Polymorphism of the Southern Green Stink Bug Nezara viridula Linnaeus, 1758 (Hemiptera: Pentatomidae) In Vietnam

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Biological Forum An International Journal 7(1): 276-281(2015) ISSN No. (Print): 0975-1130 ISSN No. (Online): 2249-3239 Polymorphism of the Southern Green Stink Bug Nezara viridula Linnaeus, 1758 (Hemiptera: Pentatomidae) In Vietnam Thai Thi Ngoc Lam *, Truong Xuan Lam ** and Tran Ngoc Lan *** * Vinh University, Nghe An province, Vietnam ** Institute of Ecology and Biological Resources, Vietnam Academy of Science and Technology (VAST) *** Ministry of Science an Technology, Vietnam (Corresponding author: Thai Thi Ngoc Lam) (Received 07 June, 2015, Accepted 01 July, 2015) (Published by Research Trend, Website: www.researchtrend.net) ABSTRACT: In Vietnam, field surveys of N. viridula (Hemiptera: Pentatomidae) on host plants (rice, corn, sesame) was collected 10 color morphs including G, O, F, R, OR, GY, OG, Y, B and C. Among them G and O types were common. The rare phenotype included B, C, Y, OR, OG and GY. Type G is the best adapted to colonize different environments. The emergence of the phenotype depends on the temperature and food (host plants). Type F and R are better adapted to higher temperatures in summer crop on sesame. Low temperatures are the good conditions for the emergence of O type. The 4 th and 5 th instars belong to both in GxG and GxO, had two colors: green and black; among colors, green was the most common. GxO pair created 4 phenotypes: G, O, F, R. The GxG pairs created 3 phenotypes: G, O, F. Key words: Nezara viridula, polymorphism, host plant, distribution INTRODUCTION The southern green stink bug, Nezara viridula (L.) is known to be polymorphic, including 12 different color morphs (Kiritani & Yukawa 1963, Yukawa & Kiritani 1965, Kiritani 1970, Vivan & Panizzi 2002). Although the color polymorphisms are manifested in both nymphal and adult stages, previous studies have focused only on the more obvious adult traits. The four main color forms were distinguished and given names based on the color pattern on the dorsum (Kiritani and Yukawa 1963, Yukawa and Kiriani 1965). N. viridula f. smaragdula F. (G type) is the common green form, N. viridula f. torquata F. (O type) has median and lateral lobes and the anterior margin of the pronotum yellow or orange, f. viridula L. (R type) has green spots on a background of yellow or orange over the entire body, and an unnamed morph (F type) is like f. torquata except it also has yellow or orange along the margins of the convexium. Other morphs with combinations of these orange and green color traits exist, including an entirely orange morph called f. aurantiaca (Y type). The ventral and dorsal coloring of N. viridula f. aurantiaca is uniformly orange. The mix of N. viridula color polymorphisms varies with region. The greatest diversity of color polymorphisms occurs in Japan, whereas N. viridula populations in many parts of the world (Australia, Pacific Islands, United States, Central America, and the West Indies) seem to be only or primarily green (Yukawa and Kiritani 1965). N. viridula is widespread in Brazil and that type G (smaragdula) is the best adapted to colonize different environments. Type O (torquata) is favored by the conditions in the South Region, which allow the gene(s) responsible for the polymorphism to be expressed (Vivan and Panizzi 2006). The f. aurantiaca (Y type) morph is very rare in Japan, occurring in 5,000 individuals (Yukawa and Kiritani 1965). In Brazil, the frequency of f. aurantiaca is higher at 0.7% (Vivan and Panizzi 2002). The different color polymorphisms are under genetic control. Crossing studies using the main color morphs (O, G, and R types) suggested the body color polymorphisms are controlled by two sets of genes on two independent loci (Ohno and Alam 1992). N. viridula f. aurantiaca was not included in the study. McLain (1981) showed that an orange color polymorphism discovered in an N. viridula population in Florida was sex linked and recessive. Some types of N. viridula are known to be better adapted to unfavorable abiotic conditions due to their variable genetic composition, and this is an important factor that contributes to the abundance of the populations (Kiritani 1971).

The polymorphisms of N. viridula has been studied worldwide. However, few data on the color of nymphs and adult of different types, related to the different parents, are available. Therefore, studies were conducted to evaluate the effect of different host plants and seasonal crops on the distribution of the color morphs of N. viridula in Vietnam. MATERIAL AND METHODS Field surveys: N. viridula adults were surveyed, each week, in 2010 and 2011, different host plants including corn ( Zea mays), rice ( Oryza sativa) and sesame (Sesamum indicum). From each host plant, 50 samples were taken at random per week, using either a sweep net or a beat cloth, depending on the type of plants available. Usually on corn and sesame, the beat cloth was used. The samples consisted of beating plants over the beat cloth 20 times. On the rice the sweep net was used, sweeping the plants 20 times. Adults of N. viridula captured were killed, using killing jars, and were pinned and stored in the Insect Collection of Vinh university. Lam, Lam and Lan 277 The relationship between the different parents with colors of nymphs and adults: Pairs of N. viridula were selected at the adult emergence date and placed each in plastic boxes (15cm diam eter 20cm high) covered with a lid as beat cloth. Bugs were fed by fresh bean. The boxes were then placed in conditions rom (28,48 o C-61,75% RH). The number of pairs used at the different parents was, respectively, 20 GxG pairs and 20 GxO pairs. Bugs were observed daily, and the food was replaced every two days. The deverlopmen of bugs were recorded, and the mean (± SEM) rate color nymph and adult were calculated. Statistics: Data of this study were submitted to analysis of variance (ANOVA). Analyses were performed using the Statistix 9.0. RESULTS AND DISCUSSION Table 1: Morphs of Nezara viridula in Vietnam. No. Type Name Color pattern 1. G f. smaragdula F. Entirely green (Fig. 1a) The color morphs of Nezara viridula in Vietnam During two yaers (2010 and 2011), field surveys of N. viridula on host plants (rice, corn, sesame) in Vietnam was collected 5671 speciments, including 10 color morphs (G, O, F, R, OR, GY, OG, Y, B, C) (Table 1). 2. O f. torquata F. Green with anterior margin of the pronotum and head yellow (Fig. 1g) 3. F In additions to the characteristic of O type, connexivum (Fig. 1c) 4. Y f. aurantica Costa Entirely yellow (Fig. 1b) 5. R f. viridula L. Green spots on yellow ground color (Fig. 1h) 6. OR Combination of O and R types (Fig. 1l) 7. GY Green-yellowish (Combination of G and Y types) (Fig. 1e) 8. GO Anterior margins of the pronotum and head yellow, with the rest of the body green-yellowish (Fig. 1f) 9. B f. vicaria Walker f. chlorocephala Westwood Brown green with thorax and head yellow; Wings, abdomen and legs with brown red (Fig. 1k). 10. C Entirely yellow red (Fig. 1d). The southern green stink bug, Nezara viridula (L.) is known to be polymorphic, including 12 different color morphs which are derived from four basic types (Yukawa & Kiritani 1965, Kiritani 1970, Kazuno & Zinnatul 1992, Vivan & Panizz 2002). In Vietnam, Nezara viridula was diversity of phynoltype with 10 types among 12 type were recorded around the world. OY and FR-types have not been recorded in Vietnam. Distribution of the color morphs of Nezara viridula on host plants Distribution of the color morphs of Nezara viridula on host plants: rice, corn, sesame showed by Table 2. During 2010 and 2011, 5671 adults of N. viridula were collected, 72.40% of the specimens belonging to type G (smaragdula). The second most abundant type O (torquata) was 16.51% (936 individuals). The rate type F was 4.94% with 280 individuals. Type B and C were rare only occurring on rice with a individual (0.02%). There were 15 individuals belonging to type Y (0.26%). The rate of type OR, R, GY and OG were from 1.10 to 1.94% (57-110 individuals) (Table 2). Among three hots plants, collected 10 color morphs on rice, 8 morphs on corn (B, C type not recorded) and 7 morphs on sesame (no bug of B, C and OR type).

Lam, Lam and Lan 278 Fig. 1. Different morphological type of Nezara viridula occurring in Vietnam. (1a) G type; (1b) Y type; (1c) F type; (1d) C type; (1e) GY type; (1f) OG type; (1g) O type; (1h) R type; (1k) B type; (1l) OR type). Table 2: Comparision of the frequencies of colour type of adults on hot plants. Type Frequencies of colour type (%) No. of on hots plants speciments Rice Corn Sesame Avarage (%) G 74.25 73.23 65.01 4106 72.40 O 16.67 18.43 12.29 936 16.51 F 2.78 5.21 10.52 280 4.94 R 1.27 1.61 4.65 110 1.94 GY 1.23 0.55 5.32 93 1.64 OG 1.31 0.18 1.99 57 1.01 Y 0.40 0.14 0.22 15 0.26 OR 2.02 0.65-72 1.27 B 0.04 - - 1 0.02 C 0.04 - - 1 0.02

Types G and O are the two most common in all 3 host plants were investigated, respectively, 74.25% and 16.67% on rice; 73.23% and 18.43% on corn and on sesame (64.90% and 12.30%). With 2598 adults were colected on rice, there are 2 types very rare (B - and C-types) accounted 0.04% (1 individual), followed by Y type was 0.4% (10 individuals). On corn, collected 2170 individuals (including 8 phenotype). Y and OG were less common at 0.14-0.18% (3-4 individuals). GY and OR phenotype were also relatively less with 0.55-0.65% (12-14 individuals). Among 905 adults were collected on sesame (including 7 phenotype), only 2 individuals belong to Y-type accounted 0.2%. However, F-type was common with 10.50% (111 individuals) after G and O types. The distribution of phenotypes of Nezara viridula was correlated with hots plants. G-type was most common followed by O type and F type. Additionally, type G is the best adapted to colonize different environments. This result is similar to the investigation of Vivan & Panizzi (2006). Type O appeared on rice (16.67%) and maize (18.43%) was higher than on sesame (12.29%). This appearance is related to temperature. Rice and corn are grown at lower temperature (in spring crop) than sesame (in summer-autumn crop). The result is similar to Vivan & Panizzi (2005, 2006). Vivan & Panizzi (2006) showed that type O less abundant than the former, was more frequent at latitudes > 23 18 S and mean annual temperatures < 20.8 o C (Southern Region) in Brazil, indicates that this type is better adapted to cooler temperatures. Data on nymph biology and adult reproduction of type O in the laboratory, comparing different temperatures, demonstrate its better adaptability to lower temperatures than the most common type G (Vivan & Panizzi, 2005). The fact that type F is common after G and O types on sesame (10.52%). Additionally, on corn and rice, type F was rather rare (2.78-5.21%). Lam, Lam and Lan 279 Table 3: The color ratio of 4 th and 5 th instars. In Vietnam, the type F is better adapted to higher temperatures in summer crop, which allow the gene(s) responsible for the polymorphism to be expressed. On sesame, ratios of type R was 4.65% and higher than on corn and rice. There is a significant difference. This phenotype was rare and less adapted, however, its occurrence is related to temperature. High temperatures in summer are the good conditions for the emergence of R type. The proportions among the types found in Vietnam, the type B, C, Y, OR, GY, OG were being rare, are in accordance with data in the literature for other regions of the world. For example, in a survey conducted in Macau (China), 85% of the specimens were of type G, 13% of type O, and less than 0.2% of type Y (Easton & Pun 1997). Hokkanen (1986) reported one specimen of type Y of 203 individuals (0.5%) in the United States. Because this last type was rare, no relationship of its occurrence with the food or temperatures can be done. The relationship between the different parents with colors of nymphs and adults Reseaching of the relationship between the different parents (GxG và GxO) with colors of nymphs and adults showed that, 1 st, 2 nd and 3 rd instars have only one color. 4 th and 5 th instars have two colors: green and black. Green was the most common both 4 th and 5 th instars. For 4 th instar, the color ratio of GxG and GxO are similar. GxG pair had 73.3% green 4 th instar and 26.7% black 4 th instar. There were 74.8% green and 25.2% black of 4 th instar belong to GxO pair (Table 3). The results of Table 4 showed that, the colors percentage of 5 th instar moult from green 4 th instar in different parents pairs (GxG and GxO) were not significant difference. Green 5 th instar was the most common. Percentage rate of green 5 th instar reached 89.6% and 87.8% respectively GxG and GxO pairs. Black 5 th instar rate was lower, respectively, 10.4% and 12.2% in GxO and GXG pairs. The color ratio (%) Nymph GxG GxO 4 th Green 73.3 74.8 instar Black 26.7 25.2 5 th Green 86.8 82.2 instar Black 13.2 17.8 For 5 th instar, green was 86.8% with GxG pair and 82.2% with GxO. Black color of 5 th instar was 17.8 % with GxO and higher than GxG pair (13.2%) (Table 3). Table 4: The color ratio of of 5 th instars molting from 4 th instar. Pairs The color ratio (%) Molting from green 4 th instar Molting from black 4 th instar Green 5 th instar Black 5 th instar Green 5 th instar Black 5 th instar GxG 89.6 10.4 48.1 51.9 GxO 87.8 12.2 25.9 74.1

Lam, Lam and Lan 280 The color rate of 5 th instar molting from black 4 th instar is related to the different parents pairs. For GxG pair, 79.9% from green 5 th instar and 59.4% from black 5 th instar. F phenotypic is he lowest rate reached 8.0% and the colors rate of green and black 5 th instar were 12.5% respectively, molting from green and black 5 th similar, reached respectively 48.1% and 51.9%. For GxO pair, black 5 th instar was common, with 74.1%, compared to green 5 th instar was only 25.9% (Table 4). instars. GxO pair created 4 phenotypes: G, O, F, R molting from green 5 th instar, is similar to Kiritani (1970). No The color of 4 th and 5 th instars was controlled by bugs belong to R phenotype molting from black 5 th genotypes of parents pairs. This result is similar to Kiritani (1970), showed that, t he Southern green stink bug, Nezara viridula, is polymorphic both in nymph and adult. The variation in body colour observed among instar. G types are dominant and lower than GxG respectively, 67.8% and 37.9% molting from green and black 5 th instar. Percentage rate of O and F types of GxG were higher than GXG pair. Rate of R phenotype nymph of 4 th and 5 th instars. was very low (0.9%) molting from green 5 th instar The color morphs of adults molting from 5 th instar (Table 5, Fig. 2). (Table 5, Fig. 2) showed: There were three phenotypes: G, O, F were moult from both in green and black 5 th The relationship between different parents pairs to the color of the offspring showed by Fig. 2. instars for GxG pairs. G-types are common, whith Table 5: The color ratio of adult molting from 5 th instar. The color ratio (%) Pairs Molting from green 5 th instar Molting from black 5 th instar G O F R G O F R GxG 79.9 12.1 8.0 0.0 59.4 28.1 12.5 0.0 GxO 67.8 20.9 1.4 0.9 37.9 41.4 20.7 0.0 Fig. 2. The relationship between the different parents with colors of nymphs and adults. CONCLUSION Type G is the best adapted to colonize different environments. The emergence of the phenotype 1. In Vietnam, field surveys of N. viridula on host depends on the temperature and food (host plants). plants (rice, corn, sesame) was collected 10 color Type F and R are better adapted to higher temperatures morphs including G, O, F, R, OR, GY, OG, Y, B and C. in summer crop on sesame. Low temperatures are the G- and O- types were common. The rare phenotype good conditions for the emergence of O-type. included B, C, Y, OR, OG and GY.

2. The 4 th and 5 th instars belong to both in GxG and GxO, had two colors: green and black; among colors, green was the most common. GxO pair created 4 phenotypes: G, O, F, R. The GxG pairs created 3 phenotypes: G, O, F. REFERENCES Easton, E.R. & W.W. Pun. (1997). Observations on some Hemiptera/ Heteroptera of Macau, Southeast Ásia. Proc. Entomol. Soc. Wash. 99: 574-582. Hokkanen, H. (1986). Polymorphism, parasites, and the native area of Nezara viridula (Hemiptera: Pentatomidae). Ann. Entomol. Fenn. 52: 28-31. Kiritani, K. (1970). Studies in the adult polymorphism in the southern green stink bug, Nezara viridula (Hemiptera: Pentatomidae). Res. Pop. Ecol. 12: 19-34. Kiritani, K. (1971). Distribution and abundance of the southern green stink bug, Nezara viridula. TARC, Tokyo, Proc. Symp. Rice Insects, pp. 235-248. Kiritani, K. & J. Yukawa. (1963). A note on the polymorphism of N. viridula and N. antennata. Rostria 5: 19-21. Lam, Lam and Lan 281 McLain,D.K. (1981). Sperm precedence and prolonged copulation in the southern green stink bug, Nezara viridula. J. Ga. Entomol. Soc. 16: 70-77. Ohno, K. & Md. Z. Alam. (1992). Hereditary basis of adult color polymorphism in the southern green stink bug, Nezara viridula Linné (Heteroptera: Pentatomidae). Appl. Entomol. Zool. 27: 133-139. Vivan, L.M. & A.R. Panizzi. (2002). Two new morphs of the southern green stink bug, Nezara viridula (L.) (Heteroptera: Pentatomidae), in Brazil. Neotrop Entomol. 31: 475-476. Vivan, L.M. & A.R. Panizzi. (2005). Nymphal and adult performance of genetically determined types of southern green stink bug, Nezara viridula (L.), under different temperature and photoperiodic conditions. Neotrop. Entomol. 34: 911-915. Yukawa, J. & K. Kiritani. (1965). Polymorphism in the southern green stink bug. Pacific Insects 7: 639-642.