Within- and between-individual sequence variation among ITS1 copies in the meadow grasshopper Chorthippus parallelus indicates frequent intrachromosomal gene conversion

Sequencing multiple copies of the ITS1 region revealed the coexistence of two or more haplotypes within the genome of Chorthippus parallelus. Using a PCR-RFLP approach, the ITS1 numbers and frequencies of haplotypes present in each of 40 individuals were investigated, revealing a consistent lack of homogeneity. For each individual, the level of intra-individual variation was estimated from a sample of 20 ITS1 copies. The level of differentiation in haplotype frequency among individuals was then estimated by maximum likelihood using models based on the Dirichlet distribution. This confirmed the existence of significant levels of variation among individuals within each population studied. The most likely turnover mechanism that could generate this pattern of variation is gene conversion, operating at the intrachromosomal level. Furthermore, the discovery of linkage disequilibrium among the ITS1 haplotypes of C. parallelus suggests that intrachromosomal gene conversion occurs more frequently than interchromosomal recombination. Subspecies of C. parallelus showed significantly different haplotype distributions following about 0.5 Myr of divergence. With respect to the process of concerted evolution, we show that homogenization of repeats is slow relative to speciation, and the standing variation among individuals is sufficient for selection to operate.     

Development of pheromone traps for control and monitoring of sweetpotato weevils, Cylas puncticollis and C. brunneus, in Uganda

Experiments were carried out in Uganda to optimise pheromone traps for the African sweetpotato weevil species, Cylas puncticollis Bohe. and C. brunneus F. (Coleoptera: Apionidae). Various designs of funnel, water and sticky traps were compared and a 5-l plastic jerry can trap was the most appropriate design for effectiveness and practicability. A solution of Omo detergent in water was found to be the most effective trapping agent. Fewer weevils were caught in red traps than in yellow, white, green or blue traps. Catches of C. puncticollis increased when the trap was raised above crop height, but catches of C. brunneus were unaffected. When marked weevils were dropped onto the trap, 36% of C. puncticollis and 23% of C. brunneus were captured, and, of weevils placed in the trap, 88% and 92%, respectively, of the two species remained overnight. Lures for the two species showed no significant loss in attractiveness after 8 weeks in the field, and chemical analysis showed 19% of the C. puncticollis pheromone and 72% of the C. brunneus pheromone remaining after this time. Summarizing, the following trap is presently recommended for monitoring/controlling African Cylas species: a 5-l plastic jerry can trap of any colour with rectangular openings of 11×5 and 6×5 cm positioned 15 cm above the crop canopy, filled with 0.5 l Omo solution (1 g/1 l water), with 0.1 mg lures to be replaced every 8 weeks.     

Evolution and structural conservation of the control region of insect mitochondrial DNA

The control regions of mitochondrial DNA of two insects, Schistocerca gregaria and Chorthippus parallelus, have been isolated and sequenced. Their sizes are 752 by and 1,512 bp, respectively, with the presence of a tandem repeat in C. parallelus. (The sequences of the two repeats are highly conserved, having a homology of 97.5%.) Comparison of their nucleotide sequences revealed the presence of several conserved sequence blocks dispersed through the whole control region, showing a different evolutionary pattern of this region in these insects as compared to that in Drosophila. A highly conserved secondary structure, located in the 3 region near the small rRNA gene, has been identified. Sequences immediately flanking this hairpin structure rather than the sequences of this structure themselves are conserved between S. gregaria/C. parallelus and Drosophila, having a sequence consensus of TATA at 5 and GAA(A)T at 3. The motif G(A)nT is also present in the 3 flanking sequences of mammalian, amphibian, and fish mitochondrial L-strand replication origins and a potential plant mitochondrial second-strand-replication origin, indicating its universal conservation and functional importance related to replication origins. The stem-and-loop structure in S. gregaria/C. parallelus appears to be closely related to that found in Drosophila despite occupying a different position, and may be potentially associated with a second-strand-replication origin. This in turn suggests that such a secondary structure might be widely conserved across invertebrates while their location in the control region may be variable. We have looked for such a conserved structure in the control regions of two other insects, G. firmus and A. mellifera, whose DNA sequences have been published, and their possible presence is discussed.Mitochondrial control regions characterized to date in five different insect taxa (Drosophila, G. firmus, A. mellifera, S. gregaria, and C. parallelus) may be classed into two distinct groups having different evolutionary patterns. It is observed that tandem repetition of regions containing a probable replication origin occurred in some species from disjunct lineages in both groups, which would be the result of convergent evolution. We also discuss the possibility of a mechanism of parahomologous recombination by unequal crossing-over in mitochondria, which can explain the generation of such tandemly repeated sequences (especially the first critical repetition) in the control region of mtDNA, and also their convergent evolution in disjunct biological lineages during evolution. Correspondence to: G.M. Hewitt     

Testing the efficiency of three 15N-labeled nitrogen compounds for indirect labeling of grasshoppers via plants in the field

In field studies of plant–insect herbivore interactions it is often difficult to establish which herbivore has fed on a particular plant. We investigated the suitability of three different ¹⁵N‐labeled nitrogen compounds (ammonium, nitrate, and glycine) for indirect marking of three grasshopper species [Omocestus viridulus (L.), Chorthippus parallelus (Zett.), and Chorthippus biguttulus (L.) (Orthoptera: Acrididae)] through labeling their food plants in the field. In two short‐term experiments grassland plots of 1 m² were separately labeled with either one of the different nitrogen compounds. Grasshoppers were caged on three food‐plant species [Dactylis glomerata L., Holcus lanatus L. (Poaceae), and Trifolium repens L. (Fabaceae)] present in these plots for 72 h. Significantly enriched δ¹⁵N values in grasshoppers were found in all plant/grasshopper combinations. Enrichment in grasshoppers was positively correlated with the enrichment of plants and labeling with nitrate resulted in highest ¹⁵N enrichment. In a long‐term experiment, individuals of C. biguttulus were placed in a cage covering an area of 1 m² for 37 days, with sampling of grasshoppers at regular intervals. δ¹⁵N values of the grasshopper and a common food plant, D. glomerata, increased steadily over time, up to 40‐fold by the end of the experiment. Our results demonstrate that ¹⁵N‐labeling of plants is an appropriate tool for the investigation of insect–plant interactions under natural conditions.     

大兴安岭地区雏蝗属一新种(直翅目:蝗总科)

雏蝗属Ｃｈｏｒｔｈｉｐｐｕｓ Ｆｉｅｂｅｒ是网翅蝗科中一个大属,广泛分布于中国古北区。本文记述采自内蒙古大兴安岭地区的雏蝗属一新种：额左旗雏蝗Ｃｈｏｒｔｈｉｐｐｕｓ ｅｚｕｏｑｉｅｎｓｉｓ ｓｐ．ｎｏｖ．,模式标本保存于东北师范大学生物系动物标本室。     

雏蝗属二新种记述（直翅目：蝗总科）

本文山西省雏蚯属Ｃｈｏｒｔｈｉｐｐｕｓ ２新种：恒山雏蚯Ｃ．ｈｅｎｇｓｈａｎｅｎｓｉｓ ｓｐ．ｎ．及太原雏蚯Ｃ．ｔａｉｙｕａｎｅｎｓｉｓ ｓｐ．ｎ．,模式标本存山西省农牧厅植保植检总站。     

Long Repeats in a Huge Genome: Microsatellite Loci in the Grasshopper Chorthippus biguttulus

It is commonly believed that both the average length and the frequency of microsatellites correlate with genome size. We have estimated the frequency and the average length for 69 perfect dinucleotide microsatellites in an insect with an exceptionally large genome: Chorthippus biguttulus (Orthoptera, Acrididae). Dinucleotide microsatellites are not more frequent in C. biguttulus, but repeat arrays are 1.4 to 2 times longer than in other insect species. The average repeat number in C. biguttulus lies in the range of higher vertebrates. Natural populations are highly variable. At least 30 alleles per locus were found and the expected heterozygosity is above 0.95 at all three loci studied. In contrast, the observed heterozygosity is much lower (≤0.51), which could be caused by long null alleles. [Reviewing Editor: Dr. Dmitri Petrov] Sequence data from this article have been deposited with the EMBL/GenBank databases under accession numbers AY532396–AY532400.     

Hsp70 level in progeny of aging grasshoppers from variously polluted habitats and additionally exposed to zinc during diapause

The hsp70 level in the bodies of 1st instars of grasshoppers Chorthippus brunneus from unpolluted (Pilica) and polluted (Olkusz, Szopienice) sites and additionally exposed to various doses of zinc during diapause and embryonic development prior to hatching were measured by Western blotting. The main aim of our work was to assess the relationship between the age of female grasshoppers originating from variously polluted habitat and the hsp70 level in their progeny. Possible reasons for population variation in hsp70 levels were discussed. The hsp70 level in the offspring's body depended on the place of origin. The strongest expression of hsp70 was found in the bodies of larvae hatching from the eggs laid by young females from Pilica (reference site). In contrast, a low initial level of hsp70 in larvae from polluted sites, especially in young females’ progeny, was observed. The application of zinc during diapause influenced the hsp70 level in grasshopper larvae; however, the direction of the changes depended on the insects’ place of origin. In larvae from the reference site, and also (but to a lesser degree) from Olkusz, the increase in the hsp70 level after zinc treatment was most pronounced. Whereas in grasshoppers from Szopienice, zinc (in 100 μg g⁻¹ dry weight of sand) did not change the hsp70 level, or (in 500 μg g⁻¹ dry weight of sand) caused a reduction in hsp70. The differences may result from maternal effects; however, possible adaptation also cannot be excluded. To confirm this statement further studies are needed.     

Nitric oxide/cGMP signaling in the corpora allata of female grasshoppers

The corpora allata (CA) of various insects express enzymes with fixation resistant NADPHdiaphorase activity. In female grasshoppers, juvenile hormone (JH) released from the CA is necessary to establish reproductive readiness, including sound production. Previous studies demonstrated that female sound production is also promoted by systemic inhibition of nitric oxide (NO) formation. In addition, allatotropin and allatostatin expressing central brain neurons were located in close vicinity of NO generating cells. It was therefore speculated that NO signaling may contribute to the control of juvenile hormone release from the CA.This study demonstrates the presence of NO/cGMP signaling in the CA of female Chorthippus biguttulus. CA parenchymal cells exhibit NADPHdiaphorase activity, express anti NOS immunoreactivity and accumulate citrulline, which is generated as a byproduct of NO generation. Varicose terminals from brain neurons in the dorsal pars intercerebralis and pars lateralis that accumulate cGMP upon stimulation with NO donors serve as intrinsic targets of NO in the CA. Both accumulation of citrulline and cyclic GMP were inhibited by the NOS inhibitor aminoguanidine, suggesting that NO in CA is produced by NOS. These results suggest that NO is a retrograde transmitter that provides feedback to projection neurons controlling JH production. Combined immunostainings and backfill experiments detected CA cells with processes extending into the CC and the protocerebrum that expressed immunoreactivity against the pan-neural marker anti-HRP. Allatostatin and allatotropin immunopositive brain neurons do not express NOS but subpopulations accumulate cGMP upon NO-formation. Direct innervation of CA by these peptidergic neurons was not observed.     

Competition among the adults of three grasshoppers (Orthop., Acrididae) on an alpine grassland

Abstract:  The intraspecific and interspecific competition among the adults of three common grasshoppers, Angaracris rhodopa (Fischer-Waldheim), Chorthippus dubius (Zubovsky) and Chorthippus fallax (Zubovsky) was investigated on Ganjia High Mountain Grassland in north-west China. The results indicated a strong intraspecific competition within species at high densities. The interspecific competition between the larger-sized species, A. rhodopa , and the smaller-sized species, C. dubius or C. fallax , was weak, while competition between C. dubius and C. fallax was strong. An asymmetric competitive interaction in mortality and fecundity associated with the diet and size was observed among the three species. Competition was strong between the two species that had a similar diet. The large-sized species had higher mortality rates and lower fecundity than the smaller-sized species. Intraspecific competition between the three species was mainly affected by food limitation, while interspecific competition was mainly associated with food limitation as well as the grasshoppers' adaptation to the environment.