First Report of Lasiodiplodia pseudotheobromae causing Postharvest Decay of Strawberries in Florida.

Postharvest decay of strawberry (Fragaria × ananassa Duch.) is a major factor causing fruit losses. Strawberries were obtained from various harvests at cooling facilities located in Dover and Plant City, FL during the 2018-19 and 2019-20 seasons. After the fruits were incubated at 22ºC for up to 5 days (d) to promote disease development, Lasiodiplodia decay was observed at up to 3% from some harvests, exhibiting gray mycelia on small lesions that gradually covered the whole fruit. The fungus was isolated onto potato dextrose agar (PDA). Five isolates (SBD18-14, SBD18-277, SBD18-279, SBD19-02 and SBD19-57) were characterized. Fungal mycelia were initially grayish white and then gradually changed to gray to dark gray on PDA at 25oC, and later produced black pigments (Fig. S1). Pycnidia were observed from inoculated strawberries at 14 d. Isolates shared similar conidia morphology: aseptate, hyaline, ellipsoid to ovoid, measuring L × W: 24.0-34.0 (28.3) × 13.0-16.0 (14.3) µm (n =100). Mature conidia were brown, one septate, measuring L × W: 25.0-33.0 (28.8) × 13.0-16.0 (14.5) µm (n =100). The isolates were identified as Lasiodiplodia spp. morphologically (Alves et al. 2008). DNA was extracted from fungal mycelia using an OmniPrep DNA extraction kit, and PCR amplification of ITS and EF1-α genes was performed following the conditions described by White et al. (1990) with some modifications using primers ITS1F-F/ITS4-R (Gardes and Bruns, 1993; White et al., 1990) and EF1-668-F/EF1-1251-R (Alves et al., 2008), respectively. The BLASTn in GenBank showed that the sequences obtained had 99.61 to 100% homology with those of ITS (EF622077) and EF1-α (EF622057) from L. pseudotheobromae CBS116459 (an ex-type strain) (Alves et al., 2008). Sequences of the isolates have been deposited in GenBank with accessions OP326017 to OP326021 for ITS, and OP356202 to OP356206 for EF1-α. Phylogenetic analysis showed that these isolates clustered in the same clade (bootstrap value at 64) with L. pseudotheobromae (Fig. S2). Two fungal inoculum types (mycelia and conidia), two fruit inoculation methods (injury and non-injury) and five fungal isolates were used for pathogenicity tests. Fungal mycelia (2-day-old) on PDA plug (5 mm) or 10 µL of conidial suspension (106 spores/mL) was placed onto each injury (1 x 1 mm in size) or a non-injury area on the surfaces of five strawberry fruits (cv. Florida Brilliance). PDA plug alone or water drops placed on injury or non-injury areas on fruits served as respective controls. Inoculated and control fruits were incubated in a covered plastic container with 100% RH at 22ºC. The experiment was repeated twice. Decay initially appeared as soft and lightly discolored tissue at inoculation areas 2 d post-inoculation (dpi) that extended quickly thereafter. Brown to dark lesions on both injury- and non-injury fruits inoculated with conidia or mycelia were observed at 3 dpi. Decay and gray mycelia gradually developed over the whole fruit at 6 dpi, and pycnidia were observed after 14 dpi (Fig. S1). Disease incidence of 100% was observed on all tests. Control fruits did not develop decay. The results indicate that these isolates are pathogenic to strawberries and infect fruit via both non-injured and injured fruit surfaces. The inoculated fungal isolates were re-isolated, thus, fulfilling Koch's postulates. L. theobromae, Neofusicoccum parvum/N. ribis species complex causing strawberry fruit rot in Florida fields was reported (Oliveira et al., 2019), but not L. pseudotheobromae. To our knowledge, this is the first report of postharvest decay caused by L. pseudotheobromae A.J.L. Phillips, A. Alves & Crous on strawberries in Florida and in the USA, and it should be considered in strawberry disease management.

Application of Thymol Vapors to Control Postharvest Decay Caused by Penicillium digitatum and Lasiodiplodia theobromae in Grapefruit.

Two of the major postharvest diseases impacting grapefruit shelf life and marketability in the state of Florida (USA) are stem-end rot (SER) caused by Lasiodiplodia theobromae and green mold (GM) caused by Penicillium digitatum. Here, we investigated the in vitro and in vivo efficacy of vapors of thymol, a natural compound found in the essential oil of various plants and the primary constituent of thyme (Thymus vulgaris) oil, as a potential solution for the management of GM and SER. Thymol vapors at concentrations lower than 10 mg L-1 significantly inhibited the mycelial growth of both pathogens, causing severe ultrastructural damage to P. digitatum conidia. In in vivo trials, the incidence and lesion area of GM and SER on inoculated grapefruit were significantly reduced after a 5 d exposure to 50 mg L-1 thymol vapors. In addition, the in vitro and in vivo sporulation of P. digitatum was suppressed by thymol. When applied in its vapor phase, thymol had no negative effect on the fruit, neither introducing perceivable off-flavor nor causing additional weight loss. Our findings support the pursuit of further studies on the use of thymol, recognized as safe for human health and the environment, as a promising strategy for grapefruit postharvest disease management.

Variation in the Root System Architecture of Peach × (Peach × Almond) Backcrosses.

The spatial arrangement and growth pattern of root systems, defined by the root system architecture (RSA), influences plant productivity and adaptation to soil environments, playing an important role in sustainable horticulture. Florida's peach production area covers contrasting soil types, making it necessary to identify rootstocks that exhibit soil-type-specific advantageous root traits. In this sense, the wide genetic diversity of the Prunus genus allows the breeding of rootstock genotypes with contrasting root traits. The evaluation of root traits expressed in young seedlings and plantlets facilitates the early selection of desirable phenotypes in rootstock breeding. Plantlets from three peach × (peach × almond) backcross populations were vegetatively propagated and grown in rhizoboxes. These backcross populations were identified as BC1251, BC1256, and BC1260 and studied in a completely randomized design. Scanned images of the entire root systems of the plantlets were analyzed for total root length distribution by diameter classes, root dry weight by depth horizons, root morphological components, structural root parameters, and root spreading angles. The BC1260 progeny presented a shallower root system and lower root growth. Backcross BC1251 progeny exhibited a more vigorous and deeper root system at narrower root angles, potentially allowing it to explore and exploit water and nutrients in deep sandy entisols from the Florida central ridge.

Modeling the relationship between air temperature and grapefruit quality traits.

BACKGROUND: Consumers of grapefruit require consistent fruit quality with a good physical appearance and taste. The air temperature during the growing season affects both the external (external color index (ECI)) and internal (titratable acidity (TA) and total soluble solids ratio (TSS/TA)) fruit quality of grapefruit. The objective of this study was to develop computer models that encompass the relationship between preharvest air temperature and fruit quality to predict fruit quality of grapefruit at harvest. RESULTS: There was a logarithmic relationship between the number of days with a daily minimum air temperature ≤13 °C and ECI, with a greater number of days resulting in higher ECI. In addition, there was a second-order polynomial relationship between the number of hours ≥21 °C and both TA and TSS/TA, with a greater number of hours resulting in lower TA and higher TSS/TA. Model performance for predicting the ECI, TA, and TSS/TA during 2004-05 and 2005-06 growing seasons was good, with Nash and Sutcliffe coefficient of efficiency (NSE) values for each season of 0.835 and 0.917 respectively for ECI, 0.896 and 0.965 respectively for TA and 0.898 and 0.966 respectively for TSS/TA. Applying the model to statistical survey data covering 13 growing seasons demonstrated that the TSS/TA model was robust. CONCLUSION: Statistical models were developed that predicted the development of grapefruit ECI, TA, and TSS/TA. The TSS/TA model was confirmed after application to long-term statistical survey data covering 13 growing seasons. © 2022 Society of Chemical Industry.

Comparison between two cultivars of longan fruit cv. 'Dongbi' and 'Fuyan' in the metabolisms of lipid and energy and its relation to pulp breakdown.

This work studied the difference in pulp breakdown between two cultivars of longan cv. 'Dongbi' and 'Fuyan' from the aspect of metabolisms of lipid and energy. The results reflected that, compared to 'Fuyan' longan, 'Dongbi' longan had higher levels of energy charge, U/S and IUFA, and higher amounts of USFA, PC, PI, ATP and ADP. Moreover, 'Dongbi' longan exhibited lower levels of SFA, PA, AMP and cell membrane permeability. Also, lower PLD, LOX and lipase activities, but higher ATPase activity, lower pulp breakdown index, and better pulp appearance were exhibited in 'Dongbi' longan. These data revealed that the mitigated pulp breakdown in 'Dongbi' longan was due to the comprehensive coordination of metabolisms in lipid and energy through maintaining a higher level of energy, a higher unsaturation degree of fatty acids, delaying the degradation of phospholipids, and better retaining the membrane structural integrity of microsome and entire cell.

Citrus disease detection using convolution neural network generated features and Softmax classifier on hyperspectral image data.

Identification and segregation of citrus fruit with diseases and peel blemishes are required to preserve market value. Previously developed machine vision approaches could only distinguish cankerous from non-cankerous citrus, while this research focused on detecting eight different peel conditions on citrus fruit using hyperspectral (HSI) imagery and an AI-based classification algorithm. The objectives of this paper were: (i) selecting the five most discriminating bands among 92 using PCA, (ii) training and testing a custom convolution neural network (CNN) model for classification with the selected bands, and (iii) comparing the CNN's performance using 5 PCA bands compared to five randomly selected bands. A hyperspectral imaging system from earlier work was used to acquire reflectance images in the spectral region from 450 to 930 nm (92 spectral bands). Ruby Red grapefruits with normal, cankerous, and 5 other common peel diseases including greasy spot, insect damage, melanose, scab, and wind scar were tested. A novel CNN based on the VGG-16 architecture was developed for feature extraction, and SoftMax for classification. The PCA-based bands were found to be 666.15, 697.54, 702.77, 849.24 and 917.25 nm, which resulted in an average accuracy, sensitivity, and specificity of 99.84%, 99.84% and 99.98% respectively. However, 10 trials of five randomly selected bands resulted in only a slightly lower performance, with accuracy, sensitivity, and specificity of 98.87%, 98.43% and 99.88%, respectively. These results demonstrate that an AI-based algorithm can successfully classify eight different peel conditions. The findings reported herein can be used as a precursor to develop a machine vision-based, real-time peel condition classification system for citrus processing.

Rhizoboxes as Rapid Tools for the Study of Root Systems of Prunus Seedlings.

Rootstocks are fundamental for peach production, and their architectural root traits determine their performance. Root-system architecture (RSA) analysis is one of the key factors involved in rootstock selection. However, there are few RSA studies on Prunus spp., mostly due to the tedious and time-consuming labor of measuring below-ground roots. A root-phenotyping experiment was developed to analyze the RSA of seedlings from 'Okinawa' and 'Guardian'™ peach rootstocks. The seedlings were established in rhizoboxes and their root systems scanned and architecturally analyzed. The root-system depth:width ratio (D:W) throughout the experiment, as well as the root morphological parameters, the depth rooting parameters, and the root angular spread were estimated. The 'Okinawa' exhibited greater root morphological traits, as well as the other parameters, confirming the relevance of the spatial disposition and growth pattern of the root system.

Effect of steam blanching on peelability and quality of Citrus reticulata Blanco.

Peeling of citrus fruits is the key procedure in the production of citrus products such as canned citrus and citrus vinegar. In order to facilitate this peeling process, Citrus reticulata Blanco is usually heated. The treatments of steam blanching and air cooling were carried out in the citrus peeling experiment. The anatomical details of citrus peel were also observed after steam blanching. The results indicated that the duration of steam blanching had a significant influence on peelability of Citrus reticulata Blanco (P < 0.01). With the proceeding of steam blanching, the maximum stripping force and the maximum stripping length of the citrus peel decreased. Two minutes after steam blanching, parenchyma cell structure and vascular elements were destroyed. The duration of cooling treatment has no significant impact on the peelability of the citrus (P > 0.05). The content of titratable acid, total soluble solid and vitamin C were considerably influenced by steam blanching. When steam blanching duration was within 2 min, the color change ∆Eij was less than 3.0, indicating no significant color change. This study determined 2 min to be the optimal steam blanching duration for achieving the best peelability and quality of peeled citrus, and 5 min or less to be optimal cooling duration. Overall, this study provides a theoretical guideline for the application of the steam blanching treatment in the citrus production.

Effect of Propagation Systems and Indole-3-Butyric Acid Potassium Salt (K-IBA) Concentrations on the Propagation of Peach Rootstocks by Stem Cuttings.

Traditionally, peach rootstocks are propagated by seeds due to their high availability, low cost, and easy storage and handling. However, stem cuttings allow the propagation of interspecific hybrids and keep the genetic uniformity of heterozygous genotypes. This study compared the effect of four different concentrations of K-IBA (indole-3-butyric acid potassium salt) on softwood cuttings of three peach backcrosses (peach × (peach × almond)) for rootstock propagation in two propagation systems: aeroponics and germination trays. The four concentrations of K-IBA applied were: 0.0% (w/v) as a control, 0.1% (w/v), 0.2% (w/v), and 0.4% (w/v). Data were collected on the survival rate (%), rooting rate (%), and root growth parameters. The relevance of auxin for peach cuttings rooting was evidenced. K-IBA at 0.2% showed the best rooting effect for peach softwood cuttings, evidenced by its high rooting rate and higher survival rate. K-IBA at 0.4% and 0.2% produced the highest number of adventitious roots. The highest root growth parameters were obtained in germination trays, confirming the suitability of this system for root growth. However, aeroponics was demonstrated to be as efficient as the traditional germination trays for the rooting of peach cuttings, allowing for a more controlled environment with a better use of resources.

The Potential of Gaseous Chlorine Dioxide for the Control of Citrus Postharvest Stem-End Rot Caused by Lasiodiplodia theobromae.

The focus of this study was to develop technologies using chlorine dioxide (ClO2) gas to control postharvest stem-end rot of citrus caused by Lasiodiplodia theobromae. Mycelial growth of L. theobromae on potato dextrose agar (PDA) plugs was completely inhibited by a 24-h ClO2 exposure provided by 0.5 g of solid ClO2 generating granular mixture in a 7.7-liter sealed container. In vivo experiments were conducted on artificially inoculated Tango and naturally infected U.S. Early Pride mandarins. When ClO2 treatments were initiated 0 to 6 h after inoculation, decay development was significantly reduced as compared with the control, and higher ClO2 doses were more effective. A ClO2 treatment (using 3 g of generating mixture per 7.7-liter sealed container) administered 0 h after inoculation resulted in 17.6% Diplodia stem-end rot incidence compared with 95.6% in the control, whereas the same treatment administered 24 h after inoculation was much less effective, resulting in 63.0% incidence compared with 85.4% in the control. Diplodia stem-end rot incidence of naturally infected fruit after using 6 or 9 g of generating mixture per 24-liter sealed box was 23.8 or 25.7%, respectively, compared with 47.9% for control fruit. The ClO2 treatments had no negative effects on fruit quality characteristics including weight loss, firmness, puncture resistance, titratable acids (TAs), total soluble solids (TSSs), and rind color. Albedo pH at wounds was significantly reduced from 6.0 to 4.8 by the ClO2 treatments, whereas undamaged albedo remained at 5.8. In addition, no visible physiologic defects, such as peel browning and bleaching, were observed on ClO2-treated fruit. These results indicate that ClO2 gas has the potential to be developed as a component of an integrated citrus postharvest decay control system to minimize fruit losses.