Susceptibility of Different Tomato Cultivars Against Rootknot Nematode, Meloidogyne spp.

5 Egypt. J. Agronematol., Vol. 12, No.1, PP. 54 62 (2013) Susceptibility of Different Tomato Cultivars Against Rootknot Nematode, Meloidogyne spp. Mostafa, S. M. El-Ansary* *Genetic Engineering and Biotechnology Research Institute (GEBRI), Plant Biotechnology Dept., Minufiya University, Sadat City, Egypt. Abstract Four tomato cultivars Castle rock, GS, Beto86 and Super-marmand were tested for their susceptibility to root-knot nematodes, Meloidogyne spp. The nematode succeeded in development and reproduce on all previously cultivars. Castle rock cultivar sustained the highest count of juveniles in soil and roots and also, the final population number. The cultivar had the highest count of the numbers of females. So, the nematode replicated very well on such cultivar and folded ten times at the end of the experiment. While, Super-marmand was found to be the least susceptible cultivar (folded seven times). Growth response of the tested cultivars to the nematode infection was determined on the basis of relative reduction in plant growth parameters. Variable reactions were noticed among the tested cultivars. Castle rock was the most affected cultivars, while Super-marmand was the least affected one. Finally, the all tested cultivars could be rated as highly susceptible hosts. Keyword: Tomato, Susceptibility, Root-knot nematode, Meloidogyne spp., Cultivars Introduction Root-knot nematodes are one of the major pathogens of tomatoes worldwide and limit fruit production (Sikora and Fernandez, 2005). Plant resistance is one of the most environmentally safe and economically viable means of controlling rootknot nematodes. The root-knot nematodes resistant crop varieties have comparatively better crop yield than the infected susceptible crop varieties. The resistant varieties can be used as part of integrated pest management in combination with other methods such as use of chemical nematicides, organic soil amendments, heat treatment, soil solarization, and crop rotation with non hosts for controlling root-knot nematodes. So, tomato, Lycopersicon peruvianum (accession 270435) and L. glandulosum (accessions 126440) and (126443) were resistant to Meloidogyne hapla. Six L. peruvianum accessions tested were resistant to M. incognita and M. arenaria. Variations in the level of resistance to four M. incognita races were observed among L. peruvianum accessions. Variation also occurred within the same accession. Race 3 of M. incognita reproduced less than other M. incognita races on L. peruvianum and L. glandulosum accessions Ammati et al., (1985). Appleman (2003) reported that, galling increased dramatically after 42 days, with 70-80% galling being reached on the 45th day. These results confirm that

55 Mostafa, S. M. EL-Ansary lettuce can function as a rapid indicator of nematode galling and will shorten the screening time from 90 days to 45 days. Vovlas et al., (2005) in their studies on pathogenicity of the root-knot nematode, M. javanica on potato, reported that, potato cvs Cara and Spunta showed a typical susceptible reaction to M. javanica under natural and artificial infections, respectively. Five sunflower cultivars were used for screening i.e. FH-75, Beimisal-205, Hysin-33, Super-25 and Engro 9704. FH-75 proved the most tolerant against root-knot nematode infestation. It gave minimum reduction in plant height and minimum increase in fresh and dry root weight. The varieties BEIMISAL-205, Hyson-33, Super-25 and Engro-9704 exhibited tolerance respectively. Engro-9704 was the most susceptible variety to root-knot nematode infestation Rehman et al., (2006). Also, six tomato cultivars Moneymaker, Beefsteak, Roma, Summertaste, Mini Roma and Smallfry were tested for their susceptibility to root-knot nematodes at inoculum levels of 200, 400, 600 juveniles (J2) per pot. All were found to be susceptible to varying degrees as eggmasses were present in all with Moneymaker and Roma being the most susceptible and Mini Roma, the least susceptible Singh and Khurma, (2007). In addition, tomato rootstocks Beaufort and Maxifort were susceptible to M. javanica (Pf/Pi > 50 and RI > 50%). Rootstocks PG76 and He-Man, and the resistant tomato cv. Caramba showed high levels of resistance in the test conducted in summer, whereas MKT-410 and 42851 and the resistant tomato cv. Monika were moderately resistant. In the field, seven rootstocks showed high levels of resistance and one (He-Man) showed an intermediate level, whereas Beaufort and Maxifort were susceptible Cortada et al., (2008). Results also show that, the cultivars of tomato cultivars i.e. HTA-18 and HTA-31 had significantly (p<0.05) fewer gall counts per root system and per gram fresh weight of root than the other cultivars. Single degree of freedom analysis showed that HTA-18, HTA-19 and HTA-31had significantly (p<0.05) fewer galls than the locally grown cultivars. HTA-18 and HTA- 31 were moderately susceptible with gall index (GI) of 3, while is the rest of the cultivars were highly susceptible (GI>4). HTA-18, HT-Ravid244 and the wild species were considered tolerant Udo et al., (2008). Additionally, the evaluation revealed the resistance of the known resistant cowpea accession, Acc 64298, to M. incognita Race 4 with low reproduction factor (RF) of 0.74 and low root gall index (GI) of 1.4 while the same cowpea accession (Acc 64298) (RF = 0.95, GI= 2.4) together with accession Acc 67010 (RF= 0.91, GI = 2.0) showed hypersusceptibility (RF = 1, GI =>2) to M. arenaria Race 1. But the two accessions; Acc 64298 (RF=3.45, GI=4.40) and Acc 67010 (RF=3.01, GI=4.60), broke down to M. javanica. Comparatively, in relation to all the three species, M. incognita R4 (RF = 0.74) and M. arenaria R1 (RF = 0.95) showed non-significant lower reproduction than M. javanica which gave a significant higher reproduction (RF = 3.45) on the same resistant accession, Acc 64298 Olower (2009). Salawu and Darabidan, (2010) reported that, sixteen selected ornamental plants were screened for resistance against infection by a rootknot nematode, M. incognita in a screenhouse. Nerium orleander and Thunbergia

Susceptibility of Different Tomato Cultivars Against Root-knot Nematode, Meloidogyne spp. 56 erecta were resistant to infection while Acalypha torta, lxora coccinea, Hibiscus rosa-sinensis, Duranta sp., Acalypha tricolor, Duranta sp., Cordiaeum variegatum, Cordyline terminalis, Acalypha sp., Dieffenbachia picta and Quisqualis indica were moderately to highly susceptible to nematode infection. Galled roots of infected plants contained developmental stages of M. incognita. Root and shoot growth of five tomato verities like, Roma Holland, Roma v.f., Sunehra, Roma Holland and Tomato Anmol was significantly reduced when inoculated with second stage infective larvae of M. incognita as compared to un-inoculated tomato varieties. Also, number of larvae was significantly increased in tomato variety Roma v.f., Gola France followed by Roma Holland and that was decreased in Anmol Khanzada et al., (2012). The purpose of this study was determining the screening of some different tomato cultivars infected with root-knot nematode, Meloidogyne spp. under greenhouse conditions. Materials and Methods Isolation and collection of nematode in pure cultures: Eggs of Meloidogyne spp. were extracted from tomato (Lycopersicon esculentum cv. Castle rock) roots infected with the nematode using sodium hypochlorite solution (Hussey and Barker 1973). Second-stage juveniles (J2s) were collected daily from eggs and stored at 15 C. The juveniles used in the experiments were less than 5 days old. Plant material: Seeds of tomato cv. Beto86, Super-marmand, Castle rock and GS relatively susceptible to Meloidogyne spp., were surface sterilized with 1% Ca (Col)2 and washed three times with sterile MgSO4 (0.1M) and dried under a laminar flow hood. The seeds were planted in 3-diam., earthen pots containing mixture of 1:1 steamsterilized peat and sand. Six weeks old seedlings were then transplanted for use in the experiments. Evaluation of resistance in tomato cultivars against root-knot nematode Meloidogyne spp.: Six weeks old, obtained from previously cultivars tomato plants and then, were transplanted in 20 cm diameter plastic pots containing a mixture of 1:2 sterilized clay/sandy soil. Sixteen pots were inoculated with Meloidogyne spp. juveniles at the rate of 1,000 J2s per pot at the planting time and replicated four times. The remaining four pots served as untreated controls without nematode, thus for every cultivar. The pots were placed in a completely randomized design in a greenhouse of Genetic Engineering and Biotechnology Research Institute (GEBRI). Plants were allowed to grow for eight weeks and then remove from their containers. The stems were cut off and the soil was gently washed with tap water from the root system. Then these were taken to laboratory. Weight and length of root and shoot

57 Mostafa, S. M. EL-Ansary were determined. Nematode population in soil and developing stages in roots were counted. The degree of root galling on each infected plant was rated according to a modified rating scale of Taylor and Sasser (1978); 0= 0 gall, resistant; 1= 1-2 galls, moderately resistant; 2= 3-10 galls, moderately susceptible; 3= 11-30 galls, susceptible; 4= 31 galls and above, highly susceptible. Statistical analysis The responses of the treatments were compared by analysis of variance (ANOVA) (Sokal and Rohlf, 1995). Significant differences between the means of parameters were determined (P 0.05). All analysis was performed with SPSS software (Statistical Package for the Social Sciences). Results All the current tomato cultivars were susceptible to root-knot nematode, Meloidogyne spp. to varying degrees. Egg-laying females were recorded in all four tomato cultivars thus indicating that none of the tested cultivars was resistance to root-knot nematode. For instance, Castle rock cultivar sustained the highest count of juveniles in soil and roots and also, the final population number. The cultivar had the highest count of the numbers of females. So, the nematode replicated very well on such cultivar and folded 10.31 times at the end of the experiment (Table, 1&Fig., 1). Table (1): Host reaction and reproduction of Meloidogyne spp. on roots of four tomato cultivars. Cultivars Number of galls Immature stages Females Egg-laying females Number of Juveniles Total** Rate*** of build-up Beto86 80.5 47.5 66 82.5 7950 8146 8.15 Castle rock 115.5 67.5 98 118.5 10022 10306 10.31 GS 84 40 66 102.5 9492 9696.5 9.69 Super-marmand 85.5 41 71 124.5 7632 7868.5 7.87 LSD 0.05 *.001.002.000.034.001.001.427 *-Non significant 0.05 using one way analysis of variance (ANOVA) Highly significant 0.05 using one way analysis of variance. **Total population including numbers of juveniles in soil (J2s) + Immature stages + females + egg-laying females. ***Rate of build-up = pf (final population /initial population) Norton, 1978.

Susceptibility of Different Tomato Cultivars Against Root-knot Nematode, Meloidogyne spp. 58 Fig. (1): Rate of build-up of root-knot nematode, Meloidogyne spp. infecting some tomato cultivars. Consequently, the greatest percentage reduction in shoot weight compared to the control was observed for the same cultivars (45.38% reduction) table, 2 & fig., 2. While, Super-marmand was found to be the least susceptible cultivar (folded 7.87 times). Consequently, the same cultivar was the least affected cultivar in root length (recorded 10.17% reduction) while, shoot weight, shoot length and root weight at the previously cultivar was not affected by nematode (Fig., 2). Also, Beto86 and GS were found to be the most susceptible cultivar having the least galls numbers. Table (2): Percentage of reduction of plant growth criteria of four tomato cultivars to the infection with Meloidogyne spp. Cultivars State Root Shoot weight (g) length (cm) weight (g) length (cm) Infected 1.515 7 4.44 22.75 Beto86 Uninfected 1.645 9.5 7.15 39.5 LSD 0.05*.002.002.037.002 Infected 1.835 9 6.54 24 Castle rock Uninfected 3.475 8.75 10.78 30 LSD 0.05*.003.689.000.000 Infected 2.435 11 13.12 44 GS Uninfected 2.39 11 9.34 34 LSD 0.05*.830 1.000.025.001 Infected 1.935 9 9.14 25.25 Super-marmand Uninfected 2.245 9.5 10.28 29.5 LSD 0.05*.088.741.491.375 *-Non significant 0.05 using one way analysis of variance (ANOVA) Highly significant 0.05 using one way analysis of variance

59 Mostafa, S. M. EL-Ansary Finally, all the tested cultivars variability sustained the nematode reproduction and responded variably to the nematode infection. Accordingly, the tested cultivars could be rated as highly susceptible hosts (Castle rock, GS, Beto86 and Supermarmand). Fig. (2): Effect of root-knot nematode, Meloidogyne spp. on percentage of plant growth reduction of four tomato cultivars. Discussion Susceptibility of a plant to root-knot nematode, Meloidogyne spp. depends on the ability of juveniles (J2s) to penetrate the roots of the plant and cause the formation of giant cells which appears as galls on the roots (Chen et al., 2004). The juveniles feed and moult twice before developing into the adult stage (Siddiqi, 2000). So, the nematode developed and reproduces on all tested cultivars. Comparatively, Castle rock cultivar recorded the highest counts of nematode in soil and roots and consequently had the highest nematode final population. Also, the nematode behaved quite similar on the other cultivars. All previously cultivars were rated as highly susceptible host according to the number of galls. Growth response of the tested cultivars to the nematode infection was also variable. The growth parameters of all tomato cultivars were significantly reduced by nematode infection. Castle rock seemed to be the mostly among the other ones; while Super-marmand was the lowest affected ones. Therefore, all the tested cultivars have reacted as susceptible hosts. The results on the occurrence of the root-knot nematode (Meloidogyne spp.) in tomato plants are in conformity with (Ammati et al., 1985; Vovlas et al., 2005; Singh and Khurma, 2007; Salawu and Darabidan, 2010).

Susceptibility of Different Tomato Cultivars Against Root-knot Nematode, Meloidogyne spp. 60 Conclusion Root-knot nematode succeeded in development and reproduce on all the tested cultivars. Castle rock cultivar sustained the highest counts of number of galls, immature stages, egg-laying females, and consequently the nematode final population. Therefore, the nematode folded than ten times in such cultivar. Growth response of the all tested cultivars to the nematode infection was determined on the basis of reduction in plant growth parameters. So, Castle rock was the most affected cultivar, while Super-marmand was the least affected one. Finally, all tested cultivars could be rated as highly susceptible hosts. Acknowledgment The author gratefully thanks for helped with Dr. Ragaa A. Hamouda statistically analysis. Also, thanks Dr. Wahby M. Hassany and Mr. Tamer M. Mosalamy for helped in this work. References Ammati, M.; Thommason, I.J. and Roberts, P.A. (1985). Screening Lycopersicon spp. for New Genes Imparting Resistance to Root-Knot Nematodes (Meloidogyne spp.). Plant Diseases, 69(2): 112-115. Appleman, L. (2003). Screening for Root Knot Nematode (Meloidogyne hapla) Using Lettuce. Journal of Undergraduate Research, 1:1-3. Chen, Z.X.; Chen, S.Y. and Dickson, D.W. (2004). Nematology advances and perspectives Vol. 2, Nematode Management and Utilization. Tsinghua University press China, 636pp. Cortada, L.F.; Sorribas, F.J.; Ornat, C.; Kaloshian, I. and Verdejo-Lucas, S. (2008). Variability in infection and reproduction of Meloidogyne javanica on tomato rootstocks with the Mi resistance gene. Plant Pathology, 57: 1125-1135. Hussey, R.S. and Barker, R.K. (1973). A comparison of methods of collecting inocula of Meloidogyne spp. including a new technique. Plant Diseases Report, 57: 1025-1028. Khanzada, S.; Jiskani, M.M.; Khanzada, S.R.; Khanzada, M.S.; Ali, S.; Khanzada, K.A.; Saeed, N.; Anwar, S. and Khalid, M. (2012). Response of some tomato cultivars against root-knot nematode, Meloidogyne incognita (Kofoid & White) Chitwood. The Journal of Animal & Plant Sciences, 22(4): 1076-1080.

61 Mostafa, S. M. EL-Ansary Norton, D.C. (1978). Ecollogy of plant parasitic nematodes. Jon Willeg and Soms, New York, P. 238. Olower, T. (2009). Cowpea Germplasm Resistant to Meloidogyne Arenaria Race 1, Meloidogyne incognita Race 4 and Meloidogyne javanica. European Journal of Scientific Research, 28 (3):338-350. Rehman, A.; Bibi, R. and Ullah, M.H. (2006). Screening of Different Sunflower Cultivars against Root-knot Nematode (Meloidogyne incognita). Journal of Agriculture and Social Sciences, 2(3): 182-184. Salawu, E.O. and Darabidan, I.A. (2010). Screening of selected ornamental plants to Meloidogyne incognita in Nigeria. Pakistan. Journal of Nematology, 28 (2): 353-358. Sikora, R.A. and Fernandez, E. (2005). Nematode parasites of vegetables. In: Luc, M., Sikora, R.A. and Bridge, J. (Eds). Plant parasitic nematodes in subtropical and tropical agriculture. 2 nd edition, CABI publishing, pp. 319-392. Siddiqi, M.R. (2000). Tylenchida Parasites of plants and insects. 2 nd Edition, CABI Publishing, 833pp. Singh, S.K. and Khurma, U.M. (2007). Susceptibility of six tomato cultivars to the root-knot nematode, Meloidogyne incognita. The South Pacific Journal of Natural Science, 13: 73-77. Sokal R.R. and Rohlf F.J. (1995). Biometry: the principles and practice of statistics in biological research.3rd ed. W.H. Freeman and company New York, 937 pp. Taylor, A.L. and Sasser, J.N. (1978). Biology, Identification and control of root-knot nematode (Meloidogyne species). North Carolina State University Graphics, North Carolina, Raleigh. 111p. Udo, I.A.; Ugunu, M.I.; Ogbuji, R.O. and Ukeh, D.A. (2008). Source of tolerance to root-knot nematode, Meloidogyne javanica, in cultivated and wild tomato species. Plant Pathology Journal, 7(1): 40-44. Vovlas, N.; Mifsud, D.; Landa, B.B. and Castillo, P. (2005). Pathogenicity of the root-knot nematode Meloidogyne javanica on potato Plant Pathology, 54: 657-66.

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