烟草细胞质雄性不育系K326 MADS-box和SUPERMAN基因的特征

doi:10.3724/SP.J.1006.2023.24268

Abstract

Abstract:

Cytoplasmic male sterility (CMS) is an important tool for hybrid production and a good system for studying the interaction between nucleus and cytoplasm, but its mechanism is still unclear. The relationship between stamen abnormality and the relative expression levels of flower development genes in tobacco CMS line K326 was studied. The CMS line K326 was derived from a natural variation in Nicotiana tabacum and was bred by continuous backcross of flue-cured tobacco variety K326. It has not only stamens with carpelloid, but also stamens with petaloid, and the base of abnormal stamens is fused into a whole. In this study, we identified MADS-box genes and SUPERMAN genes of whole genome, which controlled the flower development in N. tabacum and analyzed their chromosomal localization and collinearity. The cis-acting elements of B genes and SUPERMAN genes were predicted, and their expression characteristics in flower buds at different stages in both CMS line and its maintainer line were studied. The results showed that 160 MADS-box genes and 5 SUPERMAN genes were identified in the genome of tobacco, including 4 PI genes, 3 AP3 genes, and 7 B genes. There were 79 MADS-box genes scattered on 22 chromosomes, and 3 SUPERMAN genes distributed on 3 chromosomes. Collinearity analysis showed that tandem and fragment DNA duplication and multiplication were the driving forces of the expansion of MADS- box gene family in tobacco. The abnormal stamens of CMS line K326 appeared at small bud stage, suggesting that the abnormal stamens in CMS line K326 were the result of early meristem development defects. The qRT-PCR showed that 7 B genes and 1 SUPERMAN gene were expressed both in CMS line K326 and its maintainers. The relative expression level of PI gene NitMADS115 in the maintainer line was higher than that in CMS line at all stages. The relative expression levels of AP3 genes NitMADS72 and NitMADS100 in maintainer lines were lower than those in CMS lines. Other genes were down-regulated at small bud stage and large bud stage, and up-regulated at middle bud stage. The SUPERMAN gene could only be detected at small bud and middle bud stage of the maintainer line, but not at bud stages of CMS line. Analysis of cis-acting elements showed that NitMADS115 had an auxin response element AuxRR. The study indicates that auxin might play an important role in cytoplasmic retrograde regulation of nucleus in tobacco.

Key words: tobacco, cytoplasmic male sterility, MADS-box, SUPERMAN, auxin

CUI Fang-Fang, MENG Lin-Feng, LIU Miao-Miao, ZHANG Jian-Qiang, WANG Jian-Ge, LIU Qi-Yuan. Characteristics of MADS-box and SUPERMAN genes in tobacco cytoplasmic male sterile line K326[J].Acta Agronomica Sinica, 2023, 49(12): 3204-3214.

Figures/Tables 11

Table 1

Fig. 1

Table S1

The MADS-box and SUPERMAN genes of Nicotiana tabacum"

基因 Gene name	转录本 Transcript	蛋白长度 Protein length (aa)	TAIR10_描述 TAIR10_description
NitMADS1	Nitab4.5_0000001g0140.1	391	FEM111, AGL80, AGAMOUS-like 80
NitMADS2	Nitab4.5_0000009g0060.1	444	FEM111, AGL80, AGAMOUS-like 80
NitMADS3	Nitab4.5_0000014g0510.1	231	SEP1, AGL2, K-box region and MADS-box family protein
NitMADS4	Nitab4.5_0000016g0430.1	217	STK, AGL11, K-box region and MADS-box family protein
NitMADS5	Nitab4.5_0000027g0070.1	209	SEP2, AGL4, K-box region and MADS-box family protein
NitMADS6	Nitab4.5_0000036g0460.1	160	AGL21
NitMADS7	Nitab4.5_0000047g0250.1	219	AGL62
NitMADS8	Nitab4.5_0000093g0040.1	177	AGL29
NitMADS9	Nitab4.5_0000131g0290.1	133	SVP, AGL22, K-box region and MADS-box family protein
NitMADS10	Nitab4.5_0000140g0130.1	210	AGL20, SOC1, ATSOC1, AGAMOUS-like 20
NitMADS11	Nitab4.5_0000146g0060.1	197	AGL62
NitMADS12	Nitab4.5_0000154g0260.1	186	SEP2, AGL4, K-box region and MADS-box family protein
NitMADS13	Nitab4.5_0000163g0010.1	239	MADS-box protein AGL71-like
NitMADS14	Nitab4.5_0000167g0220.1	270	AGL2
NitMADS15	Nitab4.5_0000214g0110.1	260	AGL8 homolog
NitMADS16	Nitab4.5_0000290g0080.1	62	AGL42
NitMADS17	Nitab4.5_0000323g0030.1	268	SEP3, AGL9, K-box region and MADS-box family protein
NitMADS18	Nitab4.5_0000354g0070.1	85	ANR1, AGL44, AGAMOUS-like 44
NitMADS19	Nitab4.5_0000365g0070.1	271	AGL2
NitMADS20	Nitab4.5_0000367g0080.1	169	AGL1
NitMADS21	Nitab4.5_0000396g0070.1	165	AGL29
NitMADS22	Nitab4.5_0000418g0050.1	212	PI
NitMADS23	Nitab4.5_0000508g0030.1	209	PI
NitMADS24	Nitab4.5_0000514g0010.1	240	AGL21
NitMADS25	Nitab4.5_0000518g0100.1	175	AGL8 homolog
NitMADS26	Nitab4.5_0000579g0010.1	249	SEP3, AGL9, K-box region and MADS-box family protein
NitMADS27	Nitab4.5_0000592g0010.1	434	vegetative cell wall protein gp1-like
NitMADS28	Nitab4.5_0000599g0020.1	89	AGL20, SOC1, ATSOC1 AGAMOUS-like 20
NitMADS29	Nitab4.5_0000622g0180.1	177	AGL29
NitMADS30	Nitab4.5_0000654g0110.1	193	AGL62
NitMADS31	Nitab4.5_0000715g0210.1	442	AGL104
NitMADS32	Nitab4.5_0000738g0160.1	221	AGL80
NitMADS33	Nitab4.5_0000748g0100.1	190	AGL12, XAL1, AGAMOUS-like 12
NitMADS34	Nitab4.5_0000772g0010.1	201	AGL62
NitMADS35	Nitab4.5_0000787g0180.1	370	AGL80
NitMADS36	Nitab4.5_0000832g0060.1	97	AGL20, SOC1, ATSOC1, AGAMOUS-like 20
NitMADS37	Nitab4.5_0000885g0080.1	71	ANR1, AGL44, AGAMOUS-like 44
NitMADS38	Nitab4.5_0000902g0340.1	186	SVP, AGL22 K-box region and MADS-box family protein
NitMADS39	Nitab4.5_0000915g0190.1	159	AGL1
NitMADS40	Nitab4.5_0000927g0060.1	90	AGL12, XAL1, AGAMOUS-like 12
NitMADS41	Nitab4.5_0000934g0190.1	327	AGL66
NitMADS42	Nitab4.5_0000935g0220.1	66	AGL21
NitMADS43	Nitab4.5_0001026g0080.1	189	SVP, AGL22, K-box region and MADS-box family protein
NitMADS44	Nitab4.5_0001030g0180.1	191	AGL62
NitMADS45	Nitab4.5_0001099g0010.1	185	AGL62
NitMADS46	Nitab4.5_0001113g0010.1	101	MAF4
NitMADS47	Nitab4.5_0001121g0240.1	260	AG
NitMADS48	Nitab4.5_0001127g0140.1	250	AGL6
NitMADS49	Nitab4.5_0001209g0020.1	532	vegetative cell wall protein gp1-like
NitMADS50	Nitab4.5_0001242g0040.1	78	AGL15
NitMADS51	Nitab4.5_0001244g0010.1	333	AGL15
NitMADS52	Nitab4.5_0001262g0020.1	164	AGL1
NitMADS53	Nitab4.5_0001339g0170.1	245	SEP2, AGL4, K-box region and MADS-box family protein
NitMADS54	Nitab4.5_0001376g0040.1	68	AGL20, SOC1 ATSOC1,AGAMOUS-like 20
NitMADS55	Nitab4.5_0001434g0020.1	227	SEP2, AGL4, K-box region and MADS-box family protein
NitMADS56	Nitab4.5_0001436g0060.1	277	MADS-box transcription factor 23-like
NitMADS57	Nitab4.5_0001463g0160.1	361	FEM111,AGL80, AGAMOUS-like 80
NitMADS58	Nitab4.5_0001468g0090.1	190	SVP, AGL22, K-box region and MADS-box family protein
NitMADS59	Nitab4.5_0001477g0020.1	222	AGL62
NitMADS60	Nitab4.5_0001541g0050.1	258	AGL8, FUL, AGAMOUS-like 8
NitMADS61	Nitab4.5_0001714g0110.1	119	AGL14
NitMADS62	Nitab4.5_0001797g0040.1	210	SEP4, AGL3, K-box region and MADS-box family protein
NitMADS63	Nitab4.5_0001932g0030.1	200	AGL62
NitMADS64	Nitab4.5_0001997g0120.1	75	AGL17
NitMADS65	Nitab4.5_0002011g0030.1	242	AGL62
NitMADS66	Nitab4.5_0002041g0030.1	245	SEP4, AGL3, K-box region and MADS-box family protein
NitMADS67	Nitab4.5_0002060g0090.1	67	AGL42
NitMADS68	Nitab4.5_0002207g0020.1	172	AGL29
NitMADS69	Nitab4.5_0002210g0020.1	179	AGL61
NitMADS70	Nitab4.5_0002228g0010.1	140	AGL20, SOC1, ATSOC1, AGAMOUS-like 20
NitMADS71	Nitab4.5_0002249g0010.1	185	AP1, AGL7, K-box region and MADS-box family protein
NitMADS72	Nitab4.5_0002266g0050.1	118	AP3, ATAP3, K-box region and MADS-box family protein
NitMADS73	Nitab4.5_0002290g0100.1	146	ANR1, AGL44, AGAMOUS-like 44
NitMADS74	Nitab4.5_0002324g0050.1	243	AGL62
NitMADS75	Nitab4.5_0002449g0010.1	256	AGL8, FUL, AGAMOUS-like 8
NitMADS76	Nitab4.5_0002467g0090.1	160	AP1, AGL7, K-box region and MADS-box family protein
NitMADS77	Nitab4.5_0002502g0010.1	185	AGL2
NitMADS78	Nitab4.5_0002516g0060.1	120	FEM111, AGL80, AGAMOUS-like 80
NitMADS79	Nitab4.5_0002583g0020.1	185	AGL61, DIA, AGAMOUS-like 61
NitMADS80	Nitab4.5_0002779g0020.1	185	AGL 2
NitMADS81	Nitab4.5_0002840g0130.1	173	MADS-box transcription factor 23-like
NitMADS82	Nitab4.5_0002860g0020.1	190	AGL61, DIA, AGAMOUS-like 61
NitMADS83	Nitab4.5_0002947g0010.1	185	AGL2
NitMADS84	Nitab4.5_0002949g0060.1	182	AGL104
NitMADS85	Nitab4.5_0003151g0030.1	228	AGL 2
NitMADS86	Nitab4.5_0003182g0070.1	214	AGL20, SOC1, ATSOC1, AGAMOUS-like 20
NitMADS87	Nitab4.5_0003184g0040.1	86	AGL16
NitMADS88	Nitab4.5_0003248g0010.1	237	MAF4
NitMADS89	Nitab4.5_0003336g0080.1	231	AGL21
NitMADS90	Nitab4.5_0003356g0030.1	78	AGL20, SOC1, ATSOC1, AGAMOUS-like 20
NitMADS91	Nitab4.5_0003390g0060.1	116	AGL29
NitMADS92	Nitab4.5_0003480g0010.1	177	AGL29
NitMADS93	Nitab4.5_0003504g0010.1	609	AGL30
NitMADS94	Nitab4.5_0003507g0010.1	232	AG
NitMADS95	Nitab4.5_0003563g0030.1	242	AGL62
NitMADS96	Nitab4.5_0003707g0020.1	80	STK, AGL11, K-box region and MADS-box family protein
NitMADS97	Nitab4.5_0003863g0050.1	242	TT16, ABS, AGL32, K-box region and MADS-box family protein
NitMADS98	Nitab4.5_0004004g0010.1	219	AGL62
NitMADS99	Nitab4.5_0004036g0060.1	183	AGL104
NitMADS100	Nitab4.5_0004086g0040.1	237	AP3, ATAP3, K-box region and MADS-box family protein
NitMADS101	Nitab4.5_0004114g0070.1	175	MADS-box protein AGL24-like
NitMADS102	Nitab4.5_0004153g0060.1	187	SVP, AGL22, K-box region and MADS-box family protein
NitMADS103	Nitab4.5_0004222g0040.1	345	AGL15
NitMADS104	Nitab4.5_0004263g0020.1	218	STK, AGL11, K-box region and MADS-box family protein
NitMADS105	Nitab4.5_0004346g0030.1	209	PI
NitMADS106	Nitab4.5_0004520g0010.1	133	SEP4, AGL3, K-box region and MADS-box family protein
NitMADS107	Nitab4.5_0004622g0050.1	225	AGL6
NitMADS108	Nitab4.5_0004673g0050.1	69	AGL16
NitMADS109	Nitab4.5_0004674g0030.1	196	AGL8, FUL, AGAMOUS-like 8
NitMADS110	Nitab4.5_0004907g0010.1	179	AGL1
NitMADS111	Nitab4.5_0004911g0040.1	121	AGL20, SOC1, ATSOC1, AGAMOUS-like 20
NitMADS112	Nitab4.5_0005046g0020.1	396	AGL62
NitMADS113	Nitab4.5_0005138g0050.1	248	AG
NitMADS114	Nitab4.5_0005163g0010.1	85	AGL21
NitMADS115	Nitab4.5_0005166g0020.1	212	PI
NitMADS116	Nitab4.5_0005183g0060.1	164	MADS-box transcription factor 23-like
NitMADS117	Nitab4.5_0005362g0010.1	123	AGL28
NitMADS118	Nitab4.5_0005468g0020.1	65	AGL15
NitMADS119	Nitab4.5_0005565g0020.1	154	MADS-box protein JOINTLESS-like isoform X4
NitMADS120	Nitab4.5_0005597g0030.1	495	AGL104
NitMADS121	Nitab4.5_0005688g0010.1	173	MADS-box transcription factor 23-like
NitMADS122	Nitab4.5_0005766g0030.1	181	AGL65
NitMADS123	Nitab4.5_0005772g0040.1	180	STK, AGL11, K-box region and MADS-box family protein
NitMADS124	Nitab4.5_0005798g0020.1	183	AGL2
NitMADS125	Nitab4.5_0006437g0030.1	96	SEP3, K-box region and MADS-box family protein
NitMADS126	Nitab4.5_0006477g0040.1	229	STK, AGL11, K-box region and MADS-box family protein
NitMADS127	Nitab4.5_0006520g0020.1	79	MADS-box transcription factor 16-like
NitMADS128	Nitab4.5_0006522g0010.1	156	AGL20, SOC1, ATSOC1, AGAMOUS-like 20
NitMADS129	Nitab4.5_0007204g0030.1	248	AG
NitMADS130	Nitab4.5_0007275g0010.1	325	AGL104
NitMADS131	Nitab4.5_0007355g0010.1	267	AGL 2
NitMADS132	Nitab4.5_0007505g0060.1	68	AGL8, FUL, AGAMOUS-like 8
NitMADS133	Nitab4.5_0007530g0020.1	208	AGL8, FUL, AGAMOUS-like 8
NitMADS134	Nitab4.5_0007721g0040.1	70	AGL16
NitMADS135	Nitab4.5_0008133g0010.1	167	AGL19, GL19, AGAMOUS-like 19
NitMADS136	Nitab4.5_0009469g0040.1	69	AGL24
NitMADS137	Nitab4.5_0009635g0020.1	177	AGL2
NitMADS138	Nitab4.5_0009637g0010.1	157	AGL80
NitMADS139	Nitab4.5_0010043g0030.1	83	ANR1, AGL44, AGAMOUS-like 44
NitMADS140	Nitab4.5_0010230g0020.1	169	AGL61, DIA, AGAMOUS-like 61
NitMADS141	Nitab4.5_0010657g0020.1	185	AGL2
NitMADS142	Nitab4.5_0010693g0010.1	180	SHP1, AGL1, K-box region and MADS-box family protein
NitMADS143	Nitab4.5_0010703g0030.1	162	AGL1
NitMADS144	Nitab4.5_0011208g0020.1	210	AGL62
NitMADS145	Nitab4.5_0011644g0010.1	623	AGL30
NitMADS146	Nitab4.5_0011705g0040.1	208	AGL62
NitMADS147	Nitab4.5_0012101g0010.1	196	SEP1, AGL2, K-box region and MADS-box transcription factor family protein
NitMADS148	Nitab4.5_0012471g0010.1	185	AGL1
NitMADS149	Nitab4.5_0012476g0010.1	185	AGL2
NitMADS150	Nitab4.5_0012816g0020.1	210	MAF4, FCL4, AGL69, K-box region and MADS-box family protein
NitMADS151	Nitab4.5_0013517g0010.1	118	AGL91
NitMADS152	Nitab4.5_0014584g0010.1	270	AP3, ATAP3, K-box region and MADS-box family protein
NitMADS153	Nitab4.5_0014719g0010.1	74	AGL16
NitMADS154	Nitab4.5_0015237g0010.1	76	SEPALLATA2
NitMADS155	Nitab4.5_0021852g0010.1	113	AGL20, SOC1, ATSOC1, AGAMOUS-like 20
NitMADS156	Nitab4.5_0023347g0010.1	151	AGL2
NitMADS157	Nitab4.5_0024332g0010.1	208	AGL62
NitMADS158	Nitab4.5_0024971g0010.1	91	AGL61, DIA, AGAMOUS-like 61
NitMADS159	Nitab4.5_0025184g0010.1	407	AGL62
NitMADS160	Nitab4.5_0027705g0010.1	201	AGL62
NitSUP1	Nitab4.5_0003721g0010.1	183	SUPERMAN
NitSUP2	Nitab4.5_0000086g0190.1	174	SUPERMAN
NitSUP3	Nitab4.5_0000342g0280.1	234	SUPERMAN
NitSUP4	Nitab4.5_0009537g0030.1	227	SUPERMAN
NitSUP5	Nitab4.5_0011666g0020.1	175	SUPERMAN

Table S1

Table 2

Fig. 2

Fig. 3

Fig. 4

Fig. 5

Table 3

Table 4

Fig. 6

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基因 Gene name	正向引物 Forward primer (5°-3°)	反向引物 Reverse primer (5°-3°)
NitMADS100	TTTGCCAATGGAGTACACA	TTAACCCCATTACCAAATTGC
NitMADS72	CAGGCAAGTCACTTACTCCA	CAAGCGTCCTCTGATACTGA
NitMADS23	AAATTGAACTCAAGCACCT	TTACGGATACTAGTTAGTCCA
NitMADS105	ATGGACTCACTAGTATCCGTA	TGGTGAAACACTTCCCCTA
NitMADS152	GCATATTAGCCTTACGCCTT	TTCACTTTGCGTAGATTGACT
NitMADS22	TGCATGAATATTGTAGCCCTT	GCTTAACCTGCATACTGTCA
NitMADS115	TGCATGAATATTGTAGCCCTT	GCTTAACCTGCATACTGTCA
NitSUP1	TCGAATAACAACGACGACAAC	GGCACATCTGAATTCCCTT
NitSUP2	AGCAGCCCATTTAAGTTACCG	GCTTAAACTCACACGCCAA
NitSUP3	CCTAGTTTTGTTTCATCCCC	CAAATCTCCATTTCTTAGCTGA
NitSUP4	CCATTAAATCACTTGAGCC	CTAAGCAATCCAATTCCCAA
NitSUP5	GGCAGCTCATTTAAGTTTCCG	CACGCCAATCATCTCCGAA
Actin	CCGCTTTTGCTATTATGTCC	GCACCTCACAAATATGCTG

基因 Gene	脱落酸响应元件 ABRE	茉莉酸甲酯响应元件 CGTCA-motif	乙烯响应元件 ERE	赤霉素响应元件 TATC-box	水杨酸响应元件 TCA-element	生长素响应元件 TGA-element	茉莉酸甲酯响应元件 TGACG-motif
NitSUP1	1	3	0	1	0	0	2
NitSUP2	1	0	0	0	1	3	0
NitSUP3	2	2	0	1	6	0	2
NitSUP4	2	3	3	0	3	1	3
NitSUP5	2	1	0	0	1	0	1

Characteristics of MADS-box and SUPERMAN genes in tobacco cytoplasmic male sterile line K326

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