Light Quality Modulates Growth, Triggers Differential Accumulation of Phenolic Compounds, and Changes the Total Antioxidant Capacity in the Red Callus of Vitis davidii.

Light quality is one of the key elicitors that directly affect plant cell growth and biosynthesis of secondary metabolites. In this study, the red callus of spine grape was cultured under nine light qualities (namely, dark, white, red, yellow, blue, green, purple, warm-yellow, and warm-white light). The effects of different light qualities were studied on callus growth, accumulation of phenolic compounds, and total antioxidant capacity of the red callus of spine grape. The results showed that blue and purple light induced increased red coloration in the callus, whereas yellow light induced the greatest callus proliferation. Among all of the light quality treatments, darkness treatment downregulated the contents of phenolic compounds, whereas blue light was the treatment most conducive to the accumulation of total phenolics. White, blue, and purple light induced increased anthocyanin accumulation. Mixed-wavelength light was beneficial to the accumulation of flavonoids. Blue and purple light were conducive to the accumulation of proanthocyanidins. A further study showed that cyanidin 3-glucoside (Cy3G) and peonidin 3-glucoside (P3G) were the main anthocyanin components in the callus, and blue, purple, and white light treatments promoted their accumulation, whereas flavan-3-ols and flavonols were the main components of non-anthocyanin phenolics, and their accumulation changed in response to not only light quality but also culture duration. The total antioxidant capacity of the callus cultures changed significantly in response to different light qualities. These results will provide evidence for an abiotic elicitor strategy to stimulate callus growth and enhance the accumulation of the main phenolic compounds in the red callus of spine grape.

The Single-Stranded DNA-Binding Gene Whirly (Why1) with a Strong Pathogen-Induced Promoter from Vitis pseudoreticulata Enhances Resistance to Phytophthora capsici.

Vitis vinifera plants are disease-susceptible while Vitis pseudoreticulata plants are disease-resistant; however, the molecular mechanism remains unclear. In this study, the single-stranded DNA- and RNA-binding protein gene Whirly (VvWhy1 and VpWhy1) were cloned from V. vinifera "Cabernet Sauvignon" and V. pseudoreticulata "HD1". VvWhy1 and VpWhy1 promoter sequences (pVv and pVp) were also isolated; however, the identity of the promoter sequences was far lower than that between the Why1 coding sequences (CDSs). Both Why1 gene sequences had seven exons and six introns, and they had a C-terminal Whirly conserved domain and N-terminal chloroplast transit peptide, which was then verified to be chloroplast localization. Transcriptional expression showed that VpWhy1 was strongly induced by Plasmopara viticola, while VvWhy1 showed a low expression level. Further, the GUS activity indicated pVp had high activity involved in response to Phytophthora capsici infection. In addition, Nicotiana benthamiana transiently expressing pVp::VvWhy1 and pVp::VpWhy1 enhanced the P. capsici resistance. Moreover, Why1, PR1 and PR10 were upregulated in pVp transgenic N. benthamiana leaves. This research presented a novel insight into disease resistance mechanism that pVp promoted the transcription of Why1, which subsequently regulated the expression of PR1 and PR10, further enhancing the resistance to P. capsici.

Structural characterization of a soluble polysaccharide SSPS1 from soy whey and its immunoregulatory activity in macrophages.

A soluble soybean polysaccharide SSPS1 with a molecular weight of 2737 kDa was extracted and purified from soy whey. SSPS1 was composed of glucose (97.3 %) and a small amount of mannose (2.7 %). Structural analysis results suggested that SSPS1 had a â†’ 6)-α-d-Glcp-(1 â†’ glucan structure, with a trace amount of α-d-Glcp-(1 â†’ connected to the main chain via O-3. In vitro immunological experiments suggested that SSPS1 enhanced the growth rate and phagocytic activity of RAW 264.7 macrophages. In addition, SSPS1 stimulated the secretion of cytokines (TNF-α, INF-ß, IL-6 and IL-1ß) as well as nitric oxide (NO) production through upregulating the expression of the related genes and proteins in RAW 264.7 cells. This study provided a new method for efficient utilization of soy whey, and the results indicate that SSPS1 extracted from soy whey could be used as a novel immunomodulator.

Identifying Plasmopara viticola resistance Loci in grapevine (Vitis amurensis) via genotyping-by-sequencing-based QTL mapping.

Downy mildew, caused by Plasmopara viticola, is a major disease that affects grapevines, and a few resistance (R) genes have been identified thus far. In order to identify R genes, we investigated F1 progeny from a cross between the downy mildew-resistant Vitis amurensis 'Shuang Hong' and the susceptible Vitis vinifera 'Cabernet Sauvignon'. The P. viticola-resistance of the progeny varied continuously and was segregated as a quantitative trait. Genotyping-by-sequencing was used to construct linkage maps. The integrated map spanned 1898.09 cM and included 5603 single nucleotide polymorphisms on 19 linkage groups (LGs). Linkage analysis identified three quantitative trait loci (QTLs) for P. viticola resistance: 22 (Rpv22) on LG 02, Rpv23 on LG15, and Rpv24 on LG18. The phenotypic variance contributed by these three QTLs ranged from 15.9 to 30.0%. qRT-PCR analysis of R-gene expression in these QTLs revealed a CC-NBS-LRR disease resistance gene RPP8, two LRR receptor-like serine/threonine-protein kinases, a serine/threonine-protein kinase BLUS1, a glutathione peroxidase 8, an ethylene-responsive transcription factor ERF038, and a transcription factor bZIP11 were induced by P. viticola, and these genes may play important role in P. viticola response.

Cgr1, a ripe rot resistance QTL in Vitis amurensis 'Shuang Hong' grapevine.

Ripe rot is a serious grapevine disease in Vitis L. and Muscadinia (Planch.) Small. However, resistance to this disease has been reported in some oriental Vitis species. To identify resistance-related Quantitative Trait Loci (QTLs) from the Chinese grape species V. amurensis, an F1 population of V. vinifera 'Cabernet Sauvignon' × V. amurensis 'Shuang Hong' was used to map the ripe rot resistance loci expected in 'Shuang Hong' grape. A total of 7598 single nucleotide polymorphisms (SNPs) between the parents were identified in our previous study, and 934 SNPs were selected for genetic map construction. These SNPs are distributed across the 19 chromosomes covering a total of 1665.31 cM in length, with an average of 1.81 cM between markers. Ripe rot resistance phenotypes among the hybrids were evaluated in vitro using excised leaves for three consecutive years from 2016 to 2018; a continuous variation was found among the F1 hybrids, and the Pearson correlation coefficients of the phenotypes scored in all three years were significant at the 0.01 level. Notably, the first QTL reported for resistance to grape ripe rot disease, named Cgr1, was identified on chromosome 14 of 'Shuang Hong' grapevine. Cgr1 could explain up to 19.90% of the phenotypic variance. In addition, a SNP named 'np19345' was identified as a molecular marker closely linked to the peak of Cgr1 and has the potential to be developed as a marker for the Cgr1 resistance haplotype.

Molecular Characterization and Overexpression of VpRPW8s from Vitis pseudoreticulata Enhances Resistance to Phytophthora capsici in Nicotiana benthamiana.

RPW8 genes are atypical broad-spectrum genes that provide resistance to powdery mildew, downy mildew, the cauliflower mosaic virus in Arabidopsis thaliana, and powdery mildew in tobacco. They play important roles in basal plant pathogen defense. They also provide insights into a novel disease resistance mechanism. In this study, we report on homologous RPW8 genes in Vitis pseudoreticulata. Five VpRPW8 genes were cloned; their Open Reading Frame (ORF) sequences ranged from 1994 base pairs to 2478 base pairs. They were comprised of five exons and four introns and shared 78.66% identity. Their proteins had typical conserved RPW8 and NB-LRR (the nucleotide-binding site and the leucine-rich repeats) domains (except VpRPW8-d, which lacked LRR domains). Prokaryotic expression results were consistent with predicted molecular weights. All five RPW8 genes were located in the cytoplasm. Quantitative real-time PCR (qRT-PCR) analysis showed that VpRPW8s in V. pseudoreticulata were induced by Plasmopara viticola, but nearly only VvRPW8-d genes were induced in Vitis vinifera. Furthermore, a VpRPW8 transgenic tobacco system was established. Overexpressed VpRPW8s enhanced resistance to Phytophthora capsici and VpRPW8s conferred varying degrees of resistance to Ph. capsici in Nicotiana benthamiana. Our study presents novel members of the plant RPW8 family and suggests that VpRPW8s are involved in enhanced resistance to P. viticola and Ph. capsici.

Genome-wide identification and characterization of the superoxide dismutase gene family in Musa acuminata cv. Tianbaojiao (AAA group).

BACKGROUND: Superoxide dismutase (SOD) is an essential enzyme of the plant antioxidant system that responds to oxidative stresses caused by adverse conditions. Banana is an important staple and economic crop in tropical and subtropical regions. However, its growth and yield are constantly affected by various abiotic stresses. To analyze the roles of distinct SOD genes under various stresses, a detailed characterization and analysis of the SOD gene family in Cavendish banana is indispensable. METHODS: The presence and structure of the SOD family genes were experimentally verified using 5'/3' RACE-PCR, reverse transcription PCR and PCR. Then, their syntenic relationships, conserved motifs and phylogenetic relationships were analyzed using software. Cis-elements present in the promoters were predicted via PlantCARE. And the expression levels under abiotic and hormonal stresses were determined using real-time quantitative polymerase chain reaction. RESULTS: In total, 25 'Tianbaojiao' SOD cDNAs (MaSODs), which encoded six Cu/ZnSODs, four MnSODs and two FeSODs, were cloned. The 12 MaSOD genes were divided into four groups based on their conserved motifs, which corroborated their classifications based on gene-structure patterns and subcellular localizations. Eleven MaSOD promoters were isolated and found to contain many cis-acting elements involved in stress responses. Gene expression analysis showed that 11 out of the 12 MaSODs were expressed in all tested tissues (leaf, pseudostem and root), whereas MaCSD2B was expressed only in leaves and roots. Specific MaSOD members exhibited different expression patterns under abiotic and hormonal treatments. Among the 12 MaSOD genes, MaCSD1D was the only one that responded to all eight treatments, suggesting that this gene plays a predominant role in reactive oxygen species scavenging caused by various stresses in banana. CONCLUSIONS: A genome-wide analysis showed that the 'Tianbaojiao' banana harbored an expanded SOD gene family. Whole genome duplication, segmental duplication and complex transcriptional regulation contributed to the gene expansion and mRNA diversity of the MaSODs. The expression patterns of distinct MaSOD genes showed that they are important responses to different abiotic and hormonal stresses in banana.

Molecular cloning and expression analysis of KIN10 and cold-acclimation related genes in wild banana 'Huanxi' (Musa itinerans).

Banana cultivars may experience chilling or freezing injury in some of their cultivated regions, where wild banana can still grow very well. The clarification of the cold-resistant mechanism of wild banana is vital for cold-resistant banana breeding. In this study, the central stress integrator gene KIN10 and some cold-acclimation related genes (HOS1 and ICE1s) from the cold-resistant wild banana 'Huanxi' (Musa itinerans) were cloned and their expression patterns under different temperature treatments were analyzed. Thirteen full-length cDNA transcripts including 6 KIN10s, 1 HOS1 and 6 ICE1s were successfully cloned. Quantitative real-time PCR (qRT-PCR) results showed that all these genes had the highest expression levels at the critical temperature of banana (13 °C). Under chilling temperature (4 °C), the expression level of KIN10 reduced significantly but the expression of HOS1 was still higher than that at the optimal temperature (28 °C, control). Both KIN10 and HOS1 showed the lowest expression levels at 0 °C, the expression level of ICE1, however, was higher than control. As sucrose plays role in plant cold-acclimation and in regulation of KIN10 and HOS1 bioactivities, the sucrose contents of wild banana under different temperatures were detected. Results showed that the sucrose content increased as temperature lowered. Our result suggested that KIN10 may participate in cold stress response via regulating sucrose biosynthesis, which is helpful in regulating cold acclimation pathway in wild banana.