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Engineered oryzacystatin‐I expressed in transgenic hairy roots confers resistance to Globodera pallida
Author(s) -
Urwin Peter E.,
Atkinson Howard J.,
Waller David A.,
McPherson Michael J.
Publication year - 1995
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1046/j.1365-313x.1995.08010121.x
Subject(s) - nematode , biology , globodera pallida , fecundity , caenorhabditis elegans , botany , solanaceae , biochemistry , gene , ecology , population , demography , sociology
The cysteine proteinase inhibitor, oryzacystatin‐I (Oc‐I), and several engineered Oc‐I variants have been tested for efficacy in inhibiting growth and development of both the free‐living nematode, Caenorhabditis elegans , and the plant parasitic nematode Globodera pallida . To assist in the design of protein engineering experiments to improve the efficacy of Oc‐I, an alignment of 28 cystatins and a molecular model of Oc‐I were generated. Inhibitory activities ( K i ) of wild‐type and variant forms of Oc‐I against both papain and the C. elegans cysteine proteinase, gcp‐1, were measured. For one variant, in which residue Asp86 was deleted (Oc‐IΔD86), the K,i was reduced by 13‐ to 14‐fold. LD 50 studies to test the effect of Oc‐I and Oc‐IΔD86 against C. elegans showed the relative median potency of Oc‐IΔD86 to be 0.76 that of wild‐type Oc‐I. When expressed in tomato hairy roots both Oc‐I and Oc‐IΔD86 had a detrimental effect on growth and development of G. pallida . This effect was significantly greater on Oc‐IΔD86‐expressing roots leading to a reduction in size of G. pallida females to a level at which fecundity is profoundly affected.