Cloning and Expression Analysis of SiHd3a Gene in Foxtail Millet

doi:10.7668/hbnxb.20190592

Abstract

Abstract: Analysis of the expression pattern of florigen gene (Hd3a)under different photoperiod conditions,aims at revealing the role of florigen gene that played in photoperiod-regulated flowering process of foxtail millet. First,a SiHd3a gene sequence(Seita.4G067600,named SiHd3a)on Chr.4 of foxtail millet was obtained by bioinformatics method and according to the transcribed sequence of Seita.4G067600,a pair of specific primers were designed successfully. Then the total RNA of millet land race Huangmaogu was extracted and after reverse transcription the cDNA sequence of SiHd3a gene was obtained by RT-PCR technology. The cDNA sequence was 792 bp which included a 537 bp CDS region that coding 178 amino acids. The molecular weight,isoelectric point of SiHd3a protein were 19.74 ku and 6.82 respectively,which was preliminarily judged as a hydrophilic protein. The proportion of irregular curl was the highest in protein secondary structure(43.26%),following it were extended chain(β-sheet,29.21%),α-helix(16.29%)and β-turn(11.24%). Subcellular localization analysis showed that SiHd3a protein was located in the cytoplasm and intercellular substance. Phylogenetic analysis based on Hd3a protein sequences showed that there was a close relationship between foxtail millet and crown,maize,sorghum,but a distant relationship between foxtail millet and rice. Semi-quantitative PCR showed that under short-day condition, SiHd3a gene gave a circadian rhythm expression pattern,with two expression peaks at six am,and twelve am respectively. While under long-day condition, SiHd3a gene gave almost stable expression level in 24 hours. It could be speculated that the diurnal expression pattern difference of SiHd3a gene between long-day and short-day might be the cause leading to photoperiod sensitivity of foxtail millet.

Key words: Foxtail millet, SiHd3a gene, Photoperiod sensitivity, Bioinformatics analysis, Diurnal expression

CLC Number:

S515

JIA Xiaoping, SANG Luman, WANG Zhenshan, ZHAO Yuan, ZHANG Xiaomei, LI Jianfeng, ZHANG Bo, ZHOU Junchao. Cloning and Expression Analysis of SiHd3a Gene in Foxtail Millet[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(1): 22-28. doi: 10.7668/hbnxb.20190592.

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