Scientia Agricultura Sinica ›› 2015, Vol. 48 ›› Issue (22): 4417-4427.doi: 10.3864/j.issn.0578-1752.2015.22.003

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Molecular Cloning and Arabidopsis Ectopic Expression of a Phytochrome B gene from Brassica oleracea

SUN Guang-hua1,2, YUAN Huan-huan1,2, FAN Xiao-cong1,2, GU Hai-ke3, SONG Mei-fang2,3, XIAO Yang4, MENG Fan-hua2, GUO Lin2, YANG Qing-hua1, ZHAN Ke-hui1, YANGJian-ping1,2   

  1. 1College of Agronomy, Henan Agricultural University, Zhengzhou 450002
    2Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081
    3 Beijing Radiation Center, Beijing 100875
    4Graduate School, Chinese Academy of Agricultural Sciences, Beijing 100081
  • Received:2015-06-22 Online:2015-11-16 Published:2015-11-16

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Abstract: 【Objective】Cabbage (Brassica oleracea L.) is one of the most widely grown vegetable crops in the world. Cabbage is grown in spring and fall, however varieties need to be made suitable for cultivation all the year round. Study on the response of cabbage phytochromes to light and temperature is an important basis for crop improvement with respect to extending growth periods. Phytochromes of interest here are the red/far-red reversible photoreceptors. Among them, phyB is an essential red light receptor that predominantly mediates seedling photomorphogenesis and shade-avoidance response. The objective of this study was to isolate the phytochrome B gene in cabbage (i.e., BoPHYB), to explore its functions on seedling de-etiolation and shade avoidance by ectopic expression in Arabidopsis, and evaluate its potential in cabbage breeding. 【Method】The cDNA sequence of BoPHYB was isolated from leaves of a self incompatible line 12C. Bioinformatic analyses were employed to predict its function domains and to build a phylognetic relationship tree among plant phyB homologs. The pJIM19-Myc-BoPHYB binary construct was electroporated into the Agrobacterium tumefaciens strain GV3101 and then introduced into Arabidopsis thaliana (both of the wild type and phyB-9 mutant) via a floral dip method. Transgenic plants were selected on germination plates containing 50 mg/mL kanamycin for Myc-BoPHYB. Homozygous T3 or T4 transgenic plants were used for testing the seedling hypocotyl elongation and shade avoidance responses. 【Result】The reading frame of BoPHYB possesses 3 507 bp and encodes 1 168 amino acid residues (128.9 kD) deduced from the DNA sequence. Similar to both phyB protein of Arabidopsis thaliana (i.e., AtphyB) and Brassica rapa (i.e., BrphyB), BophyB contains one GAF domain, two PAS domains, one HisKA domain, and one HATPase_c domain. Phylogenetic analysis indicated that BophyB belongs to the same branch as BrphyB and AtphyB, while showing low similarity to phyB proteins from monocotyledonous species, such as common wheat, corn and rice. Compared with the wild type Col-0, the transgenic lines overexpressing BophyB in A. thaliana exhibited much shorter hypocotyl elongation under red and white-light conditions. Their adult plants displayed dwarf stature with dark green leaves and short petioles under long-day and short-day conditions.【Conclusion】In this study, we cloned BoPHYB gene, and evaluated it with AtPHYB in crop improvement. Taking on similar structures and functions with AtphyB, BophyB promotes seedling de-etiolation under red and white-light conditions and inhibits shade avoidance responses in mature plants under long-day and short-day conditions.

Key words: Brassica oleracea L., Arabidopsis thaliana, phytochrome B, de-etiolation, shade avoidance

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