Protease activity and phytocystatin expression in Arabidopsis thaliana upon Heterodera schachtii infection

2016 ◽  
Vol 109 ◽  
pp. 416-429 ◽  
Author(s):  
Mateusz Labudda ◽  
Elżbieta Różańska ◽  
Joanna Szewińska ◽  
Mirosław Sobczak ◽  
Jolanta Maria Dzik
1996 ◽  
Vol 112 (4) ◽  
pp. 1421-1427 ◽  
Author(s):  
A. Bockenhoff ◽  
DAM. Prior ◽  
FMW. Grundler ◽  
K. J. Oparka

Nematology ◽  
2015 ◽  
Vol 17 (9) ◽  
pp. 1027-1043 ◽  
Author(s):  
Shahbaz Anwar ◽  
Erich Inselsbacher ◽  
Florian M.W. Grundler ◽  
Julia Hofmann

The plant-parasitic cyst nematode Heterodera schachtii induces syncytial feeding structures in the roots of host plants. These syncytia provide all required nutrients, water and solutes to the parasites. Previous studies on the composition of primary metabolites in syncytia revealed significantly increased amino acid levels. However, mainly due to technical limitations, little is known about the role of arginine in plant-nematode interactions. This free amino acid plays a central role in the plant primary metabolism and serves as substrate for metabolites involved in plant stress responses. Thus, in the present work, expression of genes coding for the enzymes of arginine metabolism were studied in nematode-induced syncytia compared to non-infected control roots of Arabidopsis thaliana. Further, amiRNA lines were constructed and T-DNA lines were isolated to test their effects on nematode development. While the silencing of genes involved in arginine synthesis increased nematode development, most T-DNA lines did not show any significant difference from the wild type. Amino acid analyses of syncytia showed that they accumulate high arginine levels. In addition, manipulating arginine cycling had a global effect on the local amino acid composition in syncytia as well as on the systemic amino acid levels in roots and shoots.


2016 ◽  
Vol 33 ◽  
pp. S157
Author(s):  
Dominik Kanigowski ◽  
Mateusz Matuszkiewicz ◽  
Joanna Dąbrowska ◽  
Anna Barczak ◽  
Marcin Filipecki

2000 ◽  
Vol 13 (3) ◽  
pp. 309-315 ◽  
Author(s):  
Dieter Hermsmeier ◽  
Jennifer K. Hart ◽  
Marina Byzova ◽  
Steven R. Rodermel ◽  
Thomas J. Baum

Gene expression changes in plant roots infected by plant-parasitic cyst nematodes are involved in the formation of nematode feeding sites. We analyzed mRNA abundance changes within roots of Arabidopsis thaliana during the early compatible interaction with Heterodera schachtii, the sugarbeet cyst nematode. Approximately 1,600 root sections, each containing a single parasitic nematode and its feeding site, and 1,600 adjacent, nematode-free root sections were excised from aseptic A. thaliana cultures 3 to 4 days after inoculation with H. schachtii. These tissue samples were termed infected and uninfected, respectively. Preparasitic nematodes were added to the uninfected tissue sample to maintain the nematode to plant tissue proportion. mRNA extracted from these two tissue samples was subjected to differential display analysis. Thirty-six cDNA clones corresponding to mRNA species with different abundance between both tissue samples were isolated. Of these clones, 24 were of A. thaliana origin and 12 were from H. schachtii. Differential display data predicted that the A. thaliana cDNA clones corresponded to 13 transcripts that were more abundant in the infected root sections and 11 transcripts that were more abundant in the uninfected root sections. H. schachtii cDNA clones were predicted to correspond to four transcripts that were more abundant in parasitic nematodes and to eight transcripts that were more abundant in preparasitic nematodes. In situ hybridization experiments confirmed the mRNA abundance changes in A. thaliana roots predicted by the differential display analyses for two A. thaliana clones.


Parasitology ◽  
1994 ◽  
Vol 109 (2) ◽  
pp. 249-255 ◽  
Author(s):  
A. Böckenhoff ◽  
F. M. W. Grundler

SUMMARYA method was developed, which enables substances to be injected into the feeding structure (syncytium) established by Heterodera schachtii in roots of Arabidopsis thaliana. The technique was used to study the uptake of nutrients by the feeding nematode. The fluorescent dye lucifer yellow CH (LYCH) and fluorescence-labelled dextrans of different molecular weights were injected into the thin and translucent roots of A. thaliana. Such roots are a feature of this plant and they provide optimal conditions for microinjection. Injected LYCH was taken up by feeding juveniles and adults, indicated by the staining of the alimentary duct and the digestive system. Fluorescent dextrans of 3, 10 and 20 kDa but not of 40 and 70 kDa were ingested, suggesting that molecules of a maximum Stokes radius of 3·2 to 4·4 nm are taken up. It is likely that the feeding tube, forming the interface between the plant cytosol and the nematode's digestive system, is responsible for this size exclusion effect. The injected fluorescent substances were not detected in plant cells adjacent to the syncytium or in the root vascular elements. Injections into parts of roots which were infested by several nematodes revealed that feeding H. schachtii individuals may share one syncytium.


2020 ◽  
Vol 69 (4) ◽  
pp. 794-803 ◽  
Author(s):  
Mateusz Labudda ◽  
Elżbieta Różańska ◽  
Ewa Muszyńska ◽  
Dorota Marecka ◽  
Maria Głowienka ◽  
...  

Nematology ◽  
2015 ◽  
Vol 17 (4) ◽  
pp. 401-407 ◽  
Author(s):  
Ritushree Jain ◽  
Catherine J. Lilley ◽  
Peter E. Urwin

Phytates are mixed cationic salts of phytic acid formed by sequential phosphorylation of myo-inositol. Phytate is a phosphorus storage molecule essential for cellular and hormonal signalling in plants but exhibits anti-nutrient properties in animals. Low phytate plants have reduced basal resistance towards microbial pathogens and reduced tolerance to environmental stresses resulting in compromised yields. We report that three mutant lines of Arabidopsis thaliana, each with altered expression of myo-inositol-3-phosphate synthase (MIPS) isoforms, show altered susceptibility towards infection by the beet cyst nematode, Heterodera schachtii. Disruption of MIPS2 accompanied by increased MIPS1 expression results in reduced cyst nematode infection. Lack of MIPS3 resulted in a higher proportion of second-stage juveniles in the early phase of infection, suggesting delayed nematode development on mips3 mutants. Reduction in total phytate by down-regulation of the inositol polyphosphate kinase gene (IPK1) resulted in higher susceptibility to cyst nematode infection but a reduced average size of adult females. However, specific down-regulation of MIPS gene expression reduces susceptibility as myo-inositol is required to feed into the myo-inositol oxygenase pathway, which has an important role in development of the cyst nematode feeding site.


Nematology ◽  
2007 ◽  
Vol 9 (3) ◽  
pp. 317-323 ◽  
Author(s):  
Florian Grundler ◽  
Julia Hofmann

AbstractSedentary plant-parasitic nematodes, such as cyst and root-knot nematodes, induce feeding structures in the host root that undergo extensive changes in the gene expression. This phenomenon has previously been studied by gene chip analysis and qRT-PCR. Housekeeping genes are often used routinely as internal references for relative qRT-PCR analyses. However, due to the strong influence of nematode infection on host cell metabolism and physiology, expression of housekeeping genes may be altered considerably, thus limiting reliability of qRT-PCR analyses. Therefore, in the present work we tested UBQ10, ACT2, EF1a, UBP22 and 18S rRNA as potential candidates for relative qRT-PCR studies of gene expression in nematode infection sites in roots of Arabidopsis thaliana. Among the tested candidates only UBP22 and 18S rRNA were stably expressed and, therefore, are reliable reference genes for studying cyst and root-knot nematode infections.


Nematology ◽  
1999 ◽  
Vol 1 (4) ◽  
pp. 363-374 ◽  
Author(s):  
Miroslaw Sobczak ◽  
Wladyslaw Golinowski ◽  
Florian M.W. Grundler

Abstract The development of feeding plugs and feeding tubes formed in syncytia induced by the cyst-forming nematode Heterodera schachtii in roots of Arabidopsis thaliana was examined at the ultrastructural level. The feeding plug was first observed 24 h after selection of the initial syncytial cell (ISC) and was present throughout the entire nematode life cycle. In later stages of nematode development the feeding plug became increasingly robust and infiltrated by fibrillar syncytial wall material while the central part, through which the nematode stylet was inserted, retained an amorphous structure. Neither the feeding plug nor the nematode stylet were observed to penetrate the plasmalemma of the syncytium. After the nematode completed the preparation phase for feeding, the first secretions were released from the stylet orifice and emitted through the plasmalemma into the cytoplasm. They formed uniformly osmiophilic wavy tubes without an electron translucent lumen. The first typical feeding tubes were found 24 h after ISC selection and were composed of an electron dense wall and an electron translucent lumen. The size of a single feeding tube was about 1 X 4 mum. No difference occurred between feeding tubes formed by male and female juveniles. Frequently, membranes of the endoplasmic reticulum were connected to the wall of feeding tubes. After the nematode completed feeding, the tubes were disassociated from the stylet orifice and were dispersed in the syncytial cytoplasm. The feeding tube lumen was filled with cytoplasm and the wall gradually degraded. Die Ultrastruktur der von Heterodera schachtii gebildeten Dichtungsstopfen und Saugrohrchen - Die Entwicklung von Dichtungsstopfen (feeding plugs) und Saugrohrchen (feeding tubes), die in den von dem Zystennematoden Heterodera schachtii induzierten Synzytien in den Wurzeln von Arabidopsis thaliana gebildet werden, wurden auf der Ebene der Ultrastruktur untersucht. Die ersten Dichtungsstopfen wurden 24 h nach der Auswahl der Synzytiuminitialzelle (ISC) durch den Nematoden beobachtet. Sie waren wahrend des gesamten Lebenszyklus des Nematoden vorhanden. In den spateren Entwicklungsstadien der Nematoden wurde der Dichtungsstopfen zunehmend robuster und von fiblillarem Material aus der Zellwand durchdrungen. Der zentrale Teil des Stopfens, durch den der Mundstachel des Nematoden in die ISC ragte, behielt eine amorphe Struktur. Es wurde nicht beobachtet, dass der Dichtungsstopfen oder der Mundstachel das Plasmalemma durchbrachen. Nachdem der Nematode die vorbereitende Phase der Nahrungsaufnahme abgeschlossen hatte, traten die ersten Sekrete aus der Mundstacheloffnung aus und wurden durch das Plasmalemma hindurch in das Zytoplasma abgegeben. Sie bildeten gleichmassig osmiophile, wellige Rohrchen ohne ein elektronendurchlassiges Lumen. Die ersten typischen Saugrohrchen wurden 24 h nach der Auswahl der ISC gefunden. Sie bestanden aus einer elektronendichten Wand und einem elektronendurchlassigen Lumen. Die Grosse eines einzelnen Saugrohrchens betrug ungefahr 1 X 4 mum. Zwischen den von mannlichen oder von weiblichen Juvenilen gebildeten Saugrohrchen traten keine Unterschiede auf. Haufig waren Membranen des endoplasmatischen Reticulums mit den Wanden von Saugrohrchen verbunden. Wenn der Nematode die Nahrungsaufnahme abgeschlossen hatte, wurden die Saugrohrchen von der Offnung des Mundstachels getrennt und im synzytialen Zytoplama verteilt. Das Lumen dieser Saugrohrchen war mit Zytoplasma gefullt, und die Wand wurde langsam abgebaut.


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