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Analele Universitii din Craiova, seria Agricultur MontanologieCadastru Vol. XL/2 2010

CERCETRI PRIVIND ACTIVITATEA CATALAZEI I PEROXIDAZEILA PLANTELE DE FLOAREA SOARELUI INFECTATE CU CIUPERCI

FITOPATOGENE

RESEARCHES ON THE CATALASE AND PEROXIDASE ACTIVITY

AT SUN FLOWER PLANTS, INFECTED BY PHYTOPATOGENIC FUNGI

*LUMINITA BUSE- DRAGOMIR, **MARIANA NICULESCU

*University of Craiova, Faculty of Horticulture, Departament of Plant Physiology, Libertii 19, Craiova

**University of Craiova, Faculty of Agriculture, Departament of Botany, Libertii 19, 200583,Craiova,

e-mail:[email protected],

**University of Craiova, Faculty of Horticulture, Departament of Plant Physiology, Libertii 19, Craiova

Key w ords: c atalase, peroxidase, sunf low er, fungus

REZUMAT

Procesele enzimatice din esutul foliar parazitat al plantelor de floarea sorelui sufern cele mai multe cazuri modificri patologice. Interpretarea modificrilor privind activitateaenzimatic a plantelor se bazeaz pe cunoaterea rolului fiziologic al enzimelor i almodului n care are loc reglarea activitii lor n planta sntoas.

Peroxidaza i catalaza au o mare importan n viaa plantelor, avnd rol nrespiraia celular i n aprarea mpotriva stresului oxidativ.

Sub aciunea ciupercilor fitopatogene, activitatea celor dou enzime sufermodificri, care difer n funcie de tipul de patogen i de stadiul de evoluie a bolii.

ABSTRACT

The enzymatic processes in the infected leaf tissue of sunflower plants are mostlydamaged by pathological changes. The interpretation of changes in the enzymatic activityof plants is based on knowing the physiological role of enzymes and the way their activityis adjusted in the healthy plant.

Peroxidase and catalase are of great importance in the plants existence, especiallyfrom the cell respiration point of view and the defense against oxidative stress.

The activity of the two enzymes changes under the action of phytopathogenic fungi. Itdiffers from the point of view of the pathogen type and the disease evolution degree.

INTRODUCTION

The interpretation of the changes on the enzymatic component of plants is based onknowing the physiological influence of the enzymes and adjusting their activity in thehealthy plant.

Although there are facts about the enzymatic changes and enzymatic etiologysymptoms at damaged plants, these facts have been elucidated, due to the interpretationsof oxidative enzymes.

Peroxidase and catalase generate an oxidation-reaction between the H2O2, as anelectron acceptor and many sub-layers: phenol substances, aromatic amine, ascorbicacid, ironcitochrom.
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Although the physical and chemical features of peroxidase and catalase are wellknown, there are little facts about their functions at sprouts.

There are surveys that point out the difference between plant varieties, regarding theactivating of the oxidative metabolism in case of plant damage. There are also facts aboutthe influence of oxidative processes disturbances on the resistance of plants. All theseacknowledge the important role oxidative plant system has in maintaining its resistance

capacity.

MATERIAL AND METHOD

The late simple sunflower hybrid was used as a biologic material, FLOROM 350,which has been recommended for the cropping areas in the southern and northern part ofthe country, with a high resistance to draught and white-rot-proof, which is produced by theSclerotinia sclerotiorum, and to brown-rot-proof, produced by the Botrytis cinerea.

When surveys were taken, artificial infections with phytopathogenic fungus on thesunflower leaves were caused on purpose:

V1-witness, healthy, uninfected plants;

V2-plants infected by Plasmopara helianthifungus;V3- plants infected by Puccinia helianthifungus;

V4- plants infected by Septoria helianthifungus;

V5- plants infected by Phomopsis/Diaporthe helianthifungus;For causing the Plasmopara helianthi infection there were used inoculums made

up of a zoospore suspension in water and agar 1%.The suspension was injected into the leaves epidermis, and, in order to maintain

the humidity plants were covered, for 72 hours with a plastic foil.In order to obtain the Puccinia helianthifungus infection, inoculums obtained from

uredopsores taken from damaged plants from the previous year were used. The

spraying technique was used; the uredospores suspension was sprayed on the leaves.The inoculation was made on four leaves, while the incubation lasted between 7and 10 days.

For the Septoria helianthiinfection picnide leaves a year old were used and keptin the fridge, at -10C.

An average temperature of 25C was chosen for generating the infection. Thetested plants were afterwards sprayed with a picnospores suspension, at a 5-7 cmdistance from the leaves. They were then covered with plastic see-through bags, for 72hours.

In order to obtain the Phomopsis/Diaporthe helianthifungus infection ascosporescollected from perietecia were used. The latter developed on the damaged stalks

(picked in the autumn and kept in the fridge, at a -10C). These stalks have beenpreviously taken to the humid room and kept at a 20C temperature, where perieteciadeveloped and grew naturally, in 10 days.

Cortical tissues have been scraped with a lancet, and then mixed andhomogenized.

The suspension that was obtained was filtered and plants with 4-5 leaves weresprayed with it.

The intensity of the catalase and peroxidase actions was determined by thegasometrical method. The results were expressed in cm3 O2/experimental materialquantity/working period.

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RESULTS AND DISCUSSIONS

The activity of the catalase in the sunflower undamaged and damagedleaves

The surveys that were taken pointed out a poor enzymatic activity at thesunflower sprouts but, to the end of the experimental period (i.e. 30 days), the

enzymatic value tripled. Thus, its value is still poor, comparing it to the values that arementioned in specialist literature.

The plants damaged by pathogenic fungi had a different dynamic than thecatalase one.

As there can be seen from gr. 1, Plasmopara helianthifungus did not essentiallychange the catalase activity. In the 12thday from the inoculation, there was noticed aconstant increase, in comparison with the values of the witness plants.

Graphic 1. The act iv i ty of th e catalase(ml O2/g /15m in ) in the leaves

damaged by the Plasmo pra fungus

The leaves of the plants that are damaged by the Puccinia helianthihad a poorerenzymatic activity than the leaves of the witness plants. Nevertheless, differences arelittle(gr. 2).

The catalase enzymes activity in the leaves of the plants that were damaged bythe Septoria helianthifungus(gr 3) had similar values with the witness plants activity. Inthe 30th day from the inoculation there was noticed an increase of the enzymaticactivity (59ml O2/g/15min, in comparison with 51ml O2/g/15 min at witness plants).

The Phomopsis helianthifungus had the highest influence on the catalase, whoseactivity decreased during the experimental period. After 30 days from the inoculationperiod the catalysis value reached 35ml O2/g/15min. This value only represents64.81% of the value that was noticed at witness plants.

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Graphic 2. The act iv i ty of th e catalase(ml O2/g /15m in ) in the leaves damaged

by the Puccin ia helianth i fungus

Graphic 3. The act iv i ty of th e catalase (ml O2/g /15m in ) in the leaves

damaged by Septoria hel ianth i

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infected plants witness

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.

Graphic 4. The act iv i ty of th e catalase (ml O2/g /15m in ) in the leaves

damaged by Phomops is he l ian th i

The peroxidase act iv i ty at undamag ed and damaged su nf low er leaves

As opposed to catalase, whose activity did not change under the influence of thefungus infection, the peroxidase activity increased, up to 100% and even more.

This fact determines us to state that the peroxidase activity, under the influence ofthe fungus infection is a specific chemical reaction, which allows us to estimate the plantsability to protect against fungi.

According to diagrams no. 5-8, the peroxidase activity at undamaged plants duringthe determination period was different from the catalysis activity. The highest values werenoticed at the beginning of the experimental period, and the least, at the end of thedetermination period.

At damaged plants, the peroxidase activity increased, especially after 30 days ofinfection.

Plasmopara helianthifungusdamage determined a 100% increase of the peroxidaseactivity, beginning with the 23rdday from the infection. After 30 days, it reached an 89mlO2/g/15min value, which represents 128%, as opposed to the value noticed at witnessplants.

Puccinia helianthi fungus caused the intensification of the peroxidase activity, butwith lower values: after 18 days- 99ml O2/g/15min, and afterwards it slightly decreases.Nevertheless, the values are higher as opposed to those of witness plants.

At plants damaged by Septoria helianthi fungus the peroxidase activity increaseduntil the 18th day the infection. After a 30 days time, the values maintained high, incomparison with the witness plants ones.

The peroxidase activity dynamics, during the experimental period at plants damagedby Phomopsis helianthiwas different, as opposed to the previous plant fungi.

After two and five days after the infection, there were not significant values incomparison with the witness plant. After 12 days, the peroxidase activity doubled, reaching

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to a 98ml O2/g/15min value; it decreased afterwards and, after 30 days from the infection,it reached a 68ml O2/g/15min value, i.e. a double value as opposed to witness plants.

Graphic 5. The act iv i ty of the peroxid ase(ml O2/g /15m in ) in the leav es

damaged by the Plasmo pra fungus

Graphic 6. The act iv i ty of th e peroxidase(ml O2/g /15m in ) in the leav es damaged

by the Puccin ia hel ianth i fung us

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Graphic 7. The act iv i ty of the peroxid ase(ml O2/g /15m in ) in the leav es

damaged by Septoria hel ianth i

Graphic 8. The act iv i ty o f the perox idase(ml O2/g /15m in ) in the leaves

damaged by Phomops is he l ian th i

The causes of the increase of peroxidase activity, under the influence of infectionhave not been completely identified. Some authors think the cause of the peroxidaseactivity intensification is the neo formation of a peroxidase protein quantity with a specificcatalytic action. In this case, a de novo synthesis of the peroxidase took place, a process

which has been induced by the toxic secretions of the parasite.The increase of the peroxidase activity could also due to the metabolism produces of

the fungus. It cannot be estimated what produces we are dealing with, because

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peroxidase works on a large amount of substances: glucose, tannins, aromatic amine,phenols, amino acid.

CONCLUSIONS

- The Plasmopara helianthifungus did not change the activity of the catalase in theleaves;

- The leaves of the plants damaged by the Puccinia helianthi fungus had a poorer

enzymatic activity in comparison with witness plants;

- The Phomopsis helianthi fungus highly influenced the catalase process, whoseactivity progressively decreased during the experimental period;

- Peroxidase increased its activity, up to 100%, even more;

- The activity of the peroxidase, under the influence of infection is a specific chemicalreaction, which allows us to estimate the plants ability to protect.

BIBLIOGRAPHY

1.Atanasiu L., ipa Liubov, 1991Imunitatea la plante. Ed. Ceres, Bucureti2.Heitefuss R., Williams P.H., 1976Encyclopedia of Plant Pathologhy. Springer

Verlag, Berlin, Heidelberg, New York.3.Hulea Ana, 1991 Principalele boli produse de paraziii vegetali la floarea

soarelui.4.Lamarque Claudine, 1980Obtention dascospores de Sclerotinia Sclerotiorum

et technique dinoculation utilisables dans la slection varitale du tournesol.CETIOM, Paris.

5.Mititiuc M., 1992 Probleme actuale privind rezistena plantelor fa de ageniipatogeni i perspectivele folosirii soiurilor rezistente n prevenirea i combatereabolilor plantelor cultivate n Romnia. Natura, nr 1, anul XLV.

6.Novotel`Nova N.S. , 1996False mildew of sunflower. Taxonomy and biology ofthe causal agent, pathogenesis of the disease. Moscow Leningrad in Rev. Apll.Mycol.7.Pilarge E., 1997Mildou du tournesol: la situation au champ, in: Les rencontresannuelles du CETIOMTournesol.Paris

8.Rdulescu E., Negru A. , 1973 Septoriozele din Romnia, Ed. Ceres,Bucureti.9.Sackston W.E., 1981Downy mildew of sunflowers in: The downy mildews. D.M. Spencer dit. Academic Press. London, New York, San Francisco.10.Snea N., Tabr V., Cszos I., Vlad Silvia, 1996Aspecte economice ale

proteciei florii-soarelui(Helianthus annus L) fa de atacul ciupercii Diaporthehelianthi Munt. Cvet., f.c. Phomopsis helianthi Munt. Cvet., n sistem integrat deproducie.Lucr. t. USAMVBT, Cultura plantelor de cmp: 103-109.11.Semal J. , 1993 - Trait de Pathologie vegetale. Press Agronomique deGembleux, cap. 8. La relations hte - parasite(pg. 249302).12.Sebanek J., 1992Plant Physiology, Elsevier, Amsterdam, Oxford, New York,Tokio.13.Timur Dken M.,1989 Plasmopara halstedii in sunflower seeds and the roleof infected seeds in producing plants witk systemic symptoms.J. Phytopathology,14.Toma Liana, Jitreanu Doina, Andrei Elena, 2002La relation entre quelquesindicateurs morfo-physiologiques et la formations de la recolte de semences de

tournesol.Lucrri tiinifice, Seria Agronomie.15.Vrnceanu V.A., 2000Floarea soarelui hibrid.Ed. Ceres, Bucureti.

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