Susceptibility of Flordaguard peach rootstock to a resistant-breaking population of Meloidogyne floridensis and two populations of Meloidogyne arenaria.

Cultivar Flordaguard is suggested as a root-knot nematode (RKN) resistant rootstock for Florida peaches, however, RKN disease has been observed on this rootstock in peach orchards. Our goal was to confirm whether the RKN resistance breaking isolates of M. floridensis and M. arenaria indeed could infect and reproduce on the peach rootstock cv. Flordaguard in both laboratory and field studies. Root galling occurred on all peach cultivars evaluated including Flordaguard, Flordaglo, Okinawa, and Lovell, in the presence of the RKN resistance-breaking isolates of M. floridensis (MfGnv14) and two M. arenaria isolates (Ma1 and Ma2). These rootstocks showed varying degrees of susceptibility (to a lesser extent in Okinawa) to these three RKN resistance-breaking isolates. The importance of nematode inoculum concentrations in differentiating between resistance and susceptible plants was demonstrated, and thus are an important factor to consider in nematode resistance breeding programs. In host differential tests the peach-originated isolates of M. floridensis and M. arenaria behaved similarly with the vegetable-originated isolates of M. floridensis on tomato, peanut, watermelon, and tobacco, but showed variable host responses on cotton and pepper. The two M. arenaria isolates from peach reproduced on pepper but not on peanut. To our knowledge this is the first report of M. arenaria race 3 infecting Flordaguard and pepper in Florida. Soil and root samples collected from cv. Flordaguard infected trees at two commercial peach orchards showed that M. floridensis and M. arenaria were established on the rootstock.

Responses of 'Flordaguard' and 'MP-29' Prunus spp. rootstocks to hypoxia and high root zone temperature.

Selecting the right rootstock is crucial for successful fruit production and managing both biotic and abiotic stresses in commercial fruit orchards. To enhance the resilience of peach orchards, this study evaluated the physiological and biochemical responses of Prunus spp. rootstocks , 'Flordaguard' and 'MP-29,' under normoxia (sufficient oxygen content) or short-term hypoxia (low-oxygen content) and ambient or high temperature (40°C) in the root zone. Physiological responses measured were net photosynthesis, stomatal conductance, transpiration, intercellular CO2 concentration, water use efficiency, the leaf chlorophyll index, and the maximum potential quantum efficiency of photosystem II. The leaf concentrations of nitrogen, phosphorus, potassium, magnesium, calcium, sulfur, boron, zinc, manganese, iron, and copper were also analyzed. Reactive oxygen species (ROS) and antioxidants analyzed were superoxide dismutase (SOD) activity, peroxidase (POD) activity, catalase (CAT) activity, ascorbate peroxidase (APX) activity, glutathione peroxidase (GPX) activity, proline content, glycine betaine content (GB), lipid peroxidation (LPO), superoxide (O2 -) concentration, and hydrogen peroxide (H2O2) concentration. When subjected to root zone hypoxia or high temperature individually, 'MP-29' performed better physiologically than 'Flordaguard'. However, when root zone hypoxia and high temperature were combined, 'MP-29' performed better biochemically with enhanced antioxidant activity, osmolyte content, and nutrient absorption. Nutrient analysis of leaves revealed that 'MP-29' had higher N, P, K, Ca, and B concentrations than 'Flordaguard'. Consequently, 'MP-29' demonstrated greater tolerance to short-term exposure to the combined effects of high root zone temperature and hypoxia. This research contributes to identifying a suitable rootstock within the Prunus genus able to withstand root zone conditions that often result from severe weather events commonly experienced in Florida and other parts of the world.

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.

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.

Engineering a Plant-Derived Astaxanthin Synthetic Pathway Into Nicotiana benthamiana.

Carotenoids have been shown to be essential for human nutrition. Consumption of carotenoid-rich fruits and vegetables can reduce the risk of many diseases. The ketocarotenoid astaxanthin has become a commercially valuable compound due to its powerful antioxidant properties compared to other carotenoids. It is naturally produced in certain algae, bacteria, and the flowers of some species of the genus Adonis, although it is produced in such small quantities in these organisms that it is costly to extract. Chemical synthesis of this compound has also shown limited success with a high proportion of esterified forms of astaxanthin being produced, which decreases antioxidant properties by the conversion of hydroxyl groups to esters. Previously, transgenic astaxanthin-producing plants have been created using a ß-carotene ketolase enzyme of either bacterial or algal origin. However, a novel astaxanthin pathway exists in the flowering plants of the genus Adonis which has not been utilized in the same manner. The pathway involves two unique enzymes, ß-ring-4-dehydrogenase and 4-hydroxy-ß-ring-4-dehydrogenase, which add the necessary hydroxyl and ketone groups to the rings of ß-carotene. In the present study, Nicotiana benthamiana plants were transformed with chimeric constructs coding for these two enzymes. The regenerated, transgenic plants accumulate astaxanthin and their growth (height and weight) was unaffected, when compared to non-transformed N. benthamiana and to plants transformed with the bacterial ß-carotene ketolase. The accumulation of astaxanthin also improved seedling survivability under harsh UV light, mitigated reactive oxygen accumulation, and provided a phenotype (color) that allowed the efficient identification and recovery of transgenic plants with and without selection.

Influence of rootstocks on fruit physical and chemical properties of peach cv. UFSun.

The subtropical peach cultivar UFSun grafted on five different rootstocks ('Flordaguard', 'Barton', 'MP-29', 'P-22', and 'Okinawa') was investigated in terms of the pomological and biochemical parameters of the fruit. Significant differences in fruit weight and size, soluble solids content, titratable acidity, and firmness were found among some rootstocks. The fruit length and diameter were different between the 'MP-29' and other rootstocks. It was also found that firmness of 'UFSun' fruit was affected by the rootstock. The highest firmness value was found when 'UFSun' was grafted on 'Flordaguard'.' 'MP-29' fruit had the highest soluble solids content, but there were no differences among the other rootstocks. 'UFSun' fruit from trees grafted on 'MP-29' were smaller and had the most intense color compared to 'UFSun' fruit from other rootstocks. Rootstock had a significant influence on total fruit phenolic compounds, anthocyanin content, and total antioxidant activity, with fruit from 'UFSun' on 'MP-29' having the highest values in all of them. A high correlation between fruit total antioxidant activity and total phenolic content was found. Overall, the results showed that 'MP-29' seems to induce the highest fruit quality, showing higher contents of total soluble solids, total titratable acidity, total phenolic compounds, total antioxidant activity, and total anthocyanin content. Selecting the right combination of the rootstock and cultivar is important for optimizing fruit quality parameters.

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.

Assessment of unconventional antimicrobial compounds for the control of 'Candidatus Liberibacter asiaticus', the causative agent of citrus greening disease.

In this study, newly identified small molecules were examined for efficacy against 'Candidatus Liberibacter asiaticus' in commercial groves of sweet orange (Citrus sinensis) and white grapefruit (Citrus paradisi) trees. We used benzbromarone and/or tolfenamic acid delivered by trunk injection. We evaluated safety and efficacy parameters by performing RNAseq of the citrus host responses, 16S rRNA gene sequencing to characterize citrus-associated microbial communities during treatment, and qRT-PCR as an indirect determination of 'Ca. L. asiaticus' viability. Analyses of the C. sinensis transcriptome indicated that each treatment consistently induced genes associated with normal metabolism and growth, without compromising tree viability or negatively affecting the indigenous citrus-associated microbiota. It was found that treatment-associated reduction in 'Ca. L. asiaticus' was positively correlated with the proliferation of several core taxa related with citrus health. No symptoms of phytotoxicity were observed in any of the treated trees. Trials were also performed in commercial groves to examine the effect of each treatment on fruit productivity, juice quality and efficacy against 'Ca. L. asiaticus'. Increased fruit production (15%) was observed in C. paradisi following twelve months of treatment with benzbromarone and tolfenamic acid. These results were positively correlated with decreased 'Ca. L. asiaticus' transcriptional activity in root samples.

Isolation of nucleotide binding site-leucine rich repeat and kinase resistance gene analogues from sugarcane (Saccharum spp.).

BACKGROUND: Resistance gene analogues (RGAs) have been isolated from many crops and offer potential in breeding for disease resistance through marker-assisted selection, either as closely linked or as perfect markers. Many R-gene sequences contain kinase domains, and indeed kinase genes have been reported as being proximal to R-genes, making kinase analogues an additionally promising target. The first step towards utilizing RGAs as markers for disease resistance is isolation and characterization of the sequences. RESULTS: Sugarcane clone US01-1158 was identified as resistant to yellow leaf caused by the sugarcane yellow leaf virus (SCYLV) and moderately resistant to rust caused by Puccinia melanocephala Sydow & Sydow. Degenerate primers that had previously proved useful for isolating RGAs and kinase analogues in wheat and soybean were used to amplify DNA from sugarcane (Saccharum spp.) clone US-01-1158. Sequences generated from 1512 positive clones were assembled into 134 contigs of between two and 105 sequences. Comparison of the contig consensuses with the NCBI sequence database using BLASTx showed that 20 had sequence homology to nuclear binding site and leucine rich repeat (NBS-LRR) RGAs, and eight to kinase genes. Alignment of the deduced amino acid sequences with similar sequences from the NCBI database allowed the identification of several conserved domains. The alignment and resulting phenetic tree showed that many of the sequences had greater similarity to sequences from other species than to one another. CONCLUSION: The use of degenerate primers is a useful method for isolating novel sugarcane RGA and kinase gene analogues. Further studies are needed to evaluate the role of these genes in disease resistance.

Putative protease inhibitor gene discovery and transcript profiling during fruit development and leaf damage in grapefruit (Citrus paradisi Macf.).

Seven putative protease inhibitor (PPI) cDNAs, representing four protein families, were isolated from a grapefruit (Citrus paradisi Macf. Cv. Marsh) immature fruit flavedo cDNA library. Cloned open reading frames encoded proteins with similarity to, and protein signatures for: legume Kuntiz inhibitors (lkiL-1, lkiL-2, lkiL-3), potato trypsin inhibitor I (ptiIL-1), serpins (serpL-1), cystatins (cystL-1), and gamma thionins (gthL-1). Response of transcript abundance to fruit development and leaf wounding was determined for all but lkiL-1 using real-time RT-PCR. Immature leaves had the highest transcript levels for all PPIs. The gthL-1 transcript in immature leaves was the most abundant transcript but was absent from healthy mature leaves. In fruit flavedo, transcripts for all PPIs were most abundant in youngest fruit (<15 mm dia. fruit), and declined during development, but displayed different patterns of developmental change. Mechanical or Diaprepes root weevil (DRW) feeding damage to leaves caused a <10-fold reduction or had no effect on transcript level with the exception of gthL-1 which, as a result of damage, increased >50-fold in mature leaves and decreased >1400-fold in immature leaves. This developmental control of transcript response to wounding in a woody perennial is opposite of what has been observed for defensive proteinase inhibitors (PIs) in other plants (typically herbaceous and/or annual plants), where younger leaves typically invoke a higher defensive proteinase inhibitor transcript accumulation than older tissues. Except for gthL-1, the PPI transcripts were minimally responsive or unresponsive to wounding. Changes in PPI transcript levels suggest diverse roles for the products of these genes in citrus, with only gthL-1 responding in a defense-like manner.