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Detection of protease and protease inhibitors during development of soybean crown nodules

Dissertation (MSc)--University of Pretoria, 2010.

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Other Authors: Kunert, Karl J.
Format: Thesis
Published: University of Pretoria 2013
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access_status_str Open Access
author2 Kunert, Karl J.
author_browse Kunert, Karl J.
author_facet Kunert, Karl J.
collection Thesis
dc_rights_str_mv © 2010 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria.
description Dissertation (MSc)--University of Pretoria, 2010.
format Thesis
id oai:repository.up.ac.za:2263/29724
institution University of Pretoria (South Africa)
last_indexed 2026-06-10T12:39:42.666Z
license_str Other — see source repository
provenance_str_mv Harvested via OAI-PMH from UPSpace — University of Pretoria Institutional Repository
publishDate 2013
publishDateRange 2013
publishDateSort 2013
publisher University of Pretoria
publisherStr University of Pretoria
record_format dspace
source_str UPSpace — University of Pretoria Institutional Repository
spelling oai:repository.up.ac.za:2263/29724 Detection of protease and protease inhibitors during development of soybean crown nodules Kunert, Karl J. Schluter, Urte lufuno_mashamba@yahoo.com Mashamba, Lufuno Abigail Protease inhibitors Legume nodule formation Soybean crown nodules UCTD Dissertation (MSc)--University of Pretoria, 2010. A symbiotic association between leguminous roots and soil fixing nitrogen bacteria is required for legume nodule formation. The primary function of nodules is the fixation of nitrogen from the atmosphere into an accessible form for plants. In this study, nodules of plants of the soybean cultivar Prima 2000 were characterized and their number and weight were determined during nodule development. Their nitrogen-fixing activity during nodule development was determined by color evaluation. A pink nodule color showed active leghemoglobin required for nitrogen fixation and a green nodule color nonfunctional leghemoglobin. Strong appearance of nonfunctional leghemoglobin in the later stages of nodule development during senescence was accompanied by an increase in protease activity within crown nodules demonstrated by gelatine-containing SDS PAGE. Cysteine protease activity was identified as a major protease activity during nodule senescence when the cysteine protease inhibitor E-64 was used to block total protease activity. Products, which may indicate the expression of cysteine protease inhibitors during nodule development, were detected with the reverse zymogram technique and Western blotting. However, these bands have not been characterized so far in more detail. Putative transgenic plants were produced using the Agrobacterium transformation technique to allow determining the activity of native and mutated papaya cysteine protease inhibitor coding sequences. These sequences will ultimately be used for soybean transformation to reduce cysteine protease activity in nodules. However, the presence of coding sequence in the genome of these putative transgenic plants could not be confirmed by gene amplification and protease activity testing. Overall, this study has contributed to establish parameters to measure nodule growth and performance during development. Plant Science unrestricted 2013-09-07T16:24:29Z 2010-11-23 2013-09-07T16:24:29Z 2010-09-02 2010-11-23 2010-11-23 Dissertation Mashamba, LA 2010, Detection of protease and protease inhibitors during development of soybean crown nodules, MSc dissertation, University of Pretoria, Pretoria, viewed yymmdd < http://hdl.handle.net/2263/29724 > D10/742/ag http://hdl.handle.net/2263/29724 http://upetd.up.ac.za/thesis/available/etd-11232010-114246/ © 2010 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. application/pdf University of Pretoria
spellingShingle Protease inhibitors
Legume nodule formation
Soybean crown nodules
UCTD
Detection of protease and protease inhibitors during development of soybean crown nodules
title Detection of protease and protease inhibitors during development of soybean crown nodules
title_full Detection of protease and protease inhibitors during development of soybean crown nodules
title_fullStr Detection of protease and protease inhibitors during development of soybean crown nodules
title_full_unstemmed Detection of protease and protease inhibitors during development of soybean crown nodules
title_short Detection of protease and protease inhibitors during development of soybean crown nodules
title_sort detection of protease and protease inhibitors during development of soybean crown nodules
topic Protease inhibitors
Legume nodule formation
Soybean crown nodules
UCTD
url http://hdl.handle.net/2263/29724
http://upetd.up.ac.za/thesis/available/etd-11232010-114246/