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CHINESE JOURNAL OF OIL CROP SCIENCES ›› 2020, Vol. 42 ›› Issue (3): 380-.doi: 10.19802/j.issn.1007-9084.2019302

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Cloning and functional analysis of uroporphyrinogen Ⅲ Synthase gene BnHemd from Brassica napus

  

  1. Oil Crops Research Institute of Chinese Academy of Agricultural Sciences, Key Laboratory of Biology and Genetic Im⁃
    provement of Oil Crops, Ministry of Agriculture and Rural Affairs, Wuhan 430062, China
  • Online:2020-06-28 Published:2020-06-08

Abstract:

The object of this study was to analyse the function of gene BnHemd encoding uroporphyrinogen III synthase (UROS) in rapeseed, and to better understand its relationship with chlorophyll synthesis for higher photosyn⁃thetic efficiency and yield. Gene editing technology was used to edit 2 copies of the gene on A9 and C8 chromosomes in Brassica napus cv Zhongshuang 11. The 2 genes were named BnA09.Hemd and BnC08.Hemd and cloned from the cultivar. Their CDS sequences were 885 bp and 876 bp respectively, composed of 9 exons, each encoding 294 and 291 amino acid residues. Protein analysis showed that the 2 copies had similar physicochemical properties, and both were unstable and hydrophobins. Phylogenetic tree showed that BnA09.Hemd and BnC08.Hemd were recently related to B. rapa and B. oleracea respectively, which is consistent with genetic evolution of B. napus. RT-PCR results showed that BnHemd was expressed in roots, stems, leaves, flower buds, seeds, and keratin peels of Zhongshuang 11.The expression levels in keratin peels and flower buds were much higher than those in roots, stems and leaves. Prelim⁃
inary functional analysis showed that plants with 2 copies of the gene double-simultaneously edited at the same time showed a significant decrease in chlorophyll content, lower photosynthetic rate and delayed growth.

doi:10.19802/j.issn.1007-9084.2019302

Key words: Brassica napus, uroporphyrinogen III synthase, gene editing, chlorophyll, photosynthetic effi?ciency

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