Skip to main content
NCBI home page
Saudi Journal of Biological Sciences logoLink to Saudi Journal of Biological Sciences
. 2021 Apr 24;28(8):4442–4445. doi: 10.1016/j.sjbs.2021.04.039

Effect of some fertilizers on hatching of cereal cysts nematode, Heterodera avenae

Saroj Yadav 1,⁎, RS Kanwar 1
PMCID: PMC8325024  PMID: 34354428

Abstract

Experiments were conducted in laboratory and pot conditions to determine the effects of urea, di-ammonium phosphate (DAP), single super phosphate (SSP), muriate of potash (MOP) and zinc sulphate (ZnSO4) on hatching of Heterodera avenae. Two concentrations of each fertilizer were tested in lab for which 10 cysts and 5 ml of each concentration were taken in 5 cm diameter Petri plates. Observations were recorded at weekly intervals up to six weeks. Urea, DAP, SSP and MOP inhibited hatching and ZnSO4 increased it. After six weeks, hatching was least (5.45%) in higher dose of urea and greatest (46.9%) in higher dose of ZnSO4. In pot experiment, two doses of urea and single dose of SSP, MOP, and ZnSO4 were applied in H. avenae-infested soil and WH-1105 wheat was sown. Observations on nematodes in roots, soil and remaining cyst contents were recorded 40 days after sowing. Among all the fertilizers, least nematodes in soil and roots were found at higher dose of urea and greatest number in ZnSO4.

Keywords: Cereal cyst nematode, Heterodera avenae, Hatching, Fertilizers

1. Introduction

Cereal cyst nematodes have worldwide distribution and are documented to be of economic importance on wheat mainly in light textured soil and where predominately cereal monoculture is practiced (Nicol and Rivoal, 2008). Organic and synthetic fertilizers had a suppressant effect on nematodes (Kaplan and Neo, 1993). Application of fertilizers affects nematode populations indirectly by increasing the nematode feeding or by providing nutrition to compensate the plant from the nematode feeding (McIntoch et al., 1999). On the other hand, nutrient deficiency may make the plant weak and more susceptible to nematode attack (Melakeberhan et al., 1997). The direct effect of fertilizers on nematodes may alter nematode behavior and reproduction which may result in either a decrease or increase of the population. The effectiveness of fertilizers in changing nematode population depends on the fertilizer components and their active ingredients (Gruzdeva et al., 2007). The fertilizer components may be lethal directly to nematodes or they may alter both pH and salinity of the soil which in turn affect nematode populations (Oka and Pivonia, 2002, Tenuta and Ferris, 2004). Some studies have shown that inorganic fertilizers affect the hatching of nematode species (Tefft and Bone, 1984, Clarke and Shepherd, 1966).

Some nutrients have been reported to inhibit nematode hatching (Habash and Al-Banna, 2011, Zhen-yue et al., 2013) while others to act as stimulant (Behm et al., 1995, Tefft and Bone, 1984). Heterodera avenae is a serious pest of cereals- wheat, barley and oat (in European conditions), however rice is also a cereal but H. avenae won’t affect it. Unlike Globodera spp., its hatching is not dependent on root exudates of host plants. But some factors do affect its hatching which continues for more than two months and all the eggs do not hatch in one year (Kanwar and Bajaj, 2011, Clarke and Shepherd, 1966). Fertilizers are commonly applied to all crops/wheat according to their requirement and soil fertility status. These fertilizers having some suppressive/toxic, stimulating/attractant and neutral effect on plant diseases or pathogens (Dordas, 2008) so, present study examined the effects of synthetic fertilizers on hatching of cereal cyst nematode in laboratory and pot conditions.

2. Materials and methods

2.1. Location

The experiments were conducted in laboratory and pot conditions during rabi season of 2016–17 in the screen house of Department of Nematology, Choudhary Charan Singh Haryana Agricultural University, Hisar, Haryana (India) which has situated at Latitude − 29°10′ N, Longitude − 75° 46′ E, Altitude − 215.2 m.

2.2. Experimental procedure

Laboratory experiment was done in 5 cm diameter Petri plates. Urea, di-ammonium phosphate (DAP), single super phosphate (SSP), muriate of potash (MOP) and zinc sulphate (ZnSO4) were included in the study. These fertilizers are commonly used by farmers as sources of nitrogen, phosphorus, potash and zinc, respectively, in India. The recommended doses of fertilizers in wheat NPK and Zn mg per kg soil were converted in to the ppm for the laboratory studies. Required PPM concentration of each fertilizer was prepared by mixing calculated fertilizers amount in one liter of water. Two concentrations of each fertilizer at single and double dose of recommended dose for wheat crop were prepared. Ten hand-picked cysts were placed in each Petri dish containing 5 ml of each fertilizer’s concentration. Cysts placed in water served as control. Each treatment was replicated four times. The cysts were incubated for six weeks at room temperature (December 2016 to January 2017, temp. 15–25 °C). The hatched J2 were counted at weekly intervals, up to six weeks. Un-hatched cyst contents were estimated by crushing the cysts under a dissecting microscope and per cent hatching (larval emergence) was calculated.

%Larval emergence=Total larval emergence(Average cyst content×10)+Totallarval emergence×100

In second experiment, H. avenae susceptible wheat cultivar WH-1105 (Anonymous, 2014) was sown in 15 cm diameter earthen pots having 1 kg soil. Initial inoculum was 13 eggs and J2/g soil (327 eggs and juveniles cyst−1). The soil was got analyzed from Department of Soil sciences, CCS HAU, Hisar. It had sand, silt and clay 89.7, 4.5 and 5.8 per cent, respectively, and initial level of N = 116 kg/ha, P = 15 kg/ha, K = 165 kg/ha and Zn = 1.31 ppm. The recommended doses of NPK and Zn in wheat has 150, 60, 60 and 25 kg ha−1 (163 mg urea, 187.5 mg SSP, 50 mg MOP and 59.5 mg ZnSO4 kg soil−1) respectively, in Haryana, India. Recommended doses of urea, SSP, MOP, ZnSO4, combination of all fertilizers, half dose of urea (split dose of urea applied at the time of sowing) and without fertilizers were taken as treatment (Table 2). The treatments with three replications were arranged in completely randomized design on a bench of screen house. After 40 days of sowing observations were recorded on soil and root population, and cyst contents. For recording root population, roots were stained by acid fuchsin-lactophenol (McBeth et al., 1941). Cysts from soil were extracted by Cobb’s method (Cobb, 1918) and juveniles by modified Baermann funnel method (Schindler, 1961).

Table 2.

Effect of fertilizers on hatching of Heterodera avenae in pots grown with wheat.

Treatments Nematodes/200 cc soil Nematodes/root system Left over contents per cyst
No fertilizer (control) 185.3e 161.0d 113.7a
Urea 163 kg /ha 68.3ab 89.7ab 212.3d
Urea 326 kg /ha 60.7a 79.0a 229.0e
SSP 375 kg /ha 95.0bc 110.0bc 182.0c
MOP 100 kg /ha 106.0c 119.3c 174.7c
ZnSO4 119 kg /ha 142.3d 147.0d 132.3b
Urea (full dose) + SSP + MOP + ZnSO4 kg /ha 108.7c 111.3bc 186.0c
Open in a new tab

Data are means of three replications randomized completely. Data were subjected to analysis of variance (ANOVA) with IBM SPSS statistics version 22. Values with same letters in a column denote non-significant difference (P ≤ 0.05).

2.3. Statistical analysis

The data were analyzed using completely randomized design by IBM SPSS statistics version 22. Data were subjected to analysis of variance (ANOVA) and the comparisons in treatments were made by critical difference (CD) at 5 per cent level of significance.

3. Results

3.1. In vitro effect of fertilizers on hatching

Data (Table1) revealed that in first week, there was no hatching in urea, DAP, SSP and MOP but a few larvae hatched in ZnSO4 and water. In second week, hatching was significantly less in all fertilizers than water, except in both doses of ZnSO4. Maximum hatching (119.25) was observed in ZnSO4 at 120 ppm which was significantly higher than the hatching in control (74.75). Minimum hatching was observed in higher concentration of urea. During third week, ZnSO4 stimulated hatching while other fertilizers inhibited it. Hatching was maximum (182.25) at higher dose of ZnSO4.

Table 1.

Effect of fertilizers on hatching of Heterodera avenae under laboratory conditions.

Treatments No. of hatched juveniles from 10 cysts
 Total Left over
eggs cyst−1
1st week 2nd week 3rd week 4th week 5th week 6th week
Urea 80 ppm 0.00a 8.75b 26.00b 57.00a 35.25a 40.25a 167b 221.75cd
Urea 160 ppm 0.00a 4.25a 19.50a 45.00b 32.25a 35.50a 136a 234.75d
DAP 65 ppm 0.00a 23.25d 44.25d 79.50cd 47.00b 53.50b 247d 216.5cd
DAP 130 ppm 0.00a 17.25c 37.50c 71.25c 43.25b 48.25b 217c 220.5cd
SSP 187 ppm 0.00a 37.75f 59.00e 86.00de 59.00c 69.25d 311e 219.25cd
SSP 375 ppm 0.00a 32.25e 55.75e 94.50e 54.50c 62.75c 299e 217.5cd
MOP 50 ppm 0.00a 50.00g 77.25g 122.75f 76.50e 82.25f 409g 206.0c
MOP 100 ppm 0.00a 46.00g 71.00f 113.50f 70.25d 77.50e 378f 216.75cd
ZnSO4 60 ppm 3.75c 99.25i 156.50i 208.00h 126.25g 141.75h 732i 113.25a
ZnSO4 120 ppm 5.50d 119.25j 182.25j 254.25i 148.75h 161.50i 866j 98.75a
Water (control) 2.50b 74.75h 134.75h 190.50g 115.50f 127.00g 642h 133.0b
Open in a new tab

Data are means of four replications randomized completely. Data were subjected to analysis of variance (ANOVA) with IBM SPSS statistics version 22. Values with same letters in a column denote non-significant difference (P ≤ 0.05).

In all the treatments maximum hatching occurred in fourth week. At this time, maximum hatching was recorded in 120 ppm ZnSO4 and minimum in urea at 160 ppm. In fifth and sixth weeks hatching trend remained same. After six weeks, average un-hatched cyst contents were highest in higher concentration of urea and lowest in higher concentration of ZnSO4. It is because urea suppressed hatching and more contents were left in the cysts. Contrary to this, due to stimulatory effect of ZnSO4, more larvae hatched and less contents were left in this treatment.

Hatching was significantly more in both concentrations of ZnSO4 as compared to control. At 120 and 60 ppm ZnSO4, 46.65 and 39.31% hatching occurred respectively, in comparison to 32.55% in water (Fig. 1). In all other fertilizers, at both concentrations, weekly as well as total hatching was much less than control showing their inhibitory effect. This trend was seen from beginning through the experimental period.

Fig. 1.

Fig. 1

Open in a new tab

In vitro effect of fertilizers on hatching of Heterodera avenae.

3.2. Effect of fertilizers on hatching under pots

Data (Table 2) revealed that all the fertilizers treatment suppressed the hatching from cysts, except ZnSO4. In all the fertilizers treatments, juveniles in soil were less than soil without fertilizers and this number was maximum in ZnSO4 and lowest in higher dose of urea. ZnSO4 increased the no. of juveniles in soil and roots but it was less than untreated check. Among all the treatments, maximum juveniles in wheat roots were recorded in water (161.0) though this number was not statistically significant compared to ZnSO4. Average cyst content left in soil was affected by presence of fertilizers. It was lowest in ZnSO4 and highest in urea at 326 kg/ha. In ZnSO4, hatching was more than the combined application of all fertilizers but less than control. It is so because in combined application, stimulatory effect of ZnSO4 was mitigated by suppressive effect of other fertilizers. Hence interactive effect of different micro and macro nutrients in soil affecting the nematode hatching and survival play important role and needs to be studied.

4. Discussion

In our laboratory study among all fertilizers, both the concentrations of ZnSO4 stimulated the hatching from cysts. In pots, ZnSO4 increased the no. of larvae in soil and roots over other fertilizers though it was less than untreated check. In pots we used only recommended dose of ZnSO4 which may be the reasons for less effect than lab. Neutral effect of ZnSO4 in pot conditions might be due to the reason that soil is a buffer medium and nematodes are not in direct contact of fertilizers as in lab experiment. Clarke and Shepherd (1966) hypothesized that soil applications of zinc would not be practical for management of cyst nematodes because zinc cations are readily adsorbed to clay particles in the soil, and free zinc cations are necessary for stimulation of egg hatch. Present results are similar with the studies of Krauskopf (1972) who found that zinc salts are effective stimulants for hatching at concentrations that approximate soil levels.

Our lab studies showed that zinc sulphate increased the larval emergence from cyst at both the doses. Similar results were obtained by Behm et al. (1995) who reported that in vitro egg hatching of Heterodera glycines in reagent-grade zinc sulfate and zinc chloride, and fertilizer-grade zinc sulfate was significantly greater than hatching in deionized water, whereas zinc chelate fertilizer significantly inhibited egg hatching relative to deionized water. Commercial zinc chelate fertilizer inhibited hatching. It is likely that the hatch inhibition was caused by the chelating agent, although their investigation did not examine the effects of the chelating agent alone. They found that zinc compounds significantly affected H. glycines egg hatching in vitro, but had no effect on hatching in natural soils. In present study only fertilizer grade zinc sulphate has been tested and found that no affects in pot conditions. Stimulatory effect of zinc salts was also reported on egg hatching in soybean cyst nematode, H. glycines (Clarke and Shepherd, 1966, Tefft and Bone, 1984 and H. avenae (Zhen-yue et al., 2013). Egg hatching of the soybean cyst nematode, Heterodera glycines, was not affected by millimolar concentrations of calcium sulfate or calcium chloride. However, zinc chloride and zinc sulfate caused strong and moderate increases in hatching, respectively (Tefft and Bone, 1984).

Results indicated that the maximum suppression of larval hatch as shown by their presence in soil and roots was observed in urea. Application of ammonium fertilizers has shown nematicidal effect on plant-parasitic nematodes (Sudirman and Webster, 1995, Rodriguez-Kabana et al., 1982, Zhen-yue et al., 2013). Egg-hatching and viability of M. javanica larvae were greatly reduced when NH4Cl, (NH4)2 SO4 and NH4NO3 were used at elevated levels of EC, and there was a reduction in rate of nematode infection after their use on cucumber under greenhouse conditions (Karajeh and Al-Nasir, 2012).

In present study higher dose of urea was more effective in reducing H. avenae larvae in soil and roots. Urea is readily converted to ammonia, a conversion which is necessary for urea to be effective both as fertilizer and as nematicide (Siddiqui et al., 2001) and it is dosage dependent (Rodriguez-Kabana and King, 1980). In wheat crop, SSP, MOP and ZnSO4 are applied at sowing time while urea is given in 2–3 split doses depending on soil type. Maximum hatching in H. avenae takes place in 30–40 days, hence split application of nitrogenous fertilizers (urea) may not be effective in inhibiting its hatching.

Single super phosphate, di-ammonium phosphate and muriate of potash also suppressed hatching of H. avenae as indicated in our lab and pot studies. Saeedizadeh et al. (2020) tested four fertilizers including a chemical fertilizer (NPK), an organic fertilizer (Humic acid), a semi-organic fertilizer (Agro-health) and a bio-fertilizer (Azotobarvar-1) in vitro and observed that the fertilizers increased second stage juveniles (J2s) mortality and decreased egg hatching of root-knot nematode. Zhen-yue et al. (2013) also reported that Zn significantly stimulated the hatching of H. avenae. In their studies zinc did not cause mortality of secondary stage juveniles but other tested elements increased mortality as compared to control. Hatching suppressive effect of the fertilizers can be directly beneficial to crop for protection against nematode. Hatching stimulating effect of ZnSO4, can be used for obtaining more larvae of H. avenae from cysts for laboratory studies. Higher doses of zinc fertilizers resulting in more hatching can be exploited for management of H. avenae through chemicals and trap crops etc.

5. Conclusion

Synthetic fertilizers urea, single super phosphate, di-ammonium phosphate, muriate of potash inhibited hatching of cereal cyst nematode in lab as well as pot conditions. Zinc sulphate stimulated it in lab but showed no effect in pot conditions. All these fertilizers are widely used in agriculture and can easily be used to control nematode without adversely affecting agricultural land or harming human health. Also, the hatching stimulating agent like zinc sulphate can be exploited as management practice for cereal cyst nematode.

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgements

Authors are thankful to Professor and Head, Department of Soil Science, CCS HAU, Hisar, Haryana for analysis of soil samples and providing fertilizers.

Footnotes

Peer review under responsibility of King Saud University.

References

  1. Anonymous, 2014. Progress report of All India Coordinated Wheat and Barley Improvement Project 2013-14, Vol. III, crop protection. In: Saharan, M.S., Kumar, Sudheer, Sevakumar, R., Katare, Subhash, Sharma, Indu (Eds.), Directorate of Wheat Research, Karnal, India. p. 242.
  2. Behm J.E., Tylka G.L., Niblack T.L., Wiebold W.J., Donald P.A. Effects of zinc fertilization of corn on hatching of Heterodera glycines in soil. J. Nematol. 1995;27:164–171. [PMC free article] [PubMed] [Google Scholar]
  3. Clarke A.J., Shepherd A.M. Inorganic ions and the hatching of Heterodera spp. Ann. Appl. Biol. 1966;58(3):497–508. [Google Scholar]
  4. Cobb, N.A., 1918. Estimating the nema population of the soil. Agricultural Technology Circular-1. Bureau of Plant Industry, U.S.D.A. California, USA, pp. 48.
  5. Dordas C. Role of nutrients in controlling plant diseases in sustainable agriculture. A review. Agron. Sustain. Develop. 2008;28(1):33–46. doi: 10.1051/agro:2007051. [DOI] [Google Scholar]
  6. Gruzdeva L.I., Matveeva E.M., Kovalenko T.E. Changes in soil nematode communities under the impact of fertilizers. Eurasian Soil Sci. 2007;40(6):681–693. [Google Scholar]
  7. Habash S., Al-Banna L. Phosphonate fertilizers suppressed root knot nematodes Meloidogyne javanica and M. incognita. J. Nematol. 2011;43:95–100. [PMC free article] [PubMed] [Google Scholar]
  8. Kanwar R.S., Bajaj H.K. Effect of storage condition, age of cyst, root exudates and temperature on larval emergence from cysts of Heterodera avenae and Heterodera filipjevi. Indian J. Nematol. 2011;41(2):163–169. [Google Scholar]
  9. Karajeh M.R., Al-Nasir F.M. Effects of nitrogen fertilizers on the Javanese root-knot nematode Meloidogyne javanica and its interaction with cucumber. Arch. Phytopathol. Plant Prot. 2012;45:2177–2188. [Google Scholar]
  10. Kaplan M., Neo J.P. Effect of Chicken-excrement amendments on Meloidogyne arenaria. J. Nematol. 1993;25:71–77. [PMC free article] [PubMed] [Google Scholar]
  11. Krauskopf K.B. Geochemistry of micronutrients. In: Martuedt J.J., Giordano P.M., Lindsay W.L., editors. Micronutrients in Agriculture. Soil Society of America; Madison, Wisconsin: 1972. pp. 7–40. [Google Scholar]
  12. McBeth C.W., Taylor A.L., Smith A.L. Note on staining nematodes in root tissues. Proc. Helminthol. Soc. Washington. 1941;16:3–6. [Google Scholar]
  13. McIntoch P.D., Gibson R.S., Saggar S., Yeates G.W., McGimpsey P. Effect of contrasting farm management on vegetation and biochemical, chemical and biological condition of moist steepland soil of the South Island high country, New Zeland. Austral. J. Soil Res. 1999;37:847–865. [Google Scholar]
  14. Melakeberhan H., Bird G.W., Gore R. Impact of plant nutrition on Pratylenchus penetrans infection of Prunus avium rootstocks. J. Nematol. 1997;29(3):381–388. [PMC free article] [PubMed] [Google Scholar]
  15. Nicol J.M., Rivoal R. Global knowledge and its application for the integrated control and management of nematodes on wheat. In: Ciancio A., Mukerji K.G., editors. Integrated Management and Biocontrol of Vegetable and Grain Crops Nematodes. Springer Academic Publishing; Dordrecht, The Netherlands: 2008. pp. 243–287. [Google Scholar]
  16. Oka Y., Pivonia S. Use of ammonia-releasing compounds for control of the root-knot nematode Meloidogyne javanica. J. Nematol. 2002;4:65–71. [Google Scholar]
  17. Rodriguez-Kabana R., King P.S. Use of mixtures of urea and blackstrap molasses for control of root-knot nematodes in soil. Nematropica. 1980;10:38–44. [Google Scholar]
  18. Rodriguez-Kabana R., Shelby R.A., King P.S., Pope M.H. Combination of anhydrous ammonia and 1, 3-dichloropropenes for control of nematodes parasitic on soybean. Nematropica. 1982;12:61–69. [Google Scholar]
  19. Saeedizadeh A., Niasti F., Baghaei M.A., Hasanpour S., Agahi K. Effects of fertilizers on development of root-knot nematode, Meloidogyne javanica. Intl. J. Agric. Biol. 2020;23:431–437. doi: 10.17957/IJAB/15.1306. [DOI] [Google Scholar]
  20. Schindler A. A simple substitute for a Baermann funnel. Plant Disease Reporter. 1961;45:747–748. [Google Scholar]
  21. Siddiqui Z.A., Iqbal A., Mahmood I. Effects of Pseudomonas fluorescens and fertilizers on the reproduction of Meloidogyne incognita and growth of tomato. Appl. Soil Ecol. 2001;16:179–185. [Google Scholar]
  22. Sudirman I., Webster J.M. Effect of ammonium ions on egg hatching and second-stage juveniles of Meloidogyne incognita in axenic tomato root culture. J. Nematol. 1995;27:346–352. [PMC free article] [PubMed] [Google Scholar]
  23. Tefft P.M., Bone L.W. Zinc mediated hatching of eggs of soybean cyst nematode, Heterodera glycines. J. Chem. Ecol. 1984;10:361–372. doi: 10.1007/BF00987863. [DOI] [PubMed] [Google Scholar]
  24. Tenuta M., Ferris H. Sensitivity of nematode life-history groups to ions and osmotic tensions of nitrogenous solutions. J. Nematol. 2004;36(1):85–94. [PMC free article] [PubMed] [Google Scholar]
  25. Zhen-yue W., Yan S., Hong-xia Y., Wei-xing Y., Hong-lian L. The effects of nutrient elements on the hatching of cyst and vitality of secondary stage juvenile of cereal cyst nematode. Acta Phytopathologica Sinica. 2013;43(3):333–336. [Google Scholar]

Articles from Saudi Journal of Biological Sciences are provided here courtesy of Elsevier

RESOURCES