FaSnRK1a mediates salicylic acid pathways to enhance strawberry resistance to Botrytis cinerea

Strawberry is a major fruit crop worldwide because its nutritional and health benefits to human health,but its productivity is limited by Botrytis cinerea.Sucrose nonfermentation 1-related protein kinase 1(SnRK1)has a defense function against pathogens,but the function of SnRK1 in the defense response to B.cinerea in plants is still unclear.In this study,FaSnRK1a-OE and RNAi fruits were constructed and then inoculated with B.cinerea.The result reveals a positive role of Fa SnRK1a in the regulation of resistance to gray mold.FaSnRK1a affects SA content by regulating FaPAL1 and FaPAL2 expressions.The genes related to the SA signaling pathway(FaTGA1 and FaTGA2.1)were significantly increased/decreased in FaSnRK1a-OE or FaSnRK1a-RNAi fruit,respectively.FaSnRK1a interacted with the FaWRKY33.2 protein and negatively regulated FaWRKY33.2 expression,and FaWRKY33.2 acts as a repressor of disease resistance to B.cinerea.Finally,FaSnRK1a regulates the expression of six PR genes and the activities of antioxidant enzymes to boost defense response after B.cinerea inoculation.Our findings showed that FaSnRK1a increases the resistance of strawberry fruit to B.cinerea via SA signaling pathway and interaction with the FaWRKY33.2 transcription factor.

Real-Time Detection and Instance Segmentation of Strawberry in Unstructured Environment

The real-time detection and instance segmentation of strawberries constitute fundamental components in the development of strawberry harvesting robots.Real-time identification of strawberries in an unstructured envi-ronment is a challenging task.Current instance segmentation algorithms for strawberries suffer from issues such as poor real-time performance and low accuracy.To this end,the present study proposes an Efficient YOLACT(E-YOLACT)algorithm for strawberry detection and segmentation based on the YOLACT framework.The key enhancements of the E-YOLACT encompass the development of a lightweight attention mechanism,pyramid squeeze shuffle attention(PSSA),for efficient feature extraction.Additionally,an attention-guided context-feature pyramid network(AC-FPN)is employed instead of FPN to optimize the architecture’s performance.Furthermore,a feature-enhanced model(FEM)is introduced to enhance the prediction head’s capabilities,while efficient fast non-maximum suppression(EF-NMS)is devised to improve non-maximum suppression.The experimental results demonstrate that the E-YOLACT achieves a Box-mAP and Mask-mAP of 77.9 and 76.6,respectively,on the custom dataset.Moreover,it exhibits an impressive category accuracy of 93.5%.Notably,the E-YOLACT also demonstrates a remarkable real-time detection capability with a speed of 34.8 FPS.The method proposed in this article presents an efficient approach for the vision system of a strawberry-picking robot.

Ellagic Acid Enhances Antioxidant System Activity and Maintains the Quality of Strawberry Fruit during Storage

Ellagic acid(EA)is a natural antioxidant,widely present in a lot of forms’soft fruits,nuts,and other plant tissues,and helpful for promoting human health;however,its protective effect on postharvest fruit and improving the quality index of postharvest fruit have rarely been studied.In this experiment,the strawberries were soaked in 0,100,200,300,400,and 500 mg L^(−1) EA,respectively,and the influential EA on fruit quality and the antioxidant system of strawberries were studied.Compared with the control,EA treatment can reduce the browning degree and rotting rate of strawberry fruit during storage and augment the soluble solid content(SSC).EA treatment can also increase the content of related stuff and enzyme activity in antioxidant systems;the gene expression level of polyphenol oxidase(PPO)in strawberries treated with EA was always down-regulated,correspondingly,the expression of other antioxidant enzyme genes was enhanced.Among the strawberry fruits treated with EA of different concentrations,300 mg L^(−1) EA had the best effect in the process of strawberry preservation.The results suggested that the proper concentration of exogenous EA at 300 mg L−1 could maintain strawberries’quality and enhance the antioxidant system by improving the activities of antioxidative enzymes and the ascorbateglutathione(AsA-GSH)cycle during storage.

Nutritional, Textural, Sensory Properties and Storage Stability Evaluation of Newly Formulated Strawberry Bar

An analytical study was conducted to develop a strawberry bar and analysis of the nutritional quality and storage stability of a newly formulated strawberry bar. In proximate analysis moisture, ash, pH, acidity, vitamin C, total phenolic content, and antioxidant of strawberry pulp were found 82%, 0.5%, 2.57, 1.38%, 49 mg/100g, 0.33%, 75.17% whereas, in the strawberry bar, it was found 18%, 0.71%, 2.10, 1.10%, 22 mg/100g, 0.14%, 56.13% respectively. For textural properties analysis, the textural profile of the strawberry bar such as hardness, factorability, viscosity, cohesiveness, adhesiveness, springiness, gumminess, chewiness, probe diameter, threshold, filtering range of strawberry bar was found 5.538 × 104 N/m, 0.000 N/m2, 1.687 × 104 N/m2, 0.6636, 2.908 × 103 J/m3, 0.8664, 3.675 × 104 N/m2, 3.184 × 104 N/m2, 20.000 mm, 5 N, 0.4 N respectively whereas the maximum level was 17.4 N at 29.628 seconds and the minimum level was -5.3 N at 30.166 seconds. The sensory evaluation of the strawberry bar was conducted based on nine-point hedonic scales. The sensory evaluation results indicated that the appearance of the strawberry bar was liked very much and the taste, texture, color, aroma, and overall acceptability of the strawberry bar were liked moderately by panelists. In microbiological analysis, the Total Viable Count (TVC) and Total Coliform Count (TCC) of the strawberry bar were found safe levels for up to third days. But on the sixth day, the TCC of the strawberry bar was not found and the TVC of the strawberry bar was found 17 × 104 cfu/g respectively which is higher than the permitted value (Gulf Standard: 105 cfu/g). From the microbial point of view, the newly formulated strawberry bar was safe to consume for up to 3 days but not for 6 days.

Metabolomic and transcriptomic analysis reveals the molecular mechanism by which blue light promotes lutein synthesis in strawberry

Carotenoids are an important component of the human diet, and fruit is a primary source of carotenoids. The synthesis and regulation of carotenoids in fruit are important contributors to the formation of fruit quality. In China, strawberry is one of the main seasonal fruits grown in the winter. Previous studies have shown that light has a significant effect on the metabolism of anthocyanins, sugars, and polyphenols in strawberry. However, the understanding of the role of light in regulating the metabolism of carotenoids in strawberry remains limited. This study investigated the effects of blue, red, yellow-green, and white light on carotenoid metabolism in strawberry.Blue light treatment promoted the synthesis of multiple carotenoids, including lutein, compared with the other three treatment groups. The RNA sequencing data revealed that blue light treatment promoted the expression of lycopene ε-cyclase(LCYE), and the transient overexpression of LCYE in strawberry fruit promoted lutein accumulation in strawberry. Overall, the results suggest that blue light can promote the synthesis of lutein in strawberry by inducing the expression of LCYE.

Study on a Bowl-based Mechanism for Transplanting Potted Strawberry Seedlings

To improve the efficiency of fetching and transplanting seedlings for the mechanization of strawberry planting,an integrated transplanting mechanism was designed with protruding,fetching and planting performance to achieve rapid fetching and pushing bowl movements.According to the working principle of the slewing mechanism,a kinematics model and the optimization goal were established,respectively.Based on visual auxiliary analysis software,optimal parameters were obtained.A three-dimensional model was established to obtain a simulation trajectory by means of a virtual simulation design analysis.Three-dimensional printing technology was used to manufacture the test prototype,and the actual working trajectories of the test prototype were extracted using high-speed photography technology,which verified the consistency of the actual trajectory with the theoretical and simulated trajectories.A prototype transplanting experiment was performed with the success rate of seedling extraction of 91.2%and excellent planting rate of 82.8%,which met the requirements for integrated strawberry harvesting,planting and transplanting.The experimental results verified the correctness and feasibility of the design of integrated transplanting mechanism.

Rapid Screening of Pesticide Residues in Strawberry Based on HPLC-QTOF/MS SWATH Acquisition Mode

[Objectives]The paper was to establish a rapid screening method of pesticide residues in strawberry based on HPLC-QTOF/MS SWATH acquisition mode.[Methods]The strawberry sample was extracted with acetonitrile,purified with anhydrous magnesium sulfate,sodium chloride and primary secondary amine(PSA)powder,and then loaded on the machine.The full information spectrum was collected by positive ion mode and screened by comparing with the spectrum library,and 20 kinds of pesticide residues were quantitatively analyzed.[Results]The detection limits of 20 pesticide residues were 0.0002-0.0100 mg/kg,and they showed good linearity in the range of 0.0005-0.1000μg/mL(R 2≥0.995).The recoveries ranged from 62.8%to 106.3%.[Conclusions]This method is rapid,simple,sensitive and non-targeted,and can be used for rapid screening of multiple pesticide residues in strawberry.

In Vitro Propagation of Three Strawberry Varieties and Field Evaluation

A study was done to produce a rapid in vitro propagation of three strawberry genotypes and tested in the field under Bangladeshi circumstances. Festival, RABI-3, and Neho strawberry genotypes’ runner tips were cultivated in vitro to induce root induction and multiple shoot proliferation. MS (Murashige and Skoog) media that were basally containing three different concentrations at 1.0, 1.5, and 2.0 of BA (6-benzyl adenine), KIN (6-furfuryl amino purine), or GA3 (gibberellic acid) at 0.5 mg/L increasing tips of the runner was attained. The culture grew on the medium provided with 1.5 mg/L 6-benzyl adenine and 0.5 mg/L 6-furfuryl amino acid to increase shoot at the best level. Micro-cuttings were rooted on MS media at half strength combined with 0.5 mg/L - 1.5 mg/L IBA (indole butyric acid) or IAA (indole acetic acid). IBA attained 4 - 9 roots and 91% - 96% rooting at 1.0 mg/L. The resulting plantlets grew into hardy plants and took root in the earth. The genotype festival had the highest response rate, followed by RABI-3 and Neho.

Facility Cultivation Techniques of ‘Kaorino’ Strawberry

Strawberry‘Kaorino’is one of the perfect early-maturing strawberry varieties with high quality and high yield due to its anthracnose resistance,early maturity and good quality.The variety has become more and more popular with the promotion over the last few years,but there are great differences from other varieties in cultivation.Based on its varietal characteristics and cultivation performance in production over the last few years,we summarized the seedling cultivation techniques,planting management techniques,main disease control techniques and harvesting and storage techniques,aiming to provide reference for better promotion and application of‘Kaorino’.

In Vitro Propagation of Three Strawberry Cultivars through Runner Tips Culture

At the Genetics and Plant Breeding Laboratory of the Department of Agronomy and Agricultural Extension, University of Rajshahi, Bangladesh, strawberry in vitro propagation was done. Five Benzylaminopurine (BAP) concentrations were utilized for shoot induction—0.0 mg/L (Control), 0.5 mg/L, 1.0 mg/L, 1.5 mg/L, and 2.0 mg/L and five Indole Buteric Acid (IBA) concentrations—0.0 mg/L(Control), 0.5 mg/L, 1.0 mg/L, 1.5 mg/L, and 2.0 mg/l—were used for the induction of the root. The highest mean amount of shoots (eight) and length of the shoot (3.40 cm) were observed at a concentration of BAP of 0.5 mg/L. Festival also had the highest mean amount of leaves (6) when we used the identical concentration, while RABI-3 and Camarosa did the least well. The IBA of 0.5 mg/L concentration of rooting performed the best across all metrics tested among the five concentrations. The longest (3.3 cm) roots and most roots (7) were likewise obtained from this concentration in Festival. However, RABI-3 and Camarosa performed worse than Festival in the same concentration. Half-strength MS media without IBA concentration showed no response regarding root induction for each of the three cultivars.

Application Effect of Functional Microbial Products in the Cultivation of Green Ecological Strawberry

In order to promote the transformation and high-quality development of strawberry industry and speed up the application of functional microbial products in strawberry,the application technology of functional microbial products in strawberry planting is studied and summarized.It mainly includes:the periods and methods of continuous cropping strawberry soil reduction and disinfection at high temperature,before and after strawberry planting,before plastic film mulching and whole process of plant spraying.Through multi-point test and demonstration,the purpose of advancing the season of strawberries,improving the quality,increasing production and increasing efficiency is achieved,and there is no risk of agricultural residue pollution,which meets the needs of people's consumption upgrading.Therefore,the application of functional microbial products in green ecological planting is very necessary,and it is worth further speeding up the demonstration and promotion.

Changes in fungal community and diversity in strawberry rhizosphere soil after 12 years in the greenhouse

Soil fungi play a very important role in the soil ecological environment. In agricultural production, long-term monoculture and continuous cropping lead to changes in fungal community diversity. However, the effects of long-term monoculture and continuous cropping on strawberry plant health and fungal community diversity have not been elucidated. In this study, using high-throughput sequencing(HTS), we compared the fungal community and diversity of strawberry rhizosphere soil after various durations of continuous cropping(0, 2, 4, 6, 8, 10 and 12 years). The results showed that soil fungal diversity increased with consecutive cropping years. Specifically, the soil-borne disease pathogens Fusarium and Guehomyces were significantly increased after strawberry continuous cropping, and the abundance of nematicidal(Arthrobotrys) fungi decreased from the fourth year of continuous cropping. The results of correlation analysis suggest that these three genera might be key fungi that contribute to the changes in soil properties that occur during continuous cropping. In addition, physicochemical property analysis showed that the soil nutrient content began to decline after the fourth year of continuous cropping. Spearman's correlation analysis showed that soil pH, available potassium(AK) and ammonium nitrogen(NH_4^+-N) were the most important edaphic factors leading to contrasting beneficial and pathogenic associations across consecutive strawberry cropping systems.

Hot air treatment activates defense responses and induces resistance against Botrytis cinerea in strawberry fruit

The effect of hot air(HA, 45°C, 3.5 h) treatment on reducing gray mold caused by Botrytis cinerea in strawberry fruit and the possible mechanisms were investigated. The results showed that HA treatment significantly reduced lesion diameter and enhanced activities of chitinase(CHI), β-1,3-glucanase and phenylalanine ammonia-lyase(PAL) in strawberry fruit. Total phenolic contents were also increased by HA treatment. The activities of antioxidant enzymes including superoxide dismutase(SOD), catalase(CAT) and ascorbate peroxidase(APX) were higher in HA treated strawberry fruit than those in control. Expression of three defense related genes such as CAT, CCR-1 allele and PLA6 was greatly induced in HA treated strawberry fruit with or without inoculation by B. cinerea. In addition, the in vitro experiment showed that HA treatment inhibited spore germination and tube growth of B. cinerea. These results suggested that HA treatment directly activated disease resistance against B. cinerea in strawberry fruit without priming response and directly inhibiting growth of B. cinerea.

Changes of Activities in NAD Kinase and NADP PhosphataseDuring Ripening and Senescence of Tomatoand Strawberry Fruits

Activities of NAD kinase(NADK)and NADP phosphatase and relationship between the two enzymes and temperature, respiration, ethylene production and trifluoperazine(TFP) were studied during ripening and senescence of strawberry and tomato frnits after harvest at 4℃and 20℃. The activity of NAD kinase in strawberry decreased slowly during first four days, then increased gradually. The NADP phosphatase activity increased at the second day, decreased the next day,then increased again. In tomato fruit, the activities of NAD kinase and NADP phosphatase increased at the second day, decreased with the ripening and senescence of the fruit. The change trend of NAD kinase and respiration in the two fruits were similar, the same were NADP phosphatase and ethylene production. TFP enhanced the activity of NAD kinase and had little effect on NADP phosphatase. Low temperature(4℃ ) activated the NAD kinase and reduced the activity of NADP phosphatase. These results indicated that the NAD kinase and NADP phosphatase were related to the ripening and senescence of strawberry and tomato fruits. The activation of NAD kinase probably postponed the ripening and senescence of the fruits.

The Maturation and Senescence in Relation to Ca^2+,CaMContent and Ca^2+-ATPase Activity and Active OxygenMetabolism in Strawberry Fruits

The changes in content of Ca2 + and CaM, Ca2 + -ATPase activity and active oxygen metabolism during strawberry (Fragaria ananassa Duch. cv. Chunxing) fruits maturation and senescence were investigated in this study. The results showed that the soluble Ca2 + content and SOD activity in fruits tended to decline and O2.-production rate to increase, the Ca2+ -ATPase activity peaked at first and then declined during fruits maturation and senescence. There were the highest CaM content at white stage in preharvest fruits and at marked senescence stage in postharvest ones. The above biochemical changes in fruits stored at low temperature (4℃) were slower than those stored at normal temperature(25℃ ). Thus, it indicated that the stimulation of calcium messenger system and accumulation of active oxygen free radical were closely related to fruits maturation and senescence.

Effect of <i>Trichoderma harzianum</i>in Combination with Fungicides in Controlling Gray Mould Disease (<i>Botrytis cinerea</i>) of Strawberry

The effects of biofungicide formula containing the biocontrol agent Trichoderma harzianum (Jn14) as wettable powder in combination with the fungicides pyrimethanil and a mixture of cyprodinil and flydioxonil on Botrytis cinerea of strawberry in vitro, in vivo and in greenhouse were studied. The pathogen B. cinerea was more affected in vitro by low concentrations of the fungicides tested above 0.4 μg&middot;ml-1 than the bioagent T. harzianum (Jn14). The later was almost insensitive to pyrimethanil. In the same direction, gray mould disease severity on strawberry detached leaves and whole plants was reduced up to 89% by the tested fungicides, compared to the control, while the application of T. harzianum (Jn14) alone reduced disease severity up to 45% on strawberry detached leaves, compared to the control. In the integrated control approach, the combination of T. harzianum (Jn14) with higher concentrations of the tested fungicides (ED50) completely inhibited strawberry gray mould disease severity in pots and the greenhouse.

Biology and Physiology of <I>Colletotrichum acutatum</I>Strains Causing Strawberry’s Anthracnose

Seven Colletotrichum acutatum strains isolated from strawberries were cultivated on various culture media and tested in vitro and in vivo on Fragaria x ananassa for anthracnose symptoms. PDA caused an optimum growth of all isolates, MEA, ML and Strawberry allowed a good but not optimal growth. Czapeck, Sabouraud and the organic medium from potato gave the lowest growth rate of all isolates. PDA allowed a good sporulation of isolates follow-up by strawberry, MEA;in contrast, ML, Czapeck and Sabouraud gave a low sporulation. The fungal development is maximal at 25&degC and 27&degC for all the isolates studied. No growth was observed at 5&degC and 37&degC. The studied strains developed at all pH values. They didn’t develop at 0.6, 0.65 and 0.7 aw but mycelial growth was perfect at 1 and 0.95 aw. The in vitro test of the pathogenicity caused by C. acutatum strains on strawberry’s leaves showed an increasing percentage of infection with time and different infection rates among C. acutatum strains, strain Ca6 having a pathogenic power very high compared to the other isolates. After in vivo inoculation and incubation of the seedlings, all isolates caused severe symptoms related to anthracnose on leaflets and petioles of the studied strawberry plants.

Variation of Polyphenols, Anthocyanins and Antioxidant Power in the Strawberry Grape (<i>Vitis labrusca</i>) after Simulated Gastro-Intestinal Transit and Evaluation of <i>in Vitro</i>Antimicrobial Activity

The influence of a simulated digestive process on some biochemical and biological aspects of strawberry grape (Vitis labrusca) was investigated. The amount of total polyphenols and anthocyanins as well as the antioxidant power were evaluated. Results evidenced that the simulated gastrointestinal transit caused a decrease of the polyphenols content and total anthocyanins;these last, however, were more resistant than polyphenols, decreasing only of 50% respect to the initial value (31.50 μg/ml of extract). The extract exhibited an excellent antioxidant power (EC50 3.8 mg/ml), which decreased of about four times after the simulated gastrointestinal transit. The antimicrobial activity of the extract, evaluated against three Escherichia coli, Pseudomonas aeruginosa and Bacillus cereus pathogen strains was enhanced by the simulated digestion, with an increase of the inhibition halo.

A rapid, low-cost deep learning system to classify strawberry disease based on cloud service

Accurate and timely classification of diseases during strawberry planting can help growers deal with them in timely manner, thereby reducing losses. However, the classification of strawberry diseases in real planting environments is facing severe challenges, including complex planting environments, multiple disease categories with small differences, and so on. Although recent mobile vision technology based deep learning has achieved some success in overcoming the above problems, a key problem is how to construct a non-destructive, fast and convenient method to improve the efficiency of strawberry disease identification for the multi-region, multi-space and multi-time classification requirements. We develop and evaluate a rapid, low-cost system for classifying diseases in strawberry cultivation. This involves designing an easy-to-use cloudbased strawberry disease identification system, combined with our novel self-supervised multi-network fusion classification model, which consists of a Location network, a Feedback network and a Classification network to identify the categories of common strawberry diseases. With the help of a novel self-supervision mechanism, the model can effectively identify diseased regions of strawberry disease images without the need for annotations such as bounding boxes. Using accuracy, precision, recall and F1 to evaluate the classification effect, the results of the test set are 92.48, 90.68, 86.32 and 88.45%, respectively. Compared with popular Convolutional Neural Networks(CNN) and five other methods, our network achieves better disease classification effect. Currently, the client(mini program) has been released on the We Chat platform. The mini program has perfect classification effect in the actual test, which verifies the feasibility and effectiveness of the system, and can provide a reference for the intelligent research and application of strawberry disease identification.

P1 of strawberry vein banding virus, a multilocalized protein, functions as a movement protein and interacts with the coat protein

Although the complete nucleotide sequence of strawberry vein banding virus(SVBV) has been determined and bioinformatic analysis has revealed that the SVBV genome could encode seven proteins, the precise function of each protein is unclear. This study provided evidence that the P1 protein of SVBV(SVBV-P1) possesses the following features. Bioinformatic and subcellular localization analyses showed that SVBV-P1 is localized in the cytoplasm and cell walls of epidermal cells in Nicotiana benthamiana, and it forms inclusion bodies associated with microtubules and the endoplasmic reticulum. Dilution experiments demonstrated that SVBV-P1 could move from the original agro-infiltrated cells to adjacent cells in N. benthamiana leaves. Further trans-complementation experiments demonstrated that SVBV-P1 could facilitate the intercellular movement of a movement-deficient potato virus X mutant in N. benthamiana leaves. Finally, yeast twohybrid and bimolecular fluorescence complementation assays revealed that SVBV-P1 could interact with the SVBV coat protein, which is a major component of Caulimovirus virions. Results of the electrophoretic mobility shift assay indicated that SVBV-P1 lacks DNA-binding capability. In summary, the results suggest that SVBV-P1 is probably a movement protein of SVBV, providing new insights into the function of movement proteins of the Caulimovirus genus.