Nitrogen budgets of a lower subtropical forest as affected by 6 years of over-canopy and understory nitrogen additions.

Although tropical and subtropical regions have replaced temperate regions as the global-change hotspots for increased atmosphere nitrogen (N) deposition, whether the regional forests reach N saturation is still unclear. Understory or floor N addition has been commonly used in N-deposition studies, but the results of such studies have recently been challenged because they fail to account for canopy interception, assimilation, and leaching processes. Here, we conducted a field experiment to quantify the effects of over-canopy and understory N addition on N budgets in a lower subtropical monsoon evergreen broadleaved (LSMEB) forest. We found that the LSMEB forest was not N saturated after receiving additional N at 25 and 50 kg ha-1 yr-1 for 6 years. Plants were able to absorb the added N by increasing the N concentrations in their organs, with 120-412 % increasing trend of plant N pools under N-addition treatments. Canopy absorption of N resulting from over-canopy N addition led to increases in N concentrations in tree organs but not to increases in tree biomass. Understory N addition could underestimate the effects of N deposition in forests due to neglecting canopy N interception and canopy effects on N redistribution. Additional experiments using over-canopy N addition are needed to assess the true effects of N deposition on different forest ecosystems in different climate zones.

Canopy and understory nitrogen additions did not significantly change the community structure of soil fauna under a mature subtropical forest.

Increasing nitrogen (N) deposition has seriously harmed the structure and function of ecosystems throughout the world and this problem has been increasing. How N deposition affects soil faunal communities is poorly understood, compared to plant and microbial communities. Canopy and understory N additions of 25 and 50 kg ha-1 year-1 were employed to determine whether the effects of N addition on the soil fauna differ between N released to the canopy or to the understory. Specifically, we examined how the soil fauna survives when N additions produce desynchronized and complex impacts on the soil, microbes and litter quantity under mature subtropical forest ecosystems. Our results showed that no significant differences were observed between the soil faunal communities receiving canopy and understory N additions. This is consistent with our observation that the concentrations of ammonia nitrogen and nitrate nitrogen in the soil did not significantly differ under the two different methods of applying N. There were no observed effects on the litter quantity, soil microbial Phospholipid Fatty Acids or soil physical-chemical properties; therefore, it is not surprising that N treatments for 4 years did not significantly alter the community structure of soil fauna under the mature subtropical forest sites. However, the shifts in seasonal differences in the microbial communities under the N treatments had a positive effect on soil microbial development compared to control, which might also produce a time-delay influence on the relative development of the soil fauna under mature subtropical forest in the future. Further dynamic monitoring is needed to illustrate the possible effects and mechanisms by which increasing N deposition may alter soil faunal development in the future.

Overexpression of ribosome recycling factor is responsible for improvement of nucleotide antibiotic-toyocamycin in Streptomyces diastatochromogenes 1628.

Ribosome recycling factor (RRF), a product of the frr gene, is responsible for the dissociation of ribosomes from messenger RNA after the termination of translation. In order to overexpress frr gene in the toyocamycin (TM) producer Streptomyces diastatochromogenes 1628, we cloned and placed the gene under the control of the constitutive promoter PermE(*). The resulting plasmid pIB139-frr was integrated into the chromosome of S. diastatochromogenes 1628 by conducting intergeneric conjugation. The strain S. diastatochromogenes 1628 containing pIB139-frr (1628-FRR) showed a 33.3 % increase in cell growth and a 46 % increase in TM production compared to wild-type strain 1628 when cultivated in a 7 l fermentor. In addition, it was possible to shorten the fermentation time from 84 to 72 h. Furthermore, by conducting reverse transcription polymerase chain reaction (RT-PCR) analysis, we discovered that the transcriptional levels of regulatory gene adpA-sd, toyF, and toyG involved in TM biosynthesis were enhanced in S. diastatochromogenes 1628-FRR compared to S. diastatochromogenes 1628. In addition, by using a fluorescent intensity reporter system, which is based on the green fluorescent protein (GFP), and by using Western blot analysis, we revealed that overexpression of frr also strongly promoted protein biosynthesis in late growth phase. These findings confirmed that by increasing copy number of frr gene, it is a useful approach to improve antibiotic production.

Development of intergeneric conjugal gene transfer system in Streptomyces diastatochromogenes 1628 and its application for improvement of toyocamycin production.

Streptomyces diastatochromogenes 1628, capable of producing toyocamycin (TM), has exhibited a potential biocontrol effect in inhibiting the development of phytopathogens in the agriculture field. In this study, an efficient transformation system was developed using the intergeneric conjugation. This was achieved by optimization of experimental parameters. Under optimal conditions, a maximal conjugation frequency of 4.1 × 10(-4) per recipient was obtained. In order to heterologously express the gene vgb encoding Vitreoscilla hemoglobin in S. diastatochromogenes 1628, we placed vgb under the control of the constitutive promoter PermE(*) and constructed plasmid pIB139-vgb. This plasmid was integrated into the chromosome of S. diastatochromogenes 1628 using intergeneric conjugation established above. Finally, strain 1628-VHB-23 with the highest TM production was screened. Results indicated that expression of vgb gene had always significantly promoted the cell growth and TM production in S. diastatochromogenes 1628 under different dissolved oxygen conditions. In particular, under the limited aerobic condition, strain 1628-VHB-23 obtained 33.3 % more DCW and produced 210 % more TM in 7-l fermentor as compared with the wild-type strain.