The brown planthopper (Nilaparvata lugens Stål; BPH) is a specialist herbivore on rice (Oryza sativa) that ingests phloem sap from the plant through its stylet mouthparts. Electronic penetration graphs revealed that BPH insects spent more time wandering over plants carrying the resistance genes Bph14 and Bph15, but less time ingesting phloem than they did on susceptible plants. They also showed that their feeding was frequently interrupted. Tests with [ 14 C]sucrose showed that insects ingested much less phloem sap from the resistant than the susceptible plants. BPH feeding up-regulated callose synthase genes and induced callose deposition in the sieve tubes at the point where the stylet was inserted. The compact callose remained intact in the resistant plants, but genes encoding b-1,3-glucanases were activated, causing unplugging of the sieve tube occlusions in susceptible plants. Continuing ingestion led to a remarkable reduction in the susceptible plants' sucrose content and activation of the RAmy3D gene, leading to starch hydrolysis and ultimately carbohydrate deprivation in the plants. Our results demonstrate that BPH feeding induces the deposition of callose on sieve plates in rice and that this is an important defense mechanism that prevents insects from ingesting phloem sap. In response, however, the BPH can unplug sieve tube occlusions by activating b-1,3-glucanase genes in rice plants.

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Rice (Oryza sativa L.) and brown planthoppers (BPH) (Nilaparvata lugens Stål) provide an ideal system for studying molecular mechanisms involved in the interactions between plants and phloem-feeding insects. The phenotypic responses and changes in transcript profiles of seedlings representing two rice cultivars differing in resistance to the BPH were analyzed. In the BPH-compatible (susceptible) cv. MH63, BPH feeding reduced three examined plant growth parameters (leaf area expansion, height increases, and dry weight increases) and photosynthetic rates of the leaves. In the BPH-incompatible (resistant) cv. B5, BPH feeding caused slight reductions in protein and sucrose contents, but the plants maintained their photosynthetic activity and grew normally. A cDNA microarray containing 1,920 suppression subtractive hybridization clones was used to explore the transcript profiles differences in the two cultivars under control and BPH-feeding conditions. In total, 160 unique genes were detected as being significantly affected by BPH feeding in rice plants, covering a wide range of functional categories, and there were 38 genes that showed the similar transcript pattern in both genotypes. The physiological responses and transcript profiles of plants represented in both genotypes suggested that multiple pathways might be involved in reprogramming of BPH-infested rice plants. The differences in transcript levels between the compatible and incompatible interactions revealed in this study were not only the reaction of resistance and susceptibility but also reflections of different damage rates and genotypic backgrounds of the rice cultivars.

The New Delhi Metallo-β-lactamase (NDM-1) was first reported in 2009 in a Swedish patient. A recent study reported that Klebsiella pneumonia NDM-1 positive strain or Escherichia coli NDM-1 positive strain was highly resistant to all antibiotics tested except tigecycline and colistin. These can no longer be relied on to treat infections and therefore, NDM-1 now becomes potentially a major global health threat.In this study, we performed modeling studies to obtain its 3D structure and NDM-1/antibiotics complex. It revealed that the hydrolytic mechanisms are highly conserved. In addition, the detailed analysis indicates that the more flexible and hydrophobic loop1, together with the evolution of more positive-charged loop2 leads to NDM-1 positive strain more potent and extensive in antibiotics resistance compared with other MBLs. Furthermore, through biological experiments, we revealed the molecular basis for antibiotics catalysis of NDM-1 on the enzymatic level. We found that NDM-1 enzyme was highly potent to degrade carbapenem antibiotics, while mostly susceptible to tigecycline, which had the ability to slow down the hydrolysis velocity of meropenem by NDM-1. Meanwhile, the mutagenesis experiments, including D124A, C208A, K211A and K211E, which displayed down-regulation on meropenem catalysis, proved the accuracy of our model.At present, there are no effective antibiotics against NDM-1 positive pathogen. Our study will provide clues to investigate the molecular basis of extended antibiotics resistance of NDM-1 and then accelerate the search for new antibiotics against NDM-1 positive strain in clinical studies.