Genetic Diversity and Symbiotic Efficiency of Nodulating Rhizobia Isolated from Root Nodules of Faba Bean in One Field.

Thirty-one nodulating rhizobium strains were collected from root nodules of spring and winter type faba bean cultivars grown in micro ecoarea, i.e. the same field in Chengdu plain, China. The symbiotic efficiency and phylogeny of these strains were studied. Effectively nitrogen fixing strains were isolated from both winter type and spring type cultivars. Based on phylogenetic analysis of 16S rRNA gene and concatenated sequence of atpD, glnII and recA genes, the isolates were assigned as Rhizobium anhuiense and a potential new Rhizobium species. The isolates were diverse on symbiosis related gene level, carrying five, four and three variants of nifH, nodC and nodD, respectively. Strains carrying similar gene combinations were trapped by both winter and spring cultivars, disagreeing with the specificity of symbiotic genotypes to reported earlier faba bean ecotypes.

Diversity and phylogeny of rhizobia associated with Desmodium spp. in Panxi, Sichuan, China.

Thirty-four rhizobial isolates were obtained from root nodules of four wild Desmodium species growing in Panxi, Sichuan, China. According to the combined ARDRA and IGS-RFLP (CACAI) cluster analysis, Rhizobium, Pararhizobium and Mesorhizobium isolates outnumbered Bradyrhizobium isolates. In general, the isolates representing the same species from the same site clustered together. Furthermore, the four Desmodium species were all nodulated by more than one rhizobial species. AFLP and phenotypic analyses showed that the 34 isolates represented at least 32 distinct strains. None of the strains were found from more than one site or host, indicating a high degree of rhizobial diversity in Panxi. In the multilocus sequence analysis, the isolates were assigned to Pararhizobium giardinii, Bradyrhizobium japonicum, Mesorhizobium septentrionale, and to undescribed species of the genera Rhizobium, Bradyrhizobium and Agrobacterium.

Symbiotic effectiveness and phylogeny of rhizobia isolated from faba bean (Vicia faba L.) in Sichuan hilly areas, China.

A total of 54 rhizobial strains were isolated from faba bean root nodules in 21 counties of Sichuan hilly areas in China, and their symbiotic effectiveness, genetic diversity and phylogeny were assessed. Only six strains increased the shoot dry mass of the host plant significantly (P ≤ 0.05). Based on the cluster analysis of combined 16S rDNA and intergenic spacer region (IGS) PCR-RFLP, the strains were divided into 31 genotypes in 11 groups, indicating a high degree of genetic diversity among the strains. The sequence analysis of three housekeeping genes (atpD, glnII and recA) and 16S rDNA indicated that the strains represented two R. leguminosarum, two Rhizobium spp., R. mesosinicum, Agrobacterium sp. and A. tumefaciens. The strains representing four Rhizobium species were divided into two distinct nodC and nifH genotypes. However, the phylogeny of housekeeping genes and symbiotic genes was not congruent, implying that the strains had been shaped by vertical evolution of the housekeeping genes and lateral evolution of the symbiotic genes.

Polyphasic characterization of rhizobia isolated from Leucaena leucocephala from Panxi, China.

Leucaena leucocephala was introduced into Panxi, Sichuan, China, in the 1980s and 1990s for afforestation and preventing water loss and soil erosion in this area. The co-introduction of rhizobial symbionts of introduced plants has drawn attention since they may influence local soil communities. We studied the phylogenetic position of the L. leucocephala isolates and assessed if the rhizobia were introduced together with the host to Panxi, Sichuan, China. The glnII and atpD genes of fifteen representative isolates were sequenced and analyzed, and applied multilocus sequence analyses in which the housekeeping genes recA, glnII and atpD were included. Furthermore, we estimated the within species diversity directly with 23S rDNA and IGS RFLP and indirectly through phenotypic analysis of forty L. leucocephala isolates. The isolates represented seven species and 38 diversified strains in the genera Ensifer, Mesorhizobium, Bradyrhizobium and Rhizobium. The within species diversity of the Ensifer isolates was large, proposing a potential to occupy novel niches. There was not conclusive evidence to show that any of the strains would have been co-introduced with L. leucocephala. On the contrary, we came to a conclusion that the possible introduction should not be inferred from sequence data alone.

Symbiotic efficiency and phylogeny of the rhizobia isolated from Leucaena leucocephala in arid-hot river valley area in Panxi, Sichuan, China.

In search of effective nitrogen-fixing strains for inoculating Leucaena leucocephala, we assessed the symbiotic efficiency of 41 rhizobial isolates from root nodules of L. leucocephala growing in the arid-hot river valley area in Panxi, China. The genetic diversity of the isolates was studied by analyzing the housekeeping genes 16S rRNA and recA, and the symbiotic genes nifH and nodC. In the nodulation and symbiotic efficiency assay, only 11 of the 41 isolates promoted the growth of L. leucocephala while the majority of the isolates were ineffective in symbiotic nitrogen fixation. Furthermore, one fourth of the isolates had a growth slowing effect on the host. According to the 16S rRNA and recA gene analyses, most of the isolates were Ensifer spp. The remaining isolates were assigned to Rhizobium, Mesorhizobium and Bradyrhizobium. The sequence analyses indicated that the L. leucocephala rhizobia had undergone gene recombination. In contrast to the promiscuity observed as a wide species distribution of the isolates, the results implied that L. leucocephala is preferentially nodulated by strains that share common symbiosis genes. The symbiotic efficiency was not connected to chromosomal background of the symbionts and isolates carrying a similar nifH or nodC showed totally different nitrogen fixation efficiency.

[Effects of phosphorus fertilization on leaf area index, biomass accumulation and allocation, and phosphorus use efficiency of intercropped maize].

A 2-year field experiment was conducted in 2011 and 2012 to investigate the effects of phosphorus (P) fertilization on the leaf area index (LAI), dry matter accumulation (DMA), and P use efficiency (PUE) of maize in wheat/maize/soybean intercropping system. Five P fertilization rates were installed, i.e., 0, 45, 90, 135, and 180 kg P2O5 x hm(-2) for wheat, marked as WP0, WP1, WP2, WP3, and WP4, respectively, and 0, 37.5, 75, 112.5, and 150 kg P2O5 x hm(-2) for maize, marked as MP0, MP1, MP2, MP3, and MP4, respectively. During the coexisted growth periods of wheat and maize, P fertilization increased the LAI, leaf area duration (LAD), and stem and leaf DMA of maize significantly. After the jointing stage of maize, the maize LAI, LAD, DMA, and crop growth rate (CGR) all decreased after an initial increase with the increasing P rate, with the maximum growth in treatment MP2 or MP3. During the reproductive stage of maize, the maize dry mass translocation from vegetative to reproductive organ increased with increasing P fertilization rate, and the grain yield of both maize and whole cropping system increased firstly and decreased then, with the maximum grain yield of maize and whole cropping system being 6588 and 11955 kg x hm(-2) in treatment P3, respectively. The P apparent recovery efficiency of maize was the highest (26.3%) in treatment MP2, being 82.6%, 38.4%, and 152.9% higher than that in MP1 (14.4%), MP3 (19.0%), and MP4 (10.4%), respectively. In sum, for the wheat/maize/soybean intercropping system, applying appropriate amount of P fertilizer could promote maize growth, alleviate the impact of wheat on maize, and consequently, increase the P apparent recovery efficiency of maize. In this study, the appropriate P fertilization rate was 75-112.5 kg P2O5 x hm(-2).