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Acta Agronomica Sinica ›› 2023, Vol. 49 ›› Issue (7): 1860-1870.doi: 10.3724/SP.J.1006.2023.24259

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Transcriptome analysis reveals the regulatory role of the transcription factor StMYB113 in light-induced chlorophyll synthesis of potato tuber epidermis

ZHAO Xi-Juan1,2(), LIU Sheng-Xuan2, LIU Teng-Fei2, ZHENG Jie2, DU Juan2, HU Xin-Xi1, SONG Bo-Tao2,*(), HE Chang-Zheng1,*()   

  1. 1Key Laboratory for Vegetable Biology of Hunan Province / ERC for Germplasm Innovation and New Variety Breeding of Horticultural Crops / Hunan Agricultural University, Changsha 410128, Hunan, China
    2Key Laboratory of Potato Biology and Biotechnology, Ministry of Agriculture and Rural Affairs / Key Laboratory of Horticultural Plant Biology, Ministry of Education / Huazhong Agricultural University, Wuhan 430070, Hubei, China
  • Received:2022-11-21 Accepted:2023-02-24 Online:2023-07-12 Published:2023-03-02
  • Contact: *E-mail: hecz@hotmail.com; E-mail: songbotao@mail.hzau.edu.cn E-mail:704986451@qq.com;songbotao@mail.hzau.edu.cn;hecz@hotmail.com
  • Supported by:
    The Third Batch of Modern Agricultural Industrial Technology System Project in Hubei Province(鄂采计[2020]-09533号);The China Agriculture Research System of MOF and MARA(CARS-09-P07)

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Abstract:

Light-induced greening of tubers seriously affects its safety and economic benefits, but the mechanism of light-induced chlorophyll synthesis in potato tubers remains unclear. In this study, the related metabolites of potato tubers with different light durations were analyzed. The results were as follows: When the light duration prolonged, the chlorophyll content of the tubers gradually increased. Moreover, the chlorophyll content increased significantly at 36 hours, with which the tuber skin turned green obviously. Transcriptome sequencing and bioinformatics analysis were carried out on the samples taken at 0 hour, 6 hours, and 36 hours, and 5646 differentially expressed genes (DEGs) were identified. According to the co-expression cluster analysis and quantitative RT-PCR verification results, 9 major structure genes of chlorophyll biosynthesis pathway (StGAS1, StCHLD, StCrd1, StHEMA, StGUN4, StPORA, StUROD, StCHLM, and StCHLG) and 6 transcription factors (StSBP, StLSD, StGATA, StWRKY, StMYB-like, and StMYB113) were significant induced. Predictions of the cis-acting elements of the promoter sequences of these 9 structural genes indicated that they all contained multiple MYB binding sites. Promoter element analysis and transcriptional activation validation further revealed that StMYB113 had a light-responsive element and could activate the expression of StUROD in tobacco. Therefore, StMYB113 may be light-responsive and be able to regulate potato tuber greening in the light. This study provides a reference for the research on the regulatory mechanism of light-induced chlorophyll synthesis in potato tubers, which is important for reducing the loss caused by greening of potato tubers.

Key words: potato, light-induced greening, chlorophyll synthesis, StMYB113, transcriptional activation

Table 1

Primer sequences for qRT-PCR used in this study"

基因名称 正向引物 反向引物 基因ID
Gene name Forward primer (5°-3°) Reverse primer (5°-3°) Gene ID
GSA1 GTGGTGGGTTCATAAGGGGG TGGAGTGTGTGCTAAGCTGG Soltu.DM.04G005660.2
CHLD GACGCTAGTGGGAGTATGGC CCCCACCACAGGGAAGTTTT Soltu.DM.04G010510.1
Crd1 TGGTGCCAAGATGAGAACCG TGTGACATAAACCGAAAGGCA Soltu.DM.10G030260.1
HEMA GAGTGATTCATGGGAGAGATCCA GCAGGGTTCCAAAGTGAATGA Soltu.DM.04G031570.1
GUN4 TCGTTTTAGCCGGAGAAGCA TAACCGAATTTGCCGTCGCT Soltu.DM.06G028640.1
PORA GAGGGCAAAGCTAGTGCAAC TGACACCAGGAGAAGCAACC Soltu.DM.10G002270.1
UROD AGCTAGGCTTTCTCATTGTGT AAGCAAACCACAGAGACGGA Soltu.DM.06G011840.1
CHLM AACGCTAACGAATGGCAACG CGGCGAGGAGAAAGCCATAA Soltu.DM.03G032490.1
CHLG ACAACAACCCACTTCGCTCT TTATCCGGTGCCTGTGCTTT Soltu.DM.09G000240.1
SBP TCTGTCAACAATGCAGCCGA CTTCGCCTTTCGTTGTGTCC Soltu.DM.07G020030.1
LSD GCAGTAACAGCAGTACCACCT CACGAACATTGCACGCTTGT Soltu.DM.08G022900.1
GATA CAGCAGCGGAGAAAGTACGA GCCATCAACAAGAATGCTGCT Soltu.DM.12G001590.1
WRKY ACAGGGGTGAACACAACCAC GTCATTGCATACGGCTGCTC Soltu.DM.10G023760.1
MYB-like AAGACCTCAACCTCGGAACT TGCAACGTTTGTCGTCTTTGT Soltu.DM.10G020840.1
MYB113 TGGAACACACACCTACACAGG TCGAGGTCGTGGTCGTAGTA Soltu.DM.10G020780.1

Fig. 1

Change in phenotype and chlorophyll content of potato tubers of the RM-210 under light condition for 0-120 hours (A): the photographs of potato tubers were taken after 0, 6, 12, 36, 48, and 120 h of light exposure. (B): contents of chlorophyll a and chlorophyll b during 120 hours light exposure. Data are the averages of three replicates ± SDs. Different lowercase letters indicate the statistical differences between different groups by ANOVA (Tukey’s Method, P < 0.05)."

Fig. 2

Cluster diagrams and enrichment analysis of DEGs in tubers exposed at uninterrupted light for 0, 6, and 36 hours (A): principal component analysis of transcriptome data. (B): Volcano plot of DEGs for 6 h vs. 0 h and 36 h vs. 0 h. Blue rots represent genes of significant expression and gray rots were genes of nonsignificant expression. (C): GO enrichment analysis of DEGs. (D): the KEGG pathway analysis of DEGs."

Fig. 3

Cluster analysis of DEGs in tuber exposed to light for 0, 6, and 36 h"

Fig. 4

Relative expression pattern of genes involved in the synthesis of chlorophyll in greening potato tubers derived from RM-210 determined by qPCR (A): structural genes in the chlorophyll biosynthesis pathway; (B): RNA was extracted from the tuber epidermis under light at 22℃ for 0, 6, 36, and 48 h. The relative expression level of nine structural genes were determined by qRT-PCR. Different letters indicate significant difference between the different groups by ANOVA (Tukey’s Method, P < 0.05)."

Fig. 5

Verification of transcription factor expression by qPCR (A): qPCR results of 6 TFs selected in the transcriptome sequencing. (B): RNA-seq and qPCR data. (C): the correlation analysis of RNA-seq and qPCR results (Pearson Correlation Analysis). Different letters indicate significant difference between the different groups by ANOVA (Tukey’s Method, P < 0.05)."

Fig. 6

Prediction of cis-elements in promoters of structural genes (A) and transcription factors (B)"

Fig. 7

Transcriptional activation of structural gene promoter by MYB transcription factor EV: the co-injection of tobacco with the empty vector recombinant promoter of the structural gene; TF: the co-injection of tobacco with the transcription factor recombinant vector and the promoter recombinant vector of the structural gene. Asterisks indicate significant difference between EV and respective treatments by Student’s t-test (*: P < 0.05; **: P < 0.01)."

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