Assessment of Arabidopsis thaliana CENH3 promoter in Brassica juncea for development of haploid inducer lines.
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Date
2016-07
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Abstract
Centromeres are epigenetically specified by the centromeric histone H3 protein (CENH3). The timing and level of expression of CENH3 is tightly regulated to match the demands of the host cell. So far in plants, only CENH3 promoter of Arabidopsis thaliana (L.) Heynh. has been characterized. However, whether CENH3 promoters retain their characteristic mode of regulation in other species remains to be established. In the present study, activity of AtCENH3 promoter was investigated using reporter gene assay in Brassica juncea (L.) Czern. A 1156 bp promoter fragment of AtCENH3 gene (At1g01370) including the first 111 nucleotides of the coding sequence was amplified and cloned into the pORE-R2 binary vector to ensure translation fusion with the uidA coding sequences. The Agrobacterium tumefaciens strain GV3101 harbouring the recombinant construct was used to transform B. juncea cv. RLM198 hypocotyl explants. Histochemical assay of T0 and T1 transgenics showed GUS expression in shoot apical meristem, leaf, sepal, flower pedicel and root tip. Intense GUS expression was observed in meristematic tissues, particularly at shoot and root apices. However, mature leaves, flowers, pollen and ovules exhibited very low or no GUS expression. Our results showed that AtCENH3 promoter regulates cognate gene expression in Brassica juncea as it does in A. thaliana, and hence a suitable candidate for developing haploid inducer line in B. juncea.
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Keywords
AtCENH3, Brown mustard GUS, Haploid induction, Indian mustard, Promoter
Citation
Watts, Anshul Bhadouria, Jyoti Kumar, Vajinder Bhat, Shripad Ramachandra. Assessment of Arabidopsis thaliana CENH3 promoter in Brassica juncea for development of haploid inducer lines. Indian Journal of Experimental Biology. 2016 July; 54(7): 425-430.