GrapevineXL reliably predicts multi-annual dynamics of vine water status, berry growth, and sugar accumulation in vineyards

Climate and water availability greatly affect each season’s grape yield and quality.Using models to accurately predict environment impacts on fruit productivity and quality is a huge challenge.We calibrated and validated the functional-structural model,GrapevineXL,with a data set including grapevine seasonal midday stem water potential(�xylem),berry dry weight(DW),fresh weight(FW),and sugar concentration per volume([Sugar])for a wine grape cultivar(Vitis vinifera cv.Cabernet Franc)in field conditions over 13 years in Bordeaux,France.Our results showed that the model could make a fair prediction of seasonal�xylem and good-to-excellent predictions of berry DW,FW,[Sugar]and leaf gas exchange responses to predawn and midday leaf water potentials under diverse environmental conditions with 14 key parameters.By running virtual experiments to mimic climate change,an advanced veraison(i.e.the onset of ripening)of 14 and 28 days led to significant decreases of berry FW by 2.70%and 3.22%,clear increases of berry[Sugar]by 2.90%and 4.29%,and shortened ripening duration in 8 out of 13 simulated years,respectively.Moreover,the impact of the advanced veraison varied with seasonal patterns of climate and soil water availability.Overall,the results showed that the GrapevineXL model can predict plant water use and berry growth in field conditions and could serve as a valuable tool for designing sustainable vineyard management strategies to cope with climate change.

New insights into the heat responses of grape leaves via combined phosphoproteomic and acetylproteomic analyses

Heat stress is a serious and widespread threat to the quality and yield of many crop species,including grape(Vitis vinifera L.),which is cultivated worldwide.Here,we conducted phosphoproteomic and acetylproteomic analyses of leaves of grape plants cultivated under four distinct temperature regimes.The phosphorylation or acetylation of a total of 1011 phosphoproteins with 1828 phosphosites and 96 acetyl proteins with 148 acetyl sites changed when plants were grown at 35℃,40℃,and 45℃in comparison with the proteome profiles of plants grown at 25℃.The greatest number of changes was observed at the relatively high temperatures.Functional classification and enrichment analysis indicated that phosphorylation,rather than acetylation,of serine/arginine-rich splicing factors was involved in the response to high temperatures.This finding is congruent with previous observations by which alternative splicing events occurred more frequently in grapevine under high temperature.Changes in acetylation patterns were more common than changes in phosphorylation patterns in photosynthesis-related proteins at high temperatures,while heat-shock proteins were associated more with modifications involving phosphorylation than with those involving acetylation.Nineteen proteins were identified with changes associated with both phosphorylation and acetylation,which is consistent with crosstalk between these posttranslational modification types.

Micropropagation of the Moroccan Endemic Plant <i>Thymus broussonetii</i>Boiss. with Aromatic-Medicinal Value and Conservation Concern

Micropropagation from shoot tips and nodal segments was carried out for the conservation and domestication of spontaneous Moroccan thyme, Thymus broussonetii Boiss. subsp. broussonetii (endemic threatened). The mineral composition of the culture medium, as well as the succession of different growth regulators, influenced the in vitro growth of this species. Sterilized achenes of T. broussonetii were able to germinate on an agar medium containing Gautheret macronutrients with a rate of 25% and a degree of contamination of less than 4%. Shoot apices of 15-day seedlings (two cotyledon leaves) were cultivated on SD + 0.46 μM Kin medium and the explants obtained were transplanted every month. Six macronutrients (MS, B5, SH, SD, MSm and N30K) were tested and N30K was chosen for the following experiments. Seven cytokinins (Kin, BAP, 2iP, DPU, adenine, Zeat and TDZ) at 0.46, 0.93 and 2.32 μM/l were evaluated and the addition of 0.93 μM adenine to N30K medium favored significantly the induction of buds and the elongation of explants. Three polyamines (putrescine, spermidine and spermine) at 2, 5, 10 and 20 μM/l were tested. A better multiplication of buds, shoots and roots was noted for N30K + 10 μM spermine. Cytokinin-auxin combinations led to better root multiplication and an increase in the number of buds and the length of explants, particularly for 0.46 μM Kin + 2.85 μM IAA. Acclimatization was successfully carried out using vitroplants developing a good root system. One month after the start of acclimatization, 97% of T. broussonetii plantlets were healthy. Three months later, they were transplanted into larger pots. 100% of the acclimatized plants developed flowers in the 2nd year between June and August. Re-initiation of the in vitro culture was carried out from sterilized twig segments collected from the acclimatized plants of T. broussonetii with 1 - 2 nodes on the medium N30K + 0.46 μM Kin, and 52.1% of the explants healthily proliferated. Finally, two micropropagation prototypes were developed: shoot tip culture from seedlings obtained after germination of achenes and node culture from acclimatized plants.

Water limitation and rootstock genotype interact to alter grape berry metabolism through transcriptome reprogramming

Grapevine is a perennial crop often cultivated by grafting a scion cultivar on a suitable rootstock.Rootstocks influence scions,particularly with regard to water uptake and vigor.Therefore,one of the possibilities to adapt viticulture to the extended drought stress periods is to select rootstocks conferring increased tolerance to drought.However,the molecular mechanisms associated with the ability of rootstock/scion combination to influence grape berry metabolism under drought stress are still poorly understood.The transcriptomic changes induced by drought stress in grape berries(cv.Pinot noir)from vines grafted on either 110R(drought-tolerant)or 125AA(drought-sensitive)rootstock were compared.The experiments were conducted in the vineyard for two years and two grape berry developmental stages(50%and 100%veraison).The genome-wide microarray approach showed that water stress strongly impacts gene expression in the berries,through ontology categories that cover cell wall metabolism,primary and secondary metabolism,signaling,stress,and hormones,and that some of these effects strongly depend on the rootstock genotype.Indeed,under drought stress,berries from vines grafted on 110R displayed a different transcriptional response compared to 125AA-concerning genes related to jasmonate(JA),phenylpropanoid metabolism,and pathogenesis-related proteins.The data also suggest a link between JA and secondary metabolism in water-stressed berries.Overall,genes related to secondary metabolism and JA are more induced and/or less repressed by drought stress in the berries grafted on the drought-sensitive rootstock 125AA.These rootstock-dependent gene expression changes are relevant for berry composition and sensory properties.

Micropropagation and Acclimatization of Common Oregano (Origanum vulgare L. Subsp. vulgare) by Shoot Tip Culture

Origanum vulgare L. is a commercially valued species with remarkable biological properties. It is subject to over-exploitation practices that seriously threaten its sustainability for future generations. Thus, micropropagation serves as a tool for the protection and domestication of this species. In this study, we established an in vitro vegetative propagation protocol for Origanum vulgare. This is done through the axillary bud technique by carrying out various tests. Six culture media (MS, MSm, N30K, SD, SH and B5) were tested. Therefore, SD was chosen for the following experiments. Seven cytokinins (adenine (Ad), N6-(2-isopentenyl) (2ip), zeatin (Zeat), kinetin (Kin), benzyladenine (BAP), 1,3-diphenylurea (DPU) and thidiazuron (TDZ) at 5 concentrations (0.44, 1.33, 2.22, 3.11 and 4.44 μM/L) were evaluated. Thus, Kin at 3.11 μM allowed high regeneration of vitroplants, optimal elongation, total rooting of explants, maximum bud multiplication, and absence of hyperhydric explants. In fact, the integration of auxins (indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), and 1-naphthaleneacetic acid (NAA)) into the culture medium and their combinations with 3.11 μM Kinetin contributed to the optimization of the root part. Thus, it was improved in particular in the case of 3.11 μM Kin and 6.27 μM IBA. Three polyamines (Putrescine, Spermidine and Spermine) at different concentrations (1.134, 3.402, 5.67, 7.938 and 11.34 μM/L) combined at 3.11 μM Kin and 6.27 μM IBA were tested. In fact, 1.304 μM putrescine was considered to be the most suitable for in vitro culture of explants, since it allowed optimal propagation of buds and roots, also a high rate of regeneration and rhizogenesis. GA3 at 1.15 μM combined with 3.11 μM Kin and 6.27 μM IBA permitted maximum bud multiplication. The acclimatization was carried out successfully using vitroplants showing good foliar and root development. Thus, three months after acclimatization, the seedlings were transferred into large pots under natural light and temperature conditions. Almost all acclimatized plants developed flowers in the first year between May and July.

Chasing the queens of the alien predator of honeybees: A water drop in the invasiveness ocean

The Yellow-legged hornet, Vespa velutina , was accidentally introduced in France in 2004, and then spread rapidly through the French territory but also to adjacent European countries (Spain, Portugal, and Belgium). During summer and autumn, V. velutina workers hunt domestic honeybees, Apis mellifera, for feeding their larvae. The impact of this alien species is mainly economic, beekeepers experiencing heavy colony losses, but also ecological, V. velutina hunting other pollinators. In their year-round life cycle, nest initiation by single queen during spring is the critical stage. In invaded areas, spring queen trapping using food baits has been promoted by apicultural unions in order to limit V. velutina population expansion. The goals of this work were 1) to evaluate the yield of this method, 2) to identify appropriate sites for trapping, 3) to identify potential optimal climatic windows of capture, and 4) to quantify the impact on local entomofauna. Our results showed that water proximity enhances trapping but not beehives proximity, and that trapping is inefficient if average week temperatures are below 10°C. Although the trapping effect on biodiversity should be studied more carefully, spring queen trapping is highly questionable unless specific attractants could be proposed.

The role of auxin during early berry development in grapevine as revealed by transcript profiling from pollination to fruit set

Auxin is a key phytohormone that modulates fruit formation in many fleshy fruits through the regulation of cell division and expansion.Auxin content rapidly increases after pollination and the manipulation in its levels may lead to the parthenocarpic development.ln Vitis vinifera L.,little is known about the early fruit development that encompasses from pollination to fruit set.Pollination/fertilization events trigger fruit formation,and auxin treatment mimics their effect in grape berry set.However,the role of auxin in this process at the molecular level is not well understood.To elucidate the participation of auxin in grapevine fruit formation,morphological,reproductive,and molecular events from anthesis to fruit set were described in sequential days after pollination.Exploratory RNA-seq analysis at four time points from anthesis to fruit set revealed that the highest percentage of genes induced/repressed within the hormone-related gene category were auxin-related genes.Transcript profiling showed significant transcript variations in auxin signaling and homeostasis-related genes during the early fruit development.Indole acetic acid and several auxin metabolites were present during this period.Finally,application of an inhibitor of auxin action reduced cell number and the mesocarp diameter,similarly to unpollinated berries,further confirming the key role of auxin during early berry development.This work sheds light into the molecular features of the initial fruit development and highlights the auxin participation during this stage in grapevine.

Effect of Macronutrients, Cytokinins and Auxins, on <i>in Vitro</i>Organogenesis of <i>Thymus vulgaris</i>L.

The present study reports an efficient protocol for in vitro propagation of Thymus vulgaris L., an aromatic and medicinal plant in Morocco. Initially, we performed in vitro multiplication of Thymus vulgaris explants existing in the laboratory and obtained from micropropagation by shoot tip culture. Afterwards, we have evaluated the effect of six macronutrients. After that, seven cytokinins (Kin, BAP, 2iP, DPU, Adenine, Zeatine and TDZ) in three different concentrations (0.46, 0.93, 2.32 μM) have been evaluated to optimize cultures multiplication and elongation. Moreover, the effect of three auxins (IAA, IBA and NAA) at 0.57 μM, combined to 4 cytokinins (Kin, BAP, DPU and Ad.) at 0.46 μM, on shoot rooting has been studied. Thereby, MS medium has been proved the most favorable for plantlets growing. Also, we found that the addition of certain cytokinins, specifically 0.46 Kin, 0.46 and 0.93 BAP, 0.46 2iP, 0.46 DPU, 0.46 Ad. and 0.46 Zeat., ensures better multiplication and growth of vitroplants. In addition, multiplication and rooting of cultures were well optimized after addition 0.46 Kin + 0.57 IAA or NAA, 0.46 DPU + 0.57 IBA and 0.46 Ad. + 0.57 IBA combinations to the culture medium. Lastly, plantlets with roots were successfully acclimatized to ex-vitro conditions and these latter served as a source to establish in vitro culture again.

Assessment of the Effect of Environmental Factors on the Antagonism of <i>Bacillus amyloliquefaciens</i>and <i>Trichoderma harzianum</i>to <i>Colletotrichum acutatum</i>

The effect of temperature (18°C - 30°C), water activity (0.85 - 1) and pH (4 - 9) was studied by dual culture technique on the antagonism of Bacillus amyloliquefaciens and Trichoderma harzianum to Colletotrichum acutatum, responsible of strawberry (Fragaria x ananassa (Weston) Duchesne ex Rozier) anthracnose. The antagonistic bacteria’s strains behave significantly and differently according to the parameters studied. These results reveal useful information about the applicability of their biocontrol in agricultural culture with the change of environmental factors.

Efficient Plant Regeneration from Explants of Compact Oregano (Origanum compactum Bentham)

Origanum compactum Benth., a species endemic to Morocco characterized by its biological activities, is overexploited because of its commercial value and threatened with extinction. Accordingly, measures for its conservation are needed. Micropropagation serves as a solution for the protection and the domestication of this species. In this investigation, we established a protocol for vegetative multiplication in vitro of Origanum compactum by the axillary bud technique. Six culture media (SH, SD, N30K, MS, MSm, B5) were tested to determine the most suitable mineral medium for growth and development of explants. Four cytokinins: Kinetin, Zeatin, BAP (6-Benzylaminopurine), 2ip (2-Isopentenyladenine) and three compounds with cytokinin activity: Adénine, 1,3-Diphenylurea (DPU) and Thidiazuron (TDZ) at five concentrations (0.44, 1.33, 2.22, 3.11, 4.44 μM) were tested on budding, growth, hyperhydria and rooting. Then three auxins, Indole-3-acetic acid (IAA), Naphthalene acetic acid (NAA) and Indole butyric acid (IBA) at four concentrations (1.14, 2.85, 4.56, 6.27 μM) in presence of 2.22 μM BAP were evaluated. The combination of Gibberellic acid (0.29, 1.5, 2.60, 2.89 μM GA3) and three polyamines (Putrescine, Spermidine, Spermine) at five concentrations (1.134, 3.402, 5.67, 7.938, 11.34 μM) with cytokinins and auxins were considered. Our results show that Margara medium is the most efficient and BAP at 2.22 μM is the best cytokinin for development of the aerial parts, with a regeneration rate of 88, 90%;rhizogenesis is successful with the combination of 6.27 μM IAA and 2.22 μM BAP. Moreover, the integration of 2.89 μM GA3 with 2.22 μM BAP and 6.27 μM IAA promotes vitroplant growth, bud and shoot multiplication and elongation of the aerial part. The addition of polyamines with 2.22 μM BAP and 6.27 μM IAA does not improve the root part, but Spermine at 5.67 μM promotes bud and shoot multiplication with a high percentage of regeneration, and spermidine at the same concentration gives long explants. Finally, plantlets with good root development were successfully acclimatized to natural conditions and served as a source to establish in vitro culture again.

Effect of Cytokinins on the Micropropagation of Carob(Ceratonia siliqua L.)through Shoot Tip Culture

In order to determine the most suitable cytokinin for the micropropagation of carob (Ceratonia siliqua L.), the effect of four cytokinins: BAP, zeatin, kinetin and 2-iP, was tested on explants derived from young seedlings of seven days. Organogenesis is better in the presence of BAP (0.5 mg/l), while buds growth is favored by zeatin (0.5 mg/l). The combination of the most suitable cytokinin (BAP at 0.5 mg/l) with IBA improves the caulogenesis when the concentration of this latter is low (0.1 mg/l);high concentrations of IBA have an inhibitory effect on elongation and neoformation of shoots and leaves. The multiplication and growth of shoots are more favorable on WPM medium in the presence of BAP (0.5 and 1 mg/l) alone or combined with 0.5 mg/l with GA3, while rooting is mainly favored by IBA, especially at 2 mg/l.

Determination of Polyphenols Content in Carob Pulp from Wild and Domesticated Moroccan Trees

Carob (Ceratonia siliqua L.) fruit pulp from 12 wild and 8 domesticated trees from 15 Moroccan localities were investigated for their polyphenols contents: total polyphenols content (53.22 - 118.04 mg and 57.46 - 183.31 mg), total flavonoids (1.41 - 4.83 mg and 1.62 - 7.46 mg) and condensed tannins (1.47 - 7.36 mg and 1.85 - 6.66 mg) in one carob fresh pulp for wild and domesticated trees, respectively. Fruit pulp from trees in the same region shows variable contents in polyphenols, flavonoids and condensed tannins.

Evaluation of Plant Growth Promoting Ability of Bacillus amyloliquefaciens Bc2 and Trichoderma harzianum TR In Vivo

Bacillus amyloliquefaciens Bc2 and Trichderma harzianum TR were used to evaluate their growth-promoting activity on cultivated strawberries, under laboratory and field conditions, and we have noticed that the percentage of achene germination is important for ones treated with TR (=97%) followed by those treated with Bc2 strain (=90%) and the control (=84%). Inoculations on field showed that on untreated soil with insecticide, TR is effective and allows the development of plants and extends the duration of flowering and fruiting. On treated soil, Bc2 clearly promotes the growth and development of strawberry seedlings and its role as plant growth promoting microorganisms has been proved.

Evidences of hydroxytyrosol as an anti-inflammatory agent in Parkinson’s disease:insights into the mechanisms of action

Neuroinflammation in neurodegenerative diseases:Inflammatory processes play a critical role in neurodegenerative diseases,such as Parkinson’s disease(PD).Thus,neuroinflammation is involved in the progression and development of these diseases,becoming an important pathological hallmark.Microglial cells,the“macrophages”from central nervous system,are the initiating cells of the innate immune response against different stimuli in the brain.

Biocontrol Potential of Bacillus amyloliquefaciens Bc2 and Trichoderma harzianum TR against Strawberry Anthracnose under Laboratory and Field Conditions

The increasingly strict regulation of the use of phytosanitary products and the will of the populations to move towards sustainable development allow bio-sourced products and more particularly for biocontrol in the field of pesticides to progress. The study carried out here concerns the evaluation of the potency of Bacillus amyloliquefaciens Bc2 and Trichoderma harzianum to fight against strawberry cultures infected with anthracnose (Fragaria × ananassa Duch.). The studies were carried out in the laboratory and in the field. The results indicate that B. amyloliquefaciens Bc2 and Trichoderma harzianum TR are effective for the biological control of anthracnose, gray mold and powdery mildew on strawberries grown in field conditions. The increase in plant size and the number of fruits produced with these control agents has also been observed.

Proteomic analysis of early-stage incompatible and compatible interactions between grapevine and P.viticola

Wild grapevines can show strong resistance to the downy mildew pathogen P.viticola,but the associated mechanisms are poorly described,especially at early stages of infection.Here,we performed comparative proteomic analyses of grapevine leaves from the resistant genotype V.davidii“LiuBa-8”(LB)and susceptible V.vinifera“Pinot Noir”(PN)12h after inoculation with P.viticola.By employing the iTRAQ technique,a total of 444 and 349 differentially expressed proteins(DEPs)were identified in LB and PN,respectively.The majority of these DEPs were related to photosynthesis,respiration,cell wall modification,protein metabolism,stress,and redox homeostasis.Compared with PN,LB showed fewer downregulated proteins associated with photosynthesis and more upregulated proteins associated with metabolism.At least a subset of PR proteins(PR10.2 and PR10.3)was upregulated upon inoculation in both genotypes,whereas HSP(HSP70.2 and HSP90.6)and cell wall-related XTH and BXL1 proteins were specifically upregulated in LB and PN,respectively.In the incompatible interaction,ROS signaling was evident by the accumulation of H_(2)O_(2),and multiple APX and GST proteins were upregulated.These DEPs may play crucial roles in the grapevine response to downy mildew.Our results provide new insights into molecular events associated with downy mildew resistance in grapevine,which may be exploited to develop novel protection strategies against this disease.

In Vitro Antagonistic Activity of Trichoderma harzianum and Bacillus amyloliquefaciens against Colletotrichum acutatum

Biological control of plant diseases makes extensive use of the antagonists. The effectiveness of the control depends on the choice of an effective antagonist strains from criteria of implying a good knowledge of biological peculiarities of the material used. In this study, the antagonistic activities of nine isolates of Bacillus amyloliquefaciens and one isolate of Trichoderma harzianum were tested in vitro against seven strains of Colletotrichum acutatum, the causal agent of strawberry anthracnose. It was found that T. harzianum and B. amyloliquefaciens strains showed the ability to inhibit the mycelial growth of the pathogen by dual culture technique by more than 50 percent. The nonvolatile substances produced by the antagonists showed high inhibition percentages that are more than 99 percent, meanwhile their volatile compounds inhibited the mycelial growth of C. acutatum strains with low inhibition percentages starting from 30 percent except the substances produced by B. amyloliquefaciens Bc2 that showed a high inhibition percentage of more than 70 percent.

Effect of Different Concentrations of the Suspensions of B.amyloliquefaciens and T.harzianum on the Development of C.acutatum

The effect of nine isolates of Bacillus amyloliquefaciensand one strain of Trichoderma harzianum, TR, on mycelial growth and germination of Colletotrichum acutatum were studied. The nine isolates were identified as Bacillus amyloliquefaciens. The efficacy of isolates was tested, at different concentrations. Results showed that one Bacillus isolates (Bc2) and TR were more effective at the lower concentration tested (3 × 105 CFU/ml and 105 conidia/ml).

Towards an open grapevine information system

Viticulture,like other fields of agriculture,is currently facing important challenges that will be addressed only through sustained,dedicated and coordinated research.Although the methods used in biology have evolved tremendously in recent years and now involve the routine production of large data sets of varied nature,in many domains of study,including grapevine research,there is a need to improve the findability,accessibility,interoperability and reusability(FAIR-ness)of these data.Considering the heterogeneous nature of the data produced,the transnational nature of the scientific community and the experience gained elsewhere,we have formed an open working group,in the framework of the International Grapevine Genome Program(www.vitaceae.org),to construct a coordinated federation of information systems holding grapevine data distributed around the world,providing an integrated set of interfaces supporting advanced data modeling,rich semantic integration and the next generation of data mining tools.To achieve this goal,it will be critical to develop,implement and adopt appropriate standards for data annotation and formatting.The development of this system,the GrapeIS,linking genotypes to phenotypes,and scientific research to agronomical and oeneological data,should provide new insights into grape biology,and allow the development of new varieties to meet the challenges of biotic and abiotic stress,environmental change,and consumer demand.

Proteomic and metabolomic profiling underlines the stage- and time-dependent effects of high temperature on grape berry metabolism

Climate change scenarios predict an increase in mean air temperatures and in the frequency,intensity,and length of extreme temperature events in many wine-growing regions worldwide.Because elevated temperature has detrimental effects on berry growth and composition,it threatens the economic and environmental sustainability of wine production.Using Cabernet Sauvignon fruit-bearing cuttings,we investigated the effects of high temperature(HT)on grapevine berries through a label-free shotgun proteomic analysis coupled to a complementary metabolomic study.Among the 2,279 proteins identified,592 differentially abundant proteins were found in berries exposed to HT.The gene ontology categories“stress,”“protein,”“secondary metabolism,”and“cell wall”were predominantly altered under HT.High temperatures strongly impaired carbohydrate and energy metabolism,and the effects depended on the stage of development and duration of treatment.Transcript amounts correlated poorly with protein expression levels in HT berries,highlighting the value of proteomic studies in the context of heat stress.Furthermore,this work reveals that HT alters key proteins driving berry development and ripening.Finally,we provide a list of differentially abundant proteins that can be considered as potential markers for developing or selecting grape varieties that are better adapted to warmer climates or extreme heat waves.