Characterisation of Aceria massalongoi and a histopathological study of the leaf galls induced on chaste trees.

The eriophyoid mite Aceria massalongoi (Canestrini) was collected from globoid leaf galls on severely injured chaste trees, Vitex agnus-castus L. (Lamiaceae), in Bari and Bernalda (southern Italy), and on the Ionian island Leukade (Greece). Female, male and nymph were described in detail, following the current morphometric descriptive scheme, supplementing older and incomplete descriptions. Molecular characterization of A. massalongoi from Italy and Greece was conducted by amplifying and sequencing the ribosomal ITS, the D2-D3 expansion domains of the 28S rRNA gene and the mitochondrial COI, for the first time. Phylogenetic trees based on the three molecular markers showed congruent results, confirming that Italian and Greek A. massalongoi populations are the same species that cluster together with some intraspecific variability. Galls, ranging from 0.5 to 2.8 mm in diameter, were randomly distributed on both leaf surfaces, and protruded ca. 1 mm from the leaf surface. Sometimes they were closely aggregated on midrib and leaves, which, consequently, appeared strongly deformed. Close-up observations revealed that gall induction causes hyperplastic proliferation of leaf tissues around the gall chamber hosting mites. The uniserial cell lining inside this chamber provides the nutritional tissue for the mites. All feeding cells contained one or more (frequently 2-3) hypertrophied nuclei and dense granular cytoplasm.

Functional Variation of Two Novel Cellulases, Pv-eng-5 and Pv-eng-8, and the Heat Shock 90 Gene, Pv-hsp-90, in Pratylenchus vulnus and Their Expression in Response to Different Temperature Stress.

Functional characterization of two novel endoglucanase genes, Pv-eng-5 and Pv-eng-8, of the root-lesion nematode Pratylenchus vulnus was carried out. In situ-hybridization experiments revealed that Pv-eng-8 transcript was localized in the pharyngeal glands. Silencing of Pv-eng-5 and Pv-eng-8 resulted in a significant reduction of expression level (52% and 67%, respectively). Furthermore, the silencing of Pv-eng-8 determined a reduction (41%) in nematode reproduction, suggesting that treated nematodes are much less able to process food. Surprisingly, no significant difference on reproduction rate was observed with Pv-eng-5 dsRNA nematodes, suggesting a neofunctionalization of Pv-eng-5 despite the high similarity with nematode endoglucanases. Pratylenchus species are poikilothermic organisms showing close relationships with the environmental temperature. The effects of different temperature ranges revealed that the reproductive potential of P. vulnus increased with increasing temperature from 23 °C to 28 °C, but no reproduction was observed at 33 °C. In real time, increasing temperature from 23 °C to 28 °C the heat shock gene Pv-hsp-90 was differentially expressed in adult stages, while the levels of the effector genes Pv-eng-1 and Pv-eng-8 in females showed no significant differences compared to those observed at 23 °C, only in males Pv-eng-8 level decreased (45%). The upregulation of Pv-hsp-90 in both adult stages suggests a protective mechanism in order to cope with unfavorable environmental conditions.

Occurrence of Sheraphelenchus sucus (Nematoda: Aphelenchoidinae) and Panagrellus sp. (Rhabditida: Panagrolaimidae) Associated with Decaying Pomegranate Fruit in Italy.

Two different nematode species were recovered from pomegranate decaying fruit in two localities in Southern Italy: the mycetophagus nematode Sheraphelenchus sucus and a bacterial feeder nematode belonging to the Panagrolaimidae (Rhabditida) family. Morphometrics of the Italian population of S. sucus closely resemble that of the type population, whereas some differences were found when compared with another population from Iran. Molecular characterization of the Italian S. sucus using the 18S rRNA gene, D2-D3 expansion domains of the 28S rDNA, the ITS region, and the partial mitochondrial COI were carried out. Sequences of the 18S rRNA gene, the D2-D3 domains, and the ITS were analyzed using several methods for inferring phylogeny to reconstruct the relationships among Sheraphelenchus and Bursaphelenchus species. The bacterial feeder Panagrellus sp. was characterized at the molecular level only. The D2-D3 expansion domains and ITS sequences of this Italian panagrolaimid were determined. The D2-D3 sequences of the Italian panagrolaimid showed 99% similarity with the corresponding sequence of Panagrellus sp. associated with Rhynchophorus ferrugineus. This is the first report on the tritrophic association of S. sucus and Rhabditida that uses both insects and pomegranate fruit as hosts.

Rearing and Conservation of Nematode Populations Under Greenhouse or Axenic Conditions.

Once a nematode has been identified, to conduct studies for screening programs or pathogenicity tests, it is necessary a supply of large numbers of nematodes from field crops or reproduced and stored to be used in periods of the year when they are not available from fields. Therefore, nematodes must be reared in greenhouse or under in vitro conditions and stored for future needs. In this chapter, suggestions are given on how to obtain nematodes from fields and reproduce most of them on host plants in greenhouse (mainly Meloidogyne spp. and Globodera spp.) or in vitro. Reproductions in vitro include: On suitable callus of host plants (Pratylenchus spp., Ditylenchus spp.) On fungal cultures mainly of Botrytis cinerea or Alternaria spp. for Aphelenchoides spp. and other aphelenchids, including Bursaphelenchus xylophilus. On carrot disks for Pratylenchus spp. and Ditylenchus spp. Other specific media, such as garlic, potato, and sweet potato for D. destructor, and cocoyam disks for Radopholus similis. Guidelines are also given to store different nematodes for rather long times, including in vitro methods and in infected seeds, hay, and other plant parts. No information is given on how to prepare and store fixed materials.

Identifying important drivers of East African October to December rainfall season.

Monsoon rainfall plays a crucial part in Africa's socio-economic structure and its year-to-year variability has profound implications for agricultural, energy, and other societal sectors. The current study focuses on two of the major climate drivers of the east African rainy season during October-November-December (OND), which is when the season starts for a large portion of east Africa (e.g. Tanzania and Malawi). Such drivers could be different in early austral summer from the rest of the year, due to the relative positioning of the Intertropical convergence zone, which passes through this region - hence regions of east Africa and OND season are the focus here. The two drivers of Monsoon viz. El Niño Southern Oscillation (ENSO) and Indian Ocean Dipole (IOD) are considered: both independently indicate strong connections with monsoon OND rain. Not only is there a strong significant positive correlation in the OND season as a simultaneous relation, but the signal is also there even with the lag of a few months. This has been tested using various data sources, detrending data beforehand, analysing either recent time periods or earlier time periods - covering two decades each, and using regression analyses. To further strengthen the results, a compositing approach is applied that can additionally identify strong rainfall signals to that from years when opposite combinations of ENSO and IOD phases act as confounding factors. Results of precipitation anomaly for OND for compositing, when IOD and ENSO are both negative (positive) in July-August-September(JAS) indicate a deficit (excess) in rainfall in that region. The Walker circulation seems to play a major part via altering ascending to descending branch in two situations, when both drivers are in same phase. In the last near thirty years period, a total of 9 years matched the criteria when both drivers were negative and suggested a deficit in OND rainfall; more recently, that criteria occurred also in 2022 (JAS) and was again associated with a rainfall deficit in OND 2022. Based on this analysis, it is possible to deliver an estimation of cumulative rain in terms of median value, range and distribution, one season in advance, at a point location or average over a region. Some results of compositing are confirmed for longer record (1940-2021) too and further classifications based on threshold of drivers are tested. Rainfall (OND) variability at intra-decadal, decadal and multi-decadal scales are studied by applying the method of centered moving averages of 5-year, 11-year and 21-year respectively. Our results have implications for future planning in optimizing agricultural and energy outputs, mitigating severe consequences and losses associated with droughts and excess rain and will impact the livelihoods of millions of east Africans.

Morphological and molecular characterisation of Xiphinema macroacanthum Lamberti, Roca & Agostinelli, 1989 (Nematoda: Longidoridae) from olive orchards in southern Italy.

A population of Xiphinema macroacanthum Lamberti, Roca & Agostinelli, 1989 originating from olive orchards in Brindisi, Italy and containing both adults and all juvenile stages, is described and illustrated. The first juvenile stage is reported for the first time. Molecular characterisation of this species, using the D2-D3 expansion domains of the 28S rDNA and ITS region, was carried out. PCR-RFLP analyses of the ribosomal gene regions determined species-specific patterns that clearly differentiate X. macroacanthum. Sequences of the D2-D3 domains and the partial 18S-ITS1 rRNA genes were analysed using several methods for inferring phylogeny to reconstruct the relationships between X. macroacanthum and other Xiphinema species.

Nematicidal Activity of a Garlic Extract Formulation against the Grapevine Nematode Xiphinema index.

The nematicidal activity of garlic extracts is known on root-knot nematodes but never investigated on the grapevine nematode Xiphinema index. In this study, the nematicidal activity of a commercial garlic extract formulate (GEF) was assessed on X. index, both in vitro and in a pot assay. In the in vitro assays, mixed specimens of X. index were exposed to a 0-4 mL L-1 range of GEF concentrations, checking nematode immotility and mortality after 2, 4 or 8 h. In the experiments on potted grapevines, plants cultivated in soil infested by X. index were irrigated twice at a 15-day interval with 0.05, 0.2 and 0.5 mL L-1 solutions of GEF, including nontreated soil as a control. An almost complete mortality of X. index specimens occurred after a 2 h exposure to a 2 mL L-1 GEF concentration, while an 8 h exposure to even the 0.0312 and 0.0156 mL L-1 solutions resulted in about 50% and 30% mortality, respectively. Soil treatment with a 0.5 mL L-1 GEF solution significantly reduced the population of X. index and increased the grapevine root growth compared to nontreated soil or soil treated with the lower dosages. Results of this study indicated that garlic-based nematicides could be an effective tool for X. index management in organic and integrated vineyards.

Evaluation of Fungal Volatile Organic Compounds for Control the Plant Parasitic Nematode Meloidogyne incognita.

Plant parasitic nematodes are a serious threat to crop production worldwide and their control is extremely challenging. Fungal volatile organic compounds (VOCs) provide an ecofriendly alternative to synthetic nematicides, many of which have been withdrawn due to the risks they pose to humans and the environment. This study investigated the biocidal properties of two fungal VOCs, 1-Octen-3-ol and 3-Octanone, against the widespread root-knot nematode Meloidogyne incognita. Both VOCs proved to be highly toxic to the infective second-stage juveniles (J2) and inhibited hatching. Toxicity was dependent on the dose and period of exposure. The LD50 of 1-Octen-3-ol and 3-Octanone was 3.2 and 4.6 µL, respectively. The LT50 of 1-Octen-3-ol and 3-Octanone was 71.2 and 147.1 min, respectively. Both VOCs were highly toxic but 1-Octen-3-ol was more effective than 3-Octanone. Exposure of M. incognita egg-masses for 48 h at two doses (0.8 and 3.2 µL) of these VOCs showed that 1-Octen-3-ol had significantly greater nematicidal activity (100%) than 3-Octanone (14.7%) and the nematicide metham sodium (6.1%). High levels of reactive oxygen species detected in J2 exposed to 1-Octen-3-ol and 3-Octanone suggest oxidative stress was one factor contributing to mortality and needs to be investigated further.

Entomopathogenic nematodes and their symbiotic bacteria: from genes to field uses.

The term "microbial control" has been used to describe the use of microbial pathogens (bacteria, viruses, or fungi) or entomopathogenic nematodes (EPNs) to control various insect pest populations. EPNs are among the best biocontrol agents, and major developments in their use have occurred in recent decades, with many surveys having been conducted all over the world to identify EPNs that may have potential in the management of insect pests. For nematodes, the term "entomopathogenic" means "causing disease to insects" and is mainly used in reference to the bacterial symbionts of Steinernema and Heterorhabditis (Xenorhabdus and Photorhabdus, respectively), which cause EPN infectivity. A compendium of our multiannual experiences on EPN surveys and on their collection, identification, characterization, and use in agro-forestry ecosystems is presented here to testify and demonstrate once again that biological control with EPNs is possible and offers many advantages over chemicals, such as end-user safety, minimal damage to natural enemies, and lack of environmental pollution, which are essential conditions for an advanced IPM strategy.

New occurrence of Meloidogyne graminicola (Nematoda: Meloidogyninae) from rice fields in Italy: Variability and phylogenetic relationships.

Since the first detection of Meloidogyne graminicola in Piedmont, North Italy, in 2016, further inspections for the presence of the rice root-knot nematode were carried out in rice fields of neighboring regions, in accordance with the Italian NPPO (National Plant Protection Organization) to support the official phytosanitary measures, to enable the early detection of the rice pest, and to prevent its spread within the national territory. In 2018, surveys of rice fields in Lombardy region revealed a new occurrence of M. graminicola. In the present study, we confirmed the identification of the rice nematode in Lombardy using the ribosomal ITS region and the mitochondrial COI and COII genes. The sequences and phylogenetic analyses revealed that Lombardy M. graminicola population grouped in all trees in the main cluster containing Meloidogyne species belonging to graminis group, but always in a different subgroup compared to the Piedmont population of M. graminicola. These results clearly suggest that the two Italian populations have been recently and independently introduced and confirm that the geographic origin is not the main factor leading to M. graminicola population variability.

A simplified seasonal forecasting strategy, applied to wind and solar power in Europe.

We demonstrate levels of skill for forecasts of seasonal-mean wind speed and solar irradiance in Europe, using seasonal forecast systems available from the Copernicus Climate Change Service (C3S). While skill is patchy, there is potential for the development of climate services for the energy sector. Following previous studies, we show that, where there is skill, a simple linear regression-based method using the hindcast and forecast ensemble means provides a straightforward approach for producing calibrated probabilistic seasonal forecasts. This method extends naturally to using a larger-scale feature of the climate, such as the North Atlantic Oscillation, as the climate model predictor, and we show that this provides opportunities to improve the skill in some cases. We further demonstrate that, on seasonal-average and regional (e.g. national) average scales, wind and solar power generation are highly correlated with single climate variables (wind speed and irradiance). The detailed non-linear transformations from meteorological quantities to energy quantities, which are essential for detailed simulation of power system operations, are usually not necessary when forecasting gross wind or solar generation potential at seasonal-mean regional-mean scales. Together, our results demonstrate that where there is skill in seasonal forecasts of wind speed and irradiance, or a correlated larger-scale climate predictor, skilful forecasts of seasonal mean wind and solar power generation can be made based on the climate variable alone, without requiring complex transformations. This greatly simplifies the process of developing a useful seasonal climate service.

Molecular Characterization and Functional Analysis of the Hb-hsp90-1 Gene in Relation to Temperature Changes in Heterorhabditis bacteriophora.

Understanding how entomopathogenic nematodes respond to temperature changes and have adapted to the local environment is crucial to improve their potential as biocontrol agents. In order to improve understanding of Heterorhabditis bacteriophora's potential adaptability to future climate changes, full-length cDNA and the corresponding gene of heat shock protein 90 (Hsp90) were isolated and fully characterized. The reproductive potential of the Apulian strain of H. bacteriophora increased when the temperature rose from 23 to 30°C, but no reproduction was found at 12°C. Expression analyses revealed that Hb-hsp90-1 was differentially expressed in Infective Juveniles (IJs) and adults (hermaphrodites, females and males). Up-regulation of Hb-hsp90-1 was higher during the recovery process in Galleria mellonella larvae than adults, thus confirming the protective role of Hb-hsp90-1 in coping with the host environment. Silencing of Hb-hsp90-1 resulted in a significant reduction (76%) in the expression level. Silenced IJs took longer than untreated nematodes to infect G. mellonella, showing that Hb-hsp90-1 could be also involved in chemosensation. Furthermore, the number of adults and IJs recovered from G. mellonella infected with silenced nematodes and incubated at 30°C was higher than that obtained from G. mellonella infected with untreated nematodes. These data confirm the crucial role of Hb-hsp90-1 allowing acclimation to increased temperatures and modulation of the recovery process.

Pratylenchus vovlasi sp. Nov. (Nematoda: Pratylenchidae) on Raspberries in North Italy with a Morphometrical and Molecular Characterization.

Root-lesion nematode species rank third only to root-knot and cyst nematodes as having the greatest economic impact on crops worldwide. A survey of plant-parasitic nematodes associated with decaying raspberries (Rubus sp.) in northern Italy revealed that root-lesion nematodes were the most frequently occurring species among other phytonematodes. Several Pratylenchus species have been associated with Rubus sp. in Canada (Quebec, British Columbia) and USA (North Carolina, Maryland, New Jersey) including P. penetrans and P. crenatus. In the roots and rhizosphere of symptomatic raspberries, nematodes of two Pratylenchus spp. were detected. Detailed morphometrics of the two root-lesion nematode isolates were consistent with Pratylenchus crenatus and with an undescribed Pratylenchus species. The extracted nematodes were observed and measured as live and fixed materials and subsequently identified by integrative taxonomy (morphometrically and molecularly). The latter species is described herein as Pratylenchus vovlasi sp. nov., resulting morphometrically closest to P. mediterraneus and phylogenetically to P. pratensis. The molecular identification of Pratylenchus vovlasi sp. nov. was carried out by sequencing the ITS region, D2-D3 expansion domains of the 28S rRNA gene and a partial region of the nuclear hsp90 gene. ITS-RFLP and sequence analyses revealed that Pratylenchus vovlasi sp. nov. had species-specific restriction profiles with no corresponding sequences present in the database. The phylogenetic relationships with ITS and D2-D3 sequences placed the Pratylenchus vovlasi sp. nov. in a clade with P. pratensis and P. pseudopratensis. This research confirms the occurrence of cryptic biodiversity within the genus Pratylenchus as well as the need for an integrative approach to the identification of Pratylenchus species.

Effect of climate and geography on worldwide fine resolution economic activity.

Geography, including climatic factors, have long been considered potentially important elements in shaping socio-economic activities, alongside other determinants, such as institutions. Here we demonstrate that geography and climate variables satisfactorily explain the worldwide economic activity as measured by the per capita Gross Cell Product (GCP-PC) at a fine geographical resolution, typically much higher than country average. A 1° by 1° GCP-PC dataset has been key for establishing and testing a direct relationship between 'local' geography/climate and GCP-PC. Not only have we tested the geography and climate hypothesis using many possible explanatory variables, importantly we have also predicted and reconstructed GCP-PC worldwide by retaining the most significant predictors. While this study confirms that latitude is the most important predictor for GCP-PC when taken in isolation, the accuracy of the GCP-PC prediction is greatly improved when other factors mainly related to variations in climatic variables, rather than average climatic conditions as typically used, are considered. However, latitude diminishes in importance when only the wealthier parts of the globe are considered. This work points to specific features of the climate system which explain economic activity, such as the variability in air pressure. Implications of these findings range from an improved understanding of why socio-economically better-off societies are geographically placed where they are in the present, past and future to informing where new economic activities could be established in order to yield favourable economic outcomes based on geography and climate conditions.

Integrative diagnosis, biological observations, and histopathology of the fig cyst nematode Heteroderafici Kirjanova (1954) associated with Ficuscarica L. in southern Italy.

Morpho-biological notes and histopathology, based on LM and SEM observations, of the fig cyst nematode Heteroderafici isolated from Ficuscarica roots, collected in home and public gardens of Apulia region, southern Italy, are described and illustrated. Seventy-five localities throughout the Apulia region were sampled and one-quarter of the sampled localities had fig roots infested with H.fici, with population densities ranging from 44 to 180 cysts/100 ml of soil. All attempts to detect H.fici on ornamental Ficus spp. as well as on imported bonsai in Italy were unsuccessful. Morphometric characters of the Italian population conform to those of the type and re-description populations reported for H.fici. Molecular analysis using ITS, D2-D3 expansion domains of the 28S rRNA, and the partial 18S rRNA sequences of H.fici newly obtained in this study matched well with the corresponding sequences of H.fici present in the GenBank database. Phylogenetic trees confirmed and supported the grouping of H.fici in the Humuli group. Heteroderafici completes its embryogenic development in 14-16 days at 25 °C. Post-invasion development and maturity in the roots of F.carica seedlings is completed in 64-68 days at 25-28 °C with juveniles and adults showing different parasitic habits, being endoparasitic and semi-endoparasitic respectively. The establishment of permanent feeding sites that consist of the formation of large syncytia causes anatomical modification of vascular elements and general disorder in the root stelar structures. Syncytia structures associated with mature females showed different degrees of vacuolisation, numbers of syncytial cells, and contained nuclei and nucleoli which were constantly hypertrophied.

Molecular profiling of nematode associates with Rhynchophorus ferrugineus in southern Italy.

A survey of nematodes associated with the red palm weevil Rhynchophorus ferrugineus was conducted in southern Italy in 2015 and 2016 in order to create a species inventory and obtain data about nematode biodiversity. A total of 70 insect samples (pupae and adults) were collected from infested Phoenix canariensis, Phoenix dactylifera, and Chamaerops humilis palms in three Italian Regions: sampling took place at 11 locations in Apulia, 1 in Basilicata, and 1 in Sardinia regions. Individual insects were dissected to determine nematode presence, and different nematode species were also recovered from red palm weevil cocoons collected at the sites in Apulia. Individual nematodes were molecularly identified by sequencing the ITS, D2-D3 expansion domains of the 28SrRNA gene and the mitochondrial COI and inferring the phylogenetic relationships. The insect-associated nematofauna identified belonged to the families Rhabditidae, Cephalobidae, and Diplogastridae. Just two nematode species, Teratorhabditis synpapillata and Mononchoides macrospiculum, were always found in association with adult insects and cocoons taken from all sampling sites. This paper reports on the biodiversity of the nematodes associated with R. ferrugineus and on current knowledge of the specific habitat of specialized and divergent entomophilic nematodes.

Oscheius onirici sp. n. (Nematoda: Rhabditidae): a new entomopathogenic nematode from an Italian cave.

Oscheius onirici sp. n. (Nematoda: Rhabditidae) was isolated from a karst cave soil of Central Italy. Molecular and morphological analyses were performed. Total DNA was extracted from individual nematodes and the mitochondrial COI, the ITS containing region, the D2-D3 expansion domains of the 28S rRNA gene and the 18S rRNA gene were amplified and sequenced. BLAST search at NCBI by using all molecular markers revealed that this taxon is similar to Oscheius species. Phylogenetic trees of ITS, 28S and 18S rDNA revealed that O. onirici sp. n. belongs to Dolichura-group. Oscheius onirici sp. n. is characterized by small body size and stoma rhabditoid type. Female reproductive system is amphidelphic. Males are rare with peloderan bursa, spicules slender and small, nine pairs of papillae of different lengths, arranged in a 1+1+1/3+3 pattern. Entomopathogenicity bioassay revealed that this nematode is capable of infecting larvae of Galleria mellonella and Tenebrio molitor.

A New Root-Knot Nematode, Meloidogyne baetica n. sp. (Nematoda: Heteroderidae), Parasitizing Wild Olive in Southern Spain.

ABSTRACT High infection rates of wild olive (Olea europaea sp. sylvestris) feeder roots and soil infestation by a new root-knot nematode were found in sandy soil at Vejer de la Frontera (Cádiz), southern Spain. Morphometric traits and analyses of the nematode esterase electrophoretic pattern as well as of the internal transcribed spacer 1 (ITS1)-5.8S gene and D2-D3 fragment of the 28S gene of rDNA showed that specimens differed clearly from known root-knot nematodes. Studies of host-parasite relationships showed a typical susceptible reaction in naturally infected wild olive plants and in olive planting stocks (cvs. Arbequina and Picual) artificially inoculated with the nematode. However, the nematode did not reproduce in artificially inoculated chickpea, pea, and tomato. Because of the ability of this new nematode to infect wild and cultivated olives only, we suggest the common name, "Mediterranean olive root-knot nematode." The species is herein described and illustrated, and named as Meloidogyne baetica n. sp. The new root-knot nematode can be distinguished from other Meloidogyne spp. by (i) the perineal pattern, which is almost similar to that of M. artiellia, characterized by distinct inner striae forming two distinct longitudinal bands, extending throughout the perineum to just below the vulva; (ii) female excretory pore anterior to the level of stylet knobs, excretory pore distance from anterior end/length of stylet ratio extremely small (0.5 to 0.8); and (iii) second-stage juveniles with elongate-conoid tail. Phylogenetic trees derived from maximum parsimony analyses showed that M. baetica is closely related to M. artiellia, the cereal and legume root-knot nematode.

Ensemble decadal predictions from analysed initial conditions.

Sensitivity experiments using a coupled model initialized from analysed atmospheric and oceanic observations are used to investigate the potential for interannual-to-decadal predictability. The potential for extending seasonal predictions to longer time scales is explored using the same coupled model configuration and initialization procedure as used for seasonal prediction. It is found that, despite model drift, climatic signals on interannual-to-decadal time scales appear to be detectable. Two climatic states have been chosen: one starting in 1965, i.e. ahead of a period of global cooling, and the other in 1994, ahead of a period of global warming. The impact of initial conditions and of the different levels of greenhouse gases are isolated in order to gain insights into the source of predictability.