Fanglian He , Zuyang Qiu , Weiqing Dong , Lili Liu

1 Biotechnology Research Institute, Guangxi Zhuang Nationality Autonomous Region Academy of Agricultural Sciences, Nanning, 530007
2 Lipu Municipal Buresu of Agriculture and Rural Affairs, Lipu, 546600
Author    Correspondence author
Molecular Plant Breeding, 2020, Vol. 11, No. 8   
Received: 16 Jun., 2020    Accepted: 16 Jun., 2020    Published: 16 Jun., 2020

This article was first published in Molecular Plant Breeding ( ISSN1672-416X，CN46-1068/S) in Chinese, and here was authorized to translate and publish the paper in English under the terms of Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

In order to clone the cDNA sequence of water chestnut AGPase large subunit gene (EdAGPL1) and analyze its structural characteristics and its expression in different tissues and corm development of water chestnut. Using water chestnut 'Guilin water chestnut ' as research material. The EdAGPL1 gene was cloned by RT-PCR and analyzed by bioinformatics. The expression of EdAGPL1 in different tissues and corm development process was analyzed by real-time fluorescence quantitative PCR technology. The results showed that the length of the cloned EdAGPL1 gene was 1 744 bp, its open reading frame was 1 599 bp, it encoded 532 amino acids, the protein molecular mass was 58.84 kD, the isoelectric point was 7.54, and it had NTP_transferase and PbH1 domains. The secondary structure was composed of helix (13.72%), for the strand (25.19%) and the loop (61.09%). The tertiary structure consists of 17 α-helices, 34 β-sheets, and 51 β-turn (curl) composition. Homology and phylogenetic analysis showed that the homology of amino acid sequence of EdAGPL1 and other plants AGPL protein was 57.54%~61.64%, and has far evolutionary kinship with other plants. Fluorescence quantitative PCR analysis showed that the expression of EdAGPL1 in different tissues was corm> leaf stem > stolon > root. The expression of EdAGPL1 was the highest in the initial stage of corm development, and then it decreased and remainedstable. The study of this gene will help to clarify the regulation mechanism of starch synthesis in water chestnut and provide a theoretical basis for the breeding of high-starch varieties.

Eleocharis dulcis; Starch synthase; Gene cloning; Expression analysis