[Effects of Organic Fertilizer Combined with Biochar on Denitrifying Microorganisms and Enzyme Activities in Orchard Soil].

To study the effects of organic fertilizer combined with biochar on soil denitrification and denitrifying microbial community structure, this study took lemon orchard soil as the research object and adopted a pot experiment, setting up five fertilization treatments:no fertilization(CK), conventional fertilization(F), organic fertilizer(P), fertilizer+biochar(FP), and organic fertilizer+biochar(PP). The abundance and community structure of denitrifying microorganisms were studied using real-time quantitative PCR and T-RFLP. Redundancy analysis(RDA) was used to explore the environmental factors affecting the denitrifying microbial community structure, and PLS-PM analysis was used to explore the environmental factors affecting the denitrification potential of lemon orchard soil. The results showed as follows:① compared with that under the single fertilizer treatment(F), the organic fertilizer and biochar(P, FP, and PP) treatments significantly increased the denitrification potential of the soil, ranging from 147.8% to 1445.3%. The denitrification rate of soil treated with organic fertilizer combined with biochar was 23.8% lower than that treated with organic fertilizer alone. ② Compared with that in the CK treatment, fertilization treatment significantly increased the abundance of nirS and nirK denitrification microorganisms. Fertilizer treatments(F and FP) significantly reduced the abundance of nosZ denitrifying microorganisms. Biochar treatment significantly changed the diversity and uniformity of denitrifying microorganisms, but the specific law and mechanism quality remained unclear. ③ The results of RDA analysis showed that fertilization could affect the community structure of nirS, nirK, and nosZ denitrifying microorganisms by changing C/N, WC, NO3--N, SOC, AK, and AP. ④ PLS-PM analysis showed that soil denitrification was positively correlated with pH and the abundance of nirK denitrification microorganisms, and NO3--N indirectly affected soil denitrification by affecting the abundance of nirK denitrification microorganisms. In addition, the nirK microbial community was the dominant microbial community in soil denitrification in lemon orchards. In conclusion, organic fertilizer directly affected soil denitrification by regulating soil pH, whereas regulating NO3--N content affected nirK denitrification microbial abundance, indirectly affecting soil denitrification. The application of organic fertilizer combined with biochar could slow down the improvement of soil denitrification caused by single application of organic fertilizer, which is more suitable for promotion in orchards in this region.

[Response of Nitrification Potential and Ammonia Oxidation Microbial Community in Purple Soils to the Application of Biochar Combined with Chemical Fertilizer and Manure].

The effects of manure and chemical fertilizer combined with biochar on nitrification potential and ammonia oxidation microbial change characteristics of purple soils were studied to explore the effects of fertilization measures and soil environmental factors on nitrification potential and ammonia oxidation microbial change characteristics. In this study, purple soil was taken as the research object, and five treatments were set up:no fertilizer(CK), chemical fertilizer(F), manure(P), chemical fertilizer plus biochar(FP), and manure plus biochar(PP). PCR and T-RFLP methods were used to study the characteristics of soil AOA and AOB communities, and soil nitrification potential and environmental factors were measured at the same time to determine the effect of manure combined with biochar on the nitrification potential of purple soil. The results showed that:① compared with that in the CK treatment, the FP and PP treatments significantly increased soil nitrification potential(P<0.05). ② Compared with that in the CK treatment, the F, P, FP, and PP treatments significantly increased the number of copies of the AOA amoA gene in the soil by 78.17%-162.22%(P<0.05), and the F, FP, and PP treatments significantly increased the number of copies of the AOB amoA gene by 21.56%-78.32%(P<0.05). ③ Compared with that in the CK treatment, the PP treatment significantly improved the Shannon, richness, and evenness indices of the soil AOA community(P<0.05), and the combination of biochar(FP and PP treatments) could change the community structure of the soil AOB. ④ The chemical fertilizer and manure affected the AOA and AOB community structure by changing soil properties such as pH, TP, AP, C/N, SOM, NO3--N, and NH4+-N. ⑤ Stepwise regression analysis further showed that AOB amoA gene abundance was the main factor affecting soil nitrification potential. Therefore, under the condition that chemical fertilizer and manure were applied together with biochar, the AOB community was the main driver of the ammonia oxidation process in acidic purple soil, and fertilization affected the ammonia oxidation process by regulating C/N and NO3--N.

[Effect of Chemical Fertilizer and Manure Combined with Biochar on Denitrification Potential and Denitrifying Bacterial Community in Rhizosphere Soil].

To clarify the effect of chemical fertilizer and manure combined with biochar on denitrifying microorganisms and denitrification potential of rhizosphere soil, a pot experiment growing lemon was conducted involving five treatments, namely no fertilization (CK), chemical fertilizer (CF), manure (M), chemical fertilizer combined with biochar (CFBC), and manure combined with biochar (MBC). We determined the characteristics of the rhizosphere soil nirS-, nirK-, and nosZ-type denitrifying bacteria populations; denitrification potential; and soil environmental factors to clarify the effects of chemical and manure combined with biochar on denitrification. Our results showed that compared with that in CK, the CF treatment reduced the rhizosphere soil denitrification potential by 47.7%, whereas the M and MBC treatments increased the denitrification potential by 2192.7% and 1989.9%, respectively. The M and MBC treatments increased the gene copy number of nirS and nosZ, the CF and CFBC treatments decreased the gene copy number of nirS and nosZ, and all four fertilization treatments increased the gene copy number of nirK. Stepwise regression analysis showed that pH was the main factor for the abundance of nirS-type denitrifying bacteria and SOM and NH+4-N were the main factors for the abundance of nirK-type denitrifying bacteria, whereas pH, NO-3-N, and N/P were main factors for the abundance of nosZ-type denitrifying bacteria. The results of partial least squares analysis indicated that the abundance of nirS-and nosZ-type denitrifying bacteria, pH, TN, and N/P were the main factors affecting rhizosphere denitrification potential. Therefore, in acidic purple soil, nirS- and nosZ-type denitrifying bacteria were the main drivers of the soil denitrification process in lemon rhizospheres under chemical fertilizer and pig manure combined with biochar, whereas fertilizer affected the rhizosphere soil denitrification process by regulating soil pH, TN, and N/P.

[Effects of Chemical Fertilizer Reduction Substitute with Organic Fertilizer on Soil Functional Microbes and Lemon Yield and Quality].

To clarify the effects of non-rhizosphere/rhizosphere soil functional microbes (nitrifiers, denitrifiers, and phosphorus-solubilizing microorganisms) on lemon yield and quality, the lemon fruit and non-rhizosphere/rhizosphere soil were selected as subjects. To explore the correlation between non-rhizosphere/rhizosphere soil functional microbes and lemon yield and quality under a chemical fertilizer reduction substitute with organic fertilizer, traditional fruit quality determination and multiple molecular techniques were used. The results showed that:① 30% chemical fertilizer reduction substitute with organic fertilizer increased the nitrification intensity and phosphatase activity but effectively controlled the denitrifying enzyme activity. ② The chemical fertilizer reduction substitute with organic fertilizer significantly decreased the abundances of nitrifiers and nirS/nirK-harboring denitrifiers and significantly increased the abundances of nosZ-harboring denitrifier and phoD-harboring microorganisms. However, the diversities of functional microbial community structure did not have clear regularity under chemical fertilizer reduction substitute with organic fertilizer. ③ Compared with that under the application of chemical fertilizer and organic fertilizer alone, lemon yield and quality were the highest under the 30% reduction of chemical fertilizer substitute with organic fertilizer. ④ Nitrogen and its related microbes significantly affected lemon yield through internal and external quality. Phosphorus and its related microbes affected lemon yield mainly through internal quality. In addition, the influence factors of non-rhizosphere soil and rhizosphere soil on lemon intrinsic quality were obviously different. Altogether, these results showed that the 30% reduction of chemical fertilizer substitute with organic fertilizer significantly affected soil nitrogen and phosphorus functional microorganisms and further improved lemon yield and quality.

[Effects of Chemical Fertilizer Reduction Combined with Organic Fertilizer Application on Bacterial Community Structure in Rhizosphere/Non-Rhizosphere Soil of Lemon].

The aim of this study was to examine the effects of different fertilization methods on the physicochemical properties and bacterial community structure of lemon rhizosphere/non-rhizosphere soil in order to provide theoretical basis for scientific and rational fertilization of orchards. A pot experiment was carried out, and six fertilization treatments were set up:control (CK), conventional fertilization (FM), organic fertilizer (P), fresh organic fertilizer (NP), 70% chemical fertilizer+30% organic fertilizer (70FP), and 50% chemical fertilizer+50% organic fertilizer (50FP). Chemical analysis, real-time fluorescence quantitative PCR, and terminal restriction fragment length polymorphism (T-RFLP) were used to study the effects of different fertilization treatments on the physicochemical properties of rhizosphere and non-rhizosphere soils, the abundance of the bacterial 16S rRNA gene, and bacterial community structure. Redundancy analysis (RDA) was used to explore the environmental factors affecting the bacterial community structure of lemon rhizosphere/non-rhizosphere soil. The results showed the following:① the pH and contents of organic matter, alkali-hydrolyzed nitrogen, available phosphorus, available potassium, and nitrate nitrogen in rhizosphere/non-rhizosphere soil were significantly increased by reducing the amount of chemical fertilizer and applying organic fertilizer (50FP and 70FP) (P<0.05). Compared with conventional fertilization (FM) and single application of organic fertilizer (P and NP), the soil available P content, available K content, and nitrate nitrogen content increased by 24.76%-97.98%, 6.87%-45.11%, and 18.42%-55.82%, respectively. ② Fertilizer reduction combined with organic fertilizer significantly increased the abundance of soil bacteria and soil respiration intensity (P<0.05), and the abundance of soil rhizosphere bacteria and soil respiration intensity under the 50FP treatment increased by 15.83%-232.98% and 8.0%-162.5% compared with that under conventional fertilization and organic fertilizer alone, respectively. The bacterial abundance of rhizosphere soil was positively correlated with the pH and contents of organic matter, total nitrogen, and total phosphorus. ③ The PCoA and RDA analysis results showed that the single organic fertilizer and organic fertilizer and chemical fertilizer de-weighting of rhizosphere bacterial community structure and not adding fertilizer had a bigger difference between processing, and the main environmental factors influencing the rhizosphere/non rhizosphere bacterial community structure were organic matter, total nitrogen, total phosphorus, total potassium, alkali solution nitrogen, nitrate nitrogen, and available potassium. Fertilizer reduction combined with organic fertilizer could significantly increase soil nutrient content, increase soil bacterial abundance, and change the bacterial community structure of rhizosphere soil, and the 50FP treatment yielded better results. Therefore, 50% Chemical fertilizer+50% organic fertilizer (50FP) was a better fertilization method to improve the physical and chemical properties of orchard soil, increase the abundance of soil bacteria, and improve the soil respiration intensity.

[Effect of Combined Application of Biochar with Chemical Fertilizer and Organic Fertilizer on Soil Phosphatase Activity and Microbial Community].

To study changes in phosphatase activity, we examined the diversity of phoC and phoD gene microbial communities in the rhizosphere and non-rhizosphere soil of plants under the treatment of chemical fertilizer and organic fertilizer combined with biochar. These results can provide a certain theoretical guidance for the conversion of insoluble phosphorus in the soil phosphorus pool to the inorganic phosphate ion that can be absorbed by plant roots and also provide a certain experimental basis for the improvement of the availability of phosphorus in the soil and the agricultural utilization of biochar. In this study, corn stalks and rice husk stalks were used as test materials, and the pot experimental method was adopted using the following treatments:set control (CK), traditional fertilization (F), chemical fertilizer+20 t·hm-2 rice husk biochar (FP), chemical fertilizer+10 t·hm-2rice husk biochar+10 t·hm-2 corn biochar (FPM), organic fertilizer+20 t·hm-2 rice husk biochar (PP), and fresh organic fertilizer+20 t·hm-2 rice husk biochar (NPP). We determined the rhizosphere and non-rhizosphere soil acid phosphatase (ACP) activity and alkaline phosphatase (ALP) activity and used T-RFLP technology to analyze the diversity of phoC and phoD genes in order to clarify the impact of biochar on the micro-ecosystem formed by the plants, soil, and microorganisms. The results showed that:① the ALP and ACP activities of each treatment in the non-rhizosphere soil were lower than that of CK. In the rhizosphere soil, the ALP activity was significantly increased after the combined application of chemical fertilizer and organic fertilizer with biochar, and the ACP activity in the rhizosphere soil was higher than that in the non-rhizosphere soil. ② The combined application of biochar with chemical fertilizers and organic fertilizers significantly increased the diversity of phoC and phoD genes communities in rhizosphere and non-rhizosphere soils (P<0.05); the diversity and richness of microbial communities in rhizosphere soil were higher than that in non-rhizosphere soils. ③ ACP activity was negatively correlated with phoC gene microbial community, and most ALP activity was positively correlated with phoD microbial community.

[Responses of Soil PhoC and PhoD Gene Microbial Communities to the Combined Application of Biochar with Chemical Fertilizers and Organic Fertilizers].

Soil microorganisms have an important influence on the transformation of soil nutrients. As functional genes encoding phosphatase, phoC and phoD provide effective means for detecting the types, abundance, and community structure of microorganisms in the environment, and studying the changes in the diversity of phoC and phoD gene microbial communities in the rhizosphere and non-rhizosphere soil of the plant rhizosphere and non-rhizosphere soil under the treatment of chemical fertilizer and organic fertilizer combined with biochar can provide a scientific basis for the agricultural utilization of biochar. In this study, corn stalks and rice husk stalks were used as test materials, and the pot experiment method was used to set the following treatments:control (CK), traditional fertilization (F), chemical fertilizer+20 t·hm-2 rice husk biochar (FP), chemical fertilizer+10 t·hm-2 rice husk biochar+10 t·hm-2 corn biochar (FPM), organic fertilizer+20 t·hm-2 rice husk biochar (PP), and fresh organic fertilizer+20 t·hm-2 rice husk biochar (NPP). The community structure of phoC and phoD genes in rhizosphere and non-rhizosphere soil was analyzed by using T-RFLP and fluorescence quantitative PCR technology to clarify the response characteristics of phoC and phoD genes to the addition of biochar. The results showed that:① In rhizosphere soil and non-rhizosphere soil, the phoD gene community structure was more complicated than that of phoC, and the number of end restriction fragments of the phoC gene increased after chemical fertilizer and organic fertilizer were combined with biochar. ② The combined application of biochar with chemical fertilizer and organic fertilizer reduced the copy number of the phoC gene in non-rhizosphere soil compared with that in the CK. Compared with that in the CK, the copy number in the FP, FPM, PP, and NPP treatments decreased by 9.18%, 11.46%, 10.97%, and 13.76%, respectively. Organic fertilizer combined with biochar increased the copy number of the phoD gene in rhizosphere soil by 2.48% and 5.16% in the PP and NPP treatments, respectively, compared with that in the CK. ③ Total phosphorus in the soil was the main factor affecting the phoC gene microbial community structure in non-rhizosphere soil (P<0.01), whereas the phoC gene microbial community structure in rhizosphere soil was regulated by a variety of environmental factors. pH was the most critical factor affecting the phoD gene copy number, and the copy number of phoD gene was significantly correlated with soil nitrate nitrogen and pH. The combined application of biochar with chemical fertilizers and organic fertilizers can promote the growth and reproduction of microorganisms that function in soil phosphorus conversion, which is of great significance for improving the utilization of phosphorus fertilizers.

[Characteristics of Dissolved Organic Carbon Loss in Purple Soil Sloping Fields with Different Fertilization Treatments].

The characteristics of dissolved organic carbon loss with different fertilization treatments were examined to derive the best nutrient management method for sloping farmland in the Three Gorges Reservoir area where maintaining the soil carbon balance and reducing environmental pollution caused by carbon loss is crucial. Experimental runoff plots were set up at the Experimental Station for Soil and Water Conservation and Environmental Research in the Three Gorges Reservoir Region, Chinese Academy of Sciences, involving the following five treatments:No fertilization (CK), conventional fertilization (conventional), optimum fertilization (optimum), biochar combined with 85% optimum fertilizer (biochar), and straw combined with 85% optimum fertilizer (straw). The effects of the five treatments on runoff flux, sediment yield, and soil organic carbon flux were monitored and evaluated. Results show that:①Subsurface flow accounted for 52.84%-92.23% of the runoff (both surface and subsurface flow) and the loss flux of dissolved organic carbon (DOC) in the subsurface accounted for 43.64%-87.35% of the total loss flux. Thus, in this sloping farmland, subsurface flow was the main pathway of runoff and dominated dissolved organic carbon transport. ②Compared with the optimum treatment, straw treatment reduced the surface runoff flux, sediment yield, surface loss flux of DOC, and loss flux of organic carbon in the sediment by 30.39%, 39.41%, 28.71%, and 23.97%, respectively, but increased the subsurface runoff flux and loss flux of DOC. Compared with the optimum treatment, the biochar treatment significantly increased the surface and subsurface runoff flux, sediment yield, loss flux of DOC in the surface and subsurface, and the loss flux of organic carbon in the sediment. ③The loss flux of DOC accounted for 99.31%-99.94% of the loss flux of soil organic carbon, and DOC was the major species of organic carbon in the organic carbon loss in this type of sloping farmland. The loss flux of DOC under the different fertilization treatments was ranked biochar > optimum > straw > conventional > CK. ④Compared to the optimum treatment, the straw treatment and biochar treatment increased the soil organic carbon (SOC) content by 95.79% and 32.16%, respectively. Based on these results, straw combined with 85% of optimum fertilizer is the best nutrient management method for this sloping farmland as it can reduce surface runoff flux, sediment yield, and the loss flux of soil organic carbon while increasing the soil organic carbon content.

[Response of Nitrogen Loss Flux in Purple Soil Sloping Field to Reduced Fertilizer and Combining Straw].

The purple soil sloping field is considered as the main source of sediment and non-point source pollution in the Three Gorges Reservoir area. To prevent and control the non-point source pollution, it is indispensable to explore the characteristics of nitrogen loss in the overland flow and interflow of purple soil sloping field in Three Gorges Reservoir area. The purple soil sloping runoff plots, located in the Shibaozhai Experimental Station of Chengdu Institute of Chinese Academy of Sciences in Zhongxian County, Chongqing, were studied. The experiment included no fertilization treatment (CK), traditional fertilization treatment (T1), amended fertilization treatment (T2), and reduced fertilizer with straw treatment (T3). According to the data of volume of the interflow and runoff and the leach concentration and flux of nitrogen forms under rapeseed-maize rotation system, the response of nitrogen leaching flux to reduce fertilizer with straw application can be definite in purple soil sloping plots. The results show that the ratio of interflow to total runoff is 60.14%-88.56%, and the flux of nitrogen leaching in the interflow accounts for 72.88%-92.35% of total nitrogen loss flux. Ammonium was mainly leached by the overland flow. In addition, nitrate was mainly leached by the interflow and was the main form of nitrogen leaching. The fluxes of ammonium and nitrate under different treatments followed the order T1 > T2 > T3 > CK. The total nitrogen flux of T3 was 20.07 kg·(hm2·a)-1, which was 43.59% and 39.55% lower than that of T1 and T2, respectively. The reduced fertilizer with straw application significantly decreased the leaching flux of ammonium, nitrate, and total nitrogen, and weakened the effect on runoff nitrogen leaching in the purple soil sloping plots.

[The effect of EDARV370A on facial and ear morphologies in Uyghur population].

The ectodysplasinA receptor gene (EDAR) plays an important role in the development of ectoderm. The derived G allele of its key missense variant EDARV370A is prevalent in East Asians and Americans, but rare in Africans and Europeans. This leads to distinct ectodermal-derived phenotypes between different continental groups, such as the straighter and thicker hair, more eccrine sweat glands, feminine smaller breasts, shovel incisors characteristic of East Asians. At present, we know little about the association between EDARV370A and facial and ear morphology characteristics. To better understand the effect of EDARV370A on craniofacial phenotypes, we systematically examined the association between EDARV370A and 136 facial quantitative phenotypes, one chin ordinal phenotype and six ear ordinal phenotypes in 715 Uyghurs. The quantitative phenotypes were derived by applying our automated landmark annotation method to facial 3D photos and the ordinal phenotypes were manually graded from facial 2D photos. The analysis identified significant association (P<0.05 after multiple testing correction) between EDARV370A and eight facial phenotypes, one chin phenotype and three ear morphology phenotypes. Our study thus elucidated the pleotropic effect of EDARV370A on craniofacial phenotypes in a European-Asian admixed Uyghur population.