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.

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.

Leaf trait variation on Erythroxylum tortuosum (Erythroxylaceae) and its relationship with oviposition preference and stress by a host-specific leaf miner

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Oviposition of a leaf-miner on Erythroxylum tortuosum (Erythroxylaceae) leaves: hierarchical variation of physical leaf traits

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Morphology and anatomy of leaf miners in two species of Commelinaceae (Commelina diffusa Burm. f. and Floscopa glabrata (Kunth) Hassk)

In specialized literature, reports on anatomy of miners in host plants are few in number. These agents trigger excavations, or paths, by consumption of plant inner tissues by larvae of several insects. The aim of this work was to investigate leaf miner occurrence in Commelina diffusa (a cosmopolitan plant) and Floscopa glabrata (an amphibious plant) using anatomical techniques. The place where the plants were collected is subjected to seasonal floods, consequently both the species were exposed to the same weather conditions and seasonal floods. This study showed that members of Agromyzidae and Chironomidae families, which are Diptera endophytophagous larvae types, were responsible for the tunnels. Moreover, in Commelina diffusa Agromyzidae larvae were found, while in Floscopa glabrata three Chironomidae cephalic exuviae were found. The miners, as can be seen from anatomical studies, used only mesophyll parenchyma tissues for feeding, causing the formation of linear mines. In addition, in both the species, the epidermis and the medium-sized vascular units were kept intact, showing no structural modification, such as neoformation of tissues.

Variabilidade genética em famílias de meios-irmãos de cafeeiro robusta

O cultivo do café é uma das atividades do agronegócio de maior importância socioeconômica dentre as diferentes atividades ligadas ao comércio agrícola mundial. Uma das maiores contribuições da genética quantitativa para o melhoramento genético é a possibilidade de prever ganhos genéticos. Quando diferentes critérios de seleção são considerados, a predição de ganhos referentes a cada critério tem grande importância, pois indica os melhoristas sobre como utilizar o material genético disponível, visando obter o máximo de ganhos possível para as características de interesse. O presente trabalho foi instalado em julho de 2004, na Fazenda Experimental de Bananal do Norte, conduzida pelo Incaper, no distrito de Pacotuba, município de Cachoeiro de Itapemirim, região Sul do Estado, com o objetivo de selecionar as melhores plantas entre e dentro de progênies de meios- irmãos de Coffea canephora, por meio de diferentes critérios de seleção. Foram realizadas análises de variância individuais e conjuntas para 26 progênies de meios- irmãos Coffea canephora. O delineamento experimental utilizado foi em blocos ao acaso com quatro testemunhas adicionais com quatro repetições e parcela composta por cinco plantas, com o espaçamento de 3,0 m x 1,2 m. Neste trabalho, considerou-se os dados das últimas cinco colheitas. As características mensuradas foram: florescimento, maturação, tamanho do grão, peso, porte, vigor, ferrugem, mancha cercóspora, seca de ponteiros, escala geral, porcentagem de frutos boia e bicho mineiro. Todas as análises estatísticas foram realizadas com o aplicativo computacional em genética e estatística (GENES). Foram estimados os ganhos de seleção em função da porcentagem de seleção de 20% entre e dentro, sendo as mesmas mantidas para todas as características. Todas as características foram submetidas a seleção no sentido positivo, exceto para florescimento, porte, ferrugem, mancha cercóspora, seca de ponteiros, porcentagem de frutos boia e bicho mineiro, para obter decréscimo em suas médias originais. Os critérios de seleção estudados foram: seleção convencional entre e dentro das famílias, índice de seleção combinada, seleção massal e seleção massal estratificada. Esta dissertação é composta por dois capítulos, em que foram realizadas análises biométricas, como a obtenção de estimativas de parâmetros genéticos. Na maioria das características estudadas, verificaram-se diferenças significativas (P<0,05) para genótipos que, associados aos coeficientes de variação genotípicos e também ao coeficiente de determinação genotípico e à relação CVg/CVe, indicam a existência de variabilidade genética nos materiais genéticos para a maioria das características e condições favoráveis para obtenção de ganhos genéticos pela seleção. Essas características também foram correlacionadas. Os dados foram submetidos às análises de variância e multivariada, aplicando-se a técnica de agrupamento e UPGMA, teste de médias e estudo de correlações. Na técnica de agrupamento, foi utilizada a distância generalizada de Mahalanobis como medida de dissimilaridade, e na delimitação dos grupos, o método de Tocher. Foi encontrada diversidade genética para as características associadas à qualidade fisiológica, mobilização de reserva das sementes, dimensões e biomassa das plântulas. Quatro grupos de genótipos puderam ser formados. Peso de massa seca de sementes, redução de reserva de sementes e peso de massa seca de plântulas estão positivamente correlacionados entre si, enquanto a redução de reserva das sementes e a eficiência na conversão dessas reservas em plântulas estão negativamente correlacionadas. De acordo com os resultados obtidos, verificou-se que todas as características apresentaram níveis diferenciados de variabilidade genética e os critérios de seleção utilizados mostraram-se eficientes para o melhoramento, no qual o índice de seleção combinada é o critério de seleção que apresentou os melhores resultados em termos de ganhos, sendo indicado como critério mais apropriado para o melhoramento genético da população estudada. Nos estudos de correlações, em 70% dos casos, a correlação fenotípica foi superior à genotípica, mostrando maior influência dos fatores ambientais em relação aos genotípicos e condições propícias ao melhoramento dos diferentes caracteres. No estudo de divergência genética, observou-se que pelo agrupamento de genótipos, pela técnica de Tocher, indicou que os genótipos foram distribuídos em três grupos.

Identification of sap-feeder instars in Angelabella tecomae Vargas & Parra (Lepidoptera, Gracillariidae) feeding on Tecoma fulva fulva (Bignoniaceae)

The number of sap-feeding instars of the leaf-miner Angelabella tecomae Vargas & Parra, 2005 was determined by measuring the head capsule exuviae collected from mines (n = 125) on the shrub Tecoma fulva fulva D. Don (Bignoniaceae) in the Azapa Valley, northern Chile. Sap-feeding larvae went through four instars in 120 occasions while five larvae went through five instars. The possible causes for this variation and the usefulness of this tool for future studies are discussed.

A replacement name for Hualpenia Mundaca, Parra & Vargas (Lepidoptera, Gracillariidae)

A replacement name for Hualpenia Mundaca, Parra & Vargas (Lepidoptera, Gracillariidae). Vihualpenia nom. nov. is proposed as a replacement name for Hualpenia Mundaca, Parra & Vargas, 2013 (Lepidoptera, Gracillariidae), in order to remove homonymy with Hualpenia Franz, 1996 (Coleoptera, Staphylinidae).