Genome-wide characterization and expression of the TLP gene family associated with Colletotrichum gloeosporioides inoculation in Fragaria × ananassa.

Background: Colletotrichum gloeosporioides, a soil-borne fungal pathogen, causes significant yield losses in many plants, including cultivated strawberry (Fragaria × ananassa, 2n = 8x = 56). Thaumatin-like proteins (TLPs) are a large and complex family of proteins that play a vital role in plant host defense and other physiological processes. Methods: To enhance our understanding of the antifungal activity of F. × ananassa TLPs (FaTLP), we investigated the genome-wide identification of FaTLP gene families and their expression patterns in F. × ananassa plants upon pathogen infection. Moreover, we used RNA sequencing (RNA-seq) to detect the differences in the expression patterns of TLP genes between different resistant strawberry cultivars in response to C. gloeosporioides infection. Results: In total, 76 TLP genes were identified from the octoploid cultivated strawberry genome with a mean length of 1,439 bp. They were distributed on 24 F. × ananassa chromosomes. The FaTLP family was then divided into ten groups (Group I-X) according to the comparative phylogenetic results. Group VIII contained the highest number of TLP family genes. qRT-PCR analysis results indicated that FaTLP40, FaTLP41, FaTLP43, FaTLP68, and FaTLP75 were upregulated following C. gloeosporioides infection in the resistant octoploid strawberry. Conclusions: The data showed some differences in TLP gene expression patterns across different resistant strawberry cultivars, as well as faster TLP defense responses to pathogenic fungi in resistant cultivars. This study will aid in the characterization of TLP gene family members found in octoploid strawberries and their potential biological functions in plants' defenses against pathogenic fungi.

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.

Characterization and biological activities of polysaccharides from artificially cultivated Phellinus baumii.

The polysaccharide (PPB-2) was extracted and purified from the fruiting body of artificially cultivated Phellinus baumii by an acid-chlorite delignification method. The in vitro hypoglycemic and in vivo hepatoprotective activities of PPB-2 were investigated. FT-IR spectra, NMR spectral analysis, and scanning electron microscopy were used to investigate the structural characterization of PPB-2. PPB-2 exhibited higher α-glucosidase and α-amylase inhibitory activities and showed significant inhibitory effects against the diffusion of glucose. Furthermore, the administration of PPB-2 to mice significantly prevented the increase of serum alanine aminotransferase and aspartate aminotransferase activities, as well as the increase in total cholesterol, triglycerides, and total bilirubin, and hepatic malondialdehyde levels, and PPB-2 enhanced the activities of superoxide dismutase and catalase, as well as the glutathione levels. The results suggest that PPB-2 may be suitable as a functional food with hypoglycemic and hepatoprotective activities.

Identification of Genes Involved in the Responses of Tangor (C. reticulata × C. sinensis) to Drought Stress.

Drought is the major abiotic stress with adverse effects on citrus, decreasing the agronomical yield and influencing the fruit quality. In this study, cDNA-amplified fragment length polymorphism (cDNA-AFLP) technique was used to investigate the transcriptional profile changes and identify drought-responsive genes in "Amakusa" tangor (C. reticulata × C. sinensis), a hybrid citrus sensitive to water stress. The 255 out of 6,245 transcript-derived fragments (TDFs) displayed altered expression patterns including (A) induction, (B) repression, (C) upregulation, and (D) downregulation. With BLAST search, the gene products of differentially expressed fragments (DEFs) could be classified into several categories: cellular processes, transcription, transport, metabolism, stress/stimuli response, and developmental processes. Downregulated genes were highly represented by photosynthesis and basic metabolism, while upregulated ones were enriched in genes that were involved in transcription regulation, defense, energy, and transport. Present result also revealed some transient and up- and then downregulated genes such as aquaporin protein and photosystem enzyme. Expression patterns of 17 TDFs among 18 homologous to function-known genes were confirmed by qRT-PCR analysis. The present results revealed potential mechanism of drought tolerance in fruit crop and also provided candidate genes for future experiments in citrus.

Colored light-quality selective plastic films affect anthocyanin content, enzyme activities, and the expression of flavonoid genes in strawberry (Fragaria × ananassa) fruit.

The influence of colored light-quality selective plastic films (red, yellow, green, blue, and white) on the content of anthocyanin, the activities of the related enzymes and the transcripts of the flavonoid gene was studied in developing strawberry fruit. The results indicated that colored films had highly significant effects on the total anthocyanin content (TAC) and proportions of individual anthocyanins. Compared with the white control film, the red and yellow films led to the significant increase of TAC, while the green and blue films caused a decrease of TAC. Colored film treatments also significantly affected the related enzyme activity and the expression of structural genes and transcription factor genes, which suggested that the enhancement of TAC by the red and yellow films might have resulted from the activation of related enzymes and transcription factor genes in the flavonoid pathway. Treatment with red and yellow light-quality selective plastic films might be useful as a supplemental cultivation practice for enhancing the anthocyanin content in developing strawberry fruit.

Transcript Quantification by RNA-Seq Reveals Differentially Expressed Genes in the Red and Yellow Fruits of Fragaria vesca.

Fragaria vesca (2n = 2x = 14), the woodland strawberry, is a perennial herbaceous plant with a small sequenced genome (240 Mb). It is commonly used as a genetic model plant for the Fragaria genus and the Rosaceae family. Fruit skin color is one of the most important traits for both the commercial and esthetic value of strawberry. Anthocyanins are the most prominent pigments in strawberry that bring red, pink, white, and yellow hues to the fruits in which they accumulate. In this study, we conducted a de novo assembly of the fruit transcriptome of woodland strawberry and compared the gene expression profiles with yellow (Yellow Wonder, YW) and red (Ruegen, RG) fruits. De novo assembly yielded 75,426 unigenes, 21.3% of which were longer than 1,000 bp. Among the high-quality unique sequences, 45,387 (60.2%) had at least one significant match to an existing gene model. A total of 595 genes, representing 0.79% of total unigenes, were differentially expressed in YW and RG. Among them, 224 genes were up-regulated and 371 genes were down-regulated in the fruit of YW. Particularly, some flavonoid biosynthetic pathway genes, including C4H, CHS, CHI, F3H, DFR and ANS, as well as some transcription factors (TFs), including MYB (putative MYB86 and MYB39), WDR and MADS, were down-regulated in YW fruit, concurrent with a reduction in anthocyanin accumulation in the yellow pigment phenotype, whereas a putative transcription repressor MYB1R was up-regulated in YW fruit. The altered expression levels of the genes encoding flavonoid biosynthetic enzymes and TFs were confirmed by quantitative RT-PCR. Our study provides important insights into the molecular mechanisms underlying the yellow pigment phenotype in F. vesca.

Functional analysis of FLOWERING LOCUS T orthologs from spring orchid (Cymbidium goeringii Rchb. f.) that regulates the vegetative to reproductive transition.

The FLOWERING LOCUS T (FT) gene plays crucial roles in regulating the vegetative-to-reproductive phase transition. The FT-like gene of spring orchid (Cymbidium goeringii Rchb. f.), CgFT, was isolated and characterized. CgFT mRNA was detected in leaves, pseudobulb, and flowers. In flowers, CgFT was expressed more in young flower buds than in mature flowers, and was predominantly expressed in young ovary and sheath. Seasonal expression analysis in leaves of a three-year-old spring orchid showed that a large increase in transcription, which started on June 20 for CgFT. We propose that the increased transcription in the middle of June marks the beginning of flower induction in this species. The ectopic expression of CgFT in transgenic tobacco plants showed novel phenotypes by flowering earlier than wild-type plants. Further analysis of the flowering time-related genes indicated that the expression of LEAFY, APETALLA1, FRUITFULL and SEPALLATA1 were significantly upregulated in 35S::CgFT transgenic tobacco plants. These results indicated that CgFT is a putative FT homolog in spring orchid that regulates flower transition, similar to its homolog in Arabidopsis. This study provides the first information on the spring orchid floral gene to elucidate the regulation of the flowering transition in spring orchid.

Identification of two AFLP markers linked to bacterial wilt resistance in tomato and conversion to SCAR markers.

Tomato bacterial wilt (BW) incited by Ralstonia solanacearum is a constraint on tomato production in tropical, subtropical and humid regions of the world. In this paper, we present the results of a research aimed at the identification of PCR-based markers amplified fragment length polymorphism (AFLP) linked to the genes that confer resistance to tomato BW. To this purpose, bulked segregant analysis was applied to an F(2) population segregating for the BW resistant gene and derived from the pair-cross between a BW resistant cultivar T51A and the susceptible cultivar T9230. Genetic analysis indicated that tomato BW was conferred by two incomplete dominant genes. A CTAB method for total DNA extraction, developed by Murray and Thompson with some modifications was used to isolation the infected tomato leaves. Thirteen differential fragments were detected using 256 primer combinations, and two AFLP markers were linked to the BW resistance. Subsequently, the AFLP markers were converted to co-dominant SCAR markers, named TSCAR(AAT/CGA) and TSCAR(AAG/CAT). Linkage analysis showed that the two markers are on the contralateral side of TRSR-1. Genetic distance between TSCAR(AAT/CGA) and TRS-1 was estimated to 4.6 cM, while 8.4 cM between TSCAR(AAG/CAT) and TRS-1.

[Identification of AFLP markers linked to bacterial wilt resistance gene in tomato].

A cross between bacterial wilt resistant tomato variety "T51A"and susceptible variety 'T9230' was made for mapping bacterial wilt resistance gene(s). Through inoculation test of its F1 and F2 progeny, it was proved that the resistance of 'T51A' to bacterial wilt was controlled by one heterozygous gene and cytoplasm. With 64 EcoR I/Muse I primer combinations, AFLP analysis was performed on two parents and their F2 resistant and susceptible bulks. A total of about 4200 distinguishable bands were amplified, of which two were stable. Genetic linkage analysis of the two polymorphic DNA fragments with the resistance gene(s) was tested in the F2 segregating population derived from the cross between 'T51A' and 'T9230'. The DNA fragment AAG/CAT was found closely linked to one of the bacterial wilt resistant genes, with a genetic distance of 6.7 cM, that was tentatively named RRS-342. The cloned fragment AAG/CAT was sequenced and then successfully converted to a SCAR marker, which can be used more conveniently in marker-assisted selection for tomato resistance to bacterial wilt gene.