Acta Horticulturae Sinica ›› 2022, Vol. 49 ›› Issue (9): 1866-1882.doi: 10.16420/j.issn.0513-353x.2021-0573

• Research Papers • Previous Articles     Next Articles

Gene Cloning and Expression Analysis of miR408 and Its Target DlLAC12 in Globular Embryo Development and Abiotic Stress in Dimocarpus longan

XU Xiaoping1, CAO Qingying1, CAI Roudi1, GUAN Qingxu1, ZHANG Zihao1, CHEN Yukun1, XU HAN1,2, LIN Yuling1, LAI Zhongxiong1,*()   

  1. 1College of Horticulture,Fujian Agriculture and Forestry University,Fuzhou 350002,China
    2Institut de la Recherche Interdisciplinaire de Toulouse,IRIT-ARI,Toulouse 31300,France
  • Received:2021-10-01 Revised:2022-06-16 Online:2022-09-25 Published:2022-10-08
  • Contact: LAI Zhongxiong


To investigate the expression patterns of miR408 and its target DlLAC12 during longan globular embryo(GE)development and different abiotic stresses. The miR-RACE PCR and Tail-PCR were used to clone the cDNA and gDNA of dlo-miR408 and cDNA of DlLAC12,and the transcriptional start site(TSS),and promoter of MIR408(pro-MIR408)were ascertained. Characteristics of dlo-miR408 sequence,homologous and cis-acting elements of the promoter were bioinformatically analyzed. The transcriptional expression levels of the members of dlo-miR408 family at various abio-stress conditions were verified with qRT-PCR,and the expression patterns of dlo-miR408-3p with its target gene DlLAC12 were further analyzed at early somatic embryogenesis stages with various plant growth regulator treatments. pro-MIR408 sequence contains hormone signal transducing cis-acting elements such as ABA,GA3 and JA,and light and low temperature stress cis-elements. The results showed that the length of pri-miR408 cDNA,gDNA were 706 bp. The TSS was“C”. The length of DlLAC12 cDNA was 1 725 bp. The length of pro-MIR408 was 1 532 bp. dlo-miR408-3p showed dynamic expressions in responding to sucrose,Cu2+ and temperature treatments by means of qRT-PCR,whereas dlo-miR408-5p1 showed downregulation in the imbalanced Cu2+ and low temperature conditions,and maintained stable expression to sucrose changes. Moreover,dlo-miR408-3p and DlLAC12 responded to high concentration of ABA(≥ 500 μmol · L-1)with negative regulations,and GA3 treatment made them same in the expression pattern. The negative regulation at the late globular embryo(GE)developmental stage played an important role in the GE morphogenesis. The present work showed that dlo-miR408 and DlLAC12 were involved in the longan somatic embryogenesis and abiotic stress.

Key words: longan, somatic embryogenesis, miR408, DlLAC12, abiotic stress

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