Surface moisture induces microcracking in the cuticle of fruit skins. Our objective was to study the effects of surface moisture on cuticular microcracking, the permeance to water vapour and russeting in developing 'Pinova' apple fruit. • Surface moisture was applied by fixing to the fruit a plastic tube containing deionized water. Microcracking was quantified by fluorescence microscopy and image analysis following infiltration with acridine orange. Water vapour permeance was determined gravimetrically using skin segments (ES) mounted in diffusion cells. • Cumulative water loss through the ES increased linearly with time. Throughout development, surface moisture significantly increased skin permeance. The effect was largest during early development and decreased towards maturity. Recovery time courses revealed that following moisture treatment of young fruit for 12 days, skin permeance continued to increase until about 14 days after terminating the moisture treatment. Thereafter, skin permeance decreased over the next 28 days, then approaching the control level. This behaviour indicates gradual healing of the impaired cuticular barrier. Nevertheless, permeance still remained significantly higher compared with the untreated control. Similar patterns of permeance change were observed following moisture treatments at later stages of development. The early moisture treatment beginning at 23 DAFB resulted in russeting of the exposed surfaces. There was no russet in control fruit without a tube or in control fruit with a tube mounted for 12 days without water. • The data demonstrate that surface moisture increases microcracking and water vapour permeance. This may lead to the formation of a periderm and, hence, a russeted fruit surface.

Exposure of the fruit surface to moisture during early development is causal in russeting of apple (Malus × domestica Borkh.). Moisture exposure results in formation of microcracks and decreased cuticle thickness. Periderm differentiation begins in the hypodermis, but only after discontinuation of moisture exposure. Expressions of selected genes involved in cutin, wax and suberin synthesis were quantified, as were the wax, cutin and suberin compositions. Experiments were conducted in two phases. In Phase I (31 days after full bloom) the fruit surface was exposed to moisture for 6 or 12 d. Phase II was after moisture exposure had been discontinued. Unexposed areas on the same fruit served as unexposed controls. During Phase I, cutin and wax synthesis genes were down-regulated only in the moisture-exposed patches. During Phase II, suberin synthesis genes were up-regulated only in the moisture-exposed patches. The expressions of cutin and wax genes in the moisture-exposed patches increased slightly during Phase II, but the levels of expression were much lower than in the control patches. Amounts and compositions of cutin, wax and suberin were consistent with the gene expressions. Thus, moisture-induced russet is a two-step process: moisture exposure reduces cutin and wax synthesis, moisture removal triggers suberin synthesis.

Russeting (periderm formation) is a critical fruit-surface disorder in apple (Malus × domestica Borkh.). The first symptom of insipient russeting is cuticular microcracking. Humid and rainy weather increases russeting. The aim was to determine the ontogeny of moisture-induced russeting in ‘Pinova’ apple. We recorded the effects of duration of exposure to water and the stage of fruit development at exposure on microcracking, periderm formation and cuticle deposition. Early on (21 or 31 days after full bloom; DAFB) short periods (2 to 12 d) of moisture exposure induced cuticular microcracking—but not later on (66 or 93 DAFB). A periderm was not formed during moisture exposure but 4 d after exposure ended. A periderm was formed in the hypodermis beneath a microcrack. Russeting frequency and severity were low for up to 4 d of moisture exposure but increased after 6 d. Cuticle thickness was not affected by moisture for up to 8 d but decreased for longer exposures. Cuticular ridge thickness decreased around a microcrack. In general, moisture did not affect cuticular strain release. We conclude that a hypodermal periderm forms after termination of moisture exposure and after microcrack formation. Reduced cuticle deposition may cause moisture-induced microcracking and, thus, russeting.

Black spot disease, which is caused by the ascomycete Diplocarpon rosae, is the most severe disease in field-grown roses in temperate regions and has been distributed worldwide, probably together with commercial cultivars. Here, we present data indicating that muRdr1A is the active Rdr1 gene, a single-dominant TIR-NBS-LRR (Toll/interleukin-1 receptor-nucleotide binding site-leucine rich repeat) (TNL)-type resistance gene against black spot disease, which acts against a broad range of pathogenic isolates independent of the genetic background of the host genotype. Molecular analyses revealed that, compared with the original donor genotype, the multiple integrations that are found in the primary transgenic clone segregate into different integration patterns in its sexual progeny and do not show any sign of overexpression. Rdr1 provides resistance to 13 different single-spore isolates belonging to six different races and broad field mixtures of conidia; thus far, Rdr1 is only overcome by two races. The expression of muRdr1A, the active Rdr1 gene, leads to interaction patterns that are identical in the transgenic clones and the non-transgenic original donor genotype. This finding indicates that the interacting avirulence (Avr) factor on the pathogen side must be widespread among the pathogen populations and may have a central function in the rose-black spot interaction. Therefore, the Rdr1 gene, pyramided with only a few other R genes by sexual crosses, might be useful for breeding roses that are resistant to black spot because the spread of new pathogenic races of the fungus appears to be slow.

Russeting is a cosmetic defect of some fruit skins. Russeting (botanically: induction of periderm formation) can result from various environmental factors including wounding and surface moisture. The objective was to compare periderms resulting from wounding with those from exposure to moisture in developing apple fruit. Wounding or moisture exposure both resulted in cuticular microcracking. Cross-sections revealed suberized hypodermal cell walls by 4 d, and the start of periderm formation by 8 d after wounding or moisture treatment. The expression of selected target genes was similar in wound and moisture induced periderms. Transcription factors involved in the regulation of suberin (MYB93) and lignin (MYB42) synthesis, genes involved in the synthesis (CYP86B1) and the transport (ABCG20) of suberin monomers and two uncharacterized transcription factors (NAC038 and NAC058) were all upregulated in induced periderm samples. Genes involved in cutin (GPAT6, SHN3) and wax synthesis (KCS10, WSD1, CER6) and transport of cutin monomers and wax components (ABCG11) were all downregulated. Levels of typical suberin monomers (ω-hydroxy-C20, -C22 and -C24 acids) and total suberin were high in the periderms, but low in the cuticle. Periderms were induced only when wounding occurred during early fruit development (32 and 66 days after full bloom (DAFB)) but not later (93 DAFB). Wound and moisture induced periderms are very similar morphologically, histologically, compositionally and molecularly.

In plants, inosine is enzymatically introduced in some tRNAs, but not in other RNAs or DNA. Nonetheless, our data show that RNA and DNA from Arabidopsis thaliana contain (deoxy)inosine, probably derived from nonenzymatic adenosine deamination in nucleic acids and usage of (deoxy)inosine triphosphate (dITP and ITP) during nucleic acid synthesis.We combined biochemical approaches, LC-MS, as well as RNA-Seq to characterize a plant INOSINE TRIPHOSPHATE PYROPHOSPHATASE (ITPA) from A. thaliana, which is conserved in many organisms, and investigated the sources of deaminated purine nucleotides in plants.Inosine triphosphate pyrophosphatase dephosphorylates deaminated nucleoside di-and triphosphates to the respective monophosphates. ITPA loss-of-function causes inosine di-and triphosphate accumulation in vivo and an elevated inosine and deoxyinosine content in RNA and DNA, respectively, as well as salicylic acid (SA) accumulation, early senescence, and upregulation of transcripts associated with immunity and senescence. Cadmium-induced oxidative stress and biochemical inhibition of the INOSINE MONOPHOSPHATE DEHYDRO-GENASE leads to more IDP and ITP in the wild-type (WT), and this effect is enhanced in itpa mutants, suggesting that ITP originates from ATP deamination and IMP phosphorylation.Inosine triphosphate pyrophosphatase is part of a molecular protection system in plants, preventing the accumulation of (d)ITP and its usage for nucleic acid synthesis.

Inosine is deaminated adenosine. Inosine is enzymatically introduced in some plant tRNAs but not in other RNAs or DNA. Nonetheless, our data show that RNA and DNA from Arabidopsis thaliana contain (deoxy)inosine, probably derived from non-enzymatic adenosine deamination in nucleic acids and usage of (deoxy)inosine triphosphate (ITP / dITP) during nucleic acid synthesis. We identified a plant INOSINE TRIPHOSPHATE PYROPHOSPHATASE (ITPA) which is conserved in many organisms. Arabidopsis ITPA dephosphorylates deaminated nucleoside di- and triphosphates to their respective monophosphates. ITPA mutation causes inosine di- and triphosphate accumulation in vivo and an elevated (deoxy)inosine content in DNA and RNA. Cadmium-induced oxidative stress, known to foster deamination, leads to more ITP in the wildtype and especially in itpa mutants suggesting that ITP originates from ATP deamination. By contrast, erroneous IMP phosphorylation by AMP kinases seems not to be a major ITP source in vivo although these enzymes phosphorylate IMP in vitro. Mutation of ITPA causes salicylic acid (SA) accumulation, early senescence and upregulation of transcripts associated with immunity and senescence. ITPA is part of a molecular protection system, preventing accumulation of (d)ITP, its usage for nucleic acid synthesis, and probably nucleic acid stress leading to SA accumulation, stress gene induction and early senescence.