1 Running Head: Incidence and Morphometrics of Colotis amata on Salvadora persica
2 Please address correspondence to: 3 Dr. Shravan M. Haldhar, Scientist 4 Agricultural Entomology 5 ICAR-Central Institute for Arid Horticulture 6 Sri Gananagar Highway, Beechwal Industrial Area P.O. 7 Bikaner-334006, India 8 E-mail: [email protected] 9
10 First report of Colotis amata (Lepidoptera: Pieridae) on Salvadora
11 persica (Capparales: Salvadoraceae) in Rajasthan, India: incidence
12 and morphometric analysis
13
14 SM Haldhar1*, R Bhargava1, RS Singh1, H Krishna1 & SK Sharma1
15 1Central Institute for Arid Horticulture, ICAR, Bikaner, India
16
17 *Corresponding author; E-mail: [email protected]
18 19
20 Abstract
21
22 An infestation of small salmon Arab, Colotis amata (F.) (Lepidoptera: Pieridae), on pilu
23 (Salvadora persica L.; Capparales: Salvadoraceae) plants was first noticed in 2012 at the
24 Experimental Farm of the Central Institute for Arid Horticulture and other fields of Bikaner
25 District, Rajasthan, India. The maximum incidence (80%) was observed in 18 Dec and the
26 minimum (13%) in 4 Sep. The average numbers of insects ranged between 6.2 and 22.3 larvae
27 per 3 leaves. Adults of this small butterfly were yellow and white in color. Eggs were laid singly
28 on leaves or young shoots and were 0.58-0.72 mm in length, 0.38-0.43 mm in width, and white
29 in color when first laid, later developing red blotches. The length and width of 1st instar larvae
30 were 1.98-2.29 mm and 0.36-0.48 mm, respectively. The lengths of 2nd, 3rd, and 4th instar
31 larvae were 5.04 mm, 9.77 mm, and 14.27 mm, respectively. The 5th instar larvae were 19.30
32 mm long and 3.33 mm wide. Pupae were 14.10 mm long and 5.73 mm wide and laterally
33 compressed. Adults were salmon-pink in color, and females had a body length of 9.71 mm and a
34 wingspan of 33.62 mm. Bodies of males were 7.59 mm long and their wingspan measured 25.67
35 mm. The lengths of male and female antennae were 4.63 mm and 5.46 mm, respectively.
36
37 Key Words: small salmon Arab, pilu, incidence, morphometric characters
38
39
40 41
42 Introduction
43 44 The genus Salvadora belongs to the family Salvadoraceae. It comprises 3 genera (i.e.,
45 Azima, Dobera, and Salvadora) and 10 species distributed mainly in the tropical and subtropical
46 regions of Africa and Asia (Mabberley 2008). On the Indian subcontinent, this family is
47 represented by only 1 genus with 2 species viz. Salvadora persica L. and Salvadora oleoides
48 Decne (Qureshi 1972; Stewart 1972; Perveen & Qaiser 1996). Salvadora persica, or pilu, is a
49 medium-sized tree or shrub with a crooked trunk, seldom more than 1 ft (0.3 m) in diameter. It
50 has a pleasant fragrance as well as a warm and pungent taste. Fruits have a sweet, agreeable,
51 aromatic, and slightly sour flavor. They can be eaten raw, cooked, or dried and stored when
52 dried. Fruits with or without seeds are said to contain 1.7-1.9% sugars when ripe, and delicious
53 drinks can be made from them. Salvadora persica is a popular chewing stick commonly known
54 as “miswak” and is one of the most popular medicinal plants throughout the Indian subcontinent
55 as well as other parts of Asia and the Middle East (Ezmirly et al. 1978; Almas 2002; Al-Otaibi et
56 al. 2003, 2004; Sofrata et al. 2007). When S. persica occurs on river terraces, it is a preferred
57 host of Cistanche tubulosa (Schenk) R. Wight (Lamiales: Orobanchaceae), an obligate
58 phanerogamic root parasitic plant (Sher et al. 2010). Furthermore, defoliating larvae of several
59 beetles and of Colotis ephyia (Klug) (Lepidoptera: Pieridae) often attack the tree, and the mite
60 Eriophyes sp. causes leaf galls (Kumar et al. 2012). An infestation with small salmon Arab has
61 not been reported. Here, we report new information on the incidence of C. amata on pilu,
62 including its morphological characters and taxonomic identification.
63
64 65
66 Materials and Methods
67
68 Twenty pilu plants were selected randomly in each of 3 replicates at the Experimental
69 Farm of the Central Institute for Arid Horticulture (CIAH) (N 28° 06' E 73° 21'). Incidence of
70 infestation observed on each plant and numbers of larvae per 3 leaves were recorded from Sep
71 2012 to Feb 2013. The sampling was done by visual observation and manual counting. Average
72 incidence was calculated as the percentage of whole plants infested with C. amata. Average
73 numbers of larvae per 3 leaves were calculated from numbers recorded for 10 randomly selected
74 pilu plants with 3 replications. Twenty insects (10 males and 10 females) were used for
75 observation and measurements. The larvae were reared under laboratory conditions (temperature
76 at 28 ± 2 oC with a relative humidity of 60 ± 5 %) for measurement of different stages. The
77 average linear measurements of various body parts of male and female adults were obtained
78 under a binocular microscope (Radical Instruments, Ambala, Haryana, India) using Jenoptic Pro
79 2.8.0 software. The terminology used to denote different parts of the body was according to
80 Haldhar (2012) and Haldhar & Singh (2014). The butterfly samples collected from the CIAH
81 farm and other fields were preserved in 70% alcohol and deposited at the Insect Biosystematic
82 Section, Division of Entomology, Indian Agricultural Research Institute, New Delhi, for
83 taxonomic identification.
84 Before statistical analysis, data were transformed as necessary to achieve normal
85 distribution. The data on incidence and numbers of insects were analyzed by 1-way ANOVA
86 using SPSS software (O’Connor 2000). When significant differences were detected, means were 87 compared by using Turkey’s honest significant difference (HSD) tests for paired comparisons at
88 a probability level of 5%.
89 Results and Discussion
90 The butterflies observed on pilu plants were identified as C. amata. Damaging effects
91 were recorded from Sep to Feb. The percentage of plants infested with C. amata ranged from 13-
92 80%, with the maximum incidence recorded on 18 Dec and the minimum on 4 Sep. The
93 maximum average number of larvae (22.3 per 3 leaves) was recorded on 18 Dec and the
94 minimum (6.2 per 3 leaves) on 4 Sep, followed by 7.6 larvae per 3 leaves on 19 Sep (Table 1).
95 Larvae damaged the new leaves, old leaves, and new or top branches of the pilu plants (Fig. 1).
96 Due to attack by this pest, the growth of pilu plants was suppressed, and new branches and
97 leaves wilted. The data on linear measurements of the small salmon Arab are presented in Table
98 2 and Figure 2. Eggs were laid singly on leaves or young shoots. They were 0.58-0.72 mm in
99 length, 0.38-0.43 mm in width, and white in color when first laid, later developing red blotches.
100 The 1st larval instar was greenish yellow with a black head. The length and width of 1st instar
101 larvae were 1.98-2.29 mm and 0.36-0.48 mm, respectively. The lengths of 2nd, 3rd, and 4th
102 instar larvae were 5.04, 9.77, and 14.27 mm, respectively. The 5th instar larvae were 19.30 mm
103 long, 3.33 mm wide, and light greenish in color with a white stripe on the dorsal surface of the
104 body. Pupae were 14.10 mm long, 5.73 mm wide, and laterally compressed. They had a
105 moderately high dorso-thoracic keel and a pointed but short cephalic projection, slightly up-
106 curved distally. Adults had a salmon-pink color, and females had a body length of 9.71 mm and a
107 wingspan of 33.62 mm. The male body length and wingspan were 7.59 mm and 25.67 mm,
108 respectively. The costa on the forewing was black and thickly overlaid with grayish or pinkish
109 scales. The lengths of male and female antennae were 4.63 mm and 5.46 mm, respectively. 110 Colotis amata infestations were observed on pilu plants in the hot arid region (Thar Desert) of
111 northwestern India. To our knowledge, this is the first report of C. amata on S. persica. In West
112 Africa, this species is a common butterfly of the Sahel (Larsen 2005). The flying capacity of
113 Colotis species is very low and weak. Both sexes are frequently found feeding from flowers and
114 spend much of their time flying around in the vicinity of their larval host plant (S. persica),
115 resting periodically on the ground (Pringle et al. 1994). In Delhi, India, females of C. vestalis
116 were noted ovipositing single eggs on or near old leaves of the host plant, while females of C.
117 amata laid eggs in large clusters on fresh leaves of the same trees. These observations pertained
118 to 2 species of the plant genus Salvadora (Larsen 1988). This butterfly is damaging
119 economically important parts of the plant, such as leaves, flowers, and top branches. Therefore,
120 management practices need to be developed and implemented to minimize the losses caused by
121 this pest.
122 Acknowledgement
123
124 The authors thank V. V. Ramamurthy, National Coordinator, ICAR Network Project on
125 Insect Biosystematics, Division of Entomology, IARI, New Delhi, for the identification of this
126 pest and R. Swaminathan, Associate Professor, Department of Entomology, MPUAT, Udaipur,
127 for critical discussions and suggestions.
128 References
129 Almas K. 2002. The effect of Salvadora persica extract (Miswak) and chlorhexidine gluconate
130 on human dentin: a SEM study. Journal of Contemporary Dental Practice, 3: 27-35. 131 Al-Otaibi M, Al-Harthy M, Söder B, Gustafsson A & Angmar-Mansson B. 2003. Comparative
132 effect of chewing sticks and toothbrushing on plaque removal and gingival health. Oral
133 Health and Preventive Dentistry, 1: 301-307.
134 Al-Otaibi M, Al-Harthy M, Gustafsson A, Johansson A, Cleasson R & Angmar-Mansson B.
135 2004. Subgingival plaque microbiota in Saudi Arabians after use of miswak chewing
136 stick and toothbrush. Journal of Clinical Periodontology, 31: 1048-1053.
137 Ezmirly ST, Cheng JC & Wilson SR. 1978. Saudi Arabian medicinal plants: Salvadora persica.
138 Planta Medica, 35: 191-192.
139 Haldhar SM. 2012. Report of Homoeocerus variabilis (Hemiptera: Coreidae) on khejri (Prosopis
140 cineraria) in Rajasthan, India: incidence and morphometric analysis. Florida
141 Entomologist, 95: 848-853.
142 Haldhar SM & Singh RS. 2014. Report of Dictyla cheriani Drake (Hemiptera: Tingidae)
143 on Indian cherry (Cordia myxa) in Rajasthan, India: incidence and morphometric
144 analysis. Indian Journal of Agricultural Sciences, 84: 303-305.
145 Kumar S, Rani C & Mangal M. 2012. A critical review on Salvadora persica: an important
146 medicinal plant of arid zone. International Journal of Phytomedicine, 4: 292-303.
147 Larsen TB. 1988. Differing oviposition and larval feeding strategies in two Colotis butterflies
148 sharing the same food plant. Journal of the Lepidopterists’ Society, 42: 57-58.
149 Larsen TB. 2005. Butterflies of West Africa. Apollo Booksellers, Vester Skerninge, Denmark.
150 Mabberley DJ. 2008. Mabberley's Plant Book: A Portable Dictionary of Plants, their
151 Classifications, and Uses. 3rd ed. Cambridge University Press, Cambridge, United
152 Kingdom. 153 O’Connor RP. 2000. SPSS and SAS programs for determining the number of components using
154 parallel analysis and Velicer’s MAP test. Behavior Research Methods, Instruments, and
155 Computers, 32: 396-402.
156 Perveen A & Qaiser M. 1996. Pollen flora of Pakistan – VI. Salvadoraceae. Pakistan Journal of
157 Botany, 28: 151-154.
158 Pringle EL, Henning GA & Ball JB [eds]. 1994. Pennington’s Butterflies of Southern Africa, 2nd
159 ed. Struik Winchester, Cape Town, South Africa.
160 Sher H, Al-Yemeni MN, Masrahi YS & Shah AH. 2010. Ethnomedicinal and ethnoecological
161 evaluation of Salvadora persica L.: A threatened medicinal plant in Arabian Peninsula.
162 Journal of Medicinal Plants Research, 4: 1209-1215.
163 Sofrata A, Lingström P, Baljoon M & Gustafsson A. 2007. The effect of miswak extract on
164 plaque pH: an in vivo study. Caries Research, 41: 451-454.
165 166 Table 1. Mean percent of Salvadora persica plants infested by the small salmon Arab, Colotis
167 amata, and mean number of larvae per 3 leaves of plant in 2012-2013 at the experimental
168 farm of the ICAR-Central Institute for Arid Horticulture, Bikaner District, Rajasthan,
169 India.
Sampling date Mean incidence Mean no. of C. amata larvae
(%)a,b per 3 leavesb
4 Sep 2012 13.33 (21.14)a 6.20a
19 Sep 2012 16.67 (23.85)a 7.63a
4 Oct 2012 30.00 (32.99)b 10.37b
19 Oct 2012 36.67 (37.21)bc 11.93bc
3 Nov 2012 56.67 (48.83)de 13.33cd
18 Nov 2012 60.00 (50.83)de 16.30ef
3 Dec 2012 70.00 (56.98)efg 18.30gh
18 Dec 2012 80.00 (63.90)g 22.30i
2 Jan 2013 66.67 (54.76)ef 19.00hi
17 Jan 2013 63.33 (52.75)ef 16.77fg
01 Feb 2013 46.67 (43.06)cd 12.80cd
16 Feb 2013 36.67 (37.21)bc 10.80bc
170 a Parentheses contain the angular transformation value.
171 b Values followed by different letters are significantly different (Tukey’s HSD test, P < 0.05).
172 173 Table 2. Mean (± SEM) linear morphometric measurements of different life stages of the small 174 salmon Arab, Colotis amata. Life stage Length (mm) Width (mm)
Egg 0.62 ± 0.009 0.41 ± 0.007
1st instar larva 2.15 ± 0.023 0.42 ± 0.012
2nd instar larva 5.04 ± 0.027 1.15 ± 0.022
3rd instar larva 9.77 ± 0.022 1.96 ± 0.011
4th instar larva 14.27 ± 0.052 2.27 ± 0.021
5th instar larva 19.30 ± 0.100 3.33 ± 0.012
Pupa 14.10 ± 0.046 5.73 ± 0.020
Adult female with wing expansion 9.71 ± 0.007 33.62 ± 0.031
Adult male with wing expansion 7.59 ± 0.021 25.67 ± 0.035
Female antenna 5.46 ± 0.016 not applicable
Male antenna 4.63 ± 0.017 not applicable
175 a Mean of ten specimens.
176
177 178 Figure Captions
179
180 Fig. 1. Aggregation of Colotis amata on a pilu plant (Salvadora persica) during the winter
181 season at the ICAR-Central Institute for Arid Horticulture, Bikaner, India.
182
183 Fig. 2. Different developmental stages of Colotis amata. A, eggs; B, 1st instar larvae; C, 5th
184 instar larva; D, pupae; E, adult male; F, adult female.
185
186
187 188
189 Fig. 1. Aggregation of Colotis amata on a pilu plant (Salvadora persica), during the winter
190 season at the ICAR-Central Institute for Arid Horticulture, Bikaner, India.
191
192 193 194
195 Fig. 2. Different developmental stages of Colotis amata. A, eggs; B, 1st instar larvae; C, 5th
196 instar larva; D, pupae; E, adult male; F, adult female.
197