You are on page 1of 3

Pest science and management

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, India
E-mail: rkjvns_bhu@yahoo.co.in

Several workers have studied the life cycle of Meloidogyne graminicola (Golden
and Birchfield 1968) in rice roots and have reported differences in life-cycle
duration: 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 temperature
ranging from 22 to 29 °C; and later, Plowright and Bridge (1990), 13 d at ambient
temperatures of 25–35 °C. Because of these variations, we developed an
alternative method to study the life-cycle duration of this nematode species more
accurately at a given temperature.

Fig. 1. Symptoms caused by Meloidogyne graminicola on rice roots. (A) Radicle of a rice seedling with the first
root gall at the tip (7x). (B) Enlarged view of two radicles with the first root gall at the tip (12x).

2010 1
International Rice Research Notes (0117-4185)
Pest science and management

Seeds of rice variety MUT 7029 were soaked in water for 4 d, and the
water was changed daily. After 4 d, 30 sprouted seeds were each seeded in
earthen pots containing 1 kg of soil. Ten pots were used as replicates.
Immediately after sowing, a nematode suspension containing 3,000–5,000 active
second-stage juveniles (J2) was inoculated into each pot. The pots were watered
to field capacity. Within 6–8 h, the radicles developed; this was followed by
penetration 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 seedlings
were sampled daily until the eggs and the J2 were observed. Each day, the root
systems of these seedlings, one from each pot, were gently removed, thoroughly
washed, and stained with acid fuchsin in lactophenol following a method
described by Daykin and Hussey (1985). The stained roots were then transferred
into Petri dishes and the first root galls were cut and separated from the root
systems. To identify the different nematode developmental stages, each root gall
was transferred onto a glass slide containing a few drops of glycerol solution
(equal parts of glycerol and distilled water). The galls taken from the different
nematode developmental stages were separated from the root tissues with a
needle and mounted on permanent slides for microscopic observation and
photography.
We observed that the second molt of the feeding J2 began on day 4 and
third-stage juveniles (J3) were first seen on day 5 after inoculation. The J3 molted
on day 6 and fourth-stage juveniles (J4) were noted on day 7. Males were
observed 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 after
inoculation. In our experiment, the life cycle of M. graminicola was completed in
15 d at 27–37 °C. We noticed that a single gall was formed on the radicles (at the
tip) in almost all seedlings 2 d after inoculation, whereas, on lateral and seminal
roots, the galls developed after 1 d or more, resulting in a delay in the
subsequent stages of nematode development. Therefore, the first root galls that
formed on the radicles are most ideal for study. To obtain 100% infection and the
first 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 infected
radicles 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 3
d. This method also saves time as only a limited number of root galls have to be
examined for monitoring of the different nematode developmental stages. Each
gall contains enough nematodes.

2010 2
International Rice Research Notes (0117-4185)
Pest science and management

Fig. 2. Developmental stages of Meloidogyne graminicola on rice roots. (A) Second-stage juvenile (J2) (200x).
(B) Developing (J2) (210x). (C) Third-stage juvenile (J3) (285x). (D) Fourth-stage juvenile (J4) undergoing
molting (150x). (E) Adult female with eggs (130x). (F) Eggs (105x).

Acknowledgment
Dr. R.K. Jaiswal is grateful to Banaras Hindu University, Varanasi, India, for a research
fellowship grant.

References
Bridge J, Page SLJ. 1982. The rice root knot nematode, Meloidogyne graminicola, on deep water rice
(Oryza sativa subsp. indica). Rev. Nématol. 5:225-232.
Dabur AS, Taya AS, Bajaj HK. 2004. Life cycle of Meloidogyne graminicola on paddy and its host
range studies. Indian J. Nematol. 34:80-84.
Daykin ME, Hussey RS. 1985. Staining and histopathological techniques in nematology. In:
Barker KR, Carter CC, Sasser JN, editors. An advanced treatise on Meloidogyne. Vol II.
Methodology. Raleigh, NC (USA): North Carolina State University. p 39-48.
Golden AM, Birchfield W. 1968. Root knot nematode (Meloidogyne graminicola) as a new pest of
rice. Plant Dis. Rep. 52:423.
Plowright RA, Bridge J. 1990. Effect of Meloidogyne graminicola (Nematoda) on the establishment,
growth and yield of rice cv. IR 36. Nematologica 36:81-89.
Rao YS, Israel P. 1973. Life history and bionomics of Meloidogyne graminicola, the rice root-knot
nematode. Indian Phytopathol. 26:333-337.

2010 3
International Rice Research Notes (0117-4185)

You might also like