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Acta Agronomica Sinica ›› 2022, Vol. 48 ›› Issue (12): 3057-3070.doi: 10.3724/SP.J.1006.2022.11115

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

Transcription abundances of PHYA, PHYB, and PHYC genes in response to different light treatments in Secale cereale

YANG Lu-Hao1(), WANG Li-Jian1,2(), SUN Guang-Hua1, WANG Shao-Ci1, CUI Lian-Hua1, CHEN Chang1, SONG Mei-Fang3, ZHANG Yan-Pei1, JIANG Liang-Liang1(), YANG Jian-Ping1(), WANG Chen-Yang1()   

  1. 1College of Agronomy / State Key Laboratory of Wheat and Maize Crop Science, and Center for Crop Genome Engineering, Henan Agricultural University, Zhengzhou 450046, Henan, China
    2Henan Police College, Zhengzhou 450046, Henan, China
    3Beijing Radiation Center, Beijing Academy of Science and Technology, Beijing 100875, China
  • Received:2021-12-22 Accepted:2022-03-25 Online:2022-12-12 Published:2022-04-20
  • Contact: JIANG Liang-Liang,YANG Jian-Ping,WANG Chen-Yang E-mail:yangluhao226@163.com;wanglijian2012@163.com;Jiangll@henau.edu.cn;jpyang@henau.edu.cn;xmzxwang@henau.edu.cn
  • About author:First author contact:

    **Contributed equally to this work

  • Supported by:
    Lhasa Regional Science and Technology Collaborative Innovation Project in 2022(QYXTZX-LS2022-01);National Natural Science Foundation of China(31871709);Key Project of Beijing Natural Science Foundation(6151002);Young Talents Project of Henan Agricultural University(30501038);Young Talents Project of Henan Agricultural University(30500823)

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Abstract:

Cultivated rye (Secale cereal L.), belonging to the rye genus of the poaceae family, is a valuable food and feed crop that is highly resistant to biotic and abiotic stresses. The 1BL/1RS translocation line has improved wheat resistance to powdery mildew and stripe rust worldwide. Phytochromes are an red/far-red receptors that regulate seed germination, plant height, shade avoidance response, flowering, and other adaptive responses in plants. In this study, three phytochrome genes, ScPHYA, ScPHYB, and ScPHYC were cloned. The transcriptional abundance of ScPHYs in different tissues and light treatments was analyzed by qRT-PCR. The results showed that phys of rye had similar domains with those of Arabidopsis, rice, and wheat, and had higher amino acid sequence consistency with those of wheat and rice, suggesting that they might have similar function. The transcriptional abundances of ScPHYs were the highest in roots, and higher under dark and far-red light than other continuous light conditions, while the relative expression patterns of the three genes were slightly different in the transformation from dark to red light, far-red light, blue light, and white light. In photoperiodic response, the relative expression patterns of the three genes were similar under long-day conditions, but different under short-day conditions. These results indicated that ScPHYs may play an important role in the photomorphogenesis of rye. The transcriptional analysis of three rye phytochrome genes provides a reference for the study of rye optical signal system and a basis for exploring the application of phytochrome in rye genetic improvement and molecular breeding.

Key words: Secale cereale, phytochromes, gene cloning, light treatment, transcription abundance

Table 1

Primers for genes cloning"

基因名称
Gene name		 基因编号
Accession number		 正向引物序列
Forward sequence (5'-3')		 反向引物序列
Reverse sequence (5'-3')
ScPHYA ScWN4R01G009900 CGGGGGACTCTTGACCATGGCA ATGTCTTCCTCAATGCCTGCTT AGTTCTTCTCCTTTACTAGT GTGCCCCATTGCCGTT
ScPHYB ScWN7R01G236700 CGGGGGACTCTTGACCATGGCA ATGGCCTCGGGAAGCCG AGTTCTTCTCCTTTACTAGT GCTCCGATCCCTACTTTCT
ScPHYC ScWN4R01G173600 CGGGGGACTCTTGACCATGGCA
ATGTCGTCGTCGCGGTCC		 AGTTCTTCTCCTTTACTAGT
GAAGTTGCTCTTGCTCGTC

Table 2

Primers for qRT-PCR"

基因名称
Gene name		 基因编号
Accession number		 正向引物序列
Forward sequence (5'-3')		 反向引物序列
Reverse sequence (5'-3')
ScPHYA ScWN4R01G009900 CAGTGAAGTTCTCTCCTGTTG TCCCTGGTGCTTGATCC
ScPHYB ScWN7R01G236700 GCTATGGGCAAATCATTAG TGGTCCTTTAGACTGCTCTGAC
ScPHYC ScWN4R01G173600 GCAAGGAACCGAAGAGCAA CCTGTCAAATCTTGTGCTACG
ScActin ScWN1R01G374800 CGTGTTGGATTCTGGTGATG AGCCACATATGCGAGCTTCT

Fig. 1

Phylogenetic analysis of phytochrome proteins The amino acid sequences were obtained from the Uniprot website and WheatOmics website, the phylogenetic analysis was performed by MEGA Version 10. AtphyA: Arabidopsis thaliana, P14712; AtphyB: Arabidopsis thaliana, P14713; AtphyC: Arabidopsis thaliana, P14714; OsphyA: Oryza sativa, A2XLG5; OsphyB: Oryza sativa, A2XFW2; OsphyC: Oryza sativa, A2XM23; ZmphyA1: Zea mays, P19862; ZmphyA2: Zea mays, Q53ZT7; ZmphyB1: Zea mays, Q6XFQ3; ZmphyB2: Zea mays, Q6XFQ2; ZmphyC1: Zea mays, Q6XFQ1; ZmphyC2: Zea mays, Q6XFQ0; SbphyA: Sorghum bicolor, Q53YS9; SbphyB: Sorghum bicolor, Q6S525; SbphyC: Sorghum bicolor, P93528; TaphyA: Triticum aestivum, Q5K5K6; TaphyB: Triticum aestivum, A9JR06; TaphyC: Triticum aestivum, Q8VWN1; ScphyA: Secale cereale, ScWN4R01G009900; ScphyB: Secale cereale, ScWN7R01G236700; ScphyC: Secale cereale, ScWN4R01G173600; HvphyA: Hordeum vulgare, Q2I7M3; HvphyB: Hordeum vulgare, Q2I7M0; HvphyC: Hordeum vulgare, Q2I714; AetphyA: Aegilops tauschii, A0A453H8S4; AetphyB: Aegilops tauschii, A0A453I867; AetphyC: Aegilops tauschii, A0A453LU83; TuphyA: Triticum urartu, M8B484; TuphyB: Triticum urartu, TuG1812G0400002191; TuphyC: Triticum urartu, M8A187."

Fig. 2

Multiple sequence alignments and function domains of phyA proteins from Secale cereale, Arabidopsis thaliana, Oryza sativa, and Triticum aestivum Multiple sequence alignments at amino acid levels and function domains were analysed by DNAMAN version 9 software and Pfam website and WheatOmics website. AtphyA: Arabidopsis thaliana, P14712; OsphyA: Oryza sativa, A2XLG5; TaphyA: Triticum aestivum, Q5K5K6; ScphyA: Secale cereale, ScWN4R01G009900."

Fig. 3

Multiple sequence alignments and function domains of phyB proteins from Secale cereale, Arabidopsis thaliana, Oryza sativa, and Triticum aestivum Multiple sequence alignments at amino acid levels and function domains are analyzed by DNAMAN version 9 software and Pfam website and WheatOmics website. AtphyB: Arabidopsis thaliana, P14713; OsphyB: Oryza sativa, A2XFW2; TaphyB: Triticum aestivum, A9JR06; ScphyB: Secale cereale, ScWN7R01G236700."

Fig. 4

Multiple sequence alignments and function domains of phyC proteins from Secale cereale, Arabidopsis thaliana, Oryza sativa, and Triticum aestivum Multiple sequence alignments at amino acid levels and function domains are analysed by DNAMAN version 9 software and Pfam website and WheatOmics website. AtphyC: Arabidopsis thaliana, P14714; OsphyC: Oryza sativa, A2XM23; TaphyC: Triticum aestivum, Q8VWN1; ScphyC: Secale cereale, ScWN4R01G173600."

Fig. 5

Relative expression levels of ScPHYA, ScPHYB, and ScPHYC in different tissues The relative expression levels of ScPHYA, ScPHYB, and ScPHYC in different tissues of Weining rye were analyzed by qRT-PCR. The relative expression level of ScPHYB in the leaf (ScPHYB/ScActin set as 1) was set as control. The bar chart shows the relative mean value of three independent replicates. The error bar represents the standard deviation. *: P < 0.05; **: P < 0.01; ***: P < 0.001."

Fig. 6

Relative expression levels of ScPHYA, ScPHYB, and ScPHYC during different continuous light condition Weining rye was grown in continuous darkness (Dk), red light (R, 22.3 μmol m-2 s-1), far-red light (FR, 1.9 μmol m-2 s-1), blue light (B, 13.0 μmol m-2 s-1), and white light (W, 17.0 μmol m-2 s-1) for 7 days. The relative expression level of ScPHYB under the darkness (ScPHYB/ScActin set as 1) was set as control. The bar chart shows the relative mean value of three independent replicates. The error bar represents the standard deviation. **: P < 0.01; ***: P < 0.001."

Fig. 7

Relative expression levels of ScPHYA, ScPHYB, and ScPHYC genes during darkness to different light conditions The seedlings of Weining rye were grown in the dark for 7 days, then transferred to red light (R, 22.3 μmol m-2 s-1), far-red light (FR, 1.9 μmol m-2 s-1), blue light (B, 13.0 μmol m-2 s-1), and white light (W, 17.0 μmol m-2 s-1) for 0, 0.25, 0.5, 1, 2, 4, 8, 16, and 24 hour(s). The relative level of ScPHYB under the darkness (ScPHYB/ScActin set as 1) was set as control. The line chart shows the relative mean value of three independent replicates. The error bar represents the standard deviation."

Fig. 8

Relative expression levels of ScPHYA, ScPHYB, and ScPHYC under photoperiodic treatments The seedlings of Weining rye were grown in long-day (LD, 16 h light / 8 h darkness) and short-day (SD, 8 h light / 16 h darkness) for 10 days, then samples were taken every 2 hours. The relative level of every gene in the end of the darkness was set as the control (set as 1). The line chart shows the relative mean value of three independent replicates. The error bar represents the standard deviation."

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