Corresponding author: Alain Roques (
Academic editor: M. Engel
A survey of seed chalcids from woody plants in Kenya revealed 12 species belonging to the genus cytochrome oxidase subunit one –
Roques A, Copeland RS, Soldati L, Denux O, Auger-Rozenberg M-A (2016)
Globalization, the accelerating worldwide movement of goods and people, has led to an exponential increase in the transport of alien organisms across geographical barriers (
Seeds of woody plants can be infested by larvae of a number of arthropod groups, including, among others, dipteran seed midges in the family
Data from these recent studies indicate that, worldwide, the genus
To date, relatively few species have been reported from subtropical and tropical areas. In the Afrotropical region, only seven species have been recognized. Four species develop as phytophages in angiosperm seeds. Three of these,
The lower diversity of International Centre of Insect Physiology and Ecology
Over 700 insect species were reared from fruits and much of the data from the two projects has been published previously (e.g.
During the 1999–2004 wild-fruit survey, 3839 fruit collections were made throughout much of Kenya. An additional 347 wild-fruit collections were made during the 2011–2012 survey in the Taita Hills, bringing the total number of fruit samples to 4186, the great majority from woody plants. Over the course of the two surveys approximately 930 species of plants were sampled, representing 122 families. Details of fruit collection and insect rearing methods are available in
During transport, damage to fruits was minimised by placing each fruit sample in a separate polythene bag and suspending the bag above the bottom of a two-litre plastic storage container. A tight-fitting plastic lid fixed the bag and its contents in place. A large rectangular piece had been cut out of the lid and replaced with fine mesh cloth to allow for ventilation of the fruits. In the laboratory, fruits were removed from transport bags and placed within one-litre, rectangular plastic containers that had small elliptical holes cut out of the bottom. Each one-litre container (also provided with a mesh-covered lid) was nested within a fresh two-litre container, the bottom of which had a layer of heat-treated sand. Fruits were usually held for up to two months. Emerged adult insects were held for 1–3 days before being killed.
Beentje (1994) and Agnew and Agnew (1994) were used for preliminary plant identifications. Identifications made by the second author were confirmed or corrected by Quentin Luke of the East African Herbarium. Plant names and authors correspond to those found in The International Centre of Insect Physiology and Ecology
Independent of the wild-fruit surveys, at various times between 2005 and 2014, the second author surveyed Kenyan insects using Townes-style Malaise traps (
Adult morphology was examined using a MZ12 Leica stereomicroscope equipped with a Leica IC A camera. When more than three specimens were obtained per species and sex, one individual of each sex was dissected and wings of both sexes and male genitalia were mounted on glass slides using Dimethyl hydantoin formaldehyde
In a few specimens, head, antenna and thoracic sculpture were examined using a Cambridge StereoscanB 90 scanning electron microscope equipped with OrionB® image processing software. When less than three specimens were available per species and sex, these specimens were kept intact and mounted on triangular cardboards. Photographs and measurements of body and body parts (wings, antenna, head, thorax, gaster and ovipositor) were made as above without dissecting the mounted specimens. Morphological terminology follows : Ocellocular line: the minimum distance between the eye margin and the adjacent posterior ocellus : Posterior ocellar line: the minimum distance between the posterior ocelli
• DNA extraction, molecular markers, amplification and sequencing
In order to include the Kenyan species in an existing phylogeny of
In addition to the
• Phylogenetic analyses
For all datasets, sequences were aligned using Clustal W ( Kimura-2-parameter
Phylogenetic analyses were performed using maximum likelihood Bayesian inferences
Abbreviations used for insect specimen depositories or private collections cited in the study are as follows:
National Museums of Kenya
Robert Copeland personal collection, ICIPE, Nairobi, Kenya
Alain Roques personal collection, INRA Zoologie Forestière Orléans, France
International Centre of Insect Physiology and Ecology, Nairobi, Kenya
IZIKO South African Museum, Capetown, South Africa
British National History Museum, London, UK
Distribution in Kenya of the previously described African
Distribution in Kenya of the new
Fruit and Malaise trap samples producing
Family / Host plant species | Region | Location | Latitude N | Longitude E | Elevation1 (m) | |
---|---|---|---|---|---|---|
|
||||||
Eastern Arc Mountains | Sagalla Mountain |
|
|
1090 | ||
Central highlands | Machakos |
|
|
1586 | ||
Eastern Arc Mountains | Base of Kasigau Mountain |
|
|
619 | ||
Western mid-altitude | Gembe Hills |
|
|
1362 | ||
|
Western mid-altitude | Sindo-Mbita Road |
|
|
1205 | |
Western mid-altitude | Sindo-Mbita Road |
|
|
1205 | ||
|
Coastal forests | Arabuko-Sokoke Forest |
|
|
55 | |
|
|
Coastal forests | Arabuko-Sokoke Forest |
|
|
55 |
|
|
Coastal forests | Arabuko-Sokoke Forest |
|
|
98 |
|
|
Coastal forests | Shimba Hills |
|
|
285 |
Western highlands | nr. Narok on main road |
|
|
1919 | ||
|
Western highlands | nr. Narok on main road |
|
|
1830 | |
|
Northern highlands | Mt Kulal |
|
|
1639 | |
|
Western highlands | Mai Mahiu-Narok Road |
|
|
1900 | |
Central Highlands | Kikuyu Escarpment |
|
|
2100 | ||
|
|
Central Highlands | Kikuyu Escarpment |
|
|
2100 |
|
|
Central Highlands | Burguret |
|
|
1960 |
|
|
Northern mid-altitude | south of Marsabi town |
|
|
1200 |
|
|
Western mid-altitude | Gembe Hills |
|
|
1370 |
|
|
Eastern Arc Mountains | Ronge-Nyika, mixed shrub- and grassland |
|
|
1050 |
|
|
Eastern Arc Mountains | Kasigau forest |
|
|
1280 |
|
Central Highlands | Kikuyu escarpment |
|
|
2200 | |
|
Central Highlands | Nairobi |
|
|
1610 | |
|
Central highlands | Nairobi |
|
|
1610 | |
|
|
Central highlands | Nairobi |
|
|
1750 |
|
||||||
Central highlands | Mt. Kenya Forest |
|
|
2130 | ||
|
|
Northern highlands | Mt. Kulal |
|
|
1650 |
|
||||||
Northern highlands | Mathews Range |
|
|
1340 | ||
|
||||||
Coastal forests | Arabuko-Sokoke Forest |
|
|
60 | ||
|
Eastern savanna | Tsavo |
|
|
640 | |
|
Eastern Arc Mountains | Mwatate area, on small farm |
|
|
1050 | |
|
|
Eastern savanna | Kasaala |
|
|
740 |
|
|
Eastern Arc Mountains | Kasigau Mountain Forest |
|
|
1070 |
|
Central Highlands | Nairobi, |
|
|
1600 | |
|
|
Coastal island | Funzi Island |
|
|
0 |
A total of 12
Other host fruits of
Host-plant genera of
Family | Genus | Total no. of Kenyan species | No. of Kenyan species sampled | No. of sampled Kenyan species yielding |
Total number of Afrotropical species |
---|---|---|---|---|---|
|
|
10 | 5 | 3 (60) | 29 |
|
2 | 2 | 2 (100) | 44 | |
|
1 | 1 | 1 (100) | 2 | |
|
6 | 4 | 2 (50) | 106 | |
|
25 | 25 | 2 (100) | 25 | |
|
|
1 | 1 | 1 (100) | 4 |
|
|
2 | 2 | 1 (50) | 3 |
|
|
28 | 14 | 2 (14) | 122 |
1First record of genus as a host of
Of the host families,
Before our study the host of
Among non-
Table
Host fruit infestation indices for
Plant family/ Plant species | No. of fruits | No. of reared |
% of infestation by |
|
---|---|---|---|---|
|
||||
786 |
|
3 | 0,4 | |
|
338 |
|
11 | 3,3 |
|
168 |
|
1 | 0,6 |
2806 |
|
226 | 8,1 | |
|
1505 |
|
42 | 2,8 |
|
698 |
|
1 | 0,1 |
|
2442 |
|
22 | 0,9 |
2603 |
|
12 | 0,5 | |
1019 |
|
1 | 0,1 | |
|
674 |
|
41 | 6,1 |
|
524 |
|
6 | 1,1 |
|
2394 |
|
14 | 0,6 |
|
894 |
|
36 | 4,0 |
|
412 |
|
6 | 1,5 |
|
769 |
|
113 | 14,7 |
|
535 |
|
79 | 14,8 |
|
||||
|
82 |
|
8 | 9,8 |
|
||||
|
448 |
|
3 | 0,7 |
|
100 |
|
13 | 13,0 |
|
1056 |
|
29 | 2,7 |
The four Malaise trap samples containing
Our fruit collections bring to six the number of Afrotropical plant families that are hosts of
Table
MtDNA was sequenced for nine of the 12 African species defined morphologically, including five of the eight new species and the four previously described ones. Using the longest mitochondrial marker (“Clyde-Bonnie”), 810 bp- long
The inclusion of the nine Kenyan species in an already existing molecular phylogeny (
Mean genetic distances within and between groups of
|
|
|
|
|
|
|
Gall-former | unknown | outgroup | |
---|---|---|---|---|---|---|---|---|---|---|
|
0.061\0.057 | |||||||||
|
0.088\0.073 | 0.060\0.046 | ||||||||
|
0.081\0.074 | 0.084\0.074 | -\- | |||||||
|
0.085\0.074 | 0.093\0.080 | 0.086\0.077 | 0.079\0.068 | ||||||
|
0.098\0.086 | 0.089\0.080 | 0.094\0.083 | 0.102\0.087 | -\- | |||||
|
0.113\0.095 | 0.112\0.105 | 0.107\0.096 | 0.108\0.088 | 0.111\0.097 | -\- | ||||
|
0.111\0.096 | 0.112\0.099 | 0.099\0.104 | 0.108\0.100 | 0.111\0.095 | 0.107\0.078 | 0.068\0.063 | |||
Gall-former | 0.102\0.092 | 0.100\0.085 | 0.094\0.096 | 0.100\0.086 | 0.106\0.080 | 0.094\0.086 | 0.105\0.092 | -\- | ||
Unknown | -\0.094 | -\0.089 | -\0.096 | -\0.094 | -\0.096 | -\0.083 | -\0.082 | -\0.083 | ||
Outgroup | 0.157\0.127 | 0.151\0.123 | 0.158\0.135 | 0.157\0.134 | 0.161\0.132 | 0.140\0.113 | 0.146\0.115 | 0.154\0.121 | -\0.113 | -\- |
Between groups, similar values were observed between Holarctic groups, as well as between the species related to
The use of maximum likelihood and Bayesian inference methods resulted in similar information about phylogenetic relationships within the genus
Bayesian-likelihood inference phylogenies based on cytochrome oxidase I (
In the different trees, robust clades (
Despite amplifying for all markers used, the phylogenetic position of
The mtDNA sequences and nuclear sequences determined in the course of this study are registered under the following GenBank accession numbers:
In some
Therefore, we provide a key to females only, including the species associated with rose hips, conifers, and galls that were not collected during the surveys (i.e.
1 | Ovipositor sheaths shorter (0.4–0.9×) than gaster length (Figures |
|
– | Ovipositor sheaths longer (1.1×–1.8×) than gaster length (Figures |
|
2 | Ovipositor sheaths much shorter (0.4–0.6×) than gaster length (Figures |
|
-– | Ovipositor sheaths 0.8–0.9× as long as gaster length (Figures |
|
3 | Body colour nearly entirely orange-yellow; only a narrow black band along anterior suture of pronotum (Figures |
|
- | Body colour with a large longitudinal, median rufous band extending from pronotum to scutellum (Figure |
|
4 | Pilosity on head entirely pale (Figure |
|
– | Pilosity pale on face but dark on other parts of head (Figure |
|
5 | Minute species, body length less than 1.5 mm; Thorax pilosity pale; 2 pairs of hairs on scutellum (Figure |
|
– | Larger species, body length more than 2.5 mm; Thorax pilosity dark; from 3 to 8 pairs of hairs on scutellum (Figures |
|
6 | Body colour mostly black | |
– | Body colour nearly completely orange-yellow (Figure |
|
7 | Thorax colour orange or pale yellow without extensive dark markings (Figures |
|
– | Thorax with extensive dark markings (Figures |
|
8 | Ovipositor sheaths less than 1.2× as long as gaster length (Figure |
|
– | Ovipositor sheaths at least 1.4× as long as gaster length (Figures |
|
9 | Forewing with an elongate stigma, 1.7× as long as wide |
|
– | Forewing with an oval- rounded stigma, 1.2–1.3 × as long as wide (Figure |
|
10 | Ovipositor sheaths at least 1.8 × as long as gaster length and 0.8× as long as body length (Figure |
|
– | Ovipositor sheaths at most 1.6× as long as gaster length and 0.6× as long as body length (Figure |
|
11 | Forewing stigma oblong, 1.7 × as long as wide, with a very short stigmal vein, at most 0.2× as long as stigma length; gaster tergites pale yellow with translucent median longitudinal stripes, transversely infumate |
|
– | Forewing stigma elongate-oval, 2.0 × as long as wide, with an elongate stigmal vein, 0.4 × as long as stigma length (Figure |
|
12 | Ovipositor sheaths never longer than 1.5× gaster length (Figures |
|
– | Ovipositor sheaths at least 1.8× longer than gaster length (Figures |
|
13 | Pronotum with a large median dark brown band progressively decreasing in size from the anterior to posterior suture (Figure |
|
– | Pronotum yellow or brownish, without median dark band; scutellum with 7–8 pairs of hairs; in seeds of |
|
14 | Propodeum smooth; forewing stigma elongate, more than 2× as long as broad |
|
– | Propodeum rugose, with reticulate striae (Figure |
|
15 | Ovipositor sheaths at least 1.2× longer than body length. In |
|
– | Ovipositor sheaths never longer than body length. In |
|
16 | Pronotum orange with a conspicuous longitudinal black stripe on each side (Figures |
|
– | Pronotum without lateral black stripes but with a large, nearly rectangular dark brown band medially extending from anterior to posterior suture (Figures |
|
Updated synthesis of biological habits of the world
Head transverse (Figure
Head rectangular (Figure
Known only from the Matthews Range, Kenya where it was observed to emerge from 9.8 % of the collected fruits (Table
Named after its host plant.
This is the only species of
Forewing stigma brown without infuscation; basal cell indistinct, lacking setae, no setae on basal setal line and costal setal line (Figure
Head transverse (Figure
Known only from Sagalla Mountain, Taita Hills, Kenya. Specimens emerged from only 0.4% of sampled fruits (Table
Named in honour of Robert J. Smith, noted musicologist and exceptional left fielder.
Its comparatively very short ovipositor, nearly 2 × smaller than gaster length, allows to separate easily this species from the other ones developing in Anacardiacae fruits and showing resembling body colour patterns but a comparatively much longer ovipositor, quite as long as gaster length (
Head rounded, width: height ratio: 1.3×;
No males reared.
Known only from the Mwatate area, Taita Hills, Kenya.
Named after Dr. Robert Copeland, who collected the specimen.
The combination of an almost entirely orange body, pale pilosity on the head, and an ovipositor a bit shorter than gaster length readily distinguishes this species from other Afrotropical
23♀♀, 11♂♂ same collection data as holotype, except 19 Dec 2004, A&M coll. N°3077 (11♀♀, 5♂♂
Legs pale yellow except claws brownish; coxae with numerous pale hairs extending from small black dots, especially on hind coxa; femora with small black hairs; tibiae with 3 rows of long black hairs. Forewing stigma brown without infuscation; basal cell only partly closed, the disc with 5 very small setae; basal line with 2 setae; no setae on costal line; 4 setae in coastal cell (Figure
Head about 1.1 × as wide as long in front view (Figure
Legs yellow expect anterior part of coxae with large black spots (extending to the posterior part in fore coxa), and small brownish spots at femur insertion, 3 rows of large hairs on tibia. Forewing stigma brown without infuscation (Figure
Head about 1.2 × as wide as long in front view.
Known only from western Kenya, near Lake Victoria. In one collection, 8.1% of fruits were infested (Table
Named after the genus of the host plant.
Females are easily distinguished from those of other species observed to emerge from fruits of
3♂♂, collected together with the four females (2♂♂
Head about 1.2 × as wide as long in front view (Figure
Legs yellow with the two last tarsal segments and claws black. Pilosity pale on head and thorax, showing the same setal pattern as in female
Head transverse, about 1.3 × as wide as long in front view (Figure
Unknown. In the molecular study,
Only known from the collection site in a degraded meadow near a small stream.
Named after the institution on whose grounds the species was collected, the campus of the International Centre of Insect Physiology and Ecology (
The species can be easily distinguished from other tiny
Head rounded, width: height ratio: 1.0 (Figure
Known from the base of Kasigau Mountain, a Taita Hills outlier. Only 1 of 168 fruits (0.6%) was infested (Table
Named in honour of Leo Laventhal, historian, Yiddish scholar and union man.
Females can be separated easily from those of other species associated with fruits of
Kenya, 7♀♀, 1♂, same as holotype (
Legs entirely pale yellow except claws brown, coxae with conspicuous hair dots. Forewing stigma brown without infuscation; basal cell closed, with 13 setae on disc ; basal setal line with 6 long setae; costal setal line with 16 small setae, costal cell with 11 setae in 2 rows (Figure
Head rounded, width: height ratio: 1.1 × (Figure
Head subquadrate, width: height ratio: 1.1 × (Figure
Known from dry
Named after the genus of its host plant.
Females are easily separated from those of
Kenya; 7 ♀♀ same collection data as holotype; 2♀♀, same collection data as holotype except 25 Nov. 2004, ex.
Face quadrate, width: height ratio: 1.0 (Figure
Head about 1.1 × as wide as long in front view (Figure
Known only from farmland tree in Machakos area. Adults emerged from 3.3% of the collected fruits (Table
Named after the genus of its host plant.
Females can be separated easily from those of other species associated with fruits of
11♀♀, 14♂♂, Kenya, Coast Province, Shimba Hills,
This species has been first described by
Thorax yellowish-brown with blackish patterns (Figures
Face subquadrate, width: height ratio in front view: 1.2×;
Thorax mostly yellow- orange with a few black patterns (Figure
Face rounded, width: height ratio in front view: 1.0× (Figure
Relative dimensions of head and antenna similar as in pale form; sculpture of thorax and propodeum same as in pale form. Forewing stigma nearly rectangular, 1.3 × as long as wide (Figure
Body length as in pale form, from 4.6 to 5.4 mm. Some specimens with infuscate stigma show lighter coloration patterns than in the extreme dark form, with the median band on pronotum smaller, the lateral lobes of mesoscutum and axilla yellow, the scutellum with the median band limited to the anterior part, and the gaster with narrow yellow bands from T4 to T6.
Mixed bush-/grassland in western Kenya near Lake Victoria and in similar habitat in coastal areas. Adults emerged from up to 2.8% of the fruits in some collections (Table
The key from
Genitalic characters also allow separation of
3♀♀, 3♂♂, Kenya, Mount Kulal,
Male and female specimens fit the detailed description of the species by
In the molecular phylogeny of
Reared from samples collected in highland areas (1640–1920 m above sea level) of the Rift Valley, in northern and southwestern Kenya. Few fruits (0.6%) were infested (Table
Females can be separated from those of other species with a predominantly orange colour associated with fruits of
The genitalia allow one to separate males from those of other species identified in seeds of
3♀♀, 7♂♂, Kenya, Kikuyu Escarpment, Central Province,
Male and female specimens fit the description of the species by
Adults emerged from 14.8% of the fruits of
Females can be distinguished from these of other species associated with fruits of
Genitalia allows the separation of males from those of other species reared from seeds of
Holotype ♀ “Bristish Somaliland; ex. seeds of
1♀ “Abyssinia, Mulu, above Muger Valley; circa 8000 ft., 18–23. 12. 1926, Dr. H. Scott” (
Male and female specimens obtained from Kenya fit the general description of the species by
Sampled in highland forest and woodland in central and northwestern Kenya (Figure
This species, as well as the related
Is the radiation on Angiosperms more important than previously considered?
Our results increase to 16 the number of
This work was partly supported by grants awarded to R. Wharton, Texas A&M University, USA, from USAID (no. PCE-G-00-98-0048-00) and USDA/CSREES/IFAFS (no. 00-52103-9651) in collaboration with the International Centre for Insect Physiology and Ecology (
Sampling details on specimens used in the genetic study
Data type: Collection data
Explanation note: Collection data for the specimens used in the genetic study:
25 species already sequenced and used in a previous phylogenetic study (
4 species already described in the literature but sequenced and analysed for this paper.
5 new species described in this paper.