosome location	染色体重命名 Chromosome rename	长度/aa Length	分子量/kD MW	等电点 pI	α-螺旋/% α-helix	β-转角/% β-turn	无规则卷曲/% Random coil	延长链/% Extended strand
PbDof1	XM_009365818.2	Scaffold3.0	Scaff1	248	26.31	5.95	8.87	4.44	70.97	15.73
PbDof2	XM_009368130.2	Scaffold4.0	Scaff2	387	41.62	9.33	17.83	3.88	66.41	11.89
PbDof3	XM_018649999.1	Scaffold4.0	Scaff2	239	24.85	8.64	8.37	7.11	66.95	17.57
PbDof4	XM_009354127.2	Scaffold21.0	Scaff3	304	33.58	6.45	15.13	3.62	64.47	16.78
PbDof5	XM_009354128.2	Scaffold21.0	Scaff3	304	33.58	6.45	15.13	3.62	64.47	16.78
PbDof6	XM_009354499.2	Scaffold24.0	Scaff4	163	18.27	9.43	10.43	4.91	68.71	15.95
PbDof7	XM_009354615.2	Scaffold24.0	Scaff4	378	41.07	8.05	13.23	0.79	77.78	8.20
PbDof8	XM_009354693.2	Scaffold24.0	Scaff4	290	31.20	8.51	11.72	3.45	75.17	9.66
PbDof9	XM_009355416.2	Scaffold31.0	Scaff5	368	39.79	8.59	8.97	3.80	75.82	11.41
PbDof10	XM_009359983.2	Scaffold63.0	Scaff6	291	32.10	7.64	15.81	3.09	67.35	13.75
PbDof11	XM_009360389.2	Scaffold67.0	Scaff7	314	34.38	9.33	9.24	2.23	77.71	10.83
基因 Gene	ID	染色体位置 Chromosome location	染色体重命名 Chromosome rename	长度/aa Length	分子量/kD MW	等电点 pI	α-螺旋/% α-helix	β-转角/% β-turn	无规则卷曲/% Random coil	延长链/% Extended strand
PbDof12	XM_009361313.1	Scaffold75.0	Scaff8	327	36.06	9.00	7.95	1.53	74.62	15.90
PbDof13	XM_018648309.1	Scaffold83.0	Scaff9	523	55.98	5.94	13.77	2.29	76.67	7.27
PbDof14	XM_018648376.1	Scaffold84.0	Scaff10	302	33.89	5.11	25.50	1.66	66.89	5.96
PbDof15	XM_009364743.2	Scaffold106.0	Scaff11	327	36.23	6.69	12.54	2.75	66.36	18.35
PbDof16	XM_009371010.2	Scaffold180.0	Scaff12	465	50.96	5.79	12.04	1.94	79.78	6.24
PbDof17	XM_009377147.2	Scaffold275.0	Scaff13	286	31.52	7.71	5.59	1.05	80.42	12.94
PbDof18	XM_009377148.2	Scaffold275.0	Scaff13	270	29.76	8.44	6.30	3.33	77.78	12.59
PbDof19	XM_009378003.2	Scaffold292.0	Scaff14	511	55.22	5.72	10.37	1.37	80.82	7.44
PbDof20	XM_009336089.2	Scaffold384.0	Scaff15	289	31.75	7.68	8.30	1.73	80.28	9.69
PbDof21	XM_009340481.2	Scaffold499.0	Scaff16	366	38.92	9.13	9.29	3.83	77.60	9.29
PbDof22	XM_009344990.1	Scaffold698.0	Scaff17	316	35.40	6.43	12.03	2.53	74.68	10.76
PbDof23	XM_009344991.1	Scaffold698.0	Scaff17	316	34.97	6.52	13.61	2.85	72.78	10.76
PbDof24	XM_009345555.2	Scaffold727.0	Scaff18	372	40.05	8.78	8.06	2.42	79.84	9.68
PbDof25	XM_009345556.2	Scaffold727.0	Scaff18	337	36.26	8.96	6.53	2.97	81.31	9.20
PbDof26	XM_009346163.2	Scaffold765.0	Scaff19	225	24.05	8.18	6.67	3.56	79.11	10.67
PbDof27	XM_009346170.2	Scaffold765.0	Scaff19	225	24.05	8.18	6.67	3.56	79.11	10.67
PbDof28	XM_009348304.2	Scaffold915.0	Scaff20	304	33.60	6.62	18.42	1.64	64.80	15.13
PbDof29	XM_009348967.2	Scaffold973.0	Scaff21	347	37.14	8.90	20.46	2.59	66.28	10.66
PbDof30	XM_009349812.2	Scaffold1047.0	Scaff22	346	36.91	8.84	19.65	3.18	65.32	11.85
PbDof31	XM_009352168.2	Scaffold1355.0	Scaff23	303	33.81	4.94	20.79	1.65	69.64	7.92
PbDof32	XM_009336822.2	Scaffold1.0.1	Scaff24	291	32.49	7.70	15.81	2.06	74.23	7.90
PbDof33	XM_009343730.2	Scaffold1.0.1	Scaff24	378	41.13	8.58	15.08	0.53	76.19	8.20
PbDof34	XM_009351106.2	Scaffold1.0.1	Scaff24	163	18.34	9.08	14.11	6.13	63.19	16.56
PbDof35	XM_009354801.2	Scaffold1.0.1	Scaff24	291	31.31	9.30	16.84	5.50	63.23	14.43
PbDof36	XM_009339603.2	Scaffold14.0.1	Scaff25	323	34.90	7.20	15.17	2.79	69.04	13.00
PbDof37	XM_009340881.2	Scaffold14.0.1	Scaff25	463	50.48	5.48	13.17	1.51	76.89	8.42
PbDof38	XM_009345020.2	Scaffold16.0.1	Scaff26	362	39.20	8.99	9.12	4.14	73.76	12.98
PbDof39	XM_009347952.2	Scaffold30.0.1	Scaff27	325	34.81	7.21	8.31	1.85	76.00	13.85
PbDof40	XM_009351662.2	Scaffold17.0.2	Scaff28	282	31.49	6.77	10.64	3.90	74.11	11.35
PbDof41	XM_009353689.2	Scaffold20.0.1	Scaff29	320	35.00	7.23	10.94	2.50	75.94	10.62
PbDof42	XM_009359445.2	Scaffold58.0.1	Scaff30	357	38.71	8.93	10.36	4.76	71.43	13.45
PbDof43	XM_009362338.2	Scaffold80.0.1	Scaff31	539	58.63	6.66	13.17	1.67	78.11	7.05
PbDof44	XM_009362339.2	Scaffold80.0.1	Scaff31	521	56.57	6.22	13.05	2.30	77.93	6.72
PbDof45	XM_009363055.2	Scaffold87.0.1	Scaff32	316	34.47	8.16	9.18	4.43	74.05	12.34
PbDof46	XM_009365959.2	Scaffold116.0.1	Scaff33	428	45.70	9.03	16.12	2.57	72.43	8.88
PbDof47	XM_009366523.2	Scaffold122.0.1	Scaff34	384	40.83	8.75	15.10	2.08	73.96	8.85
PbDof48	XM_009367372.2	Scaffold131.0.1	Scaff35	253	26.84	6.15	14.62	7.51	55.34	22.53
PbDof49	XM_009369256.2	Scaffold155.0.1	Scaff36	221	23.59	8.13	10.86	2.71	76.02	10.41
PbDof50	XM_009376412.2	Scaffold262.0.1	Scaff37	376	40.32	9.64	13.83	3.99	74.20	7.98
PbDof51	XM_009378209.2	Scaffold294.0.1	Scaff38	247	25.39	8.18	9.31	5.67	67.61	17.41

基因 Gene	ID	染色体位置 Chromosome location	染色体重命名 Chromosome rename	长度/aa Length	分子量/kD MW	等电点 pI	α-螺旋/% α-helix	β-转角/% β-turn	无规则卷曲/% Random coil	延长链/% Extended strand
PbDof1	XM_009365818.2	Scaffold3.0	Scaff1	248	26.31	5.95	8.87	4.44	70.97	15.73
PbDof2	XM_009368130.2	Scaffold4.0	Scaff2	387	41.62	9.33	17.83	3.88	66.41	11.89
PbDof3	XM_018649999.1	Scaffold4.0	Scaff2	239	24.85	8.64	8.37	7.11	66.95	17.57
PbDof4	XM_009354127.2	Scaffold21.0	Scaff3	304	33.58	6.45	15.13	3.62	64.47	16.78
PbDof5	XM_009354128.2	Scaffold21.0	Scaff3	304	33.58	6.45	15.13	3.62	64.47	16.78
PbDof6	XM_009354499.2	Scaffold24.0	Scaff4	163	18.27	9.43	10.43	4.91	68.71	15.95
PbDof7	XM_009354615.2	Scaffold24.0	Scaff4	378	41.07	8.05	13.23	0.79	77.78	8.20
PbDof8	XM_009354693.2	Scaffold24.0	Scaff4	290	31.20	8.51	11.72	3.45	75.17	9.66
PbDof9	XM_009355416.2	Scaffold31.0	Scaff5	368	39.79	8.59	8.97	3.80	75.82	11.41
PbDof10	XM_009359983.2	Scaffold63.0	Scaff6	291	32.10	7.64	15.81	3.09	67.35	13.75
PbDof11	XM_009360389.2	Scaffold67.0	Scaff7	314	34.38	9.33	9.24	2.23	77.71	10.83
基因 Gene	ID	染色体位置 Chromosome location	染色体重命名 Chromosome rename	长度/aa Length	分子量/kD MW	等电点 pI	α-螺旋/% α-helix	β-转角/% β-turn	无规则卷曲/% Random coil	延长链/% Extended strand
PbDof12	XM_009361313.1	Scaffold75.0	Scaff8	327	36.06	9.00	7.95	1.53	74.62	15.90
PbDof13	XM_018648309.1	Scaffold83.0	Scaff9	523	55.98	5.94	13.77	2.29	76.67	7.27
PbDof14	XM_018648376.1	Scaffold84.0	Scaff10	302	33.89	5.11	25.50	1.66	66.89	5.96
PbDof15	XM_009364743.2	Scaffold106.0	Scaff11	327	36.23	6.69	12.54	2.75	66.36	18.35
PbDof16	XM_009371010.2	Scaffold180.0	Scaff12	465	50.96	5.79	12.04	1.94	79.78	6.24
PbDof17	XM_009377147.2	Scaffold275.0	Scaff13	286	31.52	7.71	5.59	1.05	80.42	12.94
PbDof18	XM_009377148.2	Scaffold275.0	Scaff13	270	29.76	8.44	6.30	3.33	77.78	12.59
PbDof19	XM_009378003.2	Scaffold292.0	Scaff14	511	55.22	5.72	10.37	1.37	80.82	7.44
PbDof20	XM_009336089.2	Scaffold384.0	Scaff15	289	31.75	7.68	8.30	1.73	80.28	9.69
PbDof21	XM_009340481.2	Scaffold499.0	Scaff16	366	38.92	9.13	9.29	3.83	77.60	9.29
PbDof22	XM_009344990.1	Scaffold698.0	Scaff17	316	35.40	6.43	12.03	2.53	74.68	10.76
PbDof23	XM_009344991.1	Scaffold698.0	Scaff17	316	34.97	6.52	13.61	2.85	72.78	10.76
PbDof24	XM_009345555.2	Scaffold727.0	Scaff18	372	40.05	8.78	8.06	2.42	79.84	9.68
PbDof25	XM_009345556.2	Scaffold727.0	Scaff18	337	36.26	8.96	6.53	2.97	81.31	9.20
PbDof26	XM_009346163.2	Scaffold765.0	Scaff19	225	24.05	8.18	6.67	3.56	79.11	10.67
PbDof27	XM_009346170.2	Scaffold765.0	Scaff19	225	24.05	8.18	6.67	3.56	79.11	10.67
PbDof28	XM_009348304.2	Scaffold915.0	Scaff20	304	33.60	6.62	18.42	1.64	64.80	15.13
PbDof29	XM_009348967.2	Scaffold973.0	Scaff21	347	37.14	8.90	20.46	2.59	66.28	10.66
PbDof30	XM_009349812.2	Scaffold1047.0	Scaff22	346	36.91	8.84	19.65	3.18	65.32	11.85
PbDof31	XM_009352168.2	Scaffold1355.0	Scaff23	303	33.81	4.94	20.79	1.65	69.64	7.92
PbDof32	XM_009336822.2	Scaffold1.0.1	Scaff24	291	32.49	7.70	15.81	2.06	74.23	7.90
PbDof33	XM_009343730.2	Scaffold1.0.1	Scaff24	378	41.13	8.58	15.08	0.53	76.19	8.20
PbDof34	XM_009351106.2	Scaffold1.0.1	Scaff24	163	18.34	9.08	14.11	6.13	63.19	16.56
PbDof35	XM_009354801.2	Scaffold1.0.1	Scaff24	291	31.31	9.30	16.84	5.50	63.23	14.43
PbDof36	XM_009339603.2	Scaffold14.0.1	Scaff25	323	34.90	7.20	15.17	2.79	69.04	13.00
PbDof37	XM_009340881.2	Scaffold14.0.1	Scaff25	463	50.48	5.48	13.17	1.51	76.89	8.42
PbDof38	XM_009345020.2	Scaffold16.0.1	Scaff26	362	39.20	8.99	9.12	4.14	73.76	12.98
PbDof39	XM_009347952.2	Scaffold30.0.1	Scaff27	325	34.81	7.21	8.31	1.85	76.00	13.85
PbDof40	XM_009351662.2	Scaffold17.0.2	Scaff28	282	31.49	6.77	10.64	3.90	74.11	11.35
PbDof41	XM_009353689.2	Scaffold20.0.1	Scaff29	320	35.00	7.23	10.94	2.50	75.94	10.62
PbDof42	XM_009359445.2	Scaffold58.0.1	Scaff30	357	38.71	8.93	10.36	4.76	71.43	13.45
PbDof43	XM_009362338.2	Scaffold80.0.1	Scaff31	539	58.63	6.66	13.17	1.67	78.11	7.05
PbDof44	XM_009362339.2	Scaffold80.0.1	Scaff31	521	56.57	6.22	13.05	2.30	77.93	6.72
PbDof45	XM_009363055.2	Scaffold87.0.1	Scaff32	316	34.47	8.16	9.18	4.43	74.05	12.34
PbDof46	XM_009365959.2	Scaffold116.0.1	Scaff33	428	45.70	9.03	16.12	2.57	72.43	8.88
PbDof47	XM_009366523.2	Scaffold122.0.1	Scaff34	384	40.83	8.75	15.10	2.08	73.96	8.85
PbDof48	XM_009367372.2	Scaffold131.0.1	Scaff35	253	26.84	6.15	14.62	7.51	55.34	22.53
PbDof49	XM_009369256.2	Scaffold155.0.1	Scaff36	221	23.59	8.13	10.86	2.71	76.02	10.41
PbDof50	XM_009376412.2	Scaffold262.0.1	Scaff37	376	40.32	9.64	13.83	3.99	74.20	7.98
PbDof51	XM_009378209.2	Scaffold294.0.1	Scaff38	247	25.39	8.18	9.31	5.67	67.61	17.41