Purification and partial characterisation of hatching factors for the potato cyst nematode Globodera rostochiensis from potato root leachate

Nematology ◽  
2000 ◽  
Vol 2 (2) ◽  
pp. 231-236 ◽  
Author(s):  
Ken Devine ◽  
Peter Jones
Keyword(s):  
Mass Spectrometry ◽  
Molecular Weight ◽  
Globodera Rostochiensis ◽  
Cyst Nematode ◽  
Optimum Concentration ◽  
Potato Root ◽  
Potato Plants ◽  
Low Concentrations ◽  
Partial Characterisation

AbstractHatching factors with activity towards Globodera rostochiensis were isolated from the root leachates produced by 1500 potato plants grown in gravel. In total, nine hatching factors, each with molecular weight 530.5 Da, were isolated. The main differences in the daughter ions produced from different hatching factors after fragmentation by mass spectrometry occurred between 245 and 500 Da. The main hatching factor, HF E, was found in extremely low concentrations in the leachate (less than 2.9 × 10­5% of recovered organic material) and was active in vitro at less than 2.1 × 10­8 M. Separation of the different hatching factors during purification was found to decrease the percentage hatch achieved in vitro at optimum concentration. Aus Wurzelablaufwasser von 1500 in grobem Sand gehaltenen Kartoffelpflanzen wurden gegenüber Globodera rostochiensis wirksame Schlüpffaktoren isoliert. Insgesamt wurden neun Schlüpffaktoren isoliert, jeder mit einem Molekulargewicht von 530.5 Da. Die grössten Unterschiede bei den Tochterionen, die bei einer Zerlegung durch Massenspektrometrie erzielt wurden, traten zwischen 245 und 500 Da auf. Der Hauptschlüpffaktor, HF E, wurde in extrem niedrigen Konzentrationen (weniger als 2.9 × 10­5% des gewonnenen organischen Materials) im Ablaufwasser gefunden. Er war in vitro noch bei weniger als 2.1 × 10­8 M wirksam. Eine Trennung der verschiedenen Schlüpffaktoren im Laufe der Reinigung verminderte den prozentualen Schlupf in vitro bei optimaler Konzentration.

Influence of components of Globodera rostochiensis cysts on the in vitro hatch of second-stage juveniles

Nematology ◽  
2007 ◽  
Vol 9 (6) ◽  
pp. 837-844 ◽  
Author(s):  
Vitaly Dzhavakhiya ◽  
Galina Petelina ◽  
Edward Masler ◽  
Mikhail Pridannikov ◽  
Marina Palchuk
Keyword(s):  
Globodera Rostochiensis ◽  
Cyst Nematode ◽  
Distilled Water ◽  
Freezing And Thawing ◽  
Potato Root ◽  
Second Stage ◽  
Repeated Freezing ◽  
Fold Induction ◽  
High Concentrations

AbstractThe effects of cyst components of the potato cyst nematode, Globodera rostochiensis, on in vitro hatching of juveniles were evaluated. Aqueous homogenates of eggs and cyst walls, and aqueous rinses of cyst walls and eggs were examined. Homogenates of cyst walls or rinsates of either cyst walls or eggs each significantly increased hatch of juveniles five-fold compared with sterile distilled water controls. Cyst or egg homogenates induced significant increases (7.7-9.0-fold) in hatch of juveniles compared with controls. These values represent approximately 24.8-29.0% of the 31-fold induction of hatch of juveniles by potato root diffusate, routinely used to stimulate hatch in G. rostochiensis. The hatch inducing factor(s) in cyst homogenates was thermostable and retained activity after repeated freezing and thawing. Filter fractionation of cyst homogenate localised activity between molecular mass 0.5 kDa and 5 kDa, with no hatch inducing activity present above 5 kDa. Apparent depression of hatch induction at high concentrations of non-fractionated cyst homogenates suggests the presence of hatch inhibitors.

Effect of tuber-borne micro-organisms on hatching activity of potato root leachate towards potato cyst nematodes

Nematology ◽  
2003 ◽  
Vol 5 (1) ◽  
pp. 55-63 ◽  
Author(s):  
N. Aileen Ryan ◽  
Peter Jones
Keyword(s):  
Liquid Chromatography ◽  
Low Pressure ◽  
Globodera Rostochiensis ◽  
Globodera Pallida ◽  
Potato Cyst Nematodes ◽  
Cyst Nematodes ◽  
Potato Root ◽  
Potato Plants ◽  
Micro Organisms

AbstractPotato cyst nematodes hatch in response to hatching factors (HF) present in potato root leachate (PRL). The much higher spontaneous hatch (hatch in the absence of potato plants or PRL) of both Globodera rostochiensis and Globodera pallida in sand (32.2 and 21.1%, respectively) compared to in vitro (6.0 and 4.8%) experiments suggested the presence of other hatching factor-producing organisms in the non-sterile sand. When sterile PRL (from aseptically cultured microplants) and non-sterile PRL (from aseptically cultured microplants grown in the presence of tuber washings) samples were collected and assayed for hatching activity, the in vitro hatch of both PCN species but particularly of G. pallida was greater in non-sterile PRL. When these samples were fractionated on Sephadex G-10 by low pressure liquid chromatography and the fractions tested for hatching activity, the non-sterile PRL produced more hatching factors (HF) than the sterile PRL; in the fractionated sterile PRL only one significant HF (active towards G. pallida) was observed, compared to six (towards G. pallida) and three (towards G. rostochiensis) HF from the non-sterile PRL, with two HF being active towards both species. The non-sterile PRL appeared to produce more hatching factor stimulants (HS) and fewer hatch inhibitors (HI) than the sterile PRL. These results suggest that soil micro-organisms play an important role in the production of hatching chemicals and it is proposed that the differences in HF profiles between sterile and non-sterile PRL were due, at least in part, to increased HS production in the non-sterile PRL.

The effect of mycorrhization of potato roots on the hatching chemicals active towards the potato cyst nematodes, Globodera pallida and G. rostochiensis

Nematology ◽  
2004 ◽  
Vol 6 (3) ◽  
pp. 335-342 ◽  
Author(s):  
Aileen Ryan ◽  
Peter Jones
Keyword(s):  
Liquid Chromatography ◽  
Cyst Nematode ◽  
Globodera Pallida ◽  
Potato Cyst Nematodes ◽  
Cyst Nematodes ◽  
Potato Root ◽  
Potato Plants ◽  
Nitrogen Concentrations ◽  
Mycorrhizal Plants ◽  
Species Specific

AbstractComparison of potato root leachates (PRL) collected from the roots of mycorrhizal (using the mixed-isolate inoculum, Vaminoc) and non-mycorrhizal potato cv. Golden Wonder confirmed that mycorrhization caused a significant increase in hatching activity towards Globodera pallida but not G. rostochiensis. After fractionating the leachates by low pressure molecular exclusion/anion exchange liquid chromatography, several potato cyst nematode (PCN) species-specific hatching factors (HF) were found only in PRL from mycorrhizal plants. Leachate from mycorrhizal plants also contained more of several of those HF common to PRL from both mycorrhizal and non-mycorrhizal plants. Significantly more hatching factor stimulants (HS) active towards both PCN species were found in the PRL from mycorrhizal than from non-mycorrhizal plants; several HS were specific to mycorrhizal plants. No differences (quantitative or qualitative) were observed in hatching inhibitor (HI) levels between PRL from mycorrhizal and non-mycorrhizal plants. Mycorrhization of potato plants resulted in a 20% increase in carbon but a 48% decrease in nitrogen concentrations of the PRL compared to that from the non-mycorrhizal plants.

scholarly journals Effects of bacterial endophytes from potato roots and tubers on potato cyst nematode (Globodera rostochiensis)

2018 ◽  
Vol 19 (1) ◽  
pp. 47-51 ◽  
Author(s):  
NOOR ISTIFADAH ◽  
NURHAYATI PRATAMA ◽  
SYAHRIZAL TAQWIM ◽  
TOTO SUNARTO
Keyword(s):  
Globodera Rostochiensis ◽  
Cyst Nematode ◽  
Potato Cyst Nematode ◽  
Potato Seed ◽  
In Vitro Test ◽  
Bacterial Endophytes ◽  
Positive Effects ◽  
Bacterial Endophyte ◽  
Potato Growth

Istifadah N, Pratama N, Taqwim S, Sunarto T. 2018. Effects of bacterial endophytes from potato roots and tubers on potato cyst nematode (Globodera rostochiensis). Biodiversitas 19: 47-51. Bacterial endophytes are bacteria that inhabit plant tissues without causing any diseases. The endophytes existence may have negative, neutral, or positive effects on the host plants. This paper discusses the effects of bacterial endophytes isolated from potato roots and tubers on potato growth and their abilities to suppress potato cyst nematode, Globodera rostochiensis. The bacterial endophytes were isolated from roots and tubers of potatoes obtained from six plantation areas in West Java. The endophyte isolates were examined for their effects on potato growth. The non-pathogenic isolates were tested for their abilities to suppress G. rostochiensis in vitro and in potato plants. The results showed that from 88 bacterial endophyte isolates obtained, 13 isolates caused rot in potato seed pieces, 22 isolates inhibited the potato growth, while, 2 isolates increased the growth, and as many as 51 isolates did not influence the growth. The in vitro test using the isolate culture filtrate revealed that there were seven isolates that caused mortality of G. rostochiensis juvenile-2 by 67.5-97.7%. These isolates, however, were not effective in damaging the nematode eggs. In the greenhouse experiment, the bacterial endophyte isolates suppressed the number of cysts by 51.7-65.4% and that of the juvenile-2 of G. rostochiensis by 48.6-76.4%.

scholarly journals Effect of Selenium-Arabinogalactan Nanocomposite on the Colonization of Potato Plants in vitro by the Ring Rot Pathogen

2020 ◽  
Vol 32 ◽  
pp. 3-17
Author(s):  
A. I. Perfileva ◽  
◽  
O. A. Nozhkina ◽  
I. A. Graskova ◽  
N. S. Zabanova ◽  
...  
Keyword(s):  
Shoot Apex ◽  
Plant Tissues ◽  
Similar Data ◽  
Potato Root ◽  
Potato Plants ◽  
Ring Rot ◽  
Negative Effect ◽  
Pathogen Colonization ◽  
Root Tissues

It has been previously shown that the chemically synthesized nanocomposite of selenium with arabinogalactan (NC Se/AG) is characterized by antibacterial effect upon the agent of ring rot – gram-positive bacterium Clavibacter sepedonicus (Cms), with the NC Se/AG having no negative effect on potato plants. In the present paper, it has been found that, 1 hour after the treatment of the NC Se/AG, a substantial elevation of lipid peroxidation products was observed in potato root tissues. This supports earlier results on the increase in reactive oxygen species (ROS) production in potato root tissues under the influence of NC Se/AG. It is proposed that the increased ROS content in potato may inhibit pathogen colonization of plants. This has been tested by seeding homogenised plant tissues of various potato zones (roots, stems, shoot apex zone) onto the nutrient medium. In plants infected with Cms and untreated with the NC, the number of colony forming units (CFUs) of Cms has been shown to be numerous both in potato culture medium and in root and stem tissues. In shoot apex zone of such plants, it has been revealed, bacteria also present, but in smaller quantities. Similar data have been obtained by seeding homogenised tissues from roots and stems of potato plants treated with the NC followed by infection with Cms. However, seeding from shoot apex zones of the plants has been given 4 times less CFUs than from potato plants not treated with the NC. The effect of the NC Se/AG upon the pathogen colonization of plants appears to depend on the titre of the microorganism. In shoot apex zone of plants, characterized with small number of CFUs of Cms, the pathogen growth has been decreased. For the first time, Cms bacteria in potato plant tissues in vitro have been visualized with the aid of scanning microscopy.

HIGH CONCENTRATIONS OF HEPARIN ARE MORE INHIBITORY TO PLATE- AGGREGATION IN-VITRO THAN ARE LOW MOLECULAR WEIGHT HEPARINS AND HEPARINOIDS AT THE SAME CONCENTRATION

1987 ◽  
Author(s):  
H Messmore ◽  
B Griffin ◽  
J Seghatchian ◽  
E Coyne
Keyword(s):  
Molecular Weight ◽  
Heparan Sulfate ◽  
Dermatan Sulfate ◽  
Platelet Rich Plasma ◽  
Inhibitory Effect ◽  
Low Molecular Weight ◽  
Low Concentrations ◽  
High Concentrations ◽  
Induced Aggregation

Other investigators have shown that heparin in the usual therapeutic range (0.1-0.5 units/ml) has an enhancing effect on ADP aggregation and an inhibitory effect on collagen and thrombin induced aggregation. The effects of low molecular weight heparin (LMWH)and heparinoids (dermatan sulfate, heparan sulfate) on platelet aggregation have not been as extensivelystudied. We have utilized citrated platelet rich plasma (3.2%citrate-whole blood 1:9) drawn in plastic and adjusted to a final platelet count of 250,000/ul. A Bio-Data 4 channgl aggregometer was utilized with constantstirring at 37 C. The reaction was allowed to run for 20 minutes. Platelet rich plasma was supplemented 1:9 with saline or heparin and various agonists were then added ifno aggregation occurred. ADP, collagen, thrombin, ristocetin and serum from patients with heparin inudced thrombocytopenia (HIT) were utilized as agonists. Heparin was substituted at concentrations of 0.1 to 500 units per ml and various LMWH and heparinoids were substituted in equivalent anti-Xa or gravimetric concentrations. At low concentrations no inhibitory effect on any ofthe agonists was observed with any of the heparins or heparinoids. At concentrations of heparin of 100 u/ml or greater, all agonists were inhibited. At equivalent concentrations of five different LMWH (Cy 216, Cy 222, Pk 10169, Kabi 2165 and pentasaccharide) inhibition did notoccur at all or at very high concentions only. Dermatan sulfate and heparan sulfate inhibited only at high concentrations. HIT serum could not aggregate platelets with dermatan sulfate or pentasaccharide atany concentrations, but it was a good agonist with the other heparins and heparinoids.

Comparison of the in-soil hatching responses of Globodera rostochiensis and G. pallida in the presence and absence of the host potato crop cv. British Queen

Nematology ◽  
2005 ◽  
Vol 7 (4) ◽  
pp. 587-597 ◽  
Author(s):  
Ken Devine ◽  
Aileen Ryan
Keyword(s):  
Host Plant ◽  
Potato Crop ◽  
Globodera Rostochiensis ◽  
Cyst Nematode ◽  
Potato Plants ◽  
Significant Difference ◽  
Insertion Tool ◽  
Plant Emergence ◽  
The Mean ◽  
Presence And Absence

AbstractExperiments were conducted in a field in Co. Cork, Ireland, to investigate the hatching behaviour of the two potato cyst nematode (PCN) species, Globodera rostochiensis and G. pallida, in soil in the presence and absence of a host potato crop cv. British Queen. In order to compare the hatching behaviour of the two PCN species in the field, standardised populations of each PCN species (produced under identical conditions for the two previous generations and extracted without desiccation) were inserted into the field in tissue-embedding cassettes by means of a novel 'harpoon' insertion tool. Inserting cassettes of both PCN species at different distances from host potato plants and destructively sampling the contents of the cassettes over the first 8 weeks after plant emergence revealed that there was no significant difference in the hatch of either PCN species around the host plant. Hatches in excess of 80% for both PCN species were recorded 8 weeks after plant emergence over a range of depths (10-40 cm) and distances (10-50 cm) around the host potato plants. Substantial hatches of G. rostochiensis (82%) and G. pallida (88%) were also recorded at a depth of 20 cm in the furrow at harvesting. Monitoring the rate of spontaneous hatching of G. rostochiensis and G. pallida in the absence of host potato plants over the spring and summer months (a 20-week period) revealed that there was a significant difference in the rate of spontaneous hatching between the two PCN species, with G. rostochiensis juveniles hatching in greater numbers over the first 14 weeks than G. pallida juveniles. However, by mid-August the mean spontaneous hatch for both PCN species within the top 30 cm of soil was found to be 37%.

scholarly journals Towards quantitative mass spectrometry-based metabolomics in microbial and mammalian systems

2016 ◽  
Vol 374 (2079) ◽  
pp. 20150363 ◽  
Author(s):  
Rahul Vijay Kapoore ◽  
Seetharaman Vaidyanathan
Keyword(s):  
Mass Spectrometry ◽  
Molecular Weight ◽  
Biological Systems ◽  
High Sensitivity ◽  
Quantitative Mass Spectrometry ◽  
Small Molecular Weight ◽  
Quantitative Changes ◽  
Analytical Approaches

Metabolome analyses are a suite of analytical approaches that enable us to capture changes in the metabolome (small molecular weight components, typically less than 1500 Da) in biological systems. Mass spectrometry (MS) has been widely used for this purpose. The key challenge here is to be able to capture changes in a reproducible and reliant manner that is representative of the events that take place in vivo . Typically, the analysis is carried out in vitro , by isolating the system and extracting the metabolome. MS-based approaches enable us to capture metabolomic changes with high sensitivity and resolution. When developing the technique for different biological systems, there are similarities in challenges and differences that are specific to the system under investigation. Here, we review some of the challenges in capturing quantitative changes in the metabolome with MS based approaches, primarily in microbial and mammalian systems. This article is part of the themed issue ‘Quantitative mass spectrometry’.

In vitro nematicidal activity of a garlic extract and salicylaldehyde on the potato cyst nematode, Globodera pallida

Nematology ◽  
2011 ◽  
Vol 13 (7) ◽  
pp. 869-885 ◽  
Author(s):  
Wiseborn B. Danquah ◽  
Matthew A. Back ◽  
Ivan G. Grove ◽  
Patrick P.J. Haydock
Keyword(s):  
Cyst Nematode ◽  
Potato Cyst Nematode ◽  
Globodera Pallida ◽  
Garlic Extract ◽  
Potato Root ◽  
Nonylphenol Ethoxylate ◽  
Leachate Solution ◽  
Series Of Experiments ◽  
Toxic Constituent

AbstractThe in vitro nematicidal effects of an aqueous garlic extract, salicylaldehyde, a nonylphenol ethoxylate surfactant and a formulation containing these constituents were evaluated against the potato cyst nematode, Globodera pallida. Newly hatched, infective second-stage juveniles (J2) were placed for 24, 48 and 72 h in solutions containing concentrations of the formulation from 30.080.0 μl l–1 with 20% (v/v) potato root leachate and sterile distilled water controls. The garlic extract, salicylaldehyde and surfactant treatments were assessed at concentrations proportional to their occurrence in the formulation. Hatching assays involved a series of experiments in which G. pallida cysts were incubated for 8 weeks in potato root leachate solution containing different concentrations of the test substances. A second set of experiments involved incubating cysts in different concentrations of the test substances for 2, 4 and 8 weeks prior to hatching in potato root leachate solution to determine how prior exposure to these substances influences hatching and in-egg viability. The formulation caused 100% mortality at 75.0 μl l–1 with an LC50 of 43.6 μl l–1 after 24 h exposure. Salicylaldehyde was the most toxic constituent of the formulation with an LC50 of 6.5 μl l–1 after 24 h, while the garlic extract achieved 50% J2 mortality at 983.0 μl l–1, demonstrating that the formulation and salicylaldehyde are more toxic to G. pallida in vitro than oxamyl but less toxic when compared with aldicarb. The surfactant showed no dose-dependent toxic effects on J2 when compared with the controls. Emergence of J2 from the cysts was significantly reduced by concentrations of the formulation above 688.0 μl l–1 and its equivalent concentration of salicylaldehyde, while concentrations of the formulation above 2752.0 μl l–1 and the corresponding salicylaldehyde concentrations resulted in complete irreversible hatch inhibition. Concentrations of the garlic extract below 137.6 μl l–1 caused 26% more J2 hatch in comparison to the potato root leachate solution. This study has shown that salicylaldehyde is more toxic to nematodes than the garlic extract, and is the first report of a hatch stimulatory effect of a garlic extract on G. pallida under in vitro conditions.

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