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Acta Agronomica Sinica ›› 2018, Vol. 44 ›› Issue (9): 1290-1300.doi: 10.3724/SP.J.1006.2018.01290

• RESEARCH PAPERS • Previous Articles     Next Articles

Transcription Abundances of CRY1b and CRY2 Genes in Response to Different Light Treatments in Maize

Hong-Dan LI1,2,Lei YAN1,2,Lei SUN1,2,Xiao-Cong FAN1,3,Shi-Zhan CHEN1,3,Yan ZHANG1,3,Lin GUO1,Guang-Xia YOU1,Zhuang LI1,2,Zong-Ju YANG1,2,Liang SU1,*,Jian-Ping YANG1,3,*   

  1. 1 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
    2 Graduate School, Chinese Academy of Agricultural Sciences, Beijing 100081, China
    3 College of Agronomy, Henan Agricultural University, Zhengzhou 450002, Henan, China
  • Received:2018-01-25 Accepted:2018-06-12 Online:2018-09-10 Published:2018-07-02
  • Contact: Liang SU,Jian-Ping YANG
  • Supported by:
    This study was supported by the Major Project of China on New Varieties of GMO Cultivation(2016ZX08010002-003-002);Key Project of Beijing Natural Science Foundation(6151002);National Natural Science Foundation of China (31570268)

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Abstract:

Light is lightly related to the important agronomic traits in maize such as plant height, flowering time, yield and quality. Cryptochromes are blue and ultraviolet-A photoreceptors generally existing in animal, plant and microbial, which mainly regulate photomorphogenesis in plants and circadian rhythms in both of plant and animal. Therefore, the expression pattern analysis of cryptochrome in maize could lay a research foundation in the photomorphogenesis in maize. The ZmCRY1b and ZmCRY2 genes were cloned by RT-PCR. Their proteins’ function domains and the phylogenetic analysis of amino acid sequences were carried out through bioinformatics analysis. The transcription abundances of ZmCRY1b and ZmCRY2 genes in different tissues of inbred line B73 under different light treatments were analyzed by qRT-PCR. We found that the function domains of ZmCRY1b or ZmCRY2 protein was consistent with CRY1 or CRY2 in Arabidopsis, rice and wheat, which contains the PHR and CCE domains or the PHR domain, respectively. Phylogenetic analysis indicated that the three gramineous CRYs from maize, wheat, and rice belonged to the same branch, while showing low similarity to other CRY1 proteins from dicotyledons. ZmCRY1b and ZmCRY2 genes highly expressed in leaf. Meanwhile, they could respond to all treatments of different continuous light conditions, the transitions from the dark to light conditions, as well as long-day and short-day conditions. The transcription abundances of ZmCRY1b in all treatments were higher than those of ZmCRY2, indicating that ZmCRY1b was more important than ZmCRY2 in maize. In conclusion, both of ZmCRY1b and ZmCRY2 genes can greatly respond to different light conditions and light cycle treatments, and play an important role in maize photomorphogenesis. Our results also provide a research basis for functional exploration of ZmCRY1b and ZmCRY2 in crop improvement.

Key words: maize, cryptochrome, photomorphogenesis, light treatment, transcription abundance

Table 1

Primers for cloning genes"

基因名称
Gene name		 基因编号
Accession number		 正向引物序列
Forward primer sequence (5'-3')		 反向引物序列
Reverse primer sequence (5'-3')
ZmCRY1b ZM02G13620 CACCGCCTGATGAACTGGA ATGGATGAGTAGTTCAGTGGACAA
ZmCRY2 ZM09G09240 TCTGGTTATTGTCATATTGCAGTTCT TGCCAAGATGTTCCCTTTGAGT

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 primer sequence (5'-3')		 反向引物序列
Reverse primer sequence (5'-3')
ZmCRY1b ZM02G13620 CACCGCCTGATGAACTGGA ATGGATGAGTAGTTCAGTGGACAA
ZmCRY2 ZM09G09240 GAACCACAGGCGAGATGCT GATCACTACAAACGCACCAGC
ZmTubulin NM_001174192 ACTTCATGCTTTCGTCCTACGCTCCA CTGGGAGGCTGGTAGTTGATTC

Fig. 1

Multiple sequence alignments and function domains of CRY1 proteins from Zea mays, Arabidopsis thaliana, Oryza sativa, and Triticum aestivumMultiple sequence alignments at amino acid levels were analyzed by NCBI and DNAMAN, and the function domains were analyzed by SMART. AtCRY1: Arabidopsis thaliana CRY1, AAB28724; OsCRY1a: Oryza sativa CRY1a, BAB70686; OsCRY1b: Oryza sativa CRY1b, BAB70686; TaCRY1a: Triticum aestivum CRY1a, ABX58028; ZmCRY1b: Zea may CRY1b, ZM02G13620. Black, dark grey, light grey, and white in the picture stand for 100%, 75%, 50%, and 0 of consistency, respectively."

Fig. 2

Multiple sequence alignments and function domains of CRY2 proteins from Zea mays, Arabidopsis thaliana, Oryza sativa, and Triticum aestivumMultiple sequence alignments at amino acid levels were analyzed by NCBI and DNAMAN, and the function domains were analyzed by SMART. AtCRY2: Arabidopsis thaliana CRY2, AT1G04400.2; OsCRY2: Oryza sativa CRY2, CAC82538.1; TaCRY2: Triticum aestivum CRY2, ABX58030.1; ZmCRY2: Zea may CRY2, ZM09G09240. Black, dark grey, light grey, and white in the picture stand for 100%, 75%, 50%, and 0 of consistency, respectively."

Fig. 3

Phylogenetic analysis of amino acid sequences of ZmCRY proteins and other related CRY proteinsThe amino acid sequences were obtained from the NCBI and the neighbor-joining tree was constructed by the DNAMAN. AtCRY1; Arabidopsis thaliana CRY1, AAB28724; AtCRY2: Arabidopsis thaliana CRY2, AT1G04400.2; BnCRY1: Brassica napus CRY1, CAG28805; BnCRY2a: Brassica napus CRY2a, AEA29690.1; BnCRY2b: Brassica napus CRY2b, AEA29691.1; GmCRY1a: Glycine max CRY1a, DQ401046; GmCRY1b1: Glycine max CRY1b1, AB498929; GmCRY1b2: Glycine max CRY1b2, AB498930; GmCRY2: Glycine max CRY2, XP_006588364.1; OsCRY1a: Oryza sativa japonica Group CRY1a, BAB70686; OsCRY1b: Oryza sativa japonica Group CRY1b, BAB70688; OsCRY2: Oryza sativa japonica Group CRY2, CAC82538.1; HvCRY1a: Hordeum vulgare subsp. Vulgare CRY1a, ABB13328; HvCRY1b: Hordeum vulgare subsp. Vulgare CRY1b, ABB13331; TaCRY1a: Triticum aestivum CRY1a, ABX58028; TaCRY2: Triticum aestivum CRY2, ABX58030.1; PsCRY1: Pisum sativum CRY1, AAS79662; SbCRY2: Sorghum bicolor CRY2, AAN37909; SlCRY1a: Solanum lycopersicum CRY1a, AAD44161; SlCRY1b: Solanum lycopersicum CRY1b, AAL02092; ZmCRY1a1: Zea may CRY1a1, ZM05G31560; ZmCRY1a2: Zea may CRY1a2, ZM04G17060; ZmCRY1b: Zea may CRY1b, ZM02G13620; ZmCRY2: Zea may CRY2, ZM09G09240."

Fig. 4

Relative expression analysis of both ZmCRY1b and ZmCRY2 genes in different organs of maizeDifferent tissues of maize inbred line B73, including root, stem, leaf, stamen, pulvinus, sheath, pedicel, young ear, pistil and bract, were harvested for qRT-PCR assays. The transcription abundance of ZmCRY1b in the root was set as the control, and the ratio of ZmCRY1b/Tubulin in the root was set as 1. Each column shows the mean expression of ZmCRY1b/Tubulin of three biological repeats. Error bars indicate the standard deviation. * P < 0.05, ** P < 0.01."

Fig. 5

Relative expression analysis of both ZmCRY1b and ZmCRY2 genes under different continuous light conditionsThe seedlings of maize inbred line B73 were grown in continuous far-red light (FR, 0.25 μmol m-2 s-1), red light (R, 22.3 μmol m-2 s-1), blue light (B, 13 μmol m-2 s-1), or white light (W, 170 μmol m-2 s-1) for 13 d. The transcription abundance of ZmCRY2 in the dark was set as the control, and the ratio of ZmCRY2/Tubulin in the dark was set as 1. Each column shows the mean expression of ZmCRY2/Tubulin of three biological repeats. Error bars indicate the standard deviation. * P < 0.05, ** P < 0.01."

Fig. 6

Relative expression of both ZmCRY1b and ZmCRY2 genes during transitions from darkness to different light conditionsThe seedlings of maize inbred line B73 were grown in the dark for 13 d, then transferred to blue light (B, 13 μmol m-2 s-1), white light (W, 17 μmol m-2 s-1), or red light (R, 22.3 μmol m-2 s-1) for 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, 12 h, or 24 h and far-red light (FR, 0.25 μmol m-2 s-1) for 0.16 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, 12 h, 24 h, or 24.08 h. The transcription abundance of ZmCRY2 in the dark was set as the control, and the ratio of ZmCRY2/Tubulin in the dark was set as 1. Each line graph shows the mean expression of ZmCRY2/Tubulin of three biological repeats. Error bars indicate the standard deviation."

Fig. 7

Relative expression of both ZmCRY1b and ZmCRY2 genes in response to photoperiod (long-day and short-day) treatmentThe seedlings of maize inbred line B73 were grown in long-day condition (LD, 16 h light/8 h dark) or short-day (SD, 8 h light/16 h dark) for 13 d, then sampled every two hours. The transcription abundance of ZmCRY2 in the dark was set as the control, and the ratio of ZmCRY2/Tubulin at the end of the dark period was set as 1. Each line graph shows the mean expression of ZmCRY2/Tubulin of three biological repeats. Error bars indicate the standard deviation."

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