Rice-frog cultivation is a traditional farming system in China and has been reintroduced as an agricultural practice in China in recent years. The microbial community in paddy rhizospheric soils has attracted much attention because many microorganisms participate in functional processes in soils. In this study, Illumina MiSeq sequencing-based techniques were used to investigate soil microbial communities and functional gene patterns across samples obtained by conventional rice cultivation (CR) and rice-frog cultivation (RF). The results showed that RF significantly affected the microbial community composition and richness, which indicated that the rhizospheric microorganisms responded to the introduction of tiger frogs into the paddy fields. Operational taxonomic units (OTUs) from Sandaracinaceae, Anaerolineaceae, Candidatus Nitrososphaera, Candidatus Nitrosotalea, Candidatus Nitrosoarchaeum and some unclassified OTUs from Euryarchaeota and Agaricomycetes were significantly enriched by RF. The abiotic parameters soil organic carbon (SOC), nitrate nitrogen (NO₃⁻-N), and available phosphorus (AP) changed under RF treatment and played essential roles in establishing the soil bacterial, archaeal, and fungal compositions. Correlations between environmental factors and microbial communities were described using network analysis. SOC was strongly correlated with Anaerolineaceae, Methanosaeta, and Scutellinia. NO₃⁻-N showed strong positive correlations with Opitutus, Geobacter, and Methanosaeta. NH₄⁺-N was strongly positively associated with Sideroxydans, and TN was strongly positively correlated with Candidatus Nitrotoga. Compared to conventional CR, RF greatly enriched specific microbial taxa. These taxa may be involved in the decomposition of complex organic matter and the transformation of soil nutrients, thus promoting plant growth by improving nutrient cycling. The unique patterns of microbial taxonomic and functional composition in soil profiles suggested functional redundancy in these paddy soils. RF could significantly affect the bacterial, archaeal, and fungal communities though changing SOC and AP levels.

Eco-agricultural systems aim to reduce the use of chemical fertilizers in order to improve sustainable production and maintain a healthy ecosystem. The aim of this study was to explore the effects of rice-frog farming on the bacterial community and N-cycling microbes in paddy rhizosphere soil. This experiment involved three rice cultivation patterns: Conventionally cultivated rice (CR), green rice-frog farming (GR), and organic rice-frog farming (OR). The rice yield, paddy soil enzyme activities, physicochemical variables and bacterial and N-cycling bacterial abundances were quantitatively analyzed. Rice-frog cultivations significantly increased soil protease, nitrate and reductase activity. Additionally, the nirS gene copy number and the relative abundance of denitrifying bacteria also increased, however urease activity and the relative abundance of nitrifying bacteria significantly decreased. The bacterial community richness and diversity of OR soil was significantly higher than that of the GR or CR soil. Nitrogen use efficiency (NUE) of GR was highest. The N-cycling bacterial community was positively correlated with the total carbon (TC), total nitrogren (TN) and carbon to nitrogen (C:N) ratio. The present work strengthens our current understanding of the soil bacterial community structure and its functions under rice-frog farming. The present work also provides certain theoretical support for the selection of rational rice cultivation patterns.

Introducing frogs into paddy fields can control pests and diseases, and organic farming can improve soil fertility and rice growth. The aim of this 2-year field study was compare the yield and elemental composition of rice between an organic farming system including frogs (ORF) and a conventional rice culture system (CR). The grain yields were almost the same in the ORF system and the CR system. The ORF significantly increased the contents of phosphorus (P), ion (Fe), zinc (Zn), molybdenum (Mo) and selenium (Se) in rice grain at one or both years. However, the ORF system decreased the calcium (Ca) content in grice grains, and increased the concentration of cadmium, which is potentially toxic. A principal components analysis showed the main impacts of ORF agro-ecosystem on the rice grain ionome was to increase the concentration of P and trace metal(loid)s. The results showed that the ORF system is an ecologically, friendly strategy to avoid excessive use of chemical fertilizers, herbicides and pesticides without decreasing yields, and to improve the nutritional status of rice by increasing the micronutrient contents. The potential risks of increasing Cd contents in rice grain should be addressed if this cultivation pattern is used in the long term.

SUMMARY: The environmental concern about diffuse pollution from nitrogen (N) fertilizers has led to increased research on the diagnosis of crop N status. The SPAD chlorophyll (Chl) meter is the most commonly used tool for rice (Oryza sativa L.) N status diagnosis, but measurements are conducted at a specific point and readings are affected by different leaf positions. Many measurements per plant must be taken in order to increase the accuracy of N status diagnosis, which limits its application. The present paper attempts to determine rice N status at the canopy level using Multiplex^®, a new hand-held optical fluorescence sensor. The fluorescence emission of rice leaves under light excitation was utilized by Multiplex^® to non-destructively assess rice leaf Chl and phenolic compound content. A field experiment was conducted in 2011 using a completely randomized split-plot design, with main-plot treatments being six N fertilizer application rates and subplot treatments being different plant densities. Leaf Chl and phenolic compounds were evaluated using the ratio of far-red fluorescence (FRF) to red fluorescence (RF) emission under red light excitation (simple fluorescence ratio, SFR_R) (R² = 0·35, P < 0·01) and the ratio of decadic logarithm of red to ultra-violet (UV) fluorescence emission (R² = 0·30, P < 0·01), respectively. Both SPAD reading and fluorescence-based indices including flavonoids (FLAV), nitrogen balance index (NBI_R) and SFR_R could be used to predict rice leaf N contents. The canopy FLAV, SFR_R and NBI_R were all highly correlated to average SPAD readings (R² > 0·70 in most cases, P < 0·01). Therefore, Multiplex^® can be used as an alternative to SPAD to determine rice N status in paddy fields.