Cloning and Identification of LHY Gene in Chinese Milk Vetch (Astragalus sinicus)
ZHANG Xian1, ZHUANG Li1, CAO Wei-Dong2, CAO Kai1, XU Jing1, SI Lin-Lin1, WANG Jian-Hong1,*
1 Institute of Environment, Resource, Soil & Fertilizer, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China; 2 Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, Beijing 100081, China
AbstractAstragalus sinicus has been identified as one of the most important green manure for paddy fields in the south of China. It is vital to the sustainable utilization of soil resources. Based on different farming systems, selecting flowering varieties is an important breeding target of A. sinicus. Flowering time is regulated by the circadian clock in response to the external environment. Late elongated hypocoty l (LHY) is a key biorhythm clock gene. In the present study, in order to explore the function of the LHY gene in A. sinicus, the Illumina and rapid-amplification of cDNA ends (RACE) technology were performed to obtain the sequence of LHY in A. sinicus; the expression profiles of the LHY gene in tissues of A. sinicus were subsequently analyzed by qRT-PCR; the function of LHY gene of A. sinicus is verified by the transformation of LHY gene into Arabidopsis. The full-length cDNA of the LHY gene (GenBank No. OP455117) was isolated in A. sinicus, which was 2 865 bp in length and containing the ORF of 2 259 bp, encoding 752 amino acid polypeptide. The encoded amino acid sequence is highly homologous to the LHY proteins of Medicago truncatula, Cicer arietinum, and Glycine soja with a more than 75% similarity. The qRT-PCR analysis showed that the LHY gene had the highest expression accumulation in leaves, flowers, and flower buds of Astragalus sinicus. The LHY-overexpression transgenic Arabidopsis lines showed a significant late flowering phenotype. The bolting days and flowering days of transgenic Arabidopsis lines were 18.36 and 20.72 d later than that of the wild-type respectively. This study provides important implications for genetic modification of flowering in A. sinicus.
ZHANG Xian,ZHUANG Li,CAO Wei-Dong, et al. Cloning and Identification of LHY Gene in Chinese Milk Vetch (Astragalus sinicus)[J]. 农业生物技术学报, 2023, 31(11): 2263-2271.
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