Nitric oxide production is associated with response to brown planthopper infestation in rice

Nilaparvata lugens Stål, the brown planthopper (BPH), is one of the most destructive phloem-feeding insects of rice (Oryza sativa L.) throughout Asia. Here, we show that BPH feeding increases the level of endogenous nitric oxide (NO) in the leaf and sheath tissue of both resistant and susceptible rice cultivars. However, in the roots, the NO level increased in the resistant cultivar, but decreased in the susceptible one. A burst of NO production occurred in the sheath within 1 h of infestation with BPH. The production of NO in response to BPH feeding appears to be dependent primarily on the activity of nitric oxide synthase. The application of exogenous NO reduced plant water loss by its effect on both stomatal opening and root architecture. It also stimulated the expression of certain drought stress-related genes, reduced plant height and delayed leaf senescence. Over the short term, NO supplementation reduced the seedling mortality caused by BPH feeding. This suggests that NO signaling plays a role in the rice tolerance response to BPH feeding.     

Testing four proposed barcoding markers for the identification of species within Ligustrum L. (Oleaceae)

DNA barcoding is a biological technique that uses short and standardized genes or DNA regions to facilitate species identification. DNA barcoding has been used successfully in several animal and plant groups. Ligustrum (Oleaceae) species occur widely throughout the world and are used as medicinal plants in China. Therefore, the accurate identification of species in this genus is necessary. Four potential DNA barcodes, namely the nuclear ribosomal internal transcribed spacer (ITS) and three chloroplast (cp) DNA regions (rbcL, matK, and trnH–psbA), were used to differentiate species within Ligustrum. BLAST, character-based method, tree-based methods and TAXONDNA analysis were used to investigate the molecular identification capabilities of the chosen markers for discriminating 92 samples representing 20 species of this genus. The results showed that the ITS sequences have the most variable information, followed by trnH–psbA, matK, and rbcL. All sequences of the four regions correctly identified the species at the genus level using BLAST alignment. At the species level, the discriminating power of rbcL, matK, trnH–psbA, and ITS based on neighbor-joining (NJ) trees was 36.8%, 38.9%, 77.8%, and 80%, respectively. Using character-based and maximum parsimony (MP) tree methods together, the discriminating ability of trnH–psbA increased to 88.9%. All species could be differentiated using ITS when combining the NJ tree method with character-based or MP tree methods. Overall, the results indicate that DNA barcoding is an effective molecular identification method for Ligustrum species. We propose the nuclear ribosomal ITS as a plant barcode for plant identification and trnH–psbA as a candidate barcode sequence.     

Designing a Soluble Near Full-length HIV-1 gp41 Trimer

The HIV-1 envelope spike is a trimer of heterodimers composed of an external glycoprotein gp120 and a transmembrane glycoprotein gp41. gp120 initiates virus entry by binding to host receptors, whereas gp41 mediates fusion between viral and host membranes. Although the basic pathway of HIV-1 entry has been extensively studied, the detailed mechanism is still poorly understood. Design of gp41 recombinants that mimic key intermediates is essential to elucidate the mechanism as well as to develop potent therapeutics and vaccines. Here, using molecular genetics and biochemical approaches, a series of hypotheses was tested to overcome the extreme hydrophobicity of HIV-1 gp41 and design a soluble near full-length gp41 trimer. The two long heptad repeat helices HR1 and HR2 of gp41 ectodomain were mutated to disrupt intramolecular HR1-HR2 interactions but not intermolecular HR1-HR1 interactions. This resulted in reduced aggregation and improved solubility. Attachment of a 27-amino acid foldon at the C terminus and slow refolding channeled gp41 into trimers. The trimers appear to be stabilized in a prehairpin-like structure, as evident from binding of a HR2 peptide to exposed HR1 grooves, lack of binding to hexa-helical bundle-specific NC-1 mAb, and inhibition of virus neutralization by broadly neutralizing antibodies 2F5 and 4E10. Fusion to T4 small outer capsid protein, Soc, allowed display of gp41 trimers on the phage nanoparticle. These approaches for the first time led to the design of a soluble gp41 trimer containing both the fusion peptide and the cytoplasmic domain, providing insights into the mechanism of entry and development of gp41-based HIV-1 vaccines.     

Schistosoma japonicum Soluble Egg Antigens Attenuate Invasion in a First Trimester Human Placental Trophoblast Model

Background

Schistosomiasis affects nearly 40 million women of reproductive age, and is known to elicit a pro-inflammatory signature in the placenta. We have previously shown that antigens from schistosome eggs can elicit pro-inflammatory cytokine production from trophoblast cells specifically; however, the influence of these antigens on other characteristics of trophoblast function, particularly as it pertains to placentation in early gestation, is unknown. We therefore sought to determine the impact of schistosome antigens on key characteristics of first trimester trophoblast cells, including migration and invasion.

Methods

First trimester HTR8/SVneo trophoblast cells were co-cultured with plasma from pregnant women with and without schistosomiasis or schistosome soluble egg antigens (SEA) and measured cytokine, cellular migration, and invasion responses.

Results

Exposure of HTR8 cells to SEA resulted in a pro-inflammatory, anti-invasive signature, characterized by increased pro-inflammatory cytokines (IL-6, IL-8, MCP-1) and TIMP-1. Additionally, these cells displayed 62% decreased migration and 2.7-fold decreased invasion in vitro after treatment with SEA. These results are supported by increased IL-6 and IL-8 in the culture media of HTR8 cells exposed to plasma from Schistosoma japonica infected pregnant women.

Conclusions

Soluble egg antigens found in circulation during schistosome infection increase pro-inflammatory cytokine production and inhibit the mobility and invasive characteristics of the first trimester HTR8/SVneo trophoblast cell line. This is the first study to assess the impact of schistosome soluble egg antigens on the behavior of an extravillous trophoblast model and suggests that schistosomiasis in the pre-pregnancy period may adversely impact placentation and the subsequent health of the mother and newborn.     

Blockage of MDM2-mediated p53 ubiquitination by yuanhuacine restrains the carcinogenesis of prostate carcinoma cells by suppressing LncRNA LINC00665

Prostate cancer (PCa) is a challenging issue for men's health worldwide due to its uncontrolled proliferation and high metastatic potential. Increasing evidence has supported plant extracts and natural plant derivatives as promising antitumor therapy with less toxic side effects. Yuanhuacine is an active component isolated from Daphne genkwa and can effectively suppress the tumorigenesis of several cancers. However, its role in PCa remains unclear. In this study, yuanhuacine dose-dependently inhibited the proliferation and induced apoptosis of PCa cells. Moreover, yuanhuacine also restrained the invasion and migration of PCa cells. Mechanically, yuanhuacine decreased the ubiquitination and degradation of p53 protein, and ultimately increased p53 levels, which was regulated by inhibiting the phosphorylation and total protein levels of mouse double minute 2 (MDM2). Moreover, elevation of MDM2 reversed the suppressive efficacy of yuanhuacine in PCa cell viability, invasion, and migration. The network pharmacologic and bioinformatics analysis confirmed that MDM2 might be a common target of D. genkwa and LINC00665. Furthermore, yuanhuacine inhibited LINC00665 expression. Upregulation of LINC00665 reversed yuanhuacine-mediated inhibition in MDM2 protein expression and suppressed p53 levels by enhancing its ubiquitination in yuanhuacine-treated cells. Importantly, the inhibitory effects of yuanhuacine on cell viability and metastatic potential were offset after LINC00665 elevation. Together, the current findings highlight that yuanhuacine may possess tumor-suppressive efficacy by inhibiting LINC00665-mediated MDM2/p53 ubiquitination signaling. Therefore, this study indicates that yuanhuacine may be a promising candidate for the treatment of PCa.     

燕麦-绿豆间作效应及氮素转移特性

为探究燕麦(Avena sativa)-绿豆(Phaseolus radiatus)间作效应及氮素转移特性, 在不施氮肥的大田试验条件下, 设置3种种植模式(燕麦单作、绿豆单作和燕麦-绿豆间作), 采用传统挖根法和¹⁵N同位素标记法进行研究。结果表明, 间作系统中燕麦侵袭力强于绿豆, 绿豆生长受到抑制。整个生育期, 间作燕麦地上部干物质积累量比单作增加14.9%-33.1%, 2年成熟期间作燕麦的氮素积累量比单作分别提高53.1%和44.8%; 间作减少了开花结荚期绿豆氮素积累量和根瘤重量, 降低了绿豆的固氮效率, 绿豆的固氮效率2年平均降低23.7%, 生物固氮量平均减少11.66%。间作绿豆向燕麦的氮素转移率2年平均值达31.7%, 氮素转移量为212.16 kg∙hm^-2。燕麦-绿豆间作降低了开花结荚期绿豆的根瘤固氮酶活性和固氮效率, 但绿豆体内氮素转移增加了燕麦对氮素的吸收利用, 实现了地上部与地下部生长的相互调节和促进, 优化了农田生态系统的氮素管理。     

火棘不同组织内生细菌群落多样性

为了解火棘不同组织内生细菌群落多样性,该研究采用高通量测序技术对火棘内生细菌16S rRNA V5～V7可变区进行测序,分析火棘不同组织部位内生细菌群落多样性。结果表明:(1)从火棘根、茎、叶组织中共获得内生细菌OTU 1 818个,其中根部754个,茎部 308 个,叶部756个,三者共有 OTU 152 个。(2)物种分类显示,不同火棘组织内生细菌具有丰富的群落多样性,火棘根部内生细菌种类隶属于23门53纲137目216科373属557种,其中异样根瘤菌属(Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium)和链霉菌属(Streptomyces)为优势属,其相对丰度分别为 10.57%和 8.00%; 茎部内生细菌种类隶属于21门32纲76目126科204属270种,其中马赛菌属(Massilia)和未知分类的丛毛单胞菌科属(unclassified_f_Comamonadaceae)为优势属,其相对丰度分别为31.10%和12.82%; 叶部内生细菌种类隶属于21门52纲130目210科380属581种,其中土芽孢杆菌属(Geobacillus)和假单胞菌属(Pseudomonas)为优势属,其相对丰度分别为12.31%和9.84%。(3)PICRUSt功能预测表明,根部内生细菌物种最丰富,参与各种代谢调控的细菌丰度最高。该研究结果为进一步探讨植物内生细菌功能,挖掘新的有益微生物资源提供了参考。     

嗜热蓝藻层理鞭枝藻（Mastigocladus laminosus）藻胆体在解离过程中荧光发射光谱和光能传递的研究

研究了层理鞭枝藻藻胆体在不同浓度磷酸缓冲溶液中解离过程中荧光发射光谱的变化和光能传递。完整藻胆体的７７Ｋ荧光光谱中只有一个峰,位于６８５ｎｍ它是末端发射体（核心－膜连接多肽和别藻蓝蛋白－Ｂ）的荧光峰。部分解离藻胆体的荧光光谱的主峰位移至６５２ｎｍ：次峰位于６８５ｎｍ;６６０ｎｍ为一弱荧光发射肩。它们依次为Ｃ－藻蓝蛋白,末端发射体和别藻蓝蛋白的荧光。严重解离藻胆体的荧光主峰移６４４ｎｍ;次峰由６８５ｎｍ移至６８２ｎｍ;６６０ｎｍ荧光发射肩消失。这表明Ｃ－藻蓝蛋白所捕获的光能已不能传递给别藻蓝蛋白,但可传递给末端发射体洞时又表明Ｃ－藻蓝蛋白不仅与别藻蓝蛋白相连接而且还与末端发射体相连接。提出该藻胆体光能传递链如下：核心－膜连接多肽藻红蓝蛋白→Ｃ－藻蓝蛋白→别藻蓝蛋白别藻蓝蛋白－Ｂ     

棉铃虫乙酰胆碱酯酶的底物专一性和发育期变化(英)

通过对河北省邯郸地区和山东省冠县的棉铃虫(Helicoverpa armigera H(?)bner)乙酰胆碱酯酶(AChE)研究,结果表明,棉铃虫乙酰胆碱酯酶对乙酰硫代胆碱(ATCh)和乙酰-β-甲基硫代胆碱(MeTCh)的比活力以及米氏常数(K_(?))值随其发育阶段呈有规律的变化,AChE比活力在幼虫期呈现两个高峰,一个在三龄,另一个在化蛹前;在蛹期AChE比活力没有明显的变化,而且比活力值比较低;到成虫期第4天有一个明显的高峰,比活力值高于其它任何虫态。K_(?)和最大反应速度(V_(max))的变化趋势基本上与比活力一致。棉铃虫AChE的K_(?)和比活力随其生长发育阶段的周期性变化对于指导用有机磷和氨基甲酸酯类杀虫药剂的化学防治具有重要的意义。不同地点采集到的棉铃虫AChE对AICh和MeTCh水解的活化能有所不同,邯郸地区的棉铃虫AChE水解MeTCh的活化能,蛹和成虫是幼虫期的3.9-4.3倍,而冠县棉铃虫水解MeTCh的活化能则变化不大。AChE水解ATCh的活化能,邯郸地区棉铃虫的AChE不同虫态间变化不大,冠县棉铃虫AChE,幼虫和成虫期约是蛹期的4倍。这说明不同生长发育时期,棉铃虫AChE对底物的水解所消耗的能量是不同的。棉铃虫幼虫AChE的最适反应条件是酶量以重量计为50-100mg,反应时间为10-20min,反应温度为35℃,反应体系的pH值为8.0。