Cloning and Expression Analysis of Alkaline/Neutral Invertase Gene <i>MiNI </i>in <i>Mangifera indica </i>L.
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Chinese Journal of Tropical Crops ›› 2020, Vol. 41 ›› Issue (9): 1741-1747.DOI: 10.3969/j.issn.1000-2561.2020.09.003

• Omics & Biotechnology • Previous Articles     Next Articles

Cloning and Expression Analysis of Alkaline/Neutral Invertase Gene MiNI in Mangifera indica L.

GUO Lijun,DENG Huidong,FENG Xuejie,LUO Zhiwen,CHEN Zhe,FAN Hongyan,HU Fuchu,HUA Min()   

  1. Tropical Fruit Trees Institute, Hainan Academy of Agricultural Sciences / Investigation Station of Tropical Fruit Trees, Ministry of Agriculture and Rural Affairs / Key Laboratory of Tropical Fruit Tree Biology of Hainan Province / Research Center of Tropical Fruit Tree Breeding Engineering Technology of Hainan Province, Haikou, Hainan 571100, China
  • Received:2019-11-08 Revised:2020-02-10 Online:2020-09-25 Published:2020-10-16
  • Contact: HUA Min

Abstract:

The cDNA sequence of alkaline/neutral invertase (NI) gene (MiNI) in mango was cloned with Reverse Transcription Polymerase Chain Reaction (RT-PCR) in this study. The molecular properties and genetic relationships of NI were compared with different organisms. MiNI contained an open reading frame of 2034 bp, encoding a protein of 677 aa with a theoretical molecular weight 76.6 kDa and an isoelectric point of 6.24. Bioinformatics analysis showed that the secondary structure of NI consisted of alpha helix accounting for 38.85%, random coil accounting for 35.45%, extended strand accounting for 18.91%, and beta turn accounting for 6.79%, of which had the conserved domain of glycoside hydrolase family 100 and the same motif locations as Citrus clementine, Dimocarpus longan, Hevea brasiliensis and Carica papaya. The amino acid sequence encoded by MiNI had the highest homology with that of Citrus clementine, Dimocarpus longan. The analysis of NI phylogenetic tree showed that the closest genetic distance between mango was Citrus clementine, the furthest were Zea mays and Lycium barbarum. qRT-PCR analysis showed that the expression of MiNI in exocarp was much higher than that in mesocarp. MiNI relative expression in exocarp decreased significantly during 10 to 40 days after flowering, while the gene relative expression was maintained at a relatively stable level during 40 to 100 days after flowering. However, MiNI expression increased significantly with fruit ripening during the period from 100 to 130 days after flowering. The gene expression in mesocarp was different from that in exocarp. The gene expression of MiNI in mesocarp decreased significantly during 10 to 40 days after flowering, while the gene relative expression remained at a very low level until the later stage of fruit development. The study would provide a foundation to understand the role of MiNI in sucrose metabolism and further dissection of the sugar metabolism mechanism from molecular perspective.

Key words: Mangifera indica L., alkaline/neutral invertase, cloning, bioinformatics, expression analysis

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