Seasonal adaptations to daylength often limit the effective range of insects used in biological control of weeds. The leaf beetle Diorhabda carinulata (Desbrochers) was introduced into North America from Fukang, China (latitude 44° N) to control saltcedars (Tamarix spp.), but failed to establish south of 38° N latitude because of a mismatched critical daylength response for diapause induction. The daylength response caused beetles to enter diapause too early in the season to survive the duration of winter at southern latitudes. Using climate chambers, we characterized the critical daylength response for diapause induction (CDL) in three ecotypes of Diorhabda beetles originating from 36, 38, and 43° N latitudes in Eurasia. In a field experiment, the timing of reproductive diapause and voltinism were compared among ecotypes by rearing the insects on plants in the field. CDL declined with latitude of origin among Diorhabda ecotypes. Moreover, CDL in southern (<39° N latitude) ecotypes was shortened by more than an hour when the insects were reared under a fluctuating 35-15°C thermoperiod than at a constant 25°C. In the northern (>42° N latitude) ecotypes, however, CDL was relatively insensitive to temperature. The southern ecotypes produced up to four generations when reared on plants in the field at sites south of 38° N, whereas northern ecotypes produced only one or two generations. The study reveals latitudinal variation in how Diorhabda ecotypes respond to daylength for diapause induction and how these responses affect insect voltinism across the introduced range.

The genus Diorhabda (Coleoptera: Chrysomelidae) was recently revised, using morphological characters, into five tamarisk-feeding species, four of which have been used in the tamarisk (Tamarix spp.) biological control program in North America and are the subject of these studies. The taxonomic revision is here supported using molecular genetic and hybridization studies. Four Diorhabda species separated into five clades using cytochrome c oxidase subunit 1 sequence data with Diorhabda elongata separating into two clades. Amplified fragment length polymorphism (AFLP) analysis using genomic DNA revealed only four clades, which corresponded to the four morphospecies. Hybridization between the four species yielded viable eggs in F1 crosses but viability was significantly lower than achieved with intraspecific crosses. Crosses involving Diorhabda carinulata and the other three species resulted in low F2 egg viability, whereas crosses between D. elongata, Diorhabda sublineata and Diorhabda carinata resulted in > 40% F2 egg viability. Crosses between D. carinulata and the other three species resulted in high mortality of D. carinulata females due to genital mismatch. AFLP patterns combined with principal coordinates analysis enabled effective separation between D. elongata and D. sublineata, providing a method to measure genetic introgression in the field.

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Establishment of the saltcedar leaf beetle (Diorhabda spp.) has been unpredictable when caged or released in the field for saltcedar (Tamarix spp.) biocontrol. It has been observed that one caged tree might be voraciously fed upon by beetles while an adjacent tree in the cage is left untouched. We hypothesized that differences in the nutrient content of individual trees may explain this behavior. We evaluated survival, development rate, and egg production of beetles fed in the laboratory on saltcedar foliage from trees that had been grown under a range of fertilizer treatments. Tissue samples from the experimental trees and from the field were analyzed for percent nitrogen, phosphorus, and potassium. There was essentially no survival of beetle larvae fed foliage from saltcedar trees at nitrogen levels below 2.0%. At levels above 2.0% N, beetle larvae had corresponding increased survival rates and shorter development times. Multiple regression analyses indicated that nitrogen and phosphorus are important for larval survival and faster development rates. Higher levels of potassium were important for increased egg cluster production. The plant tissue analysis showed that the percentage of nitrogen in the experimental trees reflected the range of trees in the field and also that there is high variability within trees in the field. Our research indicates that if beetles are released on trees with poor nutrient quality, the larvae will not survive.