Molecular characterization of a miraculin-like gene differentially expressed during coffee development and coffee leaf miner infestation

The characterization of a coffee gene encoding a protein similar to miraculin-like proteins, which are members of the plant Kunitz serine trypsin inhibitor (STI) family of proteinase inhibitors (PIs), is described. PIs are important proteins in plant defence against insects and in the regulation of proteolysis during plant development. This gene has high identity with the Richadella dulcifica taste-modifying protein miraculin and with the tomato protein LeMir; and was named as CoMir (Coffea miraculin). Structural protein modelling indicated that CoMir had structural similarities with the Kunitz STI proteins, but suggested specific folding structures. CoMir was up-regulated after coffee leaf miner (Leucoptera coffella) oviposition in resistant plants of a progeny derived from crosses between C. racemosa (resistant) and C. arabica (susceptible). Interestingly, this gene was down-regulated during coffee leaf miner herbivory in susceptible plants. CoMir expression was up-regulated after abscisic acid application and wounding stress and was prominent during the early stages of flower and fruit development. In situ hybridization revealed that CoMir transcripts accumulated in the anther tissues that display programmed cell death (tapetum, endothecium and stomium) and in the metaxylem vessels of the petals, stigma and leaves. In addition, the recombinant protein CoMir shows inhibitory activity against trypsin. According to the present results CoMir may act in proteolytic regulation during coffee development and in the defence against L. coffeella. The similarity of CoMir with other Kunitz STI proteins and the role of CoMir in plant development and plant stress are discussed.

Sex-biased hatching sequences in the cooperatively breeding Noisy Miner

The Noisy Miner Manorina melanocephala (Meliphagidae) is a cooperatively breeding bird species in which sons often remain on their natal home ranges and help one or both of their parents. In a population of Noisy Miners in SE Queensland, Australia, a molecular technique was used to explore adult and offspring sex ratios. and also hatching sequences. Among the adult population, there were 2.31 males for every female, and roughly 99% of helping was performed by males. At hatching and fledging, the population sex ratio was even, with exactly 57 males and 57 females. However, in 17 out of 18 broods the first egg to hatch was male, First-hatched males were significantly larger and heavier than their sisters just prior to fledging. Through their helping behaviour, large healthy sons could clearly enhance the future reproductive success of parents. and benefit the entire group. Sex-biased hatching sequences could potentially provide cooperatively breeding birds with a subtle and precise way of varying investment in the helping sex.

A new genus and species of leaf miner (Lepidoptera, Gracillariidae) for Chile associated to the native tree Lithraea caustica

A new genus and species of leaf miner (Lepidoptera, Gracillariidae) for Chile associated to the native tree Lithraea caustica. We propose the new genus and species of Gracillariidae (Lepidoptera) Hualpenia lithraeophaga Mundaca, Parra &Vargas gen. nov., sp. nov., leaf miner of Lithraea caustica (Mol.) H. et Arn (Anacardiaceae) occurring in southern central Chile. Aspects of the life cycle, adult and larval morphology, development and feeding habits of the new genus and species are also presented. We emphasise the uniqueness and importance of this new species for broadening the current knowledge on the Chilean fauna of Gracillariidae.

Egg laying site selection by a host plant specialist leaf miner moth at two intra-plant levels in the northern Chilean Atacama Desert

Egg laying site selection by a host plant specialist leaf miner moth at two intra-plant levels in the northern Chilean Atacama Desert. The spatial distribution of the immature stages of the leaf miner Angelabella tecomae Vargas & Parra, 2005 was determined at two intra-plant levels (shoot and leaflet) on the shrub Tecoma fulva fulva (Cav.) D. Don (Bignoniaceae) in the Azapa valley, northern Chilean Atacama Desert. An aggregated spatial pattern was detected for all the immature stages along the shoot, with an age dependent relative position: eggs and first instar larvae were clumped at apex; second, third and fourth instar larvae were mostly found at intermediate positions; meanwhile the spinning larva and pupa were clumped at basis. This pattern suggests that the females select new, actively growing leaflets for egg laying. At the leaflet level, the immature stages were found more frequently at underside. Furthermore, survivorship was higher for larvae from underside mines. All these results highlight the importance of an accurate selection of egg laying site in the life history of this highly specialized leaf miner. By contrast, eventual wrong choices in the egg laying site selection may be associated with diminished larval survivorship. The importance of the continuous availability of new plant tissue in this highly human modified arid environment is discussed in relation with the observed patterns.

A morphological reappraisal of the immature stages and life history of Elachista synethes Meyrick (Lepidoptera, Elachistidae), an Australian leaf miner alien to Chile

ABSTRACT Elachista synethes was recently recognized as an alien species in northern Chile, where its larvae mine the rescue grass Bromus catharticus (Poaceae). In order to provide the necessary information to allow field detection of E. synethes during early ontogeny, we conducted a morphological reappraisal of the immature stages of this leaf-miner moth, based on light and scanning electron microscopy, including the first descriptions of the egg and the first-instar larva. This is the first report of the existence of an apodal early larva for a species of Elachista Treitschke. The legs and prolegs are absent in the first two instars, but are well developed in the last two. Additional observations on the life history are also provided, including a description of the mine.

Study of simple sequence repeat markers from coffee expressed sequences associated to leaf miner resistance

The objective of this work was to identify expressed simple sequence repeats (SSR) markers associated to leaf miner resistance in coffee progenies. Identification of SSR markers was accomplished by directed searches on the Brazilian Coffee Expressed Sequence Tags (EST) database. Sequence analysis of 32 selected SSR loci showed that 65% repeats are of tetra-, 21% of tri- and 14% of dinucleotides. Also, expressed SSR are localized frequently in the 5'-UTR of gene transcript. Moreover, most of the genes containing SSR are associated with defense mechanisms. Polymorphisms were analyzed in progenies segregating for resistance to the leaf miner and corresponding to advanced generations of a Coffea arabica x Coffea racemosa hybrid. Frequency of SSR alleles was 2.1 per locus. However, no polymorphism associated with leaf miner resistance was identified. These results suggest that marker-assisted selection in coffee breeding should be performed on the initial cross, in which genetic variability is still significant.

Risk analysis of climate change on coffee nematodes and leaf miner in Brazil

The objective of this work was to assess the potential impact of climate change on the spatial distribution of coffee nematodes (races of Meloidogyne incognita) and leaf miner (Leucoptera coffeella), using a Geographic Information System. Assessment of the impacts of climate change on pest infestations and disease epidemics in crops is needed as a basis for revising management practices to minimize crop losses as climatic conditions shift. Future scenarios focused on the decades of the 2020's, 2050's, and 2080's (scenarios A2 and B2) were obtained from five General Circulation Models available on Data Distribution Centre from Intergovernmental Panel on Climate Change. Geographic distribution maps were prepared using models to predict the number of generations of the nematodes and leaf miner. Maps obtained in scenario A2 allowed prediction of an increased infestation of the nematode and of the pest, due to greater number of generations per month, than occurred under the climatological normal from 1961-1990. The number of generations also increased in the B2 scenario, but was lower than in the A2 scenario for both organisms.

Microsatellite markers in analysis of resistance to coffee leaf miner in Arabica coffee

The objective of this work was to analyze coffee (Coffea arabica) genotypes resistant to the coffee leaf miner (Leucoptera coffeella) using microsatellite markers. Sixty-six loci were evaluated, of which 63 were obtained from the Brazilian Coffee Expressed Sequence Tag (EST) database. These loci were amplified in bulks of individuals from F5 progenies of 'Siriema' (C. arabica x C. racemosa) resistant and susceptible to the insect, in eight samples of C. racemosa, and in a F6 population of 'Siriema' with 91 individuals segregating for resistance to the leaf miner. Polymorphisms were verified for two simple sequence repeat (SSR) loci in bulks of the susceptible progenies. The two polymorphic alleles were present in around 70% of the susceptible genotypes in F5 and in approximately 90% of the susceptible individuals in F6. However, the polymorphic EST-SSR markers among populations contrasting for resistance to leaf miner were not correlated to the evaluated characteristics. SSR markers show inter- and intraspecific polymorphism in C. arabica and C. racemosa.

Leaf miner incidence in coffee plants under different drip irrigation regimes and planting densities

The objective of this work was to evaluate the effect of different drip irrigation regimes and planting densities on the incidence of the leaf miner, Leucoptera coffeella, in arabica coffee plants for one year. The experiment was carried out in 2008, in a complete randomized block design, in a split-plot in time arrangement, with four replicates. The treatments consisted of four drip irrigation regimes - soil water balance, irrigations at 20 and 60 kPa soil tensions, and a nonirrigated treatment -, which were distributed at three plant densities: 2, 500, 5, 000, and 10, 000 plants per hectare. The evaluations were made on a monthly basis between January and December 2008. The highest pest occurrence period was from August to November, a season with low-air relative humidity preceded by a drought period. Irrigated coffee plants showed an incidence of intact mines 2.2 times lower than that of nonirrigated plants. Irrigation and increasing of plant density contribute to the reduction of coffee leaf miner occurrence.

Toxicity of organic farming‑compatible products to the coffee leaf miner

The objective of this work was to evaluate the toxicity of organic farming‑compatible products to the coffee leaf miner Leucoptera coffeella. Lime sulphur, enriched Bordeaux mixture (Viça Café Plus), and the "supermagro" biofertilizer were first tested in laboratory. The most promising product was tested afterwards under field conditions. In laboratory, different concentrations of each product were applied on L. coffeella eggs and on infested coffee‑mined leaves. Only lime sulphur had ovicidal effects at an acceptable concentration (1.6%) for field applications, but no significant effect on larvae mortality was found. Enriched Bordeaux mixture and the "supermagro" biofertilizer had no effect on L. coffeella eggs and larvae. In the field trial, biweekly or monthly sprayings of lime sulphur at different concentrations caused population decrease after 30 days; however, this effect was not significant after 60 or 90 days.