Targeting of anti-microbial proteins to the hyphal surface amplifies protection of crop plants against Phytophthora pathogens.

Phytophthora pathogens are a persistent threat to the world's commercially important agricultural crops, including potato and soybean. Current strategies aim at reducing crop losses rely mostly on disease-resistance breeding and chemical pesticides, which can be frequently overcome by the rapid adaptive evolution of pathogens. Transgenic crops with intrinsic disease resistance offer a promising alternative and continue to be developed. Here, we explored Phytophthora-derived PI3P (phosphatidylinositol 3-phosphate) as a novel control target, using proteins that bind this lipid to direct secreted anti-microbial peptides and proteins (AMPs) to the surface of Phytophthora pathogens. In transgenic Nicotiana benthamiana, soybean, and potato plants, significantly enhanced resistance to different pathogen isolates was achieved by expression of two AMPs (GAFP1 or GAFP3 from the Chinese medicinal herb Gastrodia elata) fused with a PI3P-specific binding domain (FYVE). Using the soybean pathogen P. sojae as an example, we demonstrated that the FYVE domain could boost the activities of GAFPs in multiple independent assays, including those performed in vitro, in vivo, and in planta. Mutational analysis of P. sojae PI3K1 and PI3K2 genes of this pathogen confirmed that the enhanced activities of the targeted GAFPs were correlated with PI3P levels in the pathogen. Collectively, our study provides a new strategy that could be used to confer resistance not only to Phytophthora pathogens in many plants but also potentially to many other kinds of plant pathogens with unique targets.

Evaluation of Polycyclic Aromatic Hydrocarbons (PAHs) in Bamboo Shoots from Soil.

The toxicity, carcinogenicity and persistence of polycyclic aromatic hydrocarbons (PAHs) pose a great threat to the ecological system and human health. The contamination levels, translocation and source analysis of 16 PAHs in bamboo shoot and its planted soil were investigated. The average concentrations of total PAHs were 18.80 ± 1.90 µg/kg and 123.98 ± 113.36 µg/kg in bamboo shoots and soils, respectively. The most abundant PAH was Phenanthrene (PHE), with the detected average concentrations of 5.85 µg/kg in bamboo shoots and 19.28 µg/kg in soils. The highest detected types of PAHs were 3 rings and 4 rings, with the proportions of 80.69% (bamboo shoots) and 35.23% (soils). The transfer factors of PAHs were ranged from 0.011 to 0.895, in which PAHs with 3 rings showed the strongest transfer ability. The combustion of biomass and petroleum might be the main source of PAHs in the planted soils of bamboo shoots.

Genome Analysis of Two Newly Emerged Potato Late Blight Isolates Sheds Light on Pathogen Adaptation and Provides Tools for Disease Management.

Phytophthora infestans, the causal agent of the Irish Potato Famine in the 1840s, is one of the most destructive crop pathogens that threaten global food security. Host resistance (R) genes may help to control the disease, but recognition by through the gene products can be evaded by newly emerging isolates. Such isolates are dangerous as they may cause disease outbreaks under favorable conditions. However, our lack of knowledge about adaptation in these isolates jeopardizes an apt response to resistance breakdown. Here we performed genome and transcriptome sequencing of HB1501 and HN1602, two field isolates from distinct Chinese geographic regions. We found extensive polymorphisms in these isolates, including gene copy number variations, nucleotide polymorphisms, and gene expression changes. Effector encoding genes, which contribute to virulence, show distinct expression landscapes in P. infestans isolates HB1501 and HN1602. In particular, polymorphisms at multiple effectors required for recognition (Avr loci) enabled these isolates to overcome corresponding R gene based resistance. Although the isolates evolved multiple strategies to evade recognition, we experimentally verified that several R genes such as R8, RB, and Rpi-vnt1.1 remain effective against these isolates and are valuable to potato breeding in the future. In summary, rapid characterization of the adaptation in emerging field isolates through genomic tools inform rational agricultural management to prevent potential future epidemics.

The Fate of Organophosphorus Pesticides during Camellia Oil Production.

The purpose of this study was to investigate the fate of organophosphorus pesticides (OPs) during camellia oil production process, from camellia fruit to the final oil product. The results showed that the OPs were mainly distributed in the peel of camellia fruit, basically above 40% after the pesticide application of 7 d (P < 0.05). A small amount of OPs could enter into the seed and convert to crude camellia oil, with the concentration of 19.5 to 548.2 mg/L. In addition, metabolites of OPs (25.7 to 768.9 mg/L) could be detected in the crude camellia oil. Moreover, the refining process (degumming, deacidfying, bleaching) had a significant effect on the removal of OPs from the crude camellia oil (P < 0.05), and the effect was related to the octanol-water partition constant (LogP) of pesticide. The larger the LogP, the more stable the OPs were during refining process. The final refined camellia oil was found to have no detectable levels of OPs metabolite.

Genome-Wide Identification of Differentially Expressed Genes Associated with the High Yielding of Oleoresin in Secondary Xylem of Masson Pine (Pinus massoniana Lamb) by Transcriptomic Analysis.

Masson pine is an important timber and resource for oleoresin in South China. Increasing yield of oleoresin in stems can raise economic benefits and enhance the resistance to bark beetles. However, the genetic mechanisms for regulating the yield of oleoresin were still unknown. Here, high-throughput sequencing technology was used to investigate the transcriptome and compare the gene expression profiles of high and low oleoresin-yielding genotypes. A total of 40,690,540 reads were obtained and assembled into 137,499 transcripts from the secondary xylem tissues. We identified 84,842 candidate unigenes based on sequence annotation using various databases and 96 unigenes were candidates for terpenoid backbone biosynthesis in pine. By comparing the expression profiles of high and low oleoresin-yielding genotypes, 649 differentially expressed genes (DEGs) were identified. GO enrichment analysis of DEGs revealed that multiple pathways were related to high yield of oleoresin. Nine candidate genes were validated by QPCR analysis. Among them, the candidate genes encoding geranylgeranyl diphosphate synthase (GGPS) and (-)-alpha/beta-pinene synthase were up-regulated in the high oleoresin-yielding genotype, while tricyclene synthase revealed lower expression level, which was in good agreement with the GC/MS result. In addition, DEG encoding ABC transporters, pathogenesis-related proteins (PR5 and PR9), phosphomethylpyrimidine synthase, non-specific lipid-transfer protein-like protein and ethylene responsive transcription factors (ERFs) were also confirmed to be critical for the biosynthesis of oleoresin. The next-generation sequencing strategy used in this study has proven to be a powerful means for analyzing transcriptome variation related to the yield of oleoresin in masson pine. The candidate genes encoding GGPS, (-)-alpha/beta-pinene, tricyclene synthase, ABC transporters, non-specific lipid-transfer protein-like protein, phosphomethylpyrimidine synthase, ERFs and pathogen responses may play important roles in regulating the yield of oleoresin. These DEGs are worthy of special attention in future studies.

Optimization of a one-step method for the multiresidue determination of organophosphorous pesticides in camellia oil.

UNLABELLED: Due to the widespread use and potential toxicity of organophosphorous pesticides (OPs), multiresidue monitoring of OPs in camellia oil has become increasingly important. A simple, rapid, and effective matrix solid-phase dispersion extraction for the determination of 15 organophosphorous pesticides in camellia oil is described. Related important factors influencing the extraction efficiency, such as type of sorbent material, eluting solvent, and ratio of sample/sorbent were studied and optimized. The best results were obtained using 0.5 g of camellia oil, 1.5 g of white carbon black as dispersant sorbent, and 5 mL of acetonitrile: ethyl acetate (2:1, V/V) as eluting solvent. Method validation was performed in order to study sensitivity, linearity, precision, and accuracy. Average recoveries ranged between 76.7% and 102.9% with relative standard deviation values from 2.9% to 13.7% at 2 concentration levels (10 and 100 µg/kg). The method limit of detection at or below the regulatory maximum residue limits for the pesticides was achieved. PRACTICAL APPLICATION: A simple, rapid, and effective method for multiresidue determination of organophosphorous pesticides in camellia oil was developed. The sample preparation could finish in 5 min.

Simultaneous determination of 15 multiresidue organophosphorous pesticides in camellia oil by MSPD-GC-MS.

A one step method based on matrix solid-phase dispersion for simultaneous determination of 15 organophosphorous pesticide residues in camellia oil was developed. The sample preparation could finish in 5 min without extraction procedure, and then analyzed by gas chromatograph-mass spectrometer. Average recoveries ranged between 73.2 % and 108.6 %, with relative standard deviation values (intra-day and inter-day) lower than 16 % at two concentration levels. The method limit of detection was 5 ng/g, which could meet the regulatory maximum residue limits for the pesticides.

Multiresidue determination of organophorous pesticides in camellia oil by matrix solid-phase dispersion followed by GC-FPD.

A novel analytical approach has been developed and evaluated for the quantitative analysis of 15 organophorous pesticides residues in camellia oils. The proposed methodology is based on acetonitrile/water (3:1, V/V) extraction, followed by matrix solid-phase dispersion, using aminopropyl as dispersant material. Then gas chromatography-flame photometric detection was applied for the pesticide residue analysis. The optimal sorbent quantity was studied. The results demonstrated that the method achieved acceptable quantitative recoveries of 71.5%-104.2% with relative standard deviations <19%, and the method limit of detection at or below the regulatory maximum residue limits for the pesticides were achieved.