Analysis of Differential Genes in Response to Sucrose Feeding in Disc Florets of Chrysanthemum nankingense

doi:10.16420/j.issn.0513-353x.2023-0321

Acta Horticulturae Sinica ›› 2024, Vol. 51 ›› Issue (6): 1297-1310.doi: 10.16420/j.issn.0513-353x.2023-0321

• Genetic & Breeding·Germplasm Resources·Molecular Biology • Previous Articles Next Articles

Analysis of Differential Genes in Response to Sucrose Feeding in Disc Florets of Chrysanthemum nankingense

WANG Wen, ZHANG Peng, WANG Zhimin, LIU Genzhong, BAO Zhilong, MA Fangfang^*()

State Key Laboratory of Crop Biology，College of Horticulture Science and Engineering，Shandong Agricultural University，Tai’an，Shandong 271018，China

Received:2023-02-24 Revised:2024-05-07 Online:2024-12-18 Published:2024-06-22
Contact: MA Fangfang

Abstract

Abstract:

To explore effects of sucrose feeding on the accumulation of total flavonoids in Chrysanthemum nankingense capitula and the underlying mechanism，Chrysanthemum nankingense，a unique wild diploid species of chrysanthemum in China，was used as the test material，and its detached capitula were fed with sucrose. The dynamic changes of total flavonoids and total phenols in disc and ray florets were determined at 0，12，36 and 60 h after sucrose feeding，and transcriptome sequencing of disc florets was performed. The results confirmed that sucrose feeding could increase the content of flavonoids and total phenols in disc florets within 24 h. However，in ray florets，these compounds changed slightly. A total of 9 135 differentially expressed genes（DEGs），which was also the maximum number of DEGs identified between treatments，were obtained by transcriptome analysis of disc florets sucrose-fed for 60 h and 0 h，while only 3 423 differentially expressed genes were obtained by transcriptome analysis of disc florets sucrose-fed for 60 h and no sucrose-fed for 60 h. These data indicate that the response degree of disc florets to sucrose increased with the increase of feeding time on the biochemistry level. KEGG analysis indicated that the flavonoid biosynthesis pathway was significantly enrichedand 32 flavonoid biosynthetic structural genes were differentially expressed. Correlation analysis showed that transcription factors MYBs，WRKYs and MADS-boxes had a strong co-expression relationship with flavonoid biosynthetic structural gene expression in disc florets in response to sucrose feeding. Nine DEGs were randomly selected for qRT-PCR，indicating that the transcriptome data were accurate. Spatio-temporal expression and subcellular localization of the encoding genes of two transcription factors were further analyzed to confirm the findings. Genes screened out from this study can offer potential targets for further efforts of metabolic engineering aiming to probe secondary metabolism in Chrysanthemum nankingense.

Key words: Chrysanthemum nankingense, response to sucrose, flavonoid metabolism, differentially expressed genes, disc florets

WANG Wen, ZHANG Peng, WANG Zhimin, LIU Genzhong, BAO Zhilong, MA Fangfang. Analysis of Differential Genes in Response to Sucrose Feeding in Disc Florets of Chrysanthemum nankingense[J]. Acta Horticulturae Sinica, 2024, 51(6): 1297-1310.

Figures/Tables 9

Table 1 Primers used in this experiment

引物用途 Primer usage	基因ID Gene ID	引物序列（5′-3′） Primer sequence
qRT-PCR	CHR00071024	F：CGTCGAGTTGATGGATTGCG
		R：AGAACAGTGTCACCCGTTGC
	CHR00071025	F：CGAGTTGATGGATTGCGACG
		R：AAAACTTACCCGTTGCCGGT
	CHR00072791	F：ACCAGAAAGATGGCCCAACA
		R：TCTGCCATCACCCATTCGAC
	CHR00057220	F：CTTGGCTCCGAGTGCTAAGA
		R：TCTCTTTGATCGCGAGTAGCA
	CHR00023313	F：ACGCAGGACCTTTTAGTCGG
		R：TCGAGCTCTTCTGTTGCCTT
	CHR00023554	F：CCAAAGAGTGGAGGGCAGTT
		R：GCCTCTTTCTCAAGGCCCAT
	CHR00068386	F：TGCCAGCATTTCGACACTGA
		R：GACGGGTTTCCACGTAGTCC
	CHR00001486	F：AGATTTGCCACCCCAGTCTT
		R：ATGGAGCAGTGTTCGGTTGT
	CHR00075330	F：AGTCTTCGACAGTTTCGGGC
		R：CAAACCGTTGTTGCCGGATT
	CnUbi	F：CTAATGAATGCTTACTGTGACCGAC
		R：TAGCTCATCGGGAGTCTGCT
亚细胞定位 Subcellular localization	CHR00001486	F：GCCAGTGCCAAGCTTCTATGGATTCAGCTGTTAAGAATGTGC
		R：CATGATTACGAATTCTAAGTTCATGGCATACCCTGTGTC
	CHR00075330	F：GCCAGTGCCAAGCTTCTATGGACGATGATGAAAAGCC
		R：CATGATTACGAATTCGTTGCGCATTATTGAAGGCAC

Fig. 1 Morphological changes of isolated flower buds of Chrysanthemum nankingense after sucrose feeding The red arrow points to the stamen.

Fig. 2 Contents of total flavonoids and total phenols in disc florets and ray florets of Chrysanthemum nankingense after sucrose feeding

Fig. 3 The principal component analysis（PCA）（A）of the expression genes and the comparison（B，C）of the number of differentially expressed genes in different time points of disc florets feeding with or without sucrose Su：Sucrose feeding；NS：No sucrose. The same below.

Fig. 4 Differential gene expression analysis（A，C）and KEGG enrichment analysis（B，D）of disc florets after feeding with sucrose for 60 h and feeding without sucrose for 60 h/feeding with sucrose for 0 h R1-R3 indicate three biological replicates for each time-point sample in RNAseq analysis.

Fig. 5 Effects of sucrose feeding on the expression of genes（A）related to flavonoid biosynthesis pathway and key transcription factors（B）in disc florets

Fig. 6 Correlation analysis between flavonoid biosynthesis related genes and important transcription factors

Fig. 7 Expression analysis of key differential genes after 60 h of sucrose feeding via qRT-PCR * P < 0.05.

Fig. 8 The expression of CHR00001486 and CHR00075330 in different tissues of Chrysanthemum nankingense（A）and subcellular localization of their proteins（B） Different lowercase letters indicate significant differences of genes expression in different tissues（P < 0.05）.

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Analysis of Differential Genes in Response to Sucrose Feeding in Disc Florets of Chrysanthemum nankingense

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