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Genomic origin, expression differentiation and regulation of multiple genes encoding CYP83A1, a key enzyme for core glucosinolate biosynthesis, from the allotetraploid Brassica juncea

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An Erratum to this article was published on 27 November 2014

Abstract

Main conclusion

The multiple BjuCYP83A1 genes formed as a result of polyploidy have retained cell-, tissue-, and condition-specific transcriptional sub-functionalization to control the complex aliphatic glucosinolates biosynthesis in the allotetraploid Brassica juncea.

Glucosinolates along with their breakdown products are associated with diverse roles in plant metabolism, plant defense and animal nutrition. CYP83A1 is a key enzyme that oxidizes aliphatic aldoximes to aci-nitro compounds in the complex aliphatic glucosinolate biosynthetic pathway. In this study, we reported the isolation of four CYP83A1 genes named BjuCYP83A1-1, -2, -3, and -4 from allotetraploid Brassica juncea (AABB genome), an economically important oilseed crop of Brassica genus. The deduced BjuCYP83A1 proteins shared 85.7–88.4 % of sequence identity with A. thaliana AtCYP83A1 and 84.2–95.8 % among themselves. Phylogenetic and divergence analysis revealed that the four BjuCYP83A1 proteins are evolutionary conserved and have evolved via duplication and hybridization of two relatively simpler diploid Brassica genomes namely B. rapa (AA genome) and B. nigra (BB genome), and have retained high level of sequence conservation following allopolyploidization. Ectopic over-expression of BjuCYP83A1-1 in A. thaliana showed that it is involved mainly in the synthesis of C4 aliphatic glucosinolates. Detailed expression analysis using real-time qRT-PCR in B. juncea and PromoterBjuCYP83A1–GUS lines in A. thaliana confirmed that the four BjuCYP83A1 genes have retained ubiquitous, overlapping but distinct expression profiles in different tissue and cell types of B. juncea, and in response to various elicitor treatments and environmental conditions. Taken together, this study demonstrated that transcriptional sub-functionalization and coordinated roles of multiple BjuCYP83A1 genes control the biosynthesis of aliphatic glucosinolates in the allotetraploid B. juncea, and provide a framework for metabolic engineering of aliphatic glucosinolates in economically important Brassica species.

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Abbreviations

CYP:

Cytochrome P450

DW:

Dry weight

GSL:

Glucosinolate (s)

GUS:

β-Glucuronidase

mya:

Million years ago

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Acknowledgments

The central instrumentation, microscopy facility, and plant growth facility at NIPGR are highly acknowledged. We thank Mr. Pawan Kumar for performing the divergence time analysis. Technical assistance of Mr. Vinod Kumar is duly acknowledged. The authors declare no conflict of interest exists.

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Authors and Affiliations

  1. National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi, 110067, India

    Meenu, Rehna Augustine, Manoj Majee & Naveen C. Bisht

  2. Department of Genetics, University of Delhi South Campus, Benito Juarez Marg, New Delhi, 110021, India

    Akshay K. Pradhan

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  1. Meenu
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  2. Rehna Augustine
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  3. Manoj Majee
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  4. Akshay K. Pradhan
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  5. Naveen C. Bisht
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Correspondence to Naveen C. Bisht.

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Meenu and R. Augustine contributed equally.

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Meenu, Augustine, R., Majee, M. et al. Genomic origin, expression differentiation and regulation of multiple genes encoding CYP83A1, a key enzyme for core glucosinolate biosynthesis, from the allotetraploid Brassica juncea . Planta 241, 651–665 (2015). https://doi.org/10.1007/s00425-014-2205-0

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