Comparison of phenolic profiles and antioxidant activities in skins and pulps of eleven grape cultivars(Vitis vinifera L.)

Eleven grape cultivars were analysed to explore the variety differences of fresh grape phenolic profiles. The results showed that free phenolics were predominant in grape skins and pulps, and showed the higher antioxidant activities than bound. In 11 cultivars, Muscat Kyoho extracts had the highest total phenolic content in skins(10.525 mg GAE g~(–1) FW) and pulps(1.134 mg GAE g~(–1) FW), and exhibited the highest DPPH radical scavening capacity(EC_(50)=11.7 μg mL~(–1)) and oxygen radical absorbance capacity(ORAC) value(190.57 μmol TE g~(–1) FW) of free phenolic in skin. In addition, the most abundant phenolics in grape skins were found to be flavonoids such as kaempferol in Kyoho skin(541.2 μg g~(–1) FW), rutin, catechin and epicatechin in Muscat Kyoho skin(262.3, 86.3 and 70.0 μg g~(–1) FW, respectively). Furthermore, the principal component analysis showed a strong difference of phenolic profiles with the cultivars, existing forms and distributions. Pearson correlation coefficient analysis showed a significant linear correlation between total phenolic content and antioxidant activity(P0.05). Therefore, both skins and pulps were rich sources of bioactive phenolic compounds, and Muscat Kyoho was the ideal source among all samples.     

Combined effect of shading time and nitrogen level on grain filling and grain quality in japonica super rice

There is limited information about the combined effect of shading time and nitrogen(N) on grain filling and quality of rice. Therefore, two japonica super rice cultivars, Nanjing 44 and Ningjing 3, were used to study the effect of shading time and N level on the characteristics of rice panicle and grain filling as well as the corresponding yield and quality. At a low N level(150 kg N ha~(–1), 150 N), grain yield decreased(by 21.07–26.07%) under the treatment of 20 days of shading before heading(BH) compared with the no shading(NS) treatment. These decreases occurred because of shortened panicle length, decreased number of primary and secondary branches, as well as the grain number and weight per panicle. At 150 N, in the treatment of 20 days of shading after heading(AH), grain yield also decreased(by 9.46–10.60%) due to the lower grain weight per panicle. The interaction of shading and N level had a significant effect on the number of primary and secondary branches. A high level of N(300 kg N ha~(–1), 300 N) could offset the negative effect of shading on the number of secondary branches and grain weight per panicle, and consequently increased the grain yield in both shading treatments. In superior grains, compared with 150 N NS, the time to reach 99% of the grain weight(T_(99)) was shortened by 1.6 to 1.7 days, and the grain weight was decreased by 4.18–5.91% in 150 N BH. In 150 N AH, the grain weight was 13.39–13.92% lower than that in 150 N NS due to the slow mean and the maximum grain-filling rate(GR_(mean )and GR_(max)). In inferior grains, grain weight and GR_(mean) had a tendency of 150 N NS150 N BH150 N AH. Under shaded conditions, 300 N decreased the grain weight due to lower GR_(mean) both in superior and inferior grains. Compared with 150 N NS, the milling and appearance qualities as well as eating and cooking quality were all decreased in 150 N BH and 150 N AH. Shading with the high level of 300 N improved the milling quality and decreased the number of chalky rice kernels, but the eating and cooking quality was reduced with increased chalky area and overall chalkiness. Therefore, in the case of short term shading, appropriate N fertilizer could be used to improve the yield and milling quality of rice, but limited application of N fertilizer is recommended to achieve good eating and cooking quality of rice.     

Residue management induced changes in soil organic carbon and total nitrogen under different tillage practices in the North China Plain

Crop residue retention has been considered a practicable strategy to improve soil organic carbon(SOC) and total nitrogen(TN), but the effectiveness of residue retention might be different under varied tillage practices. To evaluate the effects of residue management on the distribution and stocks of SOC and TN under different tillage practices, a bifactorial experiment with three levels for tillage practices(no-tillage, rotary tillage, and conventional tillage) and two levels for residue managements(residue retention and residue removal) was conducted in the North China Plain(NCP). Results showed that after a short experimental duration(3–4 years), concentrations of SOC and TN in the 0–10 cm layer were higher under no-tillage than under conventional tillage, no matter whether crop residues were retained or not. Residue retention increased SOC and TN concentrations in the upper layers of soil to some degree for all tillage practices, as compared with residue removal, with the greatest increment of SOC concentration occurred in the 0–10 cm layer under rotary tillage, but in the 10–30 cm layer under conventional tillage. The stocks of SOC in the 0–50 cm depth increased from 49.89 Mg ha–1 with residue removal to 53.03 Mg ha–1 with residue retention. However, no-tillage did not increase SOC stock to a depth of 50 cm relative to conventional tillage, and increased only by 5.35% as compared with rotary tillage. Thus, residue retention may contribute more towards SOC sequestration than no-tillage. Furthermore, the combination between residue retention and no-tillage has the greatest advantage in enhancing SOC and TN in the NCP region.     

Population genetic structure of Chinese Puccinia triticina races based on multi-locus sequences

Puccinia triticina, the causal agent of wheat leaf rust, is one of the most devastating rust fungi attacking wheat worldwide. Seventy-six isolates of the wheat leaf rust pathogen from Yunnan, Sichuan, Gansu and Henan provinces, China, were tested on wheat leaf rust differentials and the population structure was analyzed using four presumably neutral partial sequence markers such as elongation factor-1α(EF-1α), glyceraldehyde-3-phosphate dehydrogenase(GAPDH), β-tubulin(TUB) and the second largest RNA polymerase subunit(RPB2). The phenotypic diversity of Yunnan and Sichuan populations was higher than that of Gansu and Henan populations. The four populations were separated into two clusters based on the pathogenic data. A total of 12 single nucleotide polymorphisms(SNPs) and 32 haplotypes were identified among the four sequences. The 32 haplotypes were divided into two clusters in a neighbor-joining tree. Bayesian analyses also identified two clusters. Pairwise FST between populations in different regions were significantly different(P0.05). Analysis of molecular variance(AMOVA) indicated that 68% of the total genetic variation was within populations.     

Species-specific COI primers for rapid identification of a globally significant invasive pest,the cassava mealybug Phenacoccus manihoti Matile-Ferrero

The globally invasive cassava mealybug Phenacoccus manihoti Matile-Ferrero is a pernicious pest of cassava,and its recent introduction into Asia has raised considerable alarm.To slow or prevent further invasion,an accurate,simple,and developmental-stage-independent detection method for P.manihoti is required.In the present study,a PCR method based on a species-specific mitochondrial DNA cytochrome oxidase I(SS-COI)marker was developed for rapid identification of P.manihoti.One pair of SS-COI primers(PMSSZW-1F and PMSSZW-1R)was designed based on sequence variations in the COI gene among P.manihoti and related mealybug species.Specificity of the primer pair was validated on 21 closely related species.Sensitivity tests were performed on four immature developmental stages and female adults.Efficacy tests demonstrated that at the relatively low concentration of(135.2±14.7)pgresuspended DNA,the specific fragment was detected in all replicates.Furthermore,the SS-COI primer pair was assayed on three populations of P.manihoti from major exporting countries of cassava.The PCR assay was proved to be a rapid,simple,and reliable molecular measure for the identification of P.manihoti.This tool will be useful for quarantine,monitoring,and management of this invasive pest.     

Estimates on nitrogen uptake in the subsequent wheat by above-ground and root residue and rhizodeposition of using peanut labeled with 15N isotope on the North China Plain

Leguminous crops play a vital role in enhancing crop yield and improving soil fertility. Therefore, it can be used as an organic N source for improving soil fertility. The purpose of this study was to (i) quantify the amounts of N derived from rhizodeposition, root and above-ground biomass of peanut residue in comparison with wheat and (ii) estimate the effect of the residual N on the wheat-growing season in the subsequent year. The plants of peanut and wheat were stem fed with ¹⁵N urea using the cotton-wick method at the Wuqiao Station of China Agricultural University in 2014. The experiment consisted of four residue-returning strategies in a randomized complete-block design: (i) no return of crop residue (CR0); (ii) return of above-ground biomass of peanut crop (CR1); (iii) return of peanut root biomass (CR2); and (iv) return of all residue of the whole peanut plant (CR3). The 31.5 and 21% of the labeled ¹⁵N isotope were accumulated in the above-ground tissues (leaves and stems) of peanuts and wheat, respectively. N rhizodeposition of peanuts and wheat accounted for 14.91 and 3.61% of the BG¹⁵N, respectively. The ¹⁵N from the below-ground ¹⁵N-labeled of peanuts were supplied 11.3, 5.9, 13.5, and 6.1% of in the CR0, CR1, CR2, and CR3 treatments, respectively. Peanut straw contributes a significant proportion of N to the soil through the decomposition of plant residues and N rhizodeposition. With the current production level on the NCP, it is estimated that peanut straw can potentially replace 104 500 tons of synthetic N fertilizer per year. The inclusion of peanut in rotation with cereal can significantly reduce the use of N fertilizer and enhance the system sustainability.     

Molecular identification and enzymatic properties of laccase2 from the diamondback moth Plutella xylostella (Lepidoptera: Plutellidae)

Laccase(EC 1.10.3.2)is known to oxidize various aromatic and nonaromatic compounds via a radical-catalyzed reaction,which generally includes two types of laccase,Lac1 and Lac2.Lac1 oxidizes toxic compounds in the diet,and Lac2 is known to play an important role in melanizing the insect exoskeleton.In this study,we cloned and sequenced the cDNA of the diamondback moth,Plutella xylostella Lac2(PxLac2),from the third instar larvae using polymerase chain reaction(PCR)and rapid amplification of cDNA ends techniques.The results showed that the full-length PxLac2 cDNA was 1 944 bp long and had an open reading frame of 1 794 bp.PxLac2 encoded a protein with 597 amino acids and had a molecular weight of 66.09 kDa.Moreover,we determined the expression levels of PxLac2 in different stages by quantitative PCR(qPCR).The results indicated that PxLac2 was expressed differently in different stages.We observed the highest expression level in pupae and the lowest expression level in fourth instar larvae.We also investigated the enzymatic properties of laccase,which had optimal activity at pH 3.0 and at 35°C.Under these optimal conditions,laccase had a Michaelis constant(K_m)of 0.97 mmol L~(-1),maximal reaction speed(V_m)of 56.82 U mL~(-1),and activation energy(E_a)of 17.36 kJ mol~(-1) to oxidize2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid ammonium salt).Type II copper enhanced laccase activity below 0.8mmol L~(-1) and reduced enzyme activity above 0.8 mmol L~(-1) with an IC_(50) concentration of 1.26 mmol L~(-1).This study provides insights into the biological function of laccase.     

A review: The beneficial effects and possible mechanisms of aluminum on plant growth in acidic soil

Aluminum(Al) is the third most abundant element in the earth's crust and a major factor inhibiting plant growth and reducing crop yield in acidic soil. Although there is substantial research on the phytotoxic effects and the underlying mechanisms of Al by applying Al alone hydroponically, soil is a complex medium containing numerous mineral elements that can interact with Al and other elements and their bioavailability in plants. In this review, we describe the roles of Al in promoting plant growth, enhancing phosphorus availability and efficient use in plants, and alleviating H+, iron, and manganese toxicity in acidic conditions. Furthermore, we discuss the possible mechanisms of enhanced abiotic stress tolerance induced by Al. We also elucidate the role of Al in attracting plant growth promoting rhizo-bacteria(PGPR) and their interactions with plants by increasing organic exudates.     

Evaluating the efficacy of an attenuated Streptococcus equi ssp. zooepidemicus vaccine produced by multi-gene deletion in pathogenicity island SeseCisland_4

Streptococcus equi ssp. zooepidemicus(SEZ) is a pathogen associated with a wild range of animal species. Frequent outbreaks have occurred in recent years in pigs, horses, goats and dogs which is liable to infect humans. There is a lack of efficient vaccines against this disease and the occurrence of antibiotic resistance may render drug therapies ineffective. In this study, gene deletion mutant(ΔSEZ) in pathogenicity islands SeseCisland_4 was constructed. The mutant ΔSEZ had a 52-fold decrease in 50% lethal dose(LD_(50)) and had less capacity to adhere epithelial cells. Importantly, immunization of mice with attenuated vaccine ΔSEZ at the dose of 10~2 colony-forming units(CFU) mL~(–1) elicited a significant humoral antibody response, with an antibody titer of 1:12 800. Therefore, 10~2 CFU mL~(–1) might be used as the appropriate immune dose for the attenuated vaccine ΔSEZ, which provided mice with efficient protection against virulent SEZ. In addition, the hyperimmune sera against 10~2 CFU m L–1 attenuated vaccine ΔSEZ could confer significant protection against virulent SEZ infection in the passive immunization experiment and exhibited efficient bactericidal activity in the whole blood assay. Meanwhile, no viable bacteria was detected in blood when mice were immunized with ΔSEZ at the dose of 10~2 CFU mL~(–1) via hypodermic injection. Thereafter, the mutant ΔSEZ at the dose of 10~2 CFU mL~(–1) could confer significant protection in mice and had less negative effects on host, which could be an effective attenuated vaccine candidate for the prevention of SEZ.     

A rapid,simple, and sensitive immunoagglutination assay with silica nanoparticles for serotype identification of Pseudomonas aeruginosa

An agglutination test based on colored silica nanoparticles (colored SiNps) was established to detect serotypes of Pseudomonas aeruginosa. Monodisperse colored SiNps were used as agglutination test carriers. The colored SiNps were prepared through reverse microemulsion with reactive dyes, sensitized with 11 kinds of mono-specific antibodies against P. aeruginosa, and denoted as IgG-colored SiNps. Eleven kinds of IgG-colored SiNps were individually mixed with P. aeruginosa on a glass slide. Different serotypes of P. aeruginosa could be identified by agglutination test with evident agglutination. The P. aeruginosa could be detected in a range from 3.6 × 10⁵ to 3.6 × 10¹² cfu mL^?1. This new agglutination test was confirmed to be a specific, sensitive, fast, easy-to-perform, and cost-efficient tool for the routine diagnosis of P. aeruginosa.     

Seedling and adult plant resistance to leaf rust in 46 Chinese bread wheat landraces and 39 wheat lines with known Lr genes

Wheat leaf rust,caused by Puccinia triticina(Pt),is an important foliar disease that has an important influence on wheat yield.The most economic,safe and effective way to control the disease is growing resistant cultivars.In the present study,a total of 46 wheat landraces and 34 wheat lines with known Lr(leaf rust resistance)genes were inoculated with 16Pt pathotypes for postulating seedling resistance gene(s)in the greenhouse.These cultivars and five wheat differential lines with adult plant resistance(APR)genes(Lr12,Lr22b,Lr34,Lr35 and Lr37)were also evaluated for identification of slow rusting resistance in the field trials in Baoding,Hebei Province of China in the 2014–2015 and 2015–2016 cropping seasons.Furthermore,10 functional molecular markers closely linked to 10 known Lr genes were used to detect all the wheat genotypes.Results showed that most of the landraces were susceptible to most of the Pt pathotypes at seedling stage.Nonetheless,Lr1 was detected only in Hongtangliangmai.The field experimental test of the two environments showed that 38 landraces showed slow rusting resistance.Seven cultivars possessed Lr34 but none of the landraces contained Lr37 and Lr46.Lr genes namely,Lr9,Lr19,Lr24,Lr28,Lr29,Lr47,Lr51 and Lr53 were effective at the whole plant stage.Lr18,Lr36 and Lr45 had lost resistance to part of pathotypes at the seedling stage but showed high resistance at the adult plant stage.Lr34 as a slowing rusting gene showed good resistance in the field.Four race-specific APR genes Lr12,Lr13,Lr35 and Lr37 conferred good resistance in the field experiments.Seven race-specific genes,Lr2b,Lr2c,Lr11,Lr16,Lr26,Lr33 and LrB had lost resistance.The 38 landraces showed slow rusting resistance to wheat leaf rust can be used as resistance resources for wheat resistance breeding in China.     

Effects of leaf removal and cluster thinning on berry quality of Vitis vinifera cultivars in the region of Weibei Dryland in China

Leaf removal and cluster thinning were carried out prior to veraison to evaluate the effects on berry quality of two Vitis vinifera cultivars (Cabernet Sauvignon and Ugni Blanc) in the Weibei Dryland of China in 2013 and 2014, and comprehensive analysis of the chemical and volatile composition in berries was performed. The results showed that content of reducing sugar in both varieties was not affected while total acid was generally decreased by leaf removal and cluster thinning. The pH of berry juice was correspondingly higher in most treatment groups. Meanwhile, promoting effects on accumulation of total phenols, tannin in both varieties and total anthocyanins in Cabernet Sauvignon were found. As for monomeric anthocyanins, percentage of malvidin and its derivatives was decreased by leaf removal and cluster thinning. Besides, cinnamylated anthocyanins decreased with the intensity of cluster thinning. The accumulation of non-anthocyanin phenolics was similarly affected in the two varieties. Notably, cluster thinning was more effective on enhancing the phenolics content than leaf removal. The combination of middle level of leaf removal and cluster thinning was the most favor to the accumulation of phenolic acids. Furthermore, cluster thinning could also significantly enhance the synthesis of flavanols and stilbenes. Lastly, content and variety of aroma compounds in both grape varieties were also significantly affected by the treatments. The results provided a theoretical basis for a combination of leaf removal and cluster thinning to improve quality of grapes and wines.     

Phenotypic characterization and genetic mapping of the dwarf mutant m34 in maize

Plant height is one of the most important agronomic traits associated with yield in maize.In this study,a gibberellins(GA)-insensitive dwarf mutant,m34,was screened from inbred line Ye478 by treatment with the chemical mutagen ethylmethanesulfonate(EMS).Compared to Ye478,m34 showed a dwarf phenotype with shorter internodes,and smaller leaf length and width,but with similar leaf number.Furthermore,m34 exhibited smaller guard cells in internodes than Ye478,suggesting that smaller cells might contribute to its dwarf phenotype.Genetic analysis indicated that the m34 dwarf phenotype was controlled by a recessive nuclear gene.An F2 population derived from a cross between m34 and B73 was used for mutational gene cloning and this gene was mapped to a chromosome region between umc2189 and umc1553 in chromosome 1 bin1.10,which harbored a previously identified dwarf gene Zm VP8.Sequencing analysis showed a nucleotide substitution(G1606 to A1606)in the sixth exon of ZmVP8,which resulted in an amino acid change(E531 to K531)from Ye478 to m34.This amino acid change resulted in anα-helix changing to aβ-sheet in the secondary protein structure and the‘SPEC’domain changed to a‘BOT1NT’domain in the tertiary protein structure.Taken together,these results suggested that m34 is a novel allelic mutant originally derived from Ye478 that is useful for further ZmVP8 functional analysis in maize.     

Characterization of TaCOMT genes associated with stem lignin content in common wheat and development of a gene-specific marker

Stem lignin content(SLC) in common wheat(Triticum aestivum L.) contributes to lodging resistance. Caffeic acid 3-O-methyltransferase(COMT) is a key enzyme involved in lignin biosynthesis. Characterization of TaCOMT genes and development of gene-specific markers could enable marker-assisted selection in wheat breeding. In the present study, the full-length genomic DNA(gDNA) sequences of TaCOMT genes located on chromosomes 3 A, 3 B, and 3 D were cloned by homologous cloning. Two allelic variants, TaCOMT-3 Ba and TaCOMT-3 Bb, were identified and differed by a 222-bp insertion/deletion(InDel) in the 3′-untranslated region(3′-UTR). A co-dominant gene-specific marker based on this InDel was developed and designated as Ta COMT-3 BM. A total of 157 wheat cultivars and advanced lines grown in four environments were used to validate the associations between allelic patterns and SLC. The SLC of cultivars with TaCOMT-3 Ba was significantly(P0.01) higher than that of those with TaCOMT-3 Bb, and the marker TaCOMT-3 BM could be effectively used in wheat breeding.     

Molecular,serological and biological characterization of a novel Apple stem pitting virus strain from a local pear variety grown in China

Apple stem pitting virus (ASPV) is an important causal agent of pear diseases. Nowadays, the infection status and molecular characteristics of the virus in old pear trees have never been investigated. In this study, we provide the first complete genome sequence of an ASPV isolate LYC from an over 300-year-old tree of a local Pyrus bretschneideri cultivar ‘Chili’ specifically grown at Laiyang area in China. ASPV-LYC possesses a chimeric genome consisting of 9 273 nucleotides excluding a poly(A) tail at its 3’ end and harboring a recombination region in its open reading frame (ORF1) with Aurora-1 and KL9 identified as the major and minor parents. Western blot analysis with antisera against recombinant coat proteins (CPs) of three ASPV isolates from pear indicates that ASPV-LYC is serologically related to these ASPV isolates, but with differential activities. Further biological tests on indicator plants of Pyronia veitchii show that ASPV-LYC can induce serious leaf and stem symptoms as other ASPV isolates. The results provide an important information for understanding molecular evolution of ASPV and suggest a need to prevent dissemination of the isolate among pear trees.