Amino Acids

, Volume 42, Issue 6, pp 2307–2318 | Cite as

Arabidopsis mutants lacking asparaginases develop normally but exhibit enhanced root inhibition by exogenous asparagine

  • Ana Ivanov
  • Alexander Kameka
  • Agnieszka Pajak
  • Luanne Bruneau
  • Ronald Beyaert
  • Cinta Hernández-Sebastià
  • Frédéric Marsolais
Original Article

Abstract

Asparaginase catalyzes the degradation of l-asparagine to l-aspartic acid and ammonia, and is implicated in the catabolism of transported asparagine in sink tissues of higher plants. The Arabidopsis genome includes two genes, ASPGA1 and ASPGB1, belonging to distinct asparaginase subfamilies. Conditions of severe nitrogen limitation resulted in a slight decrease in seed size in wild-type Arabidopsis. However, this response was not observed in a homozygous T-DNA insertion mutant where ASPG genes had been inactivated. Under nitrogen-sufficient conditions, the ASPG mutant had elevated levels of free asparagine in mature seed. This phenotype was observed exclusively under conditions of low illumination, when a low ratio of carbon to nitrogen was translocated to the seed. Mutants deficient in one or both asparaginases were more sensitive than wild-type to inhibition of primary root elongation and root hair emergence by l-asparagine as a single nitrogen source. This enhanced inhibition was associated with increased accumulation of asparagine in the root of the double aspga1-1/-b1-1 mutant. This indicates that inhibition of root growth is likely elicited by asparagine itself or an asparagine-derived metabolite, other than the products of asparaginase, aspartic acid or ammonia. During germination, a fusion between the ASPGA1 promoter and beta-glucuronidase was expressed in endosperm cells starting at the micropylar end. Expression was initially high throughout the root and hypocotyl, but became restricted to the root tip after three days, which may indicate a transition to nitrogen-heterotrophic growth.

Keywords

Asparaginase Asparagine Mutants Root elongation Root hair formation Nutrient sensing 

Supplementary material

726_2011_973_MOESM1_ESM.pdf (55 kb)
Supplementary material 1 (PDF 55 kb) Table 1 C, S and N content of mature seed of wild-type and aspga1-1/-b1-1 grown under normal (115 μmol photons m−2 sec−1) or low (70 μmol photons m−2 sec−1) illumination. Table 2 Total amino acid profile of mature seeds of wild-type and aspga1-1/-b1-1 grown under low light conditions
726_2011_973_MOESM2_ESM.pdf (36 kb)
Supplementary material 2 (PDF 35 kb) Figure 1 Graphs of D × G interactions from ANOVA of values in Tables 2 (a) and 3 (b)

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Copyright information

© Her Majesty the Queen in Rights of Canada 2011

Authors and Affiliations

  • Ana Ivanov
    • 1
    • 2
  • Alexander Kameka
    • 2
  • Agnieszka Pajak
    • 2
  • Luanne Bruneau
    • 2
  • Ronald Beyaert
    • 3
  • Cinta Hernández-Sebastià
    • 2
  • Frédéric Marsolais
    • 1
    • 2
  1. 1.Department of BiologyUniversity of Western OntarioLondonCanada
  2. 2.Agriculture and Agri-Food Canada, Southern Crop Protection and Food Research CentreLondonCanada
  3. 3.Agriculture and Agri-Food CanadaDelhiCanada

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