Solving the regulation puzzle of periderm development using advances in fruit skin.

Periderm protects enlarged organs of most dicots and gymnosperms as a barrier to water loss and disease invasion during their secondary growth. Its development undergoes a complex process with genetically controlled and environmental stress-induced characters. Different development of periderm makes the full and partial russet of fruit skin, which diverges in inheritance with qualitative and quantitative characters, respectively, in pear pome. In addition to its specific genetics, fruit periderm has similar development and structure as that of stem and other organs, making it an appropriate material for periderm research. Recently, progress in histochemical as well as transcriptome and proteome analyses, and quantitative trait locus (QTL) mapping have revealed the regulatory molecular mechanism in the periderm based on the identification of switch genes. In this review, we concentrate on the periderm development, propose the conservation of periderm regulation between fruit and other plant organs based on their morphological and molecular characteristics, and summarize a regulatory network with the elicitors and repressors for the tissue development. Spontaneous programmed-cell death (PCD) or environmental stress produces the original signal that triggers the development of periderm. Spatio-temporal specific PCD produced by PyPPCD1 gene and its homologs can play a key role in the coordinated regulation of cell death related tissue development.

An ARF1-binding factor triggering programmed cell death and periderm development in pear russet fruit skin.

Plants have a cuticular membrane (CM) and periderm membrane (PM), which act as barriers to terrestrial stresses. The CM covers primary organs with a continuous hydrophobic layer of waxes embedded in cutin, while the PM stacks with suberized cells outermost to the secondary tissues. The formation of native periderm is regulated by a postembryonic meristem phellogen that produces suberized phellem (cork) outwardly. However, the mechanism controlling phellogen differentiation to phellem remains to be clarified. Here, map-based cloning in a pear F1 population with segregation for periderm development in fruit skin facilitated the identification of an aspartic acid repeat deletion in Pyrus Periderm Programmed Cell Death 1.1 (PyPPCD1.1) that triggers phellogen activity for cork formation in pear russet fruit skin. PyPPCD1.1 showed preferential expression in pear fruit skin, and the encoded protein shares a structural similarity to that of the viral capsid proteins. Asp deletion in PyPPCD1.1 weakened its nuclear localization but increased its accumulation in the chloroplast. Both PyPPCD1.1 and its recessive allele directly interact with ADP-ribosylation factor 1 (ARF1). PyPPCD1.1 triggered PCD in an ARF1-dependent manner. Thus, this study identified the switch gene for PCD and periderm development and provided a new molecular regulatory mechanism underlying the development of this trait.

Proteome and transcriptome profile analysis reveals regulatory and stress-responsive networks in the russet fruit skin of sand pear.

The epidermal tissues of the cuticular membrane (CM) and periderm membrane (PM) confer first-line protection from environmental stresses in terrestrial plants. Although PM protection is essentially ubiquitous in plants, the protective mechanism, the function of many transcription factors and enzymes, and the genetic control of metabolic signaling pathways are poorly understood. Different microphenotypes and cellular components in russet (PM-covered) and green (CM-covered) fruit skins of pear were revealed by scanning and transmission electron microscopy. The two types of fruit skins showed distinct phytohormone accumulation, and different transcriptomic and proteomic profiles. The enriched pathways were detected by differentially expressed genes and proteins from the two omics analyses. A detailed analysis of the suberin biosynthesis pathways identified the regulatory signaling network, highlighting the general mechanisms required for periderm formation in russet fruit skin. The regulation of aquaporins at the protein level should play an important role in the specialized functions of russet fruit skin and PM-covered plant tissues.

Genome-Wide Analysis of Sorbitol Dehydrogenase (SDH) Genes and Their Differential Expression in Two Sand Pear (Pyrus pyrifolia) Fruits.

Through RNA-seq of a mixed fruit sample, fourteen expressed sorbitol dehydrogenase (SDH) genes have been identified from sand pear (Pyrus pyrifolia Nakai). Comparative phylogenetic analysis of these PpySDHs with those from other plants supported the closest relationship of sand pear with Chinese white pear (P. bretschneideri). The expression levels varied greatly among members, and the strongest six (PpySDH2, PpySDH4, PpySDH8, PpySDH12, PpySDH13 and PpySDH14) accounted for 96% of total transcript abundance of PpySDHs. Tissue-specific expression of these six members was observed in nine tissues or organs of sand pear, with the greatest abundance found in functional leaf petioles, followed by the flesh of young fruit. Expression patterns of these six PpySDH genes during fruit development were analyzed in two sand pear cultivars, "Cuiguan" and "Cuiyu". Overall, expression of PpySDHs peaked twice, first at the fruitlet stage and again at or near harvest. The transcript abundance of PpySDHs was higher in "Cuiguan" than in "Cuiyu", accompanied by a higher content of sugars and higher ratio of fructose to sorbitol maintained in the former cultivar at harvest. In conclusion, it was suggested that multiple members of the SDH gene family are possibly involved in sand pear fruit development and sugar accumulation and may affect both the sugar amount and sugar composition.

Exploring candidate genes for pericarp russet pigmentation of sand pear (Pyrus pyrifolia) via RNA-Seq data in two genotypes contrasting for pericarp color.

Sand pear (Pyrus pyrifolia) russet pericarp is an important trait affecting both the quality and stress tolerance of fruits. This trait is controlled by a relative complex genetic process, with some fundamental biological questions such as how many and which genes are involved in the process remaining elusive. In this study, we explored differentially expressed genes between the russet- and green-pericarp offspring from the sand pear (Pyrus pyrifolia) cv. 'Qingxiang' × 'Cuiguan' F1 group by RNA-seq-based bulked segregant analysis (BSA). A total of 29,100 unigenes were identified and 206 of which showed significant differences in expression level (log2fold values>1) between the two types of pericarp pools. Gene Ontology (GO) analyses detected 123 unigenes in GO terms related to 'cellular_component' and 'biological_process', suggesting developmental and growth differentiations between the two types. GO categories associated with various aspects of 'lipid metabolic processes', 'transport', 'response to stress', 'oxidation-reduction process' and more were enriched with genes with divergent expressions between the two libraries. Detailed examination of a selected set of these categories revealed repressed expressions of candidate genes for suberin, cutin and wax biosynthesis in the russet pericarps.Genes encoding putative cinnamoyl-CoA reductase (CCR), cinnamyl alcohol dehydrogenase (CAD) and peroxidase (POD) that are involved in the lignin biosynthesis were suggested to be candidates for pigmentation of sand pear russet pericarps. Nine differentially expressed genes were analyzed for their expressions using qRT-PCR and the results were consistent with those obtained from Illumina RNA-sequencing. This study provides a comprehensive molecular biology insight into the sand pear pericarp pigmentation and appearance quality formation.

Pigmentation in sand pear (Pyrus pyrifolia) fruit: biochemical characterization, gene discovery and expression analysis with exocarp pigmentation mutant.

Exocarp color of sand pear is an important trait for the fruit production and has caused our concern for a long time. Our previous study explored the different expression genes between the two genotypes contrasting for exocarp color, which indicated the different suberin, cutin, wax and lignin biosynthesis between the russet- and green-exocarp. In this study, we carried out microscopic observation and Fourier transform infrared spectroscopy analysis to detect the differences of tissue structure and biochemical composition between the russet- and green-exocarp of sand pear. The green exocarp was covered with epidermis and cuticle which was replaced by a cork layer on the surface of russet exocarp, and the chemicals of the russet exocarp were characterized by lignin, cellulose and hemicellulose. We explored differential gene expression between the russet exocarp of 'Niitaka' and its green exocarp mutant cv. 'Suisho' using Illumina RNA-sequencing. A total of 559 unigenes showed different expression between the two types of exocarp, and 123 of them were common to the previous study. The quantitative real time-PCR analysis supports the RNA-seq-derived gene with different expression between the two types of exocarp and revealed the preferential expression of these genes in exocarp than in mesocarp and fruit core. Gene ontology enrichment analysis revealed divorced expression of lipid metabolic process genes, transport genes, stress responsive genes and other biological process genes in the two types of exocarp. Expression changes in lignin metabolism-related genes were consistent with the different pigmentation of russet and green exocarp. Increased transcripts of putative genes involved the suberin, cutin and wax biosynthesis in 'Suisho' exocarp could facilitate deposition of the chemicals and take a role in the mutant trait responsible for the green exocarp. In addition, the divorced expression of ATP-binding cassette transporters involved in the trans-membrane transport of lignin, cutin, and suberin precursors suggests that the transport process could also affect the composition of exocarp and take a role in the regulation of exocarp pigmentation. Results from this study provide a base for the analysis of the molecular mechanism underlying sand pear russet/green exocarp mutation, and presents a comprehensive list of candidate genes that could be used to further investigate the trait mutation at the molecular level.

The genome of the pear (Pyrus bretschneideri Rehd.).

The draft genome of the pear (Pyrus bretschneideri) using a combination of BAC-by-BAC and next-generation sequencing is reported. A 512.0-Mb sequence corresponding to 97.1% of the estimated genome size of this highly heterozygous species is assembled with 194× coverage. High-density genetic maps comprising 2005 SNP markers anchored 75.5% of the sequence to all 17 chromosomes. The pear genome encodes 42,812 protein-coding genes, and of these, ~28.5% encode multiple isoforms. Repetitive sequences of 271.9 Mb in length, accounting for 53.1% of the pear genome, are identified. Simulation of eudicots to the ancestor of Rosaceae has reconstructed nine ancestral chromosomes. Pear and apple diverged from each other ~5.4-21.5 million years ago, and a recent whole-genome duplication (WGD) event must have occurred 30-45 MYA prior to their divergence, but following divergence from strawberry. When compared with the apple genome sequence, size differences between the apple and pear genomes are confirmed mainly due to the presence of repetitive sequences predominantly contributed by transposable elements (TEs), while genic regions are similar in both species. Genes critical for self-incompatibility, lignified stone cells (a unique feature of pear fruit), sorbitol metabolism, and volatile compounds of fruit have also been identified. Multiple candidate SFB genes appear as tandem repeats in the S-locus region of pear; while lignin synthesis-related gene family expansion and highly expressed gene families of HCT, C3'H, and CCOMT contribute to high accumulation of both G-lignin and S-lignin. Moreover, alpha-linolenic acid metabolism is a key pathway for aroma in pear fruit.

[Photosynthetic characteristics of grapevine varieties with different ecological adaptabilities grown in a rain-shelter plastic greenhouse in the Yangtze River Delta region].

The study with two grapevine varieties Manicure Finger and Fujiminori grown in a rain-shelter plastic greenhouse in the Yangtze River Delta region showed that in the greenhouse, the photon flux density (PFD) decreased greatly, while air temperature (Ta), vapor pressure (Vp) and transpiration rate (Tr) had little changes, no matter in sunny or cloudy days. The leaf thickness and fresh weight per cm2 of the two varieties decreased by 28.6% and 18.4%, and 13.1% and 11.6%, respectively, total chlorophyll content and apparent quantum yield (AQY) had a slight increase, but chlorophyll a and b contents and chlorophyll a/b ratio had no obvious difference with the open-air cultivation (CK). In sunny days, the diurnal mean value of net photosynthetic rate Pn, diurnal integral value of Pn, and water use efficiency (WUE) of Manicure Finger and Fujiminori decreased by 18.7% and 13.1%, 17.2% and 11.9%, and 26.0% and 36.9%, but stomatal conductance (Gs) increased by 76.8% and 100.8%, respectively, while in cloudy days, the corresponding values of above-mentioned parameters were decreased by 48.7% and 33.5%, 34.0% and 29.8%, and 69.2% and 67.7%, and increased by 178.0% and 85.1%, respectively. The variation extents of the parameters were larger for Manicure Finger than for Fujiminori, especially in cloudy days, indicating the great differences of ecological adaptability between these two grapevine varieties.