In vineyards, the under-vine area is managed to control vegetation growth and to reduce the compe... more In vineyards, the under-vine area is managed to control vegetation growth and to reduce the competing effect of growing plants on vines and fruit development. Applied under-vine management methods are the application of herbicides, soil tillage or the growth of spontaneous vegetation or cover crops. These methods affect pedo-climatic conditions differentially as well as the soil biota and have, therefore, consequences on soil functions and ecosystem services. In the presented case study, the effects of five under-vine management methods on the activity of soil hydrolytic enzymes, the soil water content, vine photosynthetic activity, shoot pruning weight, grape yield and quality are investigated in a vineyard in Lower Austria over three consecutive seasons. Thereby, we hypothesise that a permanent under-vine vegetation cover, either mowed or without mowing, supports the soil microbial communities and soil functions in a way to enhance water and nutrients availability for vines which ...
Berry shrivel (BS) is one of the prominent and still unresolved ripening physiological disorders ... more Berry shrivel (BS) is one of the prominent and still unresolved ripening physiological disorders in grapevine. The causes of BS are unclear, and previous studies focused on the berry metabolism or histological studies, including cell viability staining in the rachis and berries of BS clusters. Herein, we studied the transcriptional modulation induced by BS in the rachis of pre-symptomatic and symptomatic clusters with a custom-made microarray qPCR in relation to a previous RNASeq study of BS berries. Gene set analysis of transcript expression in symptomatic rachis tissue determined suppression of cell wall biosynthesis, which could also be confirmed already in pre-symptomatic BS rachis by CESA8 qPCR analyses, while in BS berries, a high number of SWITCH genes were suppressed at veraison. Additionally, genes associated with the cell wall were differently affected by BS in berries. A high percentage of hydrolytic enzymes were induced in BS grapes in rachis and berries, while other gro...
Healthy soils form the basis of sustainable viticulture, where soil characteristics have a direct... more Healthy soils form the basis of sustainable viticulture, where soil characteristics have a direct impact on wine quantity and quality. Soil not only provides water and nutrients to vines, but is also a living medium containing micro- and macroorganisms that perform many ecological functions and provide ecosystem services. These organisms are involved in many processes, from decomposing organic matter to providing minerals to vine roots. They also control diseases, pests, and weeds, in addition to improving the soil structure in terms of its capacity to retain water and nutrients. Related to decomposition processes, the carbon content of vineyard soils influences fertility, erosion and biogeochemical cycles, with significant implications for the global climate. However, common agricultural practices represent strong threats to biodiversity and associated ecosystem services provided by vineyard soils. As consumers increasingly consider environmental aspects in their purchase decisions...
Sustainable vineyard management in inter-rows may improve biodiversity and ecosystem service prov... more Sustainable vineyard management in inter-rows may improve biodiversity and ecosystem service provision in landscapes with a high density of vineyards. The current work investigates the effect of three inter-row ground cover treatments (bare soil by tillage, alternating and complete vegetation cover) on carabid beetle communities and vine vitality, in relation to climatic, soil and landscape parameters. Pitfall traps were used to collect carabids in the spring and autumn of 2016 from nine Austrian vineyards, with all three ground cover treatments established in each vineyard. Additionally, grape berry samples were collected before harvest in order to determine juice quality parameters. Generalized linear mixed models revealed that complete vegetation cover, the most extensive vineyard inter-row management, decreased both carabid density and species richness. The variables hours of sunshine, vineyard cover at the landscape scale and mesofauna abundance had negative impacts on species ...
PHYLLI is a database of standardized microsatellite reference alleles to manage phylloxera genoty... more PHYLLI is a database of standardized microsatellite reference alleles to manage phylloxera genotypes. Protocols for the aspects of analysis (DNA extraction, PCR methods, visualization and allele description) are presented with appropriate flexibility. The open database is a major step towards developing strategies and devising measures to control aggressive phylloxera strains. This is the first report to introduce the PHYLLI database and discuss its advantages for both vineyard management and academic research.
Grape phylloxera is native to North America, whereVitisspp. acquired different mechanisms of resi... more Grape phylloxera is native to North America, whereVitisspp. acquired different mechanisms of resistance to leaf and root attack. Its appearance in European vineyards at the beginning of the 1860s, where the phylloxera-susceptible grapevine speciesV. viniferaL. is majorly cultivated, caused the devastation of a great number of vineyards, generating a deep crisis in the European wine production and trade industries. However, the origin and genetic structure of this pest across European vineyards still remain controversial and uncertain. Herein, we analysed the genetic structure of 1173 grape phylloxera individuals collected from 100 locations across eight European countries. Structure and phylogenetic analyses show that contemporary grape phylloxera populations in Europe are the result of at least two independent introductions from the native range that mirrors the historical records that also suggest two major outbreaks in Europe. The comparative analysis with samples from the native...
BackgroundAlthough native to North America, the invasion of the aphid-like grape phylloxeraDaktul... more BackgroundAlthough native to North America, the invasion of the aphid-like grape phylloxeraDaktulosphaira vitifoliaeacross the globe altered the course of grape cultivation. For the past 150 years, viticulture relied on grafting-resistant North AmericanVitisspecies as rootstocks, thereby limiting genetic stocks tolerant to other stressors such as pathogens and climate change. Limited understanding of the insect genetics resulted in successive outbreaks across the globe when rootstocks failed. Here we report the 294-Mb genome ofD. vitifoliaeas a basic tool to understand host plant manipulation, nutritional endosymbiosis, and enhance global viticulture.ResultsUsing a combination of genome, RNA, and population resequencing, we found grape phylloxera showed high duplication rates since its common ancestor with aphids, but similarity in most metabolic genes, despite lacking obligate nutritional symbioses and feeding from parenchyma. Similarly, no enrichment occurred in development genes ...
The process of grape berry ripening follows three phases with distinct metabolic processes and co... more The process of grape berry ripening follows three phases with distinct metabolic processes and complex regulations via phytohormones. The physiological ripening disorder berry shrivel (BS) is characterized by reduced sugar accumulation, low anthocyanin contents, and high acidity in affected berries. The processes leading to BS induction are unknown, but recent transcriptional data on reduced expression of switch genes hint towards a disturbed ripening onset. Herein we investigated the phytohormone composition throughout grape berry ripening in healthy and BS berries inVitis viniferaL. cultivar Blauer Zweigelt. Thereby we hypothesize that phytohormones are key players for BS induction and suppress the expression of switch genes at veraison. The presented metabolomics and RNAseq data describe two distinct phytohormone profiles in BS berries, differing between pre- and post-veraison with a clear ethylene precursor (aminocyclopropane-1-carboxylic acid, ACC) peak before veraison. Exogeno...
Grape phylloxera (Daktulosphaira vitifoliae Fitch) is a major pest of cultivated grapevines (Viti... more Grape phylloxera (Daktulosphaira vitifoliae Fitch) is a major pest of cultivated grapevines (Vitis spp.), occurring in virtually all viticultural regions around the world. Different grape phylloxera strains can be found at varying levels on leaves and roots on both own-rooted plants and in plants grafted onto partially resistant rootstocks. Considering its relevance for the adequate management of the pest in infested vineyards, the analysis of its genetic diversity has received considerable attention from the scientific community in the last decades. Here, we review 25 years of DNA-based molecular markers applied to the analysis of the genetic structure and the reproductive mode of grape phylloxera in its native range and in different introduced regions. The use given to RAPD, AFLP, mtDNA sequencing and microsatellite (SSR) genetic markers for the analysis of grape phylloxera diversity is discussed, and an overview of the main findings obtained after their application to different p...
AimBunch compactness is the result of a complex dynamic process in which many bunch, berry and ra... more AimBunch compactness is the result of a complex dynamic process in which many bunch, berry and rachis features interact during the whole grapevine reproductive cycle. The aim of this work is to identify the different developmental stages involved in bunch compactness determination during the second growing season, from full-flowering to harvest time.Methods and Results In this work, bunch features from ten wine grapevine cultivars with diverse bunch compactness (including very loose and very compact bunches) have been periodically described. Experimental data allowed us to quantify changes in bunch compactness from berry set until harvest time in an objective manner. Our results indicate that bunch compactness development follows a double-sigmoid model, characterized by two consecutive cycles, each one with a growing stage followed by a latent one. Differences in the slope factor of the growing stages and in the duration of each developmental phase can explain part of the bunch comp...
Low pollen viability may limit grapevine yield under certain conditions, causing relevant economi... more Low pollen viability may limit grapevine yield under certain conditions, causing relevant economic losses to grape-growers. It is usually evaluated by the quantification of the number of viable and non-viable pollen grains that are present in a sample after an adequate pollen grain staining procedure. Although the manual counting of both types of grains is the simplest and most sensitive approach, it is a laborious and time-demanding process. In this regard, novel image-based approaches can assist in the objective, accurate and cost-effective phenotyping of this trait. Here, we introduce PollenCounter, an open-source macro implemented as a customizable Fiji tool for the high-throughput phenotyping of pollen viability. This tool splits RGB images of stained pollen grains into its primary channels, retaining red and green color fractionated images (which contain information on total and only viable pollen grains, respectively) for the subsequent isolation and counting of the regions o...
A cytogenetic technique was developed to produce suitable chromosome spreads for phylloxera karyo... more A cytogenetic technique was developed to produce suitable chromosome spreads for phylloxera karyotype analysis. The karyotype for pathogenetic phylloxera was 2 n = 10. Karyotypes from haploid sex cells were found to vary between n = 5 and n = 6, the latter possibly indicating an aneuploidic aberration. Tetra- and polyploid cells were detected in somatic trophocytes. Preparation of phylloxera sex and somatic cells for chromosomal analysis reported here enables the study of genetic variation on a chromosomal scale.
Amplified fragment length polymorphism (AFLP) is widely used for DNA fingerprinting and it has be... more Amplified fragment length polymorphism (AFLP) is widely used for DNA fingerprinting and it has been broadly applied in population genetics. Since it is based on restriction digestion and PCR-based amplification it can be influenced by different chemical compounds commonly found in the isolated DNA. DNA extraction procedures may alter the AFLP banding profiles through DNA quality. Hence the DNA extraction method is crucial to produce reproducible AFLP-banding profiles. In this work two sets of AFLP analyses were performed on 62 Pinot noir, 6 Pinot blanc and 4 Pinot gris (Vitis vinifera L.) clones, and profiles obtained after three different DNA extraction methods were compared. AFLP profiles were different for the same genotypes due to the DNA extraction method used.
Phylloxera (Daktulosphaira vitifoliae FITCH) induce galls (nodosities) on young grapevine roots. ... more Phylloxera (Daktulosphaira vitifoliae FITCH) induce galls (nodosities) on young grapevine roots. Histological and histochemical methods were applied to study the gall's morphology and enzyme activities (peroxidases, leucine aminopeptidases and acidic phosphatases). Susceptible V. vinifera cv. Cabernet Sauvignon was compared to the resistant rootstock 5 BB (V. berlandieri x V. riparia) using aseptic dual culture conditions. The gall induction phase was analyzed before visible signs of potential resistance responses were detected. Elevated metabolic activity has been found in nodosities compared to uninfected roots. Starch granule incorporation was detected in young galls and was highest at the feeding site. As galls mature, the starch density decreased at the feeding site and increased towards the periphery of the gall. Peroxidase, acidic phosphatase and leucine aminopeptidase activities were highest at the incision. No differences in enzyme activities could be detected between t...
In vineyards, the under-vine area is managed to control vegetation growth and to reduce the compe... more In vineyards, the under-vine area is managed to control vegetation growth and to reduce the competing effect of growing plants on vines and fruit development. Applied under-vine management methods are the application of herbicides, soil tillage or the growth of spontaneous vegetation or cover crops. These methods affect pedo-climatic conditions differentially as well as the soil biota and have, therefore, consequences on soil functions and ecosystem services. In the presented case study, the effects of five under-vine management methods on the activity of soil hydrolytic enzymes, the soil water content, vine photosynthetic activity, shoot pruning weight, grape yield and quality are investigated in a vineyard in Lower Austria over three consecutive seasons. Thereby, we hypothesise that a permanent under-vine vegetation cover, either mowed or without mowing, supports the soil microbial communities and soil functions in a way to enhance water and nutrients availability for vines which ...
Berry shrivel (BS) is one of the prominent and still unresolved ripening physiological disorders ... more Berry shrivel (BS) is one of the prominent and still unresolved ripening physiological disorders in grapevine. The causes of BS are unclear, and previous studies focused on the berry metabolism or histological studies, including cell viability staining in the rachis and berries of BS clusters. Herein, we studied the transcriptional modulation induced by BS in the rachis of pre-symptomatic and symptomatic clusters with a custom-made microarray qPCR in relation to a previous RNASeq study of BS berries. Gene set analysis of transcript expression in symptomatic rachis tissue determined suppression of cell wall biosynthesis, which could also be confirmed already in pre-symptomatic BS rachis by CESA8 qPCR analyses, while in BS berries, a high number of SWITCH genes were suppressed at veraison. Additionally, genes associated with the cell wall were differently affected by BS in berries. A high percentage of hydrolytic enzymes were induced in BS grapes in rachis and berries, while other gro...
Healthy soils form the basis of sustainable viticulture, where soil characteristics have a direct... more Healthy soils form the basis of sustainable viticulture, where soil characteristics have a direct impact on wine quantity and quality. Soil not only provides water and nutrients to vines, but is also a living medium containing micro- and macroorganisms that perform many ecological functions and provide ecosystem services. These organisms are involved in many processes, from decomposing organic matter to providing minerals to vine roots. They also control diseases, pests, and weeds, in addition to improving the soil structure in terms of its capacity to retain water and nutrients. Related to decomposition processes, the carbon content of vineyard soils influences fertility, erosion and biogeochemical cycles, with significant implications for the global climate. However, common agricultural practices represent strong threats to biodiversity and associated ecosystem services provided by vineyard soils. As consumers increasingly consider environmental aspects in their purchase decisions...
Sustainable vineyard management in inter-rows may improve biodiversity and ecosystem service prov... more Sustainable vineyard management in inter-rows may improve biodiversity and ecosystem service provision in landscapes with a high density of vineyards. The current work investigates the effect of three inter-row ground cover treatments (bare soil by tillage, alternating and complete vegetation cover) on carabid beetle communities and vine vitality, in relation to climatic, soil and landscape parameters. Pitfall traps were used to collect carabids in the spring and autumn of 2016 from nine Austrian vineyards, with all three ground cover treatments established in each vineyard. Additionally, grape berry samples were collected before harvest in order to determine juice quality parameters. Generalized linear mixed models revealed that complete vegetation cover, the most extensive vineyard inter-row management, decreased both carabid density and species richness. The variables hours of sunshine, vineyard cover at the landscape scale and mesofauna abundance had negative impacts on species ...
PHYLLI is a database of standardized microsatellite reference alleles to manage phylloxera genoty... more PHYLLI is a database of standardized microsatellite reference alleles to manage phylloxera genotypes. Protocols for the aspects of analysis (DNA extraction, PCR methods, visualization and allele description) are presented with appropriate flexibility. The open database is a major step towards developing strategies and devising measures to control aggressive phylloxera strains. This is the first report to introduce the PHYLLI database and discuss its advantages for both vineyard management and academic research.
Grape phylloxera is native to North America, whereVitisspp. acquired different mechanisms of resi... more Grape phylloxera is native to North America, whereVitisspp. acquired different mechanisms of resistance to leaf and root attack. Its appearance in European vineyards at the beginning of the 1860s, where the phylloxera-susceptible grapevine speciesV. viniferaL. is majorly cultivated, caused the devastation of a great number of vineyards, generating a deep crisis in the European wine production and trade industries. However, the origin and genetic structure of this pest across European vineyards still remain controversial and uncertain. Herein, we analysed the genetic structure of 1173 grape phylloxera individuals collected from 100 locations across eight European countries. Structure and phylogenetic analyses show that contemporary grape phylloxera populations in Europe are the result of at least two independent introductions from the native range that mirrors the historical records that also suggest two major outbreaks in Europe. The comparative analysis with samples from the native...
BackgroundAlthough native to North America, the invasion of the aphid-like grape phylloxeraDaktul... more BackgroundAlthough native to North America, the invasion of the aphid-like grape phylloxeraDaktulosphaira vitifoliaeacross the globe altered the course of grape cultivation. For the past 150 years, viticulture relied on grafting-resistant North AmericanVitisspecies as rootstocks, thereby limiting genetic stocks tolerant to other stressors such as pathogens and climate change. Limited understanding of the insect genetics resulted in successive outbreaks across the globe when rootstocks failed. Here we report the 294-Mb genome ofD. vitifoliaeas a basic tool to understand host plant manipulation, nutritional endosymbiosis, and enhance global viticulture.ResultsUsing a combination of genome, RNA, and population resequencing, we found grape phylloxera showed high duplication rates since its common ancestor with aphids, but similarity in most metabolic genes, despite lacking obligate nutritional symbioses and feeding from parenchyma. Similarly, no enrichment occurred in development genes ...
The process of grape berry ripening follows three phases with distinct metabolic processes and co... more The process of grape berry ripening follows three phases with distinct metabolic processes and complex regulations via phytohormones. The physiological ripening disorder berry shrivel (BS) is characterized by reduced sugar accumulation, low anthocyanin contents, and high acidity in affected berries. The processes leading to BS induction are unknown, but recent transcriptional data on reduced expression of switch genes hint towards a disturbed ripening onset. Herein we investigated the phytohormone composition throughout grape berry ripening in healthy and BS berries inVitis viniferaL. cultivar Blauer Zweigelt. Thereby we hypothesize that phytohormones are key players for BS induction and suppress the expression of switch genes at veraison. The presented metabolomics and RNAseq data describe two distinct phytohormone profiles in BS berries, differing between pre- and post-veraison with a clear ethylene precursor (aminocyclopropane-1-carboxylic acid, ACC) peak before veraison. Exogeno...
Grape phylloxera (Daktulosphaira vitifoliae Fitch) is a major pest of cultivated grapevines (Viti... more Grape phylloxera (Daktulosphaira vitifoliae Fitch) is a major pest of cultivated grapevines (Vitis spp.), occurring in virtually all viticultural regions around the world. Different grape phylloxera strains can be found at varying levels on leaves and roots on both own-rooted plants and in plants grafted onto partially resistant rootstocks. Considering its relevance for the adequate management of the pest in infested vineyards, the analysis of its genetic diversity has received considerable attention from the scientific community in the last decades. Here, we review 25 years of DNA-based molecular markers applied to the analysis of the genetic structure and the reproductive mode of grape phylloxera in its native range and in different introduced regions. The use given to RAPD, AFLP, mtDNA sequencing and microsatellite (SSR) genetic markers for the analysis of grape phylloxera diversity is discussed, and an overview of the main findings obtained after their application to different p...
AimBunch compactness is the result of a complex dynamic process in which many bunch, berry and ra... more AimBunch compactness is the result of a complex dynamic process in which many bunch, berry and rachis features interact during the whole grapevine reproductive cycle. The aim of this work is to identify the different developmental stages involved in bunch compactness determination during the second growing season, from full-flowering to harvest time.Methods and Results In this work, bunch features from ten wine grapevine cultivars with diverse bunch compactness (including very loose and very compact bunches) have been periodically described. Experimental data allowed us to quantify changes in bunch compactness from berry set until harvest time in an objective manner. Our results indicate that bunch compactness development follows a double-sigmoid model, characterized by two consecutive cycles, each one with a growing stage followed by a latent one. Differences in the slope factor of the growing stages and in the duration of each developmental phase can explain part of the bunch comp...
Low pollen viability may limit grapevine yield under certain conditions, causing relevant economi... more Low pollen viability may limit grapevine yield under certain conditions, causing relevant economic losses to grape-growers. It is usually evaluated by the quantification of the number of viable and non-viable pollen grains that are present in a sample after an adequate pollen grain staining procedure. Although the manual counting of both types of grains is the simplest and most sensitive approach, it is a laborious and time-demanding process. In this regard, novel image-based approaches can assist in the objective, accurate and cost-effective phenotyping of this trait. Here, we introduce PollenCounter, an open-source macro implemented as a customizable Fiji tool for the high-throughput phenotyping of pollen viability. This tool splits RGB images of stained pollen grains into its primary channels, retaining red and green color fractionated images (which contain information on total and only viable pollen grains, respectively) for the subsequent isolation and counting of the regions o...
A cytogenetic technique was developed to produce suitable chromosome spreads for phylloxera karyo... more A cytogenetic technique was developed to produce suitable chromosome spreads for phylloxera karyotype analysis. The karyotype for pathogenetic phylloxera was 2 n = 10. Karyotypes from haploid sex cells were found to vary between n = 5 and n = 6, the latter possibly indicating an aneuploidic aberration. Tetra- and polyploid cells were detected in somatic trophocytes. Preparation of phylloxera sex and somatic cells for chromosomal analysis reported here enables the study of genetic variation on a chromosomal scale.
Amplified fragment length polymorphism (AFLP) is widely used for DNA fingerprinting and it has be... more Amplified fragment length polymorphism (AFLP) is widely used for DNA fingerprinting and it has been broadly applied in population genetics. Since it is based on restriction digestion and PCR-based amplification it can be influenced by different chemical compounds commonly found in the isolated DNA. DNA extraction procedures may alter the AFLP banding profiles through DNA quality. Hence the DNA extraction method is crucial to produce reproducible AFLP-banding profiles. In this work two sets of AFLP analyses were performed on 62 Pinot noir, 6 Pinot blanc and 4 Pinot gris (Vitis vinifera L.) clones, and profiles obtained after three different DNA extraction methods were compared. AFLP profiles were different for the same genotypes due to the DNA extraction method used.
Phylloxera (Daktulosphaira vitifoliae FITCH) induce galls (nodosities) on young grapevine roots. ... more Phylloxera (Daktulosphaira vitifoliae FITCH) induce galls (nodosities) on young grapevine roots. Histological and histochemical methods were applied to study the gall's morphology and enzyme activities (peroxidases, leucine aminopeptidases and acidic phosphatases). Susceptible V. vinifera cv. Cabernet Sauvignon was compared to the resistant rootstock 5 BB (V. berlandieri x V. riparia) using aseptic dual culture conditions. The gall induction phase was analyzed before visible signs of potential resistance responses were detected. Elevated metabolic activity has been found in nodosities compared to uninfected roots. Starch granule incorporation was detected in young galls and was highest at the feeding site. As galls mature, the starch density decreased at the feeding site and increased towards the periphery of the gall. Peroxidase, acidic phosphatase and leucine aminopeptidase activities were highest at the incision. No differences in enzyme activities could be detected between t...
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