Pseudomonas rhizoryzae sp. nov., isolated from rice.

Two yellow-pigmented, Gram-stain-negative and rod-shaped bacterial strains, designated as RY24T and ZYY160, were isolated from rice. Results of phylogenetic analysis based on 16S rRNA gene sequences showed that strains RY24T and ZYY160 belonged to the genus Pseudomonas, and the 16S rRNA gene sequence similarity was 100 % The DNA homology between the two strains was 99.7 %. The 16S rRNA and rpoD gene sequences of the two strains showed highest similarity values to Pseudomonas oryzihabitans CGMCC 1.3392T and Pseudomonas psychrotolerans DSM 15758T (sharing 99.31 and 94.34 %, respectively). The major fatty acids of two strains were identified as summed feature 8 (C18:1ω7c and/or C18:1ω6c), C16;0 and summed feature 3 (C16:1ω7c and/or C16:1ω6c), and the major respiratory quinone was identified as ubiquinone Q-9, which are typical chemotaxonomic features of members of the genus Pseudomonas. The genomic DNA G+C contents of strains RY24T and ZYY160 were determined to be 64.25 and 64.21 mol%, respectively. The DNA-DNA relatedness and average nucleotide identity values between the two strains and their closely related type strains were below 36 and 90 %, which supported that RY24T and ZYY160 represent a novel species in the genus Pseudomonas. Phylogenetic and chemotaxonomic evidence, together with phenotypic characteristics, showed that the two isolates constitute a novel species of the genus Pseudomonas. The type strain is RY24T (JCM 33201T=ACCC 61555T), for which the name Pseudomonas rhizoryzae sp. nov. is proposed.

Dickeya oryzae sp. nov., isolated from the roots of rice.

A novel Gram-stain-negative strain, designated ZYY5T, was isolated from rice roots. Results of 16S rRNA gene analysis indicated that strain ZYY5T was a member of the genus Dickeya, with a highest similarity to Dickeya zeae DSM 18068T (98.5%). The major fatty acids were summed feature 3 (C16:1 ω7c and/or C16:1 ω6c), C16:0 and summed feature 8 (C18:1 ω7c and/or C18:1 ω6c). Multi-locus sequence analysis using five concatenated genes (16S rRNA, atpD, infB, recA and gyrB) and phylogenomic analysis based on 2940 core gene sequences showed that strain ZYY5T formed a robust cluster with strains EC1, ZJU1202, DZ2Q, NCPPB 3531 and CSL RW192, while separated from the other strains of D. zeae. The orthologous average nucleotide identity (ANI) and digital DNA-DNAhybridization (dDDH) values among these six strains ranged from 96.8-99.9% and 73.7-99.8%, which supported that they were belonged to the same species. However, strain ZYY5T shared 58.4 of dDDH and 94.5% of ANI values with type strain D. zeae DSM 18068T, which were lower than the proposed species boundary cut-off for dDDH and ANI. The genomic analysis revealed that strain ZYY5T contained virulence-associated genes, which is same as the phylogenetic-related strains of the genus Dickeya. Based on the results of the polyphasic approaches, we propose that strain ZYY5T represents a novel species in the genus Dickeya, for which the name Dickeya oryzae sp. nov. (=JCM 33020 T=ACCC 61554 T) is proposed. Strains EC1, ZJU1202, DZ2Q, NCPPB 3531 and CSL RW192 should also be classified in the same genomospecies of D. oryzae same as ZYY5T.

Gracilibacillus oryzae sp. nov., isolated from rice seeds.

A Gram-stain-positive, facultatively anaerobic, endospore-forming bacterium, designated strain TD8T, was isolated from surface-sterilized rice seeds (Oryza sativa L.). Phylogenetic analysis of the 16S rRNA gene indicated that strain TD8T should be placed within the genus Gracilibacillus (95.2-99.0 % sequence similarity); it exhibited highest similarities to Gracilibacillus ureilyticus CGMCC 1.7727T (99.0 %), 'Gracilibacillus xinjiangensis' CGMCC 1.12449T (98.9 %) and Gracilibacillus dipsosauri CGMCC 1.3642T (97.5 %). Chemotaxonomic analysis showed that menaquinone-7 (MK-7) was the major isoprenoid quinone. Diphosphatidylglycerol, phosphatidylglycerol and one unidentified phospholipid were the major cellular polar lipids, and the major fatty acids were anteiso-C15 : 0, anteiso-C17 : 0, iso-C15 : 0, C16 : 0 and iso-C16 : 0, which supported the allocation of the strain to the genus Gracilibacillus. The digital DNA-DNA hybridization value between strain TD8T and Gracilibacillus ureilyticus CGMCC 1.7727T was lower than 70 % (22.60 %), and the average nucleotide identity score was 79.54±5.09 %, suggesting that strain TD8T represented a novel species in the genus Gracilibacillus. The genomic DNA G+C content was 37.5 %. Based on physiological and biochemical characteristics and genotypic data, strain TD8T represents a novel species of the genus Gracilibacillus, for which the name Gracilibacillus oryzae sp. nov. is proposed. The type strain is TD8T (=ACCC 61556T=CICC 24889T=JCM 33537T).

Bacillus oryzisoli sp. nov., isolated from rice rhizosphere.

The taxonomy of strain 1DS3-10T, a Gram-staining-positive, endospore-forming bacterium isolated from rice rhizosphere, was investigated using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences demonstrated that the novel strain was grouped with established members of the genus Bacillus and appeared to be closely related to the type strains Bacillus benzoevorans DSM 5391T (97.9 %), Bacillus circulans DSM 11T (97.7 %), Bacillus novalis JCM 21709T (97.3 %), Bacillus soli JCM 21710T (97.3 %), Bacillus oceanisediminis CGMCC 1.10115T (97.3 %) and BacillusnealsoniiFO-92T (97.1 %). The fatty acid profile of strain 1DS3-10T, which showed a predominance of iso-C15 : 0 and anteiso-C15 : 0, supported the allocation of the strain to the genus Bacillus. The predominant menaquinone was MK-7 (100 %). The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and unknown aminolipids. Cell-wall peptidoglycan contained meso-diaminopimelic acid. DNA-DNA hybridization values between strain 1DS3-10T and the type strains of closely related species were 25-33 %, which supported that 1DS3-10T represented a novel species in the genus Bacillus. The results of some physiological and biochemical tests also allowed the phenotypic differentiation of strain 1DS3-10T from the most closely related recognized species. On the basis of the phylogenetic and phenotypic evidence, strain 1DS3-10T represents a novel species of the genus Bacillus, for which the name Bacillus oryzisoli sp. nov. is proposed. The type strain of the novel species is 1DS3-10T (=ACCC 19781T=DSM 29761T).

Novosphingobium oryzae sp. nov., a potential plant-promoting endophytic bacterium isolated from rice roots.

A novel endophytic bacterium, strain ZYY112T, isolated from rice roots, was characterized by a polyphasic approach. In phylogenetic analyses based on 16S rRNA gene sequences, ZYY112T showed highest sequence similarity to Novosphingobium sediminicola HU1-AH51T (97.2 %) and less than 97 % similarity with respect to other Novosphingobium species with validly published names. The DNA G+C content of strain ZYY112T was 60.8 mol%. The level of DNA-DNA relatedness between strain ZYY112T and N. sediminicola DSM 27057T was 33.7 % (reciprocal 5.2 %), which supported the suggestion that ZYY112T represented a novel species of the genus Novosphingobium. Ubiquinone Q-10 was the unique respiratory quinone (100 %). The polar lipid profile contained diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, sphingoglycolipid, an unknown aminolipid and an unknown phospholipid. The major fatty acids of strain ZYY112T were summed feature 8 (consisting of C18 : 1ω7c and/or C18 : 1ω6c), summed feature 3 (consisting of C16 : 1ω7c and/or C16 : 1ω6c), C14 : 0 2-OH and C16 : 0. The major polyamine of ZYY112T was spermidine, which is a characteristic trait of the genus Novosphingobium. Characterization by genotypic, chemotaxonomic and phenotypic analysis indicated that strain ZYY112T represents a novel species of the genus Novosphingobium, for which the name Novosphingobium oryzae sp. nov. is proposed. The type strain is ZYY112T ( = ACCC 06131T = JCM 30537T).

Rhizobium oryzicola sp. nov., potential plant-growth-promoting endophytic bacteria isolated from rice roots.

Bacterial strains ZYY136(T) and ZYY9 were isolated from surface-sterilized rice roots from a long-term experiment of rice-rice--Astragalus sinicus rotation. The 16S rRNA gene sequences of strains ZYY136(T) and ZYY9 showed the highest similarity, of 97.0%, to Rhizobium tarimense PL-41(T). Sequence analysis of the housekeeping genes recA, thrC and atpD clearly differentiated the isolates from currently described species of the genus Rhizobium. The DNA-DNA relatedness value between ZYY136(T) and ZYY9 was 82.3%, and ZYY136(T) showed 34.0% DNA-DNA relatedness with the most closely related type strain, R. tarimense PL-41(T). The DNA G+C content of strain ZYY136(T) was 58.1 mol%. The major cellular fatty acids were summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c), C16 : 0 and C16 : 0 3-OH. Strains ZYY136(T) and ZYY9 could be differentiated from the previously defined species of the genus Rhizobium by several phenotypic characteristics. Therefore, we conclude that strains ZYY136(T) and ZYY9 represent a novel species of the genus Rhizobium, for which the name Rhizobium oryzicola sp. nov. is proposed (type strain ZYY136(T) = ACCC 05753(T) = KCTC 32088(T)).

Paenibacillus rhizoryzae sp. nov., isolated from rice rhizosphere.

A Gram-stain-positive, endospore-forming, rod-shaped bacterium, designated 1ZS3-5(T), was isolated from rice rhizosphere in Hunan Province, PR China. The isolate was identified as a member of the genus Paenibacillus on the basis of phenotypic characteristics and phylogenetic inference analysis. The 16S rRNA and rpoB gene (ß-subunit of bacterial RNA polymerase) sequences were closely related to those of Paenibacillus taihuensis CGMCC 1.10966(T) with similarities of 97.2% and 89.7%, respectively. The DNA-DNA hybridization value between 1ZS3-5(T) and P. taihuensis CGMCC 1.10966(T) was 33.4%. The DNA G+C content of 1ZS3-5(T) was 47.5 mol%. The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, unidentified aminophospholipid and unknown phospholipid. The predominant respiratory quinone was MK-7. The diamino acid found in the cell-wall peptidoglycan was meso-diaminopimelic acid. The major cellular fatty acids were anteiso-C15 : 0, iso-C15 : 0 and iso-C16 : 0. Based on these results, 1ZS3-5(T) is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus rhizoryzae sp. nov. is proposed. The type strain is 1ZS3-5(T) ( = ACCC 19782(T) = DSM 29322(T)).

Long-term rice and green manure rotation alters the endophytic bacterial communities of the rice root.

This study focuses on the effects of long-term rice rotated with milk vetch being as green manure on the composition of bacteria in rice roots. The endophytic bacterial communities in rice roots of the rice-rice-milk vetch (R-R-MV) and the rice-rice-winter fallow (R-R-WF) crop rotations with a 28-year research history were investigated using combined culture-dependent and culture-independent methods. It was found that the endophytic bacterial population in rice roots with the green manure was significantly higher than that of without it. There were 169 and 77 strains of endophytic bacteria that were isolated from rice roots of the R-R-MV and the R-R-WF, respectively. The 16S rRNA gene analysis shows that the 77 R-R-WF bacteria belong to 15 species of 14 genera while the other 169 R-R-MV bacteria belong to 21 species of 19 genera, in which Herbaspirillum and Cedecea were two mutually dominant populations and Burkholderia, Pseudomonas, Sphingomonas, and Pantoea accounted for large proportions of the endophytic bacteria in rice roots through R-R-MV rotation. The analysis of 16S rDNA clone libraries showed that the Shannon-Weaver diversity index of endophytic bacteria in R-R-MV approximates that in R-R-WF rotation, whereas the richness indexes of Chao 1 and ACE in R-R-MV rotation system were significantly higher than those in R-R-WF rotation. The diversity of endophytic bacteria was richer in R-R-MV. Both the culture-dependent and the culture-independent method revealed significant effect of long-term different tillage systems on the microbial community.

Effects of green manure planted in winter and straw returning on soil aggregates and organic matter functional groups in double cropping rice area.

To explore the mechanism of exogenous organic materials enhancing soil organic carbon and soil fertility, based on a long-term experiment located in Hengyang Red Soil Experimental Station, we examined the effects of winter green manure and straw returning patterns (CK, winter fallow; MV, winter Chinese milk vetch; S, early-season rice straw total returning; DS, early-season and late-season rice straw total returning; SMV, winter Chinese milk vetch + early-season rice straw total returning; DSMV, winter Chinese milk vetch + early-season and late-season rice straw total returning) on soil aggregates and organic functional groups. The results showed that the proportion of super aggregates (>2 mm) and macroaggregates (0.25-2 mm) in double cropping rice soil was the highest with a ratio of about 72.1%-81.8%, and the organic carbon content in the two kinds of aggregates was as high as 12.1-20.7 g·kg-1, accounting for 22.7%-59.0% of the total organic carbon. The main organic functional group in paddy soil was polysaccharides, followed by aliphatic carbon and aromatic carbon. The abundance of all those groups was affected by winter Chinese milk vetch growing and straw returning. Compared with other treatments, DSMV significantly increased the proportion of super aggregates (>2 mm) and macroaggregates (0.25-2 mm) and favored the accumulation of inert carbon such as aromatic carbon in the two kinds of aggregates. DSMV could enhance the stability of soil aggregates and organic matter, which had high potential in the real agricultural production.

[Effects of different fertilization regimes on nitrogen and phosphorus balance and eco-economic benefits in red paddy field.] / 不同施肥措施对红壤稻田氮磷平衡及生态经济效益的影响.

The nitrogen (N) and phosphorus (P) balance was studied and the eco-economic benefits of N and P were evaluated for the paddy field in a red soil area, under the fertilization treatments of no fertilization (CK), chemical fertilizer (NPK), organic fertilizer (M), and chemical fertilizers combined with organic fertilizers (NPKM, NPM, NKM), based on the long-term experiment started in 1982, to optimize fertilizer application and reduce N and P losses. The results indicated that the slight N surplus (27.10 kg·hm-2) and P deficit (-6.85 kg·hm-2) were obtained in CK, while the N and P surplus respectively with 110.94-243.98 kg·hm-2 and 19.06-67.49 kg·hm-2 in other treatments were obtained. The treatments NPK and M had no effects on the N and P balance. Under the same fertilization rates, the N surplus in treatment NPKM was lower than that in treatments NPM and NKM by 6.3% and 12.9%, while the P surplus was lower by 3.7% and 13.8%, respectively. The total-N, total-P, alkali-hydrolyzable N, and available P contents in the 0-20 cm soil layer increased under the treatments of NPKM. Comparatively, the available P contents were relatively higher in the 20-40 cm soil layer under the treatments of high P application rates. Furthermore, the NPKM treatment had the highest eco-economic benefit value of 0.762, which indicated the optimized fertilizer application for the paddy fields in the red soil area. The treatment CK had the lowest eco-economic benefit values with 0.560. Based on the observations, the N and P (in P2O5) fertilization respectively at the rates of 157.71 kg·hm-2 and 112.18 kg·hm-2 could well maintain the N and P balance for the paddy field in the red soil area.

[Variation characteristics of soil carbon sequestration under long-term different fertilization in red paddy soil].

The objective of this study was to clarify the changes of soil organic carbon (SOC) content, the saturation capacity of soil carbon sequestration and its cooperation with carbon input (crop source and organic fertilizer source carbon) under long-term (1982-2012) different fertilization in red paddy soil. The results showed that fertilization could increase SOC content. The SOC content of all the fertilization treatments demonstrated a trend of stabilization after applying fertilizer for 30 years. The SOC content in the treatments applying organic manure with mineral fertilizers was between 21.02 and 21.24 g · kg(-1), and the increase rate ranged from 0.41 to 0.59 g · kg(-1) · a(-1). The SOC content in the treatments applying mineral fertilizers only was 15.48 g · kg(-1). The average soil carbon sequestration in the treatments that applied organic manure with mineral fertilizers ranged from 43.61 to 48.43 t C · hm(-2), and the average SOC storage over the years in these treatments was significantly greater than those applying mineral fertilizers only. There was an exponentially positive correlation between C sequestration efficiency and annual average organic C input. It must input exogenous organic carbon at least at 0. 12 t C · hm(-2) · a(-1) to maintain the balance of soil organic carbon under the experimental conditions.

[Change characteristics of rice yield and soil organic matter and nitrogen contents under various long-term fertilization regimes].

A long-term (1982-2010) field experiment was conducted in the Red Soil Experiment Station of Chinese Academy of Agricultural Sciences in Qiyang County of Hunan Province, South-central China to investigate the dynamic changes of rice yield and soil organic matter (OM) and nitrogen contents under different fertilization treatments. The treatments included NPK, NPKM (M: manure), NPM, NKM, PKM, M, and CK. Fertilization increased the soil OM, total N, and alkaline-hydrolysable N contents and the rice yield. In treatment NPKM, the rice yield across the 28 years maintained at the highest level; while in treatment NPK, the yield showed a decreasing trend, being lower than that in other fertilization treatments. In the treatments applied with manure only or in combining with chemical fertilizers, the soil OM content increased rapidly in the first 16 years, and then fluctuated around a constant level (29.42-39.32 g x kg(-1)). In the treatments of chemical fertilization, the soil OM content only had a quicker increase in the first 8 years, and then fluctuated within a relatively stable range. Fertilization with manure increased the soil OM significantly, as compared to fertilization with chemical fertilizers only. The soil total N content in all fertilization treatments showed a rapid increase in the first 8 years, and the increment was the highest in treatment NPKM. The soil alkaline-hydrolysable N content in all fertilization treatments had a slower increase in the first 12 years, with an average annual increment of 0.66-2.25 mg x kg(-1) x a(-1). In 1994-1998, the soil alkaline-hydrolysable N content in fertilization treatments had a quicker increase, with an average annual increment of 6.45-32.45 mg x kg(-1) x a(-1); but after 1998, the soil alkaline-hydrolysable N content had a slight decrease. It was concluded that organic fertilization was the key measure to stably improve the physical and chemical properties and the productivity of red paddy soils by increasing their OM and nitrogen contents, and to sustain the rice production in red soil region in subtropical China.

[Effects of long-term applying sulfur- and chloride-containing chemical fertilizers on weed growth in paddy field].

An investigation was made at a double-rice paddy field in the Qiyang Red Soil Field Experimental Station, Hunan Province, China to study the species and biomass of weeds growing in rice (Oryza sativa L.) growth season after 34-year application of sulfur (SO4(2-)) and chloride (Cl(-))-containing chemical fertilizers under the same application rates of nitrogen (N), phosphorus (P), and potassium (K). Long-term application of Cl(-)-containing chemical fertilizer resulted in the greatest species number of weeds and the highest biomass of floating weeds and wet weeds, compared with long-term application of SO4(2-) and Cl(-) +SO4(2-)-containing chemical fertilizers. In early rice growth season, the biomass of weeds after applying Cl(-)-containing chemical fertilizer was 51.4% and 17.6% higher than that after applying Cl(-) + SO4(2-) and SO4(2-)-containing chemical fertilizers, respectively; in late rice growth season, the increment was 144% and 242%, respectively. More floating weeds were observed after applying Cl(-) + SO4(2-) and SO4(2-)-containing chemical fertilizers, but few of them were found after applying Cl(-)-containing chemical fertilizer. The total dry mass of weeds and the dry mass of wet weeds were positively correlated with soil Cl(-) content (r = 0.764, P < 0.01 and r = 0.948, P < 0.01, respectively), but negatively correlated with soil SO4(2-)-S content (r = 0.849, P < 0.01 and r = 0.641, P < 0.05). Soil alkali-hydrolyzable N and available P, under the co-effects of soil SO4(2-)-S, Cl(-), and pH, had indirect effects on the total dry mass of weeds. By adopting various fertilization measures to maintain proper soil pH and alkali-hydrolyzable N and available P contents, increase soil SO42(-)-S content, and decrease soil Cl(-) content, it could be possible to effectively inhibit the growth of wet weeds and to decrease the total biomass of weeds in double-rice paddy field.

[Variations of soil fertility level in red soil region under long-term fertilization].

Based on the long-term (1982-2007) field experiment of "anthropogenic mellowing of raw soil" at the Qiyang red soil experimental station under Chinese Academy of Agricultural Sciences, and by using numerical theory, this paper studied the variations of the fertility level of granite red soil, quaternary red soil, and purple sandy shale soil under six fertilization patterns. The fertilization patterns included non-fertilization (CK), straw-returning without fertilizers (CKR), chemical fertilization (NPK), NPK plus straw-return (NPKR), rice straw application (M), and M plus straw-return (MR). The soil integrated fertility index (IFI) was significantly positively correlated with relative crop yield, and could better indicate soil fertility level. The IFI values of the three soils all were in the order of NPK, NPKR > M, MR > CK, CKR, with the highest value in treatment NPKR (0.77, 0.71, and 0.71 for granite red soil, quaternary red soil, and purple sandy shale soil, respectively). Comparing with that in the treatments of no straw-return, the IFI value in the treatments of straw return was increased by 6.72%-18.83%. A turning point of the IFI for all the three soils was observed at about 7 years of anthropogenic mellowing, and the annual increasing rate of the IFI was in the sequence of purple sandy shale soil (0.016 a(-1)) > quaternary red clay soil (0.011 a(-1)) > granite red soil (0.006 a(-1)). It was suggested that a combined application of organic and chemical fertilizers and/or straw return could be an effective and fast measure to enhance the soil fertility level in red soil region.

[Influence of double rice cropping system innovation on paddy soil profile form and soil characteristics].

Field experiments were conducted on the double rice cropping paddy field in red soil area to evaluate the influence of cropping system innovation on soil profile form and related soil characteristics. Four cropping systems of rice-rice-Chinese Milkvetch (Astragalus sinicus Linn.), forage, paddy-upland rotation, and upland were substituted for the double rice cropping system. The results indicated that compared with those under double rice cropping system, the thickness of cultivated horizon under upland cropping system increased by 4 cm, that of plow pan declined by 2 cm, > 2 mm aggregates in wet-sieved particle-size fractions increased by 6.94%, wet-sieved mean-mass diameter increased by 0.37 mm, contents of humic acid carbon and fulvic acid carbon increased by 0.15 and 0.49 g kg(-1), respectively, and quotient of aggregates water stability was 0.78 times higher. Under paddy-upland rotation, the quotient of aggregates water stability was higher (95.86), while soil nutrient contents changed a little. Under rice-rice-Chinese Milkvetch system, soil organic matter content increased by 1.3 g kg(-1), quotient of aggregates water stability declined by 8.82, but other parameters had less changes. Under forage system, the thickness of cultivated and transitional horizons increased by 2 cm and 9 cm, respectively, quotient of aggregates water stability increased by 1.39, while the contents of soil organic matter and total potassium decreased by 5.6 and 2.8 g kg(-1), respectively. Among all test cropping systems, forage system had the greatest changes in soil characteristics. It was completely feasible to substitute the local double rice cropping system for paddy-upland rotation or upland cropping, particularly in the areas where full irrigation was not available. However, attention should be paid to the decrease of soil potassium content when the cropping system innovation was practiced.