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Legumes, unlike most land plants, can form symbiotic root nodules with nitrogen-fixing bacteria to secure nitrogen for growth. The formation of nitrogen-fixing nodules on legume roots requires the coordination of rhizobial infection at the root epidermis with cell division in the cortex. The nodules house the nitrogen-fixing rhizobia in organelle-like structures known as symbiosomes, which enable nitrogen fixation and facilitate the exchange of metabolites between the host and symbionts. In addition to this beneficial interaction, legumes are continuously exposed to would-be pathogenic microbes; therefore the ability to discriminate pathogens from symbionts is a major determinant of plant survival under natural conditions. Here, we summarize recent advances in the understanding of root nodule symbiosis signaling, transcriptional regulation, and regulation of plant immunity during legumerhizobium symbiosis. In addition, we propose several important questions to be addressed and provide insights into the potential for engineering the capacity to fix nitrogen in legume and nonlegume plants.

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Naphthalene has been widely used to study the role of soil fauna, but its potential non-target effects on soil enzyme activity remain unknown in subalpine forests. We added naphthalene for two years and determined the effect of such additions on the abundance of soil fauna and soil enzyme activities (β-glucosidase, cellobiohydrolase, invertase, peroxidase, polyphenol oxidase, N-acetyl-β-D-glucosaminidase, leucine arylamidase, urease, nitrate reductase and nitrite reductase) in a subalpine forest. Naphthalene could efficiently suppress the individual density and population of soil fauna in situ . The individual density and number of groups were decreased by 72.6–84.8% and 15.0–28.0%, respectively. Naphthalene significantly affected the activities of β-glucosidase, cellobiohydrolase, polyphenol oxidase, N-acetyl-β-D-glucosaminidase, leucine arylamidase and nitrite reductase and the activity increased in the first litter peak of naphthalene addition, and decreased at the later. The activities of β-glucosidase, cellobiohydrolase, polyphenol oxidase, peroxidase, N-acetyl-β-D-glucosaminidase, leucine arylamidase and nitrite reductase showed a negative correlation with the soil microbial PLFAs. Conversely, the activities of invertase, urease and nitrate reductase were positively correlated with the soil microbial PLFAs. Our results suggest that naphthalene is an effective method to reduce soil fauna in subalpine forest. The enzyme activity was influenced by soil fauna and microbial PLFAs.

Naphthalene has been widely used to test the functional roles of soil fauna, but its nontarget effects remain uncertain in various soils. To determine whether there is a potential nontarget effect on soil biochemical properties in subalpine forest soil, soils in a subalpine forest on the western Qinghai-Tibet Plateau were treated by naphthalene in microcosms. The responses of soil microbial activity and nutrients to naphthalene were studied following 52 days of incubation. The results showed that the naphthalene application obviously decreased the microbial respiration rate in the first 10 days of the incubation and then increased the rate in the following days of the incubation. Moreover, the naphthalene application did not significantly affect the microbial activities overall, measured as soil microbial phospholipid fatty acid (PLFA) abundances and biomasses, or most enzyme activities (invertase, nitrate reductase and nitrite reductase) during the whole incubation period. However, naphthalene suppressed increases in the DON, NH 4 + -N and NO 3 - -N contents and urease activity and led to the net mineralization of inorganic N (NH 4 + -N + NO 3 - -N), in contrast to the net immobilization result in the controls. These results suggest that naphthalene can exert direct nontarget effects on soil microbial respiration and N mineralization processes in subalpine soils. Caution should be taken when using naphthalene to repel soil animals in field experiments.

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Naphthalene is a biocide of soil fauna, particularly of soil arthropods, that has been widely applied to test the functional roles of soil fauna in soil processes. However, whether the use of naphthalene to expel soil fauna has a non-target effect on soil bacteria in subalpine forests remains unclear. We conducted a naphthalene treatment experiment to explore the effects of naphthalene on the soil bacterial community in subalpine forest soil. The results suggested that naphthalene treatment (at 100 g.m −2 per month) significantly increased the abundances of total bacterial, gram-positive bacterial and gram-negative bacterial phospholipid fatty acids (PLFA) and did not change the microbial biomass carbon (MBC), microbial biomass nitrogen (MBN) or MBC/MBN ratio. Moreover, a total of 1038 operational taxonomic units (OTUs) were detected by Illumina MiSeq sequencing analysis. Proteobacteria, Actinobacteria, and Acidobacteria Chloroflexi were the dominant phyla, and Bradyrhizobium was the most abundant genus. The naphthalene treatment did not affect soil bacterial diversity or community structure. Overall, these results demonstrated that the naphthalene treatment had non-target effects on the active bacterial community abundance but not the soil bacterial community structure. Thus, the non-target effects of naphthalene treatment should be considered before using it to expel soil fauna.