Additive effects of plant expressed double-stranded RNAs on root-knot nematode development

Ectopically expressed double-stranded RNAs (dsRNAs) have recently been shown to suppress parasitic success of Meloidogyne spp. in plants. We have targeted two genes from the root-knot nematode Meloidogyne incognita; a dual oxidase gene implicated in the tyrosine cross-linking of the developing cuticle and a subunit of signal peptidase, a protein complex required for the processing of secreted proteins. While these genes are involved in different aspects of nematode development, the phenotypic consequences of RNA interference (RNAi) were similar with ?50% reduction in nematode numbers in the roots and retardation of development to the egg-producing saccate females. Expression of processed dsRNA was observed, but no evidence of detectable levels of small interfering RNAs (siRNAs) was found in the transgenic plants. We show, to our knowledge for the first time, that combining expression of these dsRNAs by crossing appropriate Arabidopsis thaliana lines resulted in an additive effect that further reduced nematode numbers and developmental capacity. Combining RNAi target genes has the potential to enhance the efficacy of RNAi and may allow control of different nematode species or genera in the crop of interest.     

A plant growth promoting rhizobacterium, Paenibacillus polymyxa strain GBR-1, suppresses root-knot nematode

Exposure of root-knot nematode, Meloidogyne incognita to various concentrations (5-100%) of culture filtrate of Paenibacillus polymyxa GBR-1 under in vitro conditions significantly reduced egg hatch and caused substantial mortality of its juveniles. The increase in the exposure durations of juveniles to culture filtrate and its concentrations increased the mortality rate. Similarly, higher concentrations increased its inhibitory effect on egg hatch. In higher concentrations (25-100%) egg hatch was inhibited by 84-91% after 2 days of exposures as compared to control in sterile distilled water. Application of various concentrations of culture filtrate extract or bacterial suspension of P. polymyxa GBR-1 into potting soil infested with 2000 J2 of M. incognita, reduced the root galling and nematode populations and increased tomato plant growth and root-mass production compared with untreated control (P< or = 0.05). The beneficial effect of P. polymyxa GBR-1 into potted soil increased exponentially with the increase in dose concentrations. Root gall index was reduced from 4.8 to 1.4 and 1.8 when potting soil was treated with 10% concentrations of culture filtrate extract and bacterial suspension, respectively, compared with untreated control. Application of bacterial suspension of P. polymyxa GBR-1 into potted soil at 3 day pre-inoculation of nematode was the most effective followed by simultaneously and at 2 days post-inoculation; as root galling was reduced by 62.5%, 58.3% and 50.0%, respectively, compared with untreated control.     

Interaction of a root-knot nematode (Meloidogyne hapla) and plant intraspecific diversity in clover-grass communities

AimsSoil organisms can influence the plant diversity-productivity relationship at species level; however, little is known about their role in the relationship at an intraspecific level. This study aimed to investigate the interaction between the effects of plant intraspecific diversity and a root-knot nematode on primary production and community evenness.     

根结线虫放线菌及其生物防治活性研究

从感染植物根部的根结线虫卵和雌虫中,分离得到放线菌20株。形态、细胞壁氨基酸组分和16S rRNA基因序列分析表明:分离菌株分属于链霉菌属(Streptomyces)、诺卡氏菌属(Nocardia)和假诺卡氏菌属(Pseudonocardia),其中链霉菌占80.0%。分离菌株对根结线虫Meloidogynespp.卵的平均寄生率、卵的孵化率、幼虫死亡率分别为54.1%、40.4%和26.2%。根据体外测试的结果,选择具有高致病力的3株链霉菌(1-17,2-6,9-47)和1株诺卡氏菌(5-1)进行温室番茄防效测试,其生防效率分别为31.4%、37.7%、56.4%和42.4%。     

Effect of amino acids on root-knot nematode (Meloidogyne javanica) infecting tomato plant

Six amino acids viz. DL-methionine, DL-valine, DL-serine, DL-phenylalanine, L-proline and L-histidine were tested against root knot of tomato caused by Meloidogyne javanica. All amino acids showed significant response in plant growth characters with corresponding reduction in the number of galls, adult females, egg masses and juvenile stages within the treated plants. DL-phenylalanine gave significantly higher response in reducing the hatch of egg masses and survival of juveniles in in vitro test compared to control. The highest plant growth and maximum reduction of galling incidence of tomato were recorded in the DL-phenylalanine- treated plants followed by L-proline and L-histidine. All the amino acids gave positive response in suppressing the development of the nematode in the treated plants.     

A novel small peptide as a targeting ligand for receptor tyrosine kinase Tie2

Tie2 is an endothelium-specific receptor tyrosine kinase known to play an important role in tumor angiogenesis. We sought to identify a small peptide ligand against Tie2 for developing a delivery targeting agent. We used hydrophobic analysis and comparative sequence/structure analysis to select a minimal peptide based on angiopoietin-2 amino acid sequence. The resulting peptide named GA3(WTIIQRREDGSVDFQRTWKEYK) was synthesized and labeled with iodine-125 at the C-terminal tyrosine residue to characterize its binding capability. In in vitro binding assays, GA3 can not only specifically bind to SMMC7721-Tie2 but also compete with angiopoietin-2 in binding. Via mouse tail vein injection, 125I-labeled GA3 was found to favorably accumulate in SPC-A1 xenograft tumor tissues which positively express Tie2. These results demonstrated that GA3 may be useful as a drug or gene delivery ligand for targeted chemotherapy, radiotherapy, and gene therapy.     

Cathepsin H functions as an aminopeptidase in secretory vesicles for production of enkephalin and galanin peptide neurotransmitters

Peptide neurotransmitters function as key intercellular signaling molecules in the nervous system. These peptides are generated in secretory vesicles from proneuropeptides by proteolytic processing at dibasic residues, followed by removal of N- and/or C-terminal basic residues to form active peptides. Enkephalin biosynthesis from proenkephalin utilizes the cysteine protease cathepsin L and the subtilisin-like prohormone convertase 2 (PC2). Cathepsin L generates peptide intermediates with N-terminal basic residue extensions, which must be removed by an aminopeptidase. In this study, we identified cathepsin H as an aminopeptidase in secretory vesicles that produces (Met)enkephalin (ME) by sequential removal of basic residues from KR-ME and KK-ME, supported by in vivo knockout of the cathepsin H gene. Localization of cathepsin H in secretory vesicles was demonstrated by immunoelectron microscopy and immunofluorescence deconvolution microscopy. Purified human cathepsin H sequentially removes N-terminal basic residues to generate ME, with peptide products characterized by nano-LC-MS/MS tandem mass spectrometry. Cathepsin H shows highest activities for cleaving N-terminal basic residues (Arg and Lys) among amino acid fluorogenic substrates. Notably, knockout of the cathepsin H gene results in reduction of ME in mouse brain. Cathepsin H deficient mice also show a substantial decrease in galanin peptide neurotransmitter levels in brain. These results illustrate a role for cathepsin H as an aminopeptidase for enkephalin and galanin peptide neurotransmitter production.     

Effects of some plant extracts on larval hatch of the root-knot nematode,Meloidogyne incognita

Abstract

The inhibitory effect of water extract of seed, leaf and bark of five plants, viz., Tamarindus indica, Cassia siamea, Isoberlinia doka, Dolnix regia and Cassia sieberiana was evaluated on larval hatch of Meloidogyne incognita in the laboratory. All the plant parts inhibited larval hatch of M. incoginta Percentage inhibition was higher in the seeds followed by the leaves and bark. Degree of inhibition observed, was directly related to the concentration of the extract. The standard suspensions inhibited hatching by about 97% while dilutions of S/100 inhibited larval hatch by 3%. Nematicidal activity of the plant parts of the five plants showed that C. siamea was the most effective followed by C. sieberiana, I. doka, T. indica and D. regia.     

Exocyst complexes multiple functions in plant cells secretory pathways

Endomembrane trafficking pathways involving exocyst complexes function. The two established pathways — exocytosis of TGN/EE produced vesicles mediated by EXO70A1 harbouring exocyst and EXO70B1 dependent autophagy related transport to the vacuole — are highlighted by solid arrows; other putative pathways are marked by the dashed arrows. A, autophagosome and autophagy related GA-independent traffic; CW, cell wall; ER, endoplasmic reticulum; GA, Golgi; IVB, intravacuolar bodies; MVB, multivesicular bodies; PM, cytoplasmic membrane; TGN/EE, trans-Golgi network/early endosome; TN, tonoplast; V, vacuole. Exocyst complexes with different EXO70 subunits are symbolized by diamonds connecting the transport containers to the target membranes.

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