Assessing the efficacy and impact of management of an invasive species in a protected area: Poa annua on sub‐Antarctic Macquarie Island

Plant eradication is difficult, particularly in remote, protected areas. The Southern Ocean Islands are very isolated and highly protected, but the flora contains many alien plants. Small restricted populations have been eradicated, but eradication of established species has proven difficult. A better understanding of the efficacy of control methods at sub‐Antarctic temperatures and their off‐target impacts may increase eradication success. With interest in controlling non‐native Poa annua in the region, we aimed to determine if physical and chemical methods can control P. annua (the sub‐Antarctic biotype) in sub‐Antarctic conditions and examined their impact on native plants. We quantified the effectiveness of physical control methods on P. annua in situ on sub‐Antarctic Macquarie Island through field‐based experiments and assessed their selectivity on P. annua compared with native grasses. We also quantified the effectiveness of several herbicides on P. annua at sub‐Antarctic temperatures and assessed their selectivity on native grasses. Of the four physical disturbance methods tested, none effectively reduced P. annua cover as one‐off treatments. Of the herbicide treatments, glyphosate and trifloxysulfuron sodium were effective and were less damaging to native grass species, indicating potential selectivity. Physical control was of limited effectiveness, but did not affect native species richness. An integrated weed management programme utilising the strategic use of selective herbicides with follow‐up chemical and physical intervention may balance control and biodiversity outcomes. This research highlights the importance of site‐specific testing of control methods and understanding off‐target impacts of control when managing alien plant species in protected areas.     

Wide hybridization and introgression breeding in safflower: Effectiveness of different selection methods

Possibility of using interspecific hybridization to increase genetic diversity of drought tolerance in safflower (Carthamus tinctorius L.) and effectiveness of different selection methods is poorly understood. The main objectives of this study were to evaluate the effectiveness of (a) interspecific hybridization to expand genetic diversity in safflower for agronomic traits and drought tolerance in advance generations and (b) different selection methods (direct vs. indirect) for improving seed yield. Interspecific hybridizations were performed between species of C. tinctorius × C. palaestinus (TP), C. palaestinus × C. oxyacantha (PO) and C. tinctorius × C. oxyacantha (TO). Based on the field screening of genotypes in F3 generation, four groups of selected genotypes (derived from combination of direct and indirect selection under both drought and normal environments along with selection based on drought tolerance index (STI)) were evaluated in F4 and F5 generations. Indirect selection for seed yield was significantly more efficient than direct selection. Selection in stress environment resulted to more efficiency than selection in normal environment. No significant difference was observed between direct selection and selection based on STI. The result showed that TP was the best population to combine high seed yield and drought tolerance in safflower.     

Improvement,identification, and target prediction for miRNAs in the porcine genome by using massive,public high-throughput sequencing data

Despite the broad variety of available microRNA (miRNA) research tools and methods, their application to the identification, annotation, and target prediction of miRNAs in nonmodel organisms is still limited. In this study, we collected nearly all public sRNA-seq data to improve the annotation for known miRNAs and identify novel miRNAs that have not been annotated in pigs (Sus scrofa). We newly annotated 210 mature sequences in known miRNAs and found that 43 of the known miRNA precursors were problematic due to redundant/missing annotations or incorrect sequences. We also predicted 811 novel miRNAs with high confidence, which was twice the current number of known miRNAs for pigs in miRBase. In addition, we proposed a correlation-based strategy to predict target genes for miRNAs by using a large amount of sRNA-seq and RNA-seq data. We found that the correlation-based strategy provided additional evidence of expression compared with traditional target prediction methods. The correlation-based strategy also identified the regulatory pairs that were controlled by nonbinding sites with a particular pattern, which provided abundant complementarity for studying the mechanism of miRNAs that regulate gene expression. In summary, our study improved the annotation of known miRNAs, identified a large number of novel miRNAs, and predicted target genes for all pig miRNAs by using massive public data. This large data-based strategy is also applicable for other nonmodel organisms with incomplete annotation information.     

Cyprinid herpesvirus 3 as a potential biological control agent for carp (Cyprinus carpio) in Australia: susceptibility of non‐target species

Carp (Cyprinus carpio L.) is a pest species in Australian waterways, and cyprinid herpesvirus 3 (CyHV‐3) is being considered as a potential biological control (biocontrol) agent. An important consideration for any such agent is its target specificity. In this study, the susceptibility to CyHV‐3 of a range of non‐target species (NTS) was tested. The NTS were as follows: 13 native Australian, and one introduced, fish species; a lamprey species; a crustacean; two native amphibian species (tadpole and mature stages); two native reptilian species; chickens; and laboratory mice. Animals were exposed to 100–1000 times the approximate minimum amount of CyHV‐3 required to cause disease in carp by intraperitoneal and/or bath challenge, and then examined clinically each day over the course of 28 days post‐challenge. There were no clinical signs, mortalities or histological evidence consistent with a viral infection in a wide taxonomic range of NTS. Furthermore, there was no molecular evidence of infection with CyHV‐3, and, in particular, all RT‐PCRs for viral mRNA were negative. As a consequence, the results encourage further investigation of CyHV‐3 as a potential biocontrol agent that is specific for carp.     

Antioxidant activity and lipid peroxidation in Artemia nauplii enriched with DHA‐rich oil emulsion and the effect of adding an external antioxidant based on hydroxytyrosol

Artemia nauplii catabolize polyunsaturated fatty acids (PUFA); in particular, they retroconvert docosahexaenoic acid (DHA, 22:6n‐3), so enrichment is a continuous quest towards increasing PUFA through the use of PUFA‐rich enrichment products. However, optimal conditions during enrichment (aeration, illumination and temperatures around 28°C) tend to accelerate autoxidation of PUFA and the formation of potentially toxic oxidation products. Water‐soluble antioxidants like the polyphenolic compound hydroxytyrosol (3,4‐dihydroxyphenylethanol), a polar molecule found in the water fraction resulting after the milling process of olives, arise as promising compounds to prevent oxidation during Artemia enrichments. We investigated the antioxidant activity and lipid peroxidation in Artemia nauplii during enrichment and the effect of adding an external antioxidant based on hydroxytyrosol during the enrichment with a PUFA‐rich emulsion (M70). For this purpose, the activity of antioxidant enzymes (catalase, superoxide dismutase, glutathione‐S‐transferase, glutathione peroxidase), as well as lipid peroxidation, was determined in enriched and unenriched Artemia nauplii. To validate antioxidant activity and lipid peroxidation, in a first experiment, nauplii were enriched with microalgae (Tetraselmis suecica), yeast (Saccharomyces cerevisiae) and M70 emulsion. In a second experiment, enrichment with a commercial emulsion (DC Super Selco), M70, and a combination of M70 and hydroxytyrosol (Hytolive, HYT) added as an external antioxidant were performed. The combination of M70 with HYT produced the best results, in terms of activity of antioxidant enzymes. The analysis of the fatty acids from total lipids showed that the addition of hydroxytyrosol preserved the DHA percentage of enriched nauplii.     

富硒猕猴桃高产栽培技术

鲜嫩的果肉、独特的风味以及丰富的营养价值,使得猕猴桃被称为“维C之王”,微量元素硒对人体的健康至关重要。江苏北部自然资源丰富,适宜种植富硒猕猴桃,且临近上海等沿海经济带,有利于猕猴桃的保质保量运输和及时供应市场。本文主要分析了江苏北部地区优质猕猴桃的配套种植技术,从苗木的选择到高产措施,为高品质富硒猕猴桃的生产提供技术支持。     

Effect of organic amendment amount on soil nematode community structure and metabolic footprints in soybean phase of a soybean-maize rotation on Mollisols

It has been well documented that organic amendment affects soil nematode community structure. However, little is known about the effect of organic amendment amount on soil nematodes. To assess the effect of the amount of organic amendments on soil nematode community structure and metabolic activity, the community composition, abundance, and metabolic footprints of soil nematodes were determined in a long-term field experiment with various amounts of organic amendment in Northeast China. Fertilization treatments included an unfertilized control (CK), chemical fertilizer without manure amendment (OM₀), manure applied at 7.5 Mg ha^-1 plus chemical fertilizer (OM₁), and manure applied at 22.5 Mg ha^-1 plus chemical fertilizer (OM₂). A total of 46 nematode genera were found. Treatments with the largest amount of organic amendment had the smallest number of plant parasite genera (5), but a largest number of dominant genera (7). Soil nematodes, bacterivores, and fungivores were the most abundant in OM₂, followed by OM₁, and the lowest in OM₀ and CK. Organic amendment increased the enrichment index (EI), and the large amount of organic amendment increased the metabolic footprints of bacterivore (Baf) and fungivore (Fuf) and enrichment footprint (Ef). The relationships between Baf (or Fuf) and the increases in soil organic carbon (ΔSOC) and total nitrogen (ΔTN) were stronger than those of bacterivore (or fungivore) abundance with ΔSOC and ΔTN, except for the relationship between bacterivore abundance and ΔSOC. The EI and Ef were positively correlated with ΔSOC and ΔTN. These findings suggest that the amount of organic amendment affects soil nematode activity and function at entry levels in soil food web, and that metabolic footprints of soil nematodes may be better indicators than their abundances in assessing their relationships with soil nutrients.     

不同温度下TMV侵染枯斑三生烟的LncRNA差异表达

【目的】 筛选不同温度下烟草花叶病毒（Tobacco mosaic virus,TMV）侵染后枯斑三生烟（Nicotiana tabacum var. Samsun NN）差异表达的长链非编码RNA（long non-coding RNA,lncRNA）,研究lncRNA在枯斑三生烟抗性反应中的作用。【方法】 N基因的温度敏感性使枯斑三生烟在25℃时具备对TMV的抗性、在31℃抗性丧失,在这两个温度条件下对枯斑三生烟接种TMV和磷酸盐缓冲盐水（phosphate buffered saline,PBS）,48 h后提取系统叶总RNA,构建链特异性文库后进行深度测序。对测序结果进行过滤后利用HTSeq将有效数据与近缘品种TN90（N. tabacum var. TN90）基因组比对,筛选得到lncRNA后利用FPKM法估计lncRNA的表达水平。通过edgeR筛选差异表达lncRNA（differentially expressed lncRNA,DElncRNA）,并利用qRT-PCR技术对这一结果进行验证。通过共定位及共表达分析预测DElncRNA的靶基因,通过参考基因组注释、GO和KEGG富集分析研究靶基因的功能。【结果】 4个处理共12个样本经lncRNA-seq各测得约8 000万条clean reads,共获得4 737条已知lncRNA、40 169条新lncRNA。其中64个lncRNA在不同温度条件下TMV侵染后存在差异表达,qRT-PCR测定结果显示这些lncRNA的测序正确率在80%左右,表明本研究所得测序数据具备较高的可信度。对DElncRNA进行靶基因预测,发现一些基因同时被25℃下调和31℃上调的DElncRNA靶向。靶基因注释功能丰富,主要参与植物抗病、激素和代谢等生理过程。部分可能与激素通路相关的lncRNA,在25℃下TMV侵染时呈现下调趋势,而在31℃下TMV侵染则呈现上调趋势。GO富集分析显示靶基因主要参与构成膜、囊泡等组分,具备钙、钾离子通道抑制剂活性等分子功能,使相应离子得以转运引发随后的反应,同时也参与发病、抗原加工和呈现、细胞分裂素代谢等生理过程。KEGG分析发现靶基因显著富集在植物激素信号转导通路,25℃下调和31℃上调的DElncRNA靶基因同时富集在激素信号传导、ABC运输蛋白、苯丙烷类生物合成等通路。【结论】 不同温度（25℃和31℃）条件下TMV侵染枯斑三生烟后,长链非编码RNA差异表达,DElncRNA通过作用于激素信号传导、物质转运等过程参与寄主系统获得性抗性反应。研究结果可为揭示植物系统获得性抗性中lncRNA的调控功能以及新型抗病毒技术开发提供依据。     

设施农田土壤重金属污染评价及分区阈值研究

设施农田高强度的耕作模式使土壤重金属累积日益加重，因地制宜地确定其土壤重金属目标值与筛选值对设施农田可持续生产意义重大。本文以石家庄、衡水与唐山为研究区，基于土壤重金属污染风险和生态风险评价，用熵权法得出综合风险等级并确定重金属污染分区阈值。结果表明土壤Cd、Cu、Zn污染最严重，高风险区占比达40.57%。研究区设施农田土壤Cd、Cu、Pb、Zn全量与有效态的目标值分别为0.12、40.17、19.23、116.03 mg·kg^-1与0.02、4.19、0.59、8.80 mg·kg^-1；筛选值分别为0.40、90.27、38.33、170.68 mg·kg^-1与0.11、11.77、1.55、14.24 mg·kg^-1。基于综合评价结果反推分区阈值的方法简单便捷，重金属全量与有效态的目标值及筛选值的确定为研究区设施农田重金属污染的预防提供了多方位的指导。     

mTOR kinase inhibitor CC-223 suppresses human breast cancer cell viability

AIM: To investigate the inhibitory effect of CC-223, an inhibitor of mammalian target of rapamycin (mTOR) kinase, on the viability of human breast cancer cells and its mechanism. METHODS: The inhibitory effect of CC-223 on the viability of MCF-7 cells and MDA-MB-231 cells was measured by CCK-8 assay. The cell cycle distribution of breast cancer cells was examined by flow cytometry. The expression of cell cycle-related proteins and oncoproteins c-Myc and survivin was analyzed by Western blot. RESULTS: CC-223 significantly inhibited the viability of both MCF-7 and MDA-MB-231 cells (P<0.05). CC-223 induced cell cycle arrest in both G₁ phase and G₂/M phase in the MCF-7 cells (P<0.05). However, low concentration of CC-223 treatment resulted in the accumulation of MDA-MB-231 cell cycle in the G₂/M phase, and the cell number in G₁ phase was unaffected. Treatment with CC-223 for 24 h clearly inhibited the protein levels of cyclin B1, cyclin D1 and phosphorylated cell division cycle protein 2 in the breast cancer cells (P<0.05). CC-223 suppressed the expression of c-Myc and survivin in both MCF-7 cells and MDA-MB-231 cells (P<0.05). CONCLUSION: CC-223 inhibits cell viability by blocking cell cycle progression and down-regulating expression of c-Myc and survivin in both MCF-7 and MDA-MB-231 cells.