Exploring site-specific N application rate to reduce N footprint and increase crop production for green manure-rice rotation system in southern China.

Milk vetch (Astragalus sinicus L.) is leguminous green manure (GM) which produces organic nitrogen (N) for subsequent crops and is widely planted and utilized to simultaneously reduce the use of synthetic N fertilizer and its environmental costs in rice systems. Determination of an optimal N application rate specific to the GM-rice system is challenging because of the large temporal and spatial variations in soil, climate, and field management conditions. To solve this problem, we developed a framework to explore the site-specific N application rate for the low-N footprint rice production system in southern China based on multi-site field experiments, farmer field survey, and process-based model (WHCNS_Rice, soil water heat carbon nitrogen simulator for rice). The results showed that a process-based model can explain >83.3% (p < 0.01) of the variation in rice yield, aboveground biomass, crop N uptake, and soil mineral N. Based on the scenario analysis of the tested WHCNS_Rice model, the simple regression equation was developed to implement site-specific N application rates that considered variations in GM biomass, soil, and climatic conditions. Simulation evaluation on nine provinces in southern China showed that the site-specific N application rate reduced regional synthetic N fertilizer input by 29.6 ± 17.8% and 65.3 ± 23.0% for single and early rice, respectively; decreased their total N footprints (NFs) by 23.4% and 49.3%, respectively; and without reduction in rice yield, compared with traditional farming N practices. The reduction in total NF was attributed to the reduced emissions from ammonia volatilization by 35.2%, N leaching by 28.4%, and N runoff by 32.7%. In this study, we suggested a low NF rice production system that can be obtained by combining GM with site-specific N application rate in southern China.

[Effects of Chinese Milk Vetch Returning Incorporated with Chemical Fertilizer Reduction on the Composition and Function of Soil Bacterial Communities in Paddy Fields].

Chinese milk vetch (Astragalus sinicus L.) is an important organic nutrient resource in the southern Henan rice-growing area. Thus, the effects of Chinese milk vetch (MV) returning incorporated with reduced chemical fertilizer on the physicochemical properties and bacterial community characteristics in paddy soil were studied. These results can provide a certain theoretical basis for the improvement of soil fertility and reduction of chemical fertilizer in this area. A field experiment was conducted for 12 consecutive years, involving six fertilization treatments (blank control, CK; 100% chemical fertilizer, F100; 80% chemical fertilizer+22.5 t·hm-2 MV, MV1F80; 80% chemical fertilizer+45 t·hm-2 MV, MV2F80; 60% chemical fertilizer+22.5 t·hm-2 MV, MV1F60; and 60% chemical fertilizer+45 t·hm-2 MV, MV2F60). The high-throughput sequencing method was used to compare the effects of different fertilization treatments on soil bacterial community diversity, composition, and structural characteristics. The FAPROTAX function prediction method was used to analyze the abundance differences of functional groups between different fertilization treatments. Additionally, combined with soil physicochemical properties and bacterial community characteristics, we explored the key soil environmental factors that changed the structure and functional characteristics of the soil bacterial community. Compared with that under CK, the soil bulk density (BD) under the MV returning incorporated with reduced chemical fertilizer treatment was decreased, whereas soil organic carbon (SOC), total nitrogen (TN), total phosphorus (TP), and total potassium (TK) were increased by 12.7%-35.5%, 38.2%-65.7%, 66.7%-95.2%, and 20.3%-31.6%, respectively. Compared with that under the F100 treatment, the Sobs index and Shannon diversity index of the bacterial community under the MV returning incorporated with reduced chemical fertilizer were decreased, and the Sobs index and Shannon diversity index were significantly positively correlated with BD (P<0.05) but significantly negatively correlated with SOC and TN (P<0.05). Compared with that under the F100 treatment, the relative abundances of Firmicutes under the MV1F80 and MV2F60 treatments were significantly increased by 82.2% and 67.4% (P<0.05), but the relative abundances of Acidobacteria were significantly reduced by 32.6% and 40.5% (P<0.05), respectively. The relative abundance of Actinobacteria under the MV2F60 treatment was significantly increased by 30.0% (P<0.05) compared with that under the F100 treatment. According to RDA analysis, soil SOC, TN, and TK were the main soil environmental factors that significantly affected bacterial community (P<0.05). Compared with that under CK and the F100 treatment, the abundance of functional groups of chemoheterotrophy, nitrogen fixation, fermentation, and ureolysis under the MV returning incorporated with reduced chemical fertilizer treatment were improved, whereas the abundance of functional groups of animal parasites or symbionts, all human pathogens, and human pathogen pneumonia were reduced, particularly under MV1F80 and MV2F60. To summarize, the long-term MV returning to the field incorporated with reduced chemical fertilizer improved the soil physical and chemical properties, thus changing the structure and functional characteristics of the soil bacterial communities, contributing to the improvement in the soil fertility, stability, and health of micro-ecosystems in paddy fields, thus ensuring the green and sustainable development of regional agriculture.

Study on the relationships between the accumulation and translocation of dry matter and nitrogen and flower/pod development into seeds and seed yields in Chinese milk vetch.

Further improvements to the yield potential of Chinese milk vetch seed are essential for the planting demand of green manure. Flower and pod development directly determines the number of seeds and the seed yield of Chinese milk vetch. However, the accumulation and translocation of dry matter and nitrogen between plant organs directly affects flower and pod development and morphological formation. There are few studies that analyse the relationship between the accumulation and transport of dry matter and nitrogen and the number of flowers, pods, grains and seed yield during Chinese milk vetch's critical development period. This study aimed to determine the seed yield response to dry matter and nitrogen accumulation and translocation during the Chinese milk vetch growth period and to quantify the relationship between these factors to predict Chinese milk vetch seed yield. Experiments were performed during the 2017-2018 and 2018-2019 growing seasons at the Dayuzhuang experimental field. The first experiment involved five foliar application stages (late wintering stage, returning green stage, squaring stage, pre-flowering stage, and 5 days after flowering) and six foliar application concentrations of borate solution (0, 500, 1000, 2000, 4000, and 6000 mg L-1). Experiment 2 included five foliar application stages (late wintering stage, returning green stage, squaring stage, pre-flowering stage, and 5 days after flowering) and six foliar application concentrations of paclobutrazol (0, 200, 300, 400, 500, and 600 mg L-1). When the dry matter mass in the full flowering stage was 3500-4500 kg hm-2, the seed yield reached more than 800 kg hm-2. When the translocated assimilates were stored in the vegetative organs before flowering, the assimilate translocation rate and their contributions to seed yield were 1500-1800 kg hm-2, 30-35%, and 28-38%, respectively, and the Chinese milk vetch seed yield was predicted to reach 800-1000 kg hm-2 at maturity. When the nitrogen translocation amount in the vegetative organs before flowering, the nitrogen translocation rate, and the contribution rate to the seed yield were 68-78 kg hm-2, 65-75%, and 75-85%, respectively, the Chinese milk vetch seed yield was predicted to reach 800-1000 kg hm-2 at maturity. If the accumulation and translocation index values of dry matter and nitrogen were lower or higher than the above ranges, the seed yield was lower than 800 kg hm-2. The results of this study revealed the mechanism by which dry matter and nitrogen accumulation and translocation affect the Chinese milk vetch seed yield. These findings enrich the seed yield formation theory of Chinese milk vetch. They provide an early determination and quantitative regulation of high and stable seed yield for Chinese milk vetch in the field and aid researchers to integrate multiple production technologies for the sustainable production of Chinese milk vetch.

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Clarifying the pattern of nitrogen absorption and utilization of rice under the treatments of Astragalus sinicus combined with chemical fertilizer application and the pattern of absorption, utilization, distribution and residue of A. sinicus nitrogen in rice-soil system could provide basis to rational fertilization for rice planting area in southern Henan. In this study, undisturbed soil column simulation and isotope tracer technology of 15N were used to examine the differences of nitrogen uptake and utilization of rice, nitrogen nutrient balance of rice-soil system and nitrogen uptake, utilization, distribution and residue of A. sinicus nitrogen after mineralization and decomposition among seven treatments. The treatments involved 1) no fertilization (CK); 2) chemical fertilizer+22500 kg·hm-2 A. sinicus (FM1); 3) chemical fertilizer+30000 kg·hm-2 A. sinicus (FM2); 4) chemical fertilizer+37500 kg·hm-2 A. sinicus (FM3); 5) chemical fertilizer+22500 kg·hm-2 A. sinicus +lime (FM1+CaO); 6) chemical fertilizer+30000 kg·hm-2 A. sinicus lime (FM2+CaO); 7) chemical fertilizer+37500 kg·hm-2 A. sinicus +lime (FM3+CaO). Results showed that compared with CK, fertilization significantly increased nitrogen uptake of grain and rice stalks, apparent nitrogen loss, and nitrogen surplus. The grain nitrogen uptake, rice straw nitrogen uptake and nitrogen use efficiency of rice increased firstly and then decreased with the increasing A. sinicus application rates, while the apparent nitrogen loss and nitrogen surplus showed the opposite trend. The best performance was presented under the treatment of chemical fertilizer combined with 30000 kg·hm-2 of A. sinicus. Lime addition could increase grain nitrogen uptake, rice straw nitrogen uptake, and nitrogen use efficiency of rice, while reducing apparent nitrogen loss and nitrogen surplus, with the best performance of FM2+CaO. For all the treatments, the proportion of nitrogen absorbed by rice from A. sinicus was 6.3%-13.2%, while that from soil and chemical fertilizer was 86.8%-93.7%. The utilization ratio of A. sinicus nitrogen by rice was 23.8%-33.6%. The utilization ratio of A. sinicus nitrogen in different parts of rice was grain > stem and leaf > root. The residue rate of A. sinicus nitrogen in soil was 37.6%-62.4%. The loss rate of A. sinicus nitrogen was 7.8%-38.6%. Comprehensively considering nitrogen absorption and utilization of rice, nitrogen nutrient balance of rice-soil system, and the distribution situation of nitrogen from A. sinicus in rice, FM2+CaO was the optimum fertilization pattern in the study area.

Flower and pod development, grain-setting characteristics and grain yield in Chinese milk vetch (Astragalus sinicus L.) in response to pre-anthesis foliar application of paclobutrazol.

The number of grains per unit land area is the most important grain yield component in Chinese milk vetch. Flower and pod survival seem to be critical determinants of grain number, which is related to the number of fertile flowers and pods during the anthesis period. Flower and pod growth are frequently considered the key determinants to establish grain number. The objective of this study was to explore the influences of paclobutrazol on flower and pod development, grain-setting characteristics and grain yield in Chinese milk vetch under different concentrations of foliar spray and try to explore the physiological regulatory mechanisms. Field experiments were carried out during the 2017-2018 and 2018-2019 growing seasons at the Dayuzhuang experimental field. The experiment involved the Chinese milk vetch cultivar "Xinzi No. 1" and six levels of foliar application of paclobutrazol, 0, 200, 300, 400, 500, and 600 mg L-1, in treatments CK, T1, T2, T3, T4, and T5, respectively. Foliar spray was applied once, at the squaring stage. In comparison with the CK treatment, all of the paclobutrazol treatments yielded, to various degrees, increased values of the number of inflorescences per unit area, number of pods per unit area, grain-setting rate of pods, and number of grains per pod in all six inflorescence layers, with the largest increases observed in the T3 treatment. In the T3 treatment compared with the CK treatment, from the first to sixth inflorescence layers, the number of inflorescences per unit area was increased by 34.07-58.97%, the number of pods per unit area was increased by 39.69-68.35%, the grain number per pod was increased by 44.31-53.69%, and the grain-setting rate of pods was increased by 1.84-4.89%. An analysis of yield composition revealed that the paclobutrazol spray treatment had little impact on the grain weight of Chinese milk vetch. The correlations between the concentration of paclobutrazol spray and the grain yield of Chinese milk vetch reached a significant level. Grain yield was highest at the paclobutrazol concentration of 373.10 mg/L. The inflorescence contents of gibberellic acid 3 (GA3), indole-3-acetic acid (IAA), and abscisic acid (ABA) were reduced, whereas that of cytokinin (CTK) was increased, by foliar application of paclobutrazol (400 mg L-1, T3 treatment) relative to CK treatment during the stages of flowers and pods developing into grains.

Mesorhizobium jarvisii is a dominant and widespread species symbiotically efficient on Astragalus sinicus L. in the Southwest of China.

In order to identify rhizobia of Astragalus sinicus L. and estimate their geographic distribution in the Southwest China, native rhizobia nodulating A. sinicus were isolated and their genetic diversity were studied at 13 sites cultivated in four Chinese provinces. A total of 451 rhizobial isolates were trapped with A. sinicus plants from soils and classified into 8 different genotypes defined by PCR-based restriction fragment length polymorphism (RFLP) of 16S-23S rRNA intergenic spacer (IGS). Twenty-one representative strains were further identified into three defined Mesorhizobium species by phylogenetic analyses of 16S rRNA genes and housekeeping genes (glnII and atpD). M. jarvisii was dominant accounting for 76.3% of the total isolates, 22.8% of the isolates were identified as M. huakuii and five strains belonged to M. qingshengii. All representatives were assigned to the symbiovar astragali by sharing high nodC sequence similarities of more than 99%. Furthermore, the biogeography distribution of these rhizobial genotypes and species was mainly affected by contents of available phosphorus, available potassium, total salts and pH in soils. The most remarkable point was the identification of M. jarvisii as a widespread and predominant species of A. sinicus in southwest of China. These results revealed a novel geographic pattern of rhizobia associated with A. sinicus in China.

Rhizobium changzhiense sp. nov., isolated from effective nodules of Vicia sativa L. in North China.

Three fast-growing rhizobial strains isolated from effective nodules of common vetch (Vicia sativa L.) were characterized using a polyphasic approach. All three strains were assigned to the genus Rhizobium on the basis of the results of 16S rRNA gene sequence analysis. Phylogenetic analysis based on concatenated atpD-recA genes separated the strains into a distinct lineage represented by WYCCWR 11279T, which showed average nucleotide identity values of 95.40 and 93.61 % with the most similar phylogenetic type strains of Rhizobium sophorae CCBAU 03386T and Rhizobium laguerreae FB TT, respectively. The digital DNA-DNA hybridization relatedness values between WYCCWR 11279T and the closest related type strains were less than 70 %. Therefore, a novel rhizobial species is proposed, Rhizobium changzhiense sp. nov., and strain WYCCWR 11279T (=HAMBI 3709T=LMG 31534T) is designated as the type strain for the novel species.