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A technique for studying the life cycle of Meloidogyne graminicola in rice roots

A technique for studying the life cycle of Meloidogyne graminicola in rice roots

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Published by Grace Cañas
2010 PestSci_2010-016
2010 PestSci_2010-016

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Categories:Types, Research, Science
Published by: Grace Cañas on Jan 14, 2011
Copyright:Attribution Non-commercial


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Pest science and management
International Rice Research Notes
A technique for studying the life cycle of
Meloidogyne graminicola 
in rice roots
R.K. Jaiswal and K.P. Singh, Department of Mycology and Plant Pathology,Institute of Agricultural Sciences, Banaras Hindu University, Varanasi 221005, IndiaE-mail: rkjvns_bhu@yahoo.co.in
Several workers have studied the life cycle of
 Meloidogyne graminicola
(Goldenand Birchfield 1968) in rice roots and have reported differences in life-cycleduration: Rao and Israel (1973) reported 26–51 d, depending on the season;Dabur et al (2004), 24 d; Bridge and Page (1982), 19 d at ambient temperatureranging from 22 to 29 °C; and later, Plowright and Bridge (1990), 13 d at ambienttemperatures of 25–35 °C. Because of these variations, we developed analternative method to study the life-cycle duration of this nematode species moreaccurately at a given temperature.
Fig. 1. Symptoms caused by
Meloidogyne graminicola 
on rice roots. (A) Radicle of a rice seedling with the firstroot gall at the tip (7x). (B) Enlarged view of two radicles with the first root gall at the tip (12x).
Pest science and management
International Rice Research Notes
 Seeds of rice variety MUT 7029 were soaked in water for 4 d, and thewater was changed daily. After 4 d, 30 sprouted seeds were each seeded inearthen pots containing 1 kg of soil. Ten pots were used as replicates.Immediately after sowing, a nematode suspension containing 3,000–5,000 activesecond-stage juveniles (J2) was inoculated into each pot. The pots were wateredto field capacity. Within 6–8 h, the radicles developed; this was followed bypenetration of the active J2. One gall per radicle was formed 2 d after inoculation.To determine life-cycle duration, the first root galls of the 10 seedlingswere sampled daily until the eggs and the J2 were observed. Each day, the rootsystems of these seedlings, one from each pot, were gently removed, thoroughlywashed, and stained with acid fuchsin in lactophenol following a methoddescribed by Daykin and Hussey (1985). The stained roots were then transferredinto Petri dishes and the first root galls were cut and separated from the rootsystems. To identify the different nematode developmental stages, each root gallwas transferred onto a glass slide containing a few drops of glycerol solution(equal parts of glycerol and distilled water). The galls taken from the differentnematode developmental stages were separated from the root tissues with aneedle and mounted on permanent slides for microscopic observation andphotography.We observed that the second molt of the feeding J2 began on day 4 andthird-stage juveniles (J3) were first seen on day 5 after inoculation. The J3 moltedon day 6 and fourth-stage juveniles (J4) were noted on day 7. Males wereobserved on day 9 and young females with vulval slits were seen on day 10. Egg-laying was first observed on day 11 and J2 were detected on day 15 afterinoculation. In our experiment, the life cycle of
 M. graminicola
was completed in15 d at 27–37 °C. We noticed that a single gall was formed on the radicles (at thetip) in almost all seedlings 2 d after inoculation, whereas, on lateral and seminalroots, the galls developed after 1 d or more, resulting in a delay in thesubsequent stages of nematode development. Therefore, the first root galls thatformed on the radicles are most ideal for study. To obtain 100% infection and thefirst root galls on the radicles, more inoculum should be used (for instance,3,000–5,000 active J2 per kilogram of soil).The use of active J2 as inoculum and the daily sampling of infectedradicles enabled us to get more accurate results than those of Dabur et al (2004),who used infested soil, and of Rao and Israel (1973), who sampled only every 3d. This method also saves time as only a limited number of root galls have to beexamined for monitoring of the different nematode developmental stages. Eachgall contains enough nematodes.

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