Excavation of key genes for yellow flower formation in Meconopsis integrifolia

doi:10.3969/j.issn.1004-1524.20230535

Acta Agriculturae Zhejiangensis ›› 2024, Vol. 36 ›› Issue (4): 811-824.DOI: 10.3969/j.issn.1004-1524.20230535

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Excavation of key genes for yellow flower formation in Meconopsis integrifolia

CHEN Xiaojuan¹(), LUO Jun¹, WANG Fumin¹, LI Tuojian¹, QU Yan¹^,²^,³^,^*()

1. College of Landscape Architecture and Horticulture, Southwest Forestry University, Kunming 650224, China
2. Southwest Engineering and Technology Research Center of Landscape Architecture (National Forestry and Grassland Administration), Kunming 650224, China
3. Yunnan Engineering Research Center for Functional Flower Resources and Industrialization, Kunming 650224, China

Received:2023-04-26 Online:2024-04-25 Published:2024-04-29
Contact: QU Yan

Abstract

Abstract:

To find out the key genes of yellow flower formation in Meconopsis integrifolia and to study the regulatory mechanism of flower color formation at the transcriptional level, in this study, the petal tissues of M. integrifolia were selected for RNA sequencing at three flower developmental stages, and the absolute quantification of flavonoids in petal tissue were measured at blooming period. The results showed that: (1) A total of 38 flavonoids were detected in LC-MS/MS, among which quercetin and its derivatives were the main ones, and their content exceeded 80% of the total content of flavonoids. (2) A total of 171 902 Unigenes were obtained from RNA sequencing, and 14 906 DEGs were obtained after screening, which were significantly enriched in 48 KEGG pathways (P<0.05), among which the phenylpropanoid biosynthesis and flavonoid biosynthesis were significantly enriched in the three flower development stages (P<0.05). (3) At the same time, it was also found that genes related to flavonoid synthesis were expressed differently in the three flower development periods, and the expression of MiFLSs (Cluster-28469.86, Cluster-28469.89, Cluster-28469.91) was 1.2-4.7 times higher than that of MiDFR(Cluster-49617.1) in the same period. The expression of MiFG3 (Cluster-15071.0) related to the synthesis of quercetin derivatives showed a gradual up-regulation trend. The expression of transcription factor MiMYB4 also increased gradually, and it showed a strong negative correlation with MiMYB38, MiMYB39, MiDFR(Cluster-49617.1) and MiANS(Cluster-29740.1). (4) The qRT-PCR results of the selected genes correlated well with RNA-seq (R²=0.820 4, P<0.05), indicating that the transcriptome data was reliable. In summary, the main component of the yellow petals of M. integrifolia may be quercetin and its derivatives. The stronger competitiveness of MiFLSs compare with MiDFR drives more substrates toward flavonol synthesis, providing precursors for the bulk synthesis of flavonols, and MiFG3 consumption of precursors promotes the bulk synthesis of quercetin derivatives. Meanwhile, the transcription factor MiMYB4 may indirectly promote substrate flow toward flavonol synthesis by repressing structural genes and transcription factors associated with anthocyanin synthesis, which together with transcription factors regulate the process of flower color formation in M. integrifolia. In this study, 6 key genes regulating the flower color formation of M. integrifolia are screened, and the molecular mechanism of its yellow flower formation is preliminarily explored, which provides a theoretical reference for further research on the flower color variation of Meconopsis.

Key words: Meconopsis integrifolia, flower color, flavonol, key gene

CLC Number:

S681.9

CHEN Xiaojuan, LUO Jun, WANG Fumin, LI Tuojian, QU Yan. Excavation of key genes for yellow flower formation in Meconopsis integrifolia[J]. Acta Agriculturae Zhejiangensis, 2024, 36(4): 811-824.

Figures/Tables 14

References 29

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实验材料 Experimental materials	开花时期 Flowering period	编号 Number	皇家比色卡色号 Royal color card number	采集地 Collection location	经纬度 Latitude and Longitude
全缘叶绿绒蒿	花蕾期Budding period	P1	YELLOW-GREEN 145D	云南省丽江市玉龙雪山	27°0'10″N,
M. integrifolia	初开期Initial opening period	P2	YELLOW-GREEN 150D	Jade Dragon Snow Mountain,	100°10'56″E
	盛花期Blooming period	P3	YELLOW-GREEN 150B	Lijiang City, Yunnan Province

实验材料 Experimental materials	开花时期 Flowering period	编号 Number	皇家比色卡色号 Royal color card number	采集地 Collection location	经纬度 Latitude and Longitude
全缘叶绿绒蒿	花蕾期Budding period	P1	YELLOW-GREEN 145D	云南省丽江市玉龙雪山	27°0'10″N,
M. integrifolia	初开期Initial opening period	P2	YELLOW-GREEN 150D	Jade Dragon Snow Mountain,	100°10'56″E
	盛花期Blooming period	P3	YELLOW-GREEN 150B	Lijiang City, Yunnan Province

样本 Sample	原始序列 Raw reads	过滤后序列 Clean reads	过滤后的碱基 Clean bases/GB	错误率 Error rate/%	Q20/%	Q30/%	GC/%
QYY-P1-1	46 105 634	45 312 642	6.80	0.03	97.27	92.42	41.41
QYY-P1-2	46 446 078	45 552 808	6.83	0.03	97.53	93.02	41.65
QYY-P1-3	46 078 118	45 182 696	6.78	0.03	97.26	92.44	41.62
QYY-P2-1	44 529 162	43 227 950	6.48	0.03	97.30	92.56	41.13
QYY-P2-2	47 350 222	46 244 742	6.94	0.03	97.30	92.42	41.16
QYY-P2-3	48 324 836	47 314 210	7.10	0.03	97.72	93.41	40.90
QYY-P3-1	48 823 172	47 635 926	7.15	0.03	97.34	92.59	41.23
QYY-P3-2	46 197 126	44 753 484	6.71	0.03	97.21	92.39	41.18
QYY-P3-3	48 061 136	46 960 540	7.04	0.03	97.51	92.97	41.03

样本 Sample	原始序列 Raw reads	过滤后序列 Clean reads	过滤后的碱基 Clean bases/GB	错误率 Error rate/%	Q20/%	Q30/%	GC/%
QYY-P1-1	46 105 634	45 312 642	6.80	0.03	97.27	92.42	41.41
QYY-P1-2	46 446 078	45 552 808	6.83	0.03	97.53	93.02	41.65
QYY-P1-3	46 078 118	45 182 696	6.78	0.03	97.26	92.44	41.62
QYY-P2-1	44 529 162	43 227 950	6.48	0.03	97.30	92.56	41.13
QYY-P2-2	47 350 222	46 244 742	6.94	0.03	97.30	92.42	41.16
QYY-P2-3	48 324 836	47 314 210	7.10	0.03	97.72	93.41	40.90
QYY-P3-1	48 823 172	47 635 926	7.15	0.03	97.34	92.59	41.23
QYY-P3-2	46 197 126	44 753 484	6.71	0.03	97.21	92.39	41.18
QYY-P3-3	48 061 136	46 960 540	7.04	0.03	97.51	92.97	41.03

数据库 Database	注释到的基因数量 Number of genes	占比 Percentage/%
KEGG	76 662	44.60
Nr	97 732	56.85
SwissProt	70 402	40.95
TrEMBL	103 886	60.43
KOG	61 267	35.64
GO	87 982	51.18
Pfam	51 596	30.01
Annotated in at least	106 907	62.19
one Database
Total Unigenes	171 902	100

Excavation of key genes for yellow flower formation in Meconopsis integrifolia

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Figures/Tables 14

References 29

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基因 Gene name	基因ID Gene ID	蛋白长度 Protein length/aa	等电点 Isoelectric point	分子量 Molecular weight/u	所属拟南芥亚族 The subgroup of Arabidopsis to which the gene belongs
MiMYB1	Cluster-13873.16	1 185	9.35	131 836.98	—
MiMYB2	Cluster-73976.7	570	6.33	63 623.76	—
MiMYB3	Cluster-74122.1	501	5.89	55 023.13	—
MiMYB4	Cluster-46647.219	271	8.59	30 734.68	S4
MiMYB5	Cluster-176.1	257	9.20	29 366.95	—
MiMYB6	Cluster-24930.0	352	7.76	39 557.77	S9
MiMYB7	Cluster-26024.6	371	9.37	42 556.11	—
MiMYB8	Cluster-26024.7	354	9.32	40 353.59	—
MiMYB9	Cluster-26960.10	357	6.01	39 008.3	—
MiMYB10	Cluster-28521.4	297	5.58	33 656.54	S11
MiMYB11	Cluster-29061.0	355	5.70	40 225.53	S11
MiMYB12	Cluster-41024.1	255	9.00	28 447.39	—
MiMYB13	Cluster-42663.2	429	9.35	47 588.58	—
MiMYB14	Cluster-43725.2	130	6.17	14 861.21	S19
MiMYB15	Cluster-43725.4	209	6.59	23 973.62	S19
MiMYB16	Cluster-74557.6	791	5.47	86 693.98	—
MiMYB17	Cluster-48749.21	294	8.85	32 860.93	—
MiMYB18	Cluster-54127.0	165	4.38	18 843.61	—
MiMYB19	Cluster-54127.1	250	4.90	28 578.83	—
MiMYB20	Cluster-54127.5	165	4.38	18 843.61	—
MiMYB21	Cluster-56396.9	514	6.76	52 138.66	S13
MiMYB22	Cluster-56419.0	414	4.87	45 932.63	—
MiMYB23	Cluster-58089.0	220	9.97	24 934.41	—
MiMYB24	Cluster-59054.5	629	6.47	71 492.12	—
MiMYB25	Cluster-59054.7	583	5.65	66 005.51	—
MiMYB26	Cluster-60970.0	420	8.72	41 583.2	S22
MiMYB27	Cluster-61134.0	209	8.36	23 333.70	—
MiMYB28	Cluster-73421.2	310	5.28	34 749.97	—
MiMYB29	Cluster-61134.9	137	5.61	14 979.42	—
MiMYB30	Cluster-61178.1	266	5.13	30 023.11	S16
MiMYB31	Cluster-64347.0	244	8.38	27 986.6	S18
MiMYB32	Cluster-6447.11	115	5.88	13 429.9	S20
MiMYB33	Cluster-6447.2	349	5.00	40 058.43	S20
MiMYB34	Cluster-6447.5	115	5.88	13 429.90	S20
MiMYB35	Cluster-66523.6	508	9.27	56 339.69	—
MiMYB36	Cluster-67045.0	479	9.15	53 421.65	—
MiMYB37	Cluster-67577.0	265	7.22	30 531.08	—
MiMYB38	Cluster-67577.1	372	9.30	42 471.57	S6
MiMYB39	Cluster-67577.2	243	9.72	28 100.98	S6
MiMYB40	Cluster-69149.2	144	4.67	16 543.61	—
MiMYB41	Cluster-69645.1	278	9.30	31 487.15	—
MiMYB42	Cluster-69645.5	277	9.37	31 357.03	—
MiMYB43	Cluster-70083.7	208	9.27	22 811.18	—
MiMYB44	Cluster-69949.7	302	7.20	32 368.05	—
MiMYB45	Cluster-70083.0	208	9.42	22 879.34	—