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Acta Agronomica Sinica ›› 2023, Vol. 49 ›› Issue (12): 3250-3260.doi: 10.3724/SP.J.1006.2023.34002

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

Transcriptomic profile of key stages of sex differentiation in cassava flowers and discovery of candidate genes related to female flower differentiation

CHEN Hui-Xian(), LIANG Zhen-Hua, HUANG Zhen-Ling, WEI Wan-Ling, ZHANG Xiu-Fen, YANG Hai-Xia, LI Heng-Rui*(), HE Wen*(), LI Tian-Yuan, LAN Xiu, RUAN Li-Xia, CAI Zhao-Qin, NONG Jun-Xin   

  1. Guangxi Institute of Sub-tropical Agricultural Sciences, Longzhou 532415, Guangxi, China
  • Received:2023-01-05 Accepted:2023-05-24 Online:2023-12-12 Published:2023-06-06
  • Contact: * E-mail: lihengrui88@163.com; E-mail: 675559090@qq.com
  • Supported by:
    National Key Research and Development Program of China(2020YFD1000603-10);Guangxi Youth Science Foundation Project(2023GXNSFBA026150);Guangxi Academy of Agricultural Sciences Development Fund Project(Guinongke 2022JM96);Science and Technology Plan Project of Special Project for Basic Scientific Research of Chongzuo(Chongkegong 2021ZC18);Guangxi Academy of Agricultural Sciences(Guinongke 2023YM44)

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Abstract:

To solve the breeding problem of the serious shortage of female cassava flowers, cassava variety ‘Xinxianxuan 048’ was used as the experimental material. Transcriptional sequencing technology was used to analyze the biological information of differentially expressed genes in female and male flowers during the critical period of cassava flower sex differentiation, explore the functions of differentially expressed genes and possible regulatory pathways involved, excavate candidate genes related to female differentiation, and verify the sequencing results by qRT-PCR method. The results showed that: There were 545 differentially expressed genes between male and female cassava flowers at the critical stage of gender differentiation. Among them, 48.63% of the differential genes were enriched in GO pathway of floral organ morphogenesis and development, and the genes enriched in plant phenylpropanol biosynthesis and plant hormone signal transduction were the most. AGL11, YABBY4, CRC, SUP, and other flower sex differentiation genes were significantly up-regulated in female flowers. Four genes of the cytokinin signaling pathway, including HK4, HPt4, ARR8, and ARR12, were significantly up-regulated in female flowers, while IAA14, GH3, SAUR22, and SAURR20, the early auxin response genes, were significantly down-regulated in female flowers. Therefore, the sexual differentiation of cassava flowers mainly involved the biological pathways of pollen, gametophyte, floral organ morphogenesis and development, and two metabolic pathways of plant phenylpropanoid biosynthesis and plant hormone signal transduction. Cytokinin and auxin might be the main hormones in cassava female flower differentiation. AGL11, YABBY4, CRC, SUP, HK4, HPt4, ARR8, and ARR12 may be positive regulators of cassava female flower differentiation.

Key words: cassava, key stages of sex differentiation, transcriptome, female flowers, gene

Table 1

Primer sequences of real-time fluorescence quantitative PCR"

基因名称
Gene name		 正向引物
Forward primer (5°-3°)		 反向引物
Reverse primer (5°-3°)		 基因登录号
Gene ID
AGL11 TATGAATTGTCAGTCCTTTGTG TCTAACAGGCATCTAATGGGT LOC110616214
CRC CACAGTTCTTGCGGTTGG TTGGCCTTGAAAAGGAGGT LOC110615357
YABBY4 CAGATTTGCTACGTCCAATGTG AATGGCACGGTTCACTGGG LOC110627391
MADS9 CACCTGAAAGGGGAGGAT TGACGAACACTGGCAAGG LOC110607164
TM6 TTAGCCCTACCATAACGA ATAATGCGTGCTCCACAG LOC110630551
ARR12 TGCCGAGGTTGAGAAATC TGCTCCTGCCGTTTGATC LOC110631158
HPt4 GCAGGGCAGGAAATGGAG CTTGGGGCGACATGCAGT LOC110627048
HK4 GCGGAGTAGCCTATGCAC TTCCCGGTAGCTCTAGCC LOC110603154
TGA9 CTTCACCAACTTCGCCGGATAC AGGACGGGATGCCCAGAGG LOC110626841
TGA10 GCTTATGACTTGGGAGAA CATGGACGGCTGATTTGA LOC110620640

Table 2

Statistical table of base information"

样本
Sample		 干净序列读数(个)
Read number		 碱基数
Base number (G)		 Q20 (%) Q30 (%)
A1-1 50,981,844 7.25 95.680 87.990
A1-2 35,421,366 4.83 97.975 93.795
A1-3 39,490,380 5.40 97.780 93.375
A2-1 33,138,892 4.48 97.765 93.350
A2-2 31,980,834 4.38 96.455 89.915
A2-3 42,204,558 5.79 98.130 94.145

Fig. 1

Statistical graph of differential expressed genes (female flower)"

Fig. 2

Venn diagram of differential expressed genes"

Fig. 3

Volcano plot of male flowers vs female Red areas represent up-regulated genes in male flowers, and blue areas represent down-regulated genes in male flowers."

Fig. 4

The first 30 GO items enriched significantly"

Fig. 5

GO items of differential expressed genes significantly"

Fig. 6

The first 20 KEEG pathways enriched significantly"

Fig. 7

Heat map of candidate genes related to flower sex differentiation The color represents the FPKM value of the response. The larger the value, the redder the color. From left to right, the samples are female flower-1 (A1-1), female flower-2 (A1-2), female flower-3 (A1-3), male flower-1 (A2-1), male flower-2 (A2-2), and male flower-3 (A2-3)."

Fig. 8

Model of gene expression patterns of cytokinin signaling pathway The color represents the FPKM value of the response, and the sample labels from left to right refer to those given in Fig. 7."

Fig. 9

Relative expression patterns of auxin responsive genes The color represents the FPKM value of the response, and the sample labels from left to right refer to those given in Fig. 7."

Fig. 10

Relative expression level of differentially expressed genes verified by qRT-PCR Different lowercase letters on the histogram indicate that there are significantly different in the relative expression levels of 10 candidate genes between female and male flowers at P < 0.05."

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