378 Invasive Stink Bugs and Related Species (Pentatomoidea)
7.4.4.5 Diapause Maintenance in the Field: Dynamics of the Physiological
Condition of Adults during the Overwintering Period
In central Japan, adults of Nezara viridula essentially cease all movement by early December, although
they will slowly respond if disturbed or if the temperature increases sharply. At this time, the fat bodies
have the highest degree of development/accumulation. Adults remain in such a state until February, when
depletion of the fat bodies becomes noticeable (Takeda et al. 2010; also see Figure 11.12). During most
of the overwintering period, the majority of adults of both sexes remain dark-colored; the percentage of
green adults varies, and increases as spring approaches (Figure 7.11; Takeda et al. 2010).
Previous field records indicate that many or all Nezara viridula males collected from hibernacula have
reproductive organs that are fully developed (van Heerden 1934) or contain active sperm in their testes
(Kiritani 1963). When collected in the field in January and February, females do not mate under long-
day conditions in the laboratory, whereas males will mate when paired with nondiapausing laboratory
females (Kiritani and Hokyo 1970). Such observations suggest that, in contrast to females who spend
winter in diapause, males overwinter in a state of quiescence. This finding is used to explain the higher
winter survival of females (Kiritani et al. 1966). However, regular dissections during the winter demon-
strate that all adults of both sexes have reproductive organs and fat bodies in a state characteristic of true
diapause until late March (Takeda et al. 2010; also see Figure 11.12). Thus, it is likely that both females
and males overwinter in a state of true diapause, not quiescence. It appears that more research is needed
to better understand the differences in diapause maintenance between the sexes.
7.4.4.6 Diapause Termination in the Laboratory
When diapausing adults are kept for an extended period under intermediate (L:D 13:11) or short-day
conditions (L:D 10:14 and 12:12) at 25°C, they remain russet for varying periods of time but then start to
change color spontaneously to the intermediate grade and then to green. A few adults, mostly those kept
under L:D 13:11, go through a cycle of color change (i.e., from green to intermediate and russet, and then
back to intermediate and green) more than once. The shorter the day length, the longer the adults remain
russet, and the longer they need to reach the final stable green color (Figure 7.12; Musolin et al. 2007).
Completion of the final body color change from russet and intermediate to green might be considered a
marker of diapause termination because the attainment of the final green color almost always precedes
the start of reproduction. In laboratory experiments, almost 94% of females already are green on the day
of the first copulation (all three photoperiodic conditions combined), and only one russet female was
found in the laboratory experiment to have copulated with a green male. Males show a similar trend. All
females are green on the first day of oviposition (Musolin et al. 2007).
These data show that adult diapause in Nezara viridula can end spontaneously under laboratory constant
short-day conditions without any preceding low temperature treatment, although in the absence of chilling
and change of photoperiodic conditions from short-day to long-day, the termination of diapause and start of
reproduction are poorly synchronized (Musolin et al. 2007). The time to first oviposition ranges from 106 to
158 days under different short-day conditions at 25°C (Musolin et al. 2007). At the same temperature, this
timing is only 43 days under nondiapausing long-day conditions, and even less (23 days) in adults that have
overwintered and started reproduction outdoors (Musolin and Numata 2003b, 2004).
From a physiological standpoint, the timing of the end of insect diapause (= the end of the termina-
tion phase, according to Koštál 2006; also see Chapter 11) is difficult to detect precisely, perhaps not
possible at all, and the indirect markers visible externally vary between different species (Hodek 1996).
The onset of copulation and oviposition are indicative that diapause has terminated and postdiapause
development has commenced. The observation that almost all adults of Nezara viridula show a reversion
of body color to green prior to copulation, and that all females are green on the day of first oviposition,
suggests that the change of body color from russet to intermediate and then to green is the closest and
most easily observed event indicating the end of diapause.
There has been discussion on whether the russet coloration might serve as a reliable indicator of
diapause in Nezara viridula (Harris et al. 1984, Seymour and Bowman 1994). Results of several
experiments suggest this is true in most cases: a great majority of adults that emerge in autumn are