Novel root nodule bacteria belonging to the genus Caulobacter

Aims: Aim of this study is to determine the genetic variation of rhizobia associated with horse gram [Macrotyloma uniflorum (Lam.) Verdc.] plants grown in different regions of Andhra Pradesh, India. Methods and Results: Four representative isolates having most representative characters from the previous characterization were selected for 16S rRNA sequence. The sequences were submitted to the NCBI GenBank and Ribosomal Database Project (RDP). The isolates HGR‐4, 6 and 13 showed more than 99% homology between them and they were grouped with Rhizobium reference strains where as the isolate HGR‐25 showed 87·1, 87·4 and 87·2% homology with the isolates HGR‐4, 6 and 13, respectively, and were grouped with reference strains for Caulobacter. The nodulation ability of these isolates on horse gram was confirmed by inoculation tests. Conclusions: The isolate HGR‐25 was identified as Caulobacter isolated from the plants growing in soil samples collected from Khareemnagar district, Andhra Pradesh, India. Inoculation tests revealed that Caulobacter formed nodules on horse gram. It was also confirmed by RDP. Significance and Impact of the Study: This is the first report that a legume was nodulated by a member of the genus Caulobacter, which belongs to the family Caulobacteriaceae in the order Caulobacterales of Alphaproteobacteria.     

Phenotypical and genotypical characteristics of root-nodulating bacteria isolated from annual Medicago spp. in Soummam Valley (Algeria)

AIMS: In the framework of agro-pastoral system management using local annual medics coupled with their native root-nodulating bacteria to extend pasture zones, increase forage yields and improve ovine and bovine breeding in Algeria, we investigated diversity of rhizobia from annual Medicago spp. (Medicago arabica, Medicago polymorpha, Medicago minima and Medicago orbicularis). METHODS AND RESULTS: Ten nodulating-isolates were characterized by morphological, cultural, physiological and biochemical features, SDS-PAGE analysis and PCR-RFLP of 16S rDNA. The results show some degree of genetic diversity among the isolates; three can be affiliated to Sinorhizobium meliloti, one to Rhizobium galegae and six were separate. CONCLUSIONS: Local annual medics would have a high degree of specificity in their symbiotic interaction. Furthermore, our results support the presence of Rh. galegae in the Mediterranean region. SIGNIFICANCE AND IMPACT OF THE STUDY: This work is a preliminary step towards selection of efficient symbiotic Medicago-rhizobia to develop inoculants for management of agro-pastoral systems using local annual medics in Algeria.     

秸秆覆盖配施氮肥根际土真菌群落及其与小麦产量的关系

为研究秸秆覆盖与施氮条件下土壤真菌群落变化及其驱动因素与冬小麦产量的关系,试验采用二因素裂区设计,主区为秸秆覆盖(SM)和不覆盖(NSM);副区为两种施氮量0(N0)和180(N1)kg/hm²。于小麦开花期采集土壤样品测定土壤养分及采用Illumina Miseq高通量测序技术分析根际真菌群落结构和多样性。结果表明,SM较NSM处理小麦产量提高40.3%,差异显著。NSMN1较NSMN0处理小麦产量显著提高75%;SMN1处理比SMN0处理小麦产量显著提高92%。SM处理较NSM处理土壤有机碳(SOC)、全氮(TN)、碱解氮(AN)、氨态氮(NH⁺₄-N)、速效磷(AP)和速效钾(AK)的含量显著增加,土壤硝态氮(NO^-₃-N)含量降低。无论秸秆覆盖与否,施氮显著提高了土壤TN、AN、NH⁺₄-N、NO^-₃-N、AP和AK的含量。秸秆覆盖根际真菌群落多样性(Chao1与Shannon指数)...     

小麦类根瘤胞间根瘤菌对分生细胞核的影响

在小麦类根瘤中,分生细胞核很大,而且较圆整,一般为一个核仁,有的核仁还有核仁泡,但通常一个核仁只有一个核仁泡,染色质较少,呈团块状分布。若胞间根瘤菌离分生细胞核较远,分生细胞核变化不大,只是染色质开始由团块状变为长条形,但仍有核仁。当胞间根瘤菌靠近分生细胞核时,细胞核变小,染色质密度增加并形成网状,而且无核仁出现。在上述这些无核仁的分生细胞核内有一种特殊的管状内含体,它们位于网状染色质附近的核基质中。本文还讨论了分生细胞核的变化规律及其与胞间根瘤菌存在的关系。     

Root nodule development: origin, function and regulation of nodulin genes

The symbiotic root nodule, an organ formed on leguminous plants, is a product of successful interactions between the host plant and the soil bacteria, Rhizobium spp. Plant hormones play an important role in the genesis of this organ. The hormonal balance appears to be modulated by the signals produced by bacteria. Many host genes induced during nodule organogenesis and the symbiotic state have been identified and characterized from several legumes. These genes encode nodule-specific proteins (nodulins) which perform diverse functions in root nodule development and metabolism. Formation of a subcellular compartment housing the bacteria is essential to sustain the symbiotic state, and several nodulins are involved in maintaining the integrity and function of this compartment. The bacteroid enclosed in the perbacteroid membrane behaves as an 'organelle,'completely dependent on the host for all its requirements for carbon, nitrogen and other essential elements. Thus it seems likely that the nodulins in the peribacteroid membrane perform specific transport functions. While the function of a few other nodulins is known (e.g. nodulin-100, nodulin-35), a group of uncharacterized nodulins exists in soybean root nodules. These nodulins share structural similarities and seem to have been derived from a common ancestor. Induction of nodulin genes occurs prior to and independent of nitrogen fixation, and thus is a prelude to symbiosis. Although some of the early nodulin genes are induced prior to or during infection, induction of late nodulins requires endocytotic release of bacteria.     

小麦叶片气孔性状与产量和抗旱性的关系

以小麦旱选10号/鲁麦14 DH群体为试材,对干旱胁迫和正常灌溉条件下花后10 d和20d旗叶中部气孔密度、气孔大小、气孔导度、光合速率、蒸腾速率与产量和抗旱系数的关系进行相关和通径分析.结果表明:在两种水分条件下,花后10 d气孔密度、气孔大小、气孔导度、光合速率、蒸腾速率与产量和抗旱系数的相关性大多不显著;但花后20 d与千粒重呈极显著正相关,与穗粒数、单株产量和抗旱系数的相关性仍大多不显著.气孔导度、光合速率和蒸腾速率是影响单株产量和抗旱系数的主要因素,不仅对单株产量和抗旱系数的直接作用较大,间接作用也较大;气孔密度与气孔大小对单株产量和抗旱系数的直接作用和间接作用均较小.在两种水分条件下,花后10 d和20 d,气孔密度与气孔长度之间,气孔长度与气孔宽度、气孔导度、光合速率和蒸腾速率之间的相关性均显著或极显著,但气孔密度、气孔长度与气孔导度、光合速率和蒸腾速率间的相关性在不同水分条件和不同生育阶段存在差异,表明水分条件、生长发育阶段对这些性状之间的相关性影响较大.通过育种手段以叶片气孔密度和气孔大小为选择目标,来改善小麦气孔导度、光合速率和蒸腾速率,进而提高产量并不总是一种有效手段.     

Isolation and characterization of bacteria from the rhizosphere of wheat

In this study, we isolated bacteria from rhizosphere and endorhizophere of wheat crops of the central region of Argentina. The isolates were phenotypically characterized and the restriction patterns of 16S rDNA (ARDRA) using endonuclease AluI were analysed. Representative isolates were used to evaluate the effect of the inoculation on the growth of wheat under greenhouse conditions. The effects of plant growth-promoting bacteria on wheat plants were studied by evaluating shoot fresh and dry weights and root fresh and dry weights. One native strain increased the shoot and root dry biomass by 23% and 45% respectively. Other strains increased the shoot dry biomass. A 1.5 kb fragment of the 16S rRNA gene of one isolate was sequenced. This isolate showed high identity with different species of Pseudomonas.     

Genetics of transgressive segregation for yield and yield components in wheat

Three crosses of spring wheat (Triticum aestivum L. em Thell) involving six cultivars (WC29, WH291, SGP 14, RAJ. 1972, WH377 and HD 2329) were selected on the basis of combining ability analysis to study genetics of transgressive segregation for tillers/plant, grains/spike, 1000 grain weight and grain yield/plant using various mating designs. Diallel analysis indicated that both additive and non- additive components were significant for all the characters. On the basis of general combining ability and specific combining ability effects, the parents WH 291 and WH 377 were found to be good general combiners for tillers/ plant, 1000 grain weight and grain yield/plant. For grains/spike SGP 14 was found to be a good general combiner. The cross WH 377 × HD 2329 for tillers/plant, SGP 14 × Raj. 1972 for grains/spike and grain yield/plant and WC 29 × WH 291 for 1000 grain weight were found to be good cross combinations. Generation mean analysis indicated that the additive-dominance model was inadequate for all the characters in all the crosses except for 1000 grain weight in WC29 × WH291. Additive component was more pronounced than non-additive components for all the characters in all the crosses except for tillers/ plant in WH 377 × HD 2329. Predictions for transgressive segregants from F₃ was more accurate than that from generation mean analysis. However, prediction from both the sources were equally efficient if additive-dominance model was adequate. In general, observed frequencies of transgressive segregants were more in F₂ and BIPi than F₄ but the majority of them were discarded on progeny testing. Biparental mating had an impact in increasing the frequencies of transgressive segregants for different characters in all the crosses. The crosses, WH 377 × HD 2329 for grain yield/ plant, SGP 14 × Raj. 1972 for tillers/plant and WC 29 × WH 291 for grains/spike and 1000 grain weight were found to be potential crosses for transgressive segregants. A comparison of combining ability of parents and crosses, and observed and predicted frequencies of transgressive segregants indicated that the potential crosses for transgressive segregants were those that had high sea effects and involved high and low general combiners. The crosses involving low general combiners irrespective of their sea effects showed poor performance with respect to transgressive segregation.     

Plant growth promotion and suppression of charcoal‐rot fungus (Macrophomina phaseolina) in velvet bean (Mucuna pruriens L.) by root nodule bacteria

Root‐nodulating bacteria are intimate associates of legumes. From a pool of rhizobia isolated from root nodules of Mucuna pruriens (Velvet bean/Kaunch), RMP66 and BMP17 were found to be capable of promoting siderophore and IAA production and phosphate solubilization (insoluble tri‐calcium). Both symbionts were studied further to determine their abilities to promote plant growth and to control root‐rot in Mucuna pruriens caused by the pathogenic plant fungus Macrophomina phaseolina. RMP66 and BMP17 were selected based on their excellent inhibitory activities against M. phaseolina (by 78% and 71%, respectively) in dual culture and in agar‐well assays using cell‐free culture filtrate (CFCF) (by 76% and 62%, respectively). Both strains inhibited fungal growth to a greater extent in iron‐deficient medium (51% and 69%) than in iron‐supplemented medium (37% and 0%), respectively. CFCFs of RMP66 and BMP17 obtained from Pikovskaya's broth and tryptophan‐amended YEM broth inhibited fungal growth by 80%‐55% and 70%‐43%, respectively, and were identified as Sinorhizobium meliloti RMP66 and Bradyrhizobium diazoefficiens BMP17 by 16S rDNA sequencing. Centrifuged and pelleted cells harvested from exponentially grown cultures of S. meliloti RMP66 and B. diazoefficiens BMP17 were used to bacterize seeds of M. pruriens, which then showed enhanced seed germination (by up to 17% and 12%, respectively), and subsequent increases in other plant growth parameters in field trials. Considerable increases in seedling vigour indices (62%: 53% and 110%: 130%) and biomass (8%: 13% and 25%: 28%) were also observed for bacterial treatments. Tn5‐mediated antibiotic‐resistant marker strains showed enhanced nodule occupancy by up to 72% and 68%, respectively. This study describes a multifunctional legume nodule rhizobia that could be utilized in multicropping systems under different agroclimatic conditions as a bioinoculant and alternative to fertilizers.     

大穗型小麦产量形成过程中光合特性的动态变化

在大田条件下,对8个大穗型小麦新品系和多穗型品种西农979(对照)的产量性状以及不同生育期叶片光合速率、叶绿素荧光参数、叶绿素含量(Chl)和叶面积指数(LAI)进行研究.结果表明:除单位面积穗数低于对照外,8个新品系的穗粒数、穗粒重和千粒重均显著高于对照,大穗型小麦新品系2036、2037、2038、2039和2040的产量显著高于对照;8个新品系的平均光合速率(Pn)与对照差异不显著,而PSⅡ最大光能转换效率、PSⅡ实际量子效率、光化学猝灭系数和PSⅡ反应中心活性均高于对照;品系2037、2040、2039、2038和2036的Chl比对照分别提高17.5％、19.1％、15.3％、13.9％和7.9％;大穗型小麦品系的LAI明显高于对照,且在生育后期下降缓慢.     

黄土高原塬区旱地长期施肥对小麦产量的影响

以18年长期肥料定位试验为背景,研究了长期施肥条件下小麦的增产效应18年单施氮肥平均增产418．1kg·hm^-2,增产率达28．4％,其中干旱年减产率为13．9％,常态年、丰水年增产率达30．3％、58．9％;单施磷肥平均减产率为9．3％,干旱年增产率达6．5％,常态年、丰水年减产率为15．4％、10．0％;有机肥平均增产率达82．8％;NP、PM、NM、NPM平均增产率分别达127．8％、118．9％、144．4％,169．3％,不同降水年型施肥对小麦的产量结构影响不同,普遍表现为干旱年成穗数、穗粒数、千粒重减少,丰水年增加,不同降水年型穗粒数、成穗数、千粒重的变化是不同施肥处理对小麦产量结构的调控措施。     

Grain protein and grain yield of tritordeum in comparison to wheat and triticale

The new species of cereal × Tritordeum Ascherson et Graebner (Hordeum chilense Roem. et Shultz × Triticum ssp.) has a grain protein concentration (GPC) of up to 25%. The relationship between GPC and yield, and the factors responsible for the high GPC of tritordeum were examined and compared in field experiments. Three experimental tritordeum lines, two early and a later released (recombined and secondary tritordeums) were compared to wheat (cv. Cajeme) and triticale cultivars (cv. Trujillo). GPC's were 19%–22% for recombined tritordeums, 16% for the secondary tritordeum, 12–15% for wheats and 11% for triticale. Grain yields of the recombined and secondary tridordeum were 17–33% and 45–57% that of the wheats and triticale, respectively. Reducing grain sink size by spikelet removal resulted in an increased GPC of remaining grains. Considering all species together there were a strong inverse relationship between GPC and grain yield (GY) per main ear (GPC=26–4.76 ln GY; r²=0.82). In another experiment, frost damage to an early sown treatment of wheat reduced sink size. Harvest index (HI) of early sown wheat was reduced from 0.45 to 0.19, values comparable to that of tritordeum. Having similar HI, the GPC of the early sown wheat was the same as an early sown tritordeum (around 18%). Data for total N uptake and the N concentration of plant tissue during the growing season indicated that enhanced N uptake and remobilisation were not responsible for tritordeum's high GPC. These results suggest that the high GPC of the early lines of tritordeum is a consequence of the small grain yield concentrating the grain protein.     

农田秸秆覆盖对冬小麦水氮效应的影响

通过田间试验研究了杨凌红油土农田秸秆覆盖条件下冬小麦的水氮效应.结果表明,秸秆覆盖使土壤水分状况明显改善,从而使补充灌水的效果相应减小,氮肥的作用更加突出;无覆盖条件下,水氮交互效应均为负值,而秸秆覆盖时水氮有正的交互作用.旱地秸秆覆盖条件下更应重视养分的投入.无秸秆覆盖条件下小麦取得高产的基础是良好的土壤底墒和充足的氮肥.秸秆覆盖使小麦达到最高产量所需要的灌水量降低,灌水时期后延.不论有无秸秆覆盖,拔节期灌水对冬小麦籽粒产量没有显著影响.     

Survival of potentially pathogenic human-associated bacteria in the rhizosphere of hydroponically grown wheat

Plants may serve as reservoirs for human-associated bacteria (H-AB) in long-term space missions containing bioregenerative life support systems. The current study examined the abilities of five human-associated potential pathogens, Pseudomonas aeruginosa, Pseudomonas cepacia, Staphylococcus aureus, Streptococcus pyogenes and Escherichia coli, to colonize and grow in the rhizosphere of hydroponically grown wheat, a candidate crop for life support. All of these bacteria have been recovered from past NASA missions and present potential problems for future missions. The abilities of these organisms to adhere to the roots of axenic five-day-old wheat (Triticum aestivum L. cv. Yecora rojo) were evaluated by enumeration of the attached organisms after a one hour incubation of roots in a suspension (approximately 10(8) cfu ml-1) of the H-AB. Results showed that a greater percentage of P. aeruginosa cells adhered to the wheat roots than the other four H-AB. Similarly incubated seedlings were also grown under attempted axenic conditions for seven days to examine the potential of each organism to proliferate in the rhizosphere (root colonization capacity). P. cepacia and P. aerogiunosa showed considerable growth, E. coli and S. aureus showed no significant growth, and S. pyogenes died off in the wheat rhizosphere. Studies examining the effects of competition on the survival of these microorganisms indicated that P. aeruginosa was the only organism that survived in the rhizosphere of hydroponically grown wheat in the presence of different levels of microbial competition.     

QTL mapping for yield and yield contributing traits in two mapping populations of bread wheat

In bread wheat, single-locus and two-locus QTL analyses were conducted for seven yield and yield contributing traits using two different mapping populations (P I and P II). Single-locus QTL analyses involved composite interval mapping (CIM) for individual traits and multiple-trait composite interval mapping (MCIM) for correlated yield traits to detect the pleiotropic QTLs. Two-locus analyses were conducted to detect main effect QTLs (M-QTLs), epistatic QTLs (E-QTLs) and QTL × environment interactions (QE and QQE). Only a solitary QTL for spikelets per spike was common between the above two populations. HomoeoQTLs were also detected, suggesting the presence of triplicate QTLs in bread wheat. Relatively fewer QTLs were detected in P I than in P II. This may be partly due to low density of marker loci on P I framework map (173) than in P II (521) and partly due to more divergent parents used for developing P II. Six QTLs were important which were pleiotropic/coincident involving more than one trait and were also consistent over environments. These QTLs could be utilized efficiently for marker assisted selection (MAS).     

Elucidating the genetic basis of biomass accumulation and radiation use efficiency in spring wheat and its role in yield potential

One of the major challenges for plant scientists is increasing wheat (Triticum aestivum) yield potential (YP). A significant bottleneck for increasing YP is achieving increased biomass through optimization of radiation use efficiency (RUE) along the crop cycle. Exotic material such as landraces and synthetic wheat has been incorporated into breeding programmes in an attempt to alleviate this; however, their contribution to YP is still unclear. To understand the genetic basis of biomass accumulation and RUE, we applied genome‐wide association study (GWAS) to a panel of 150 elite spring wheat genotypes including many landrace and synthetically derived lines. The panel was evaluated for 31 traits over 2 years under optimal growing conditions and genotyped using the 35K wheat breeders array. Marker‐trait association identified 94 SNPs significantly associated with yield, agronomic and phenology‐related traits along with RUE and final biomass (BM_PM) at various growth stages that explained 7%–17% of phenotypic variation. Common SNP markers were identified for grain yield, BM_PM and RUE on chromosomes 5A and 7A. Additionally, landrace and synthetic derivative lines showed higher thousand grain weight (TGW), BM_PM and RUE but lower grain number (GM2) and harvest index (HI). Our work demonstrates the use of exotic material as a valuable resource to increase YP. It also provides markers for use in marker‐assisted breeding to systematically increase BM_PM, RUE and TGW and avoid the TGW/GM2 and BM_PM/HI trade‐off. Thus, achieving greater genetic gains in elite germplasm while also highlighting genomic regions and candidate genes for further study.     

Screening plant growth-promoting rhizobacteria for improving growth and yield of wheat

AIMS: Plant growth promoting rhizobacteria (PGPR) are commonly used as inoculants for improving the growth and yield of agricultural crops, however screening for the selection of effective PGPR strains is very critical. This study focuses on the screening of effective PGPR strains on the basis of their potential for in vitro auxin production and plant growth promoting activity under gnotobiotic conditions. METHODS AND RESULTS: A large number of bacteria were isolated from the rhizosphere soil of wheat plants grown at different sites. Thirty isolates showing prolific growth on agar medium were selected and evaluated for their potential to produce auxins in vitro. Colorimetric analysis showed variable amount of auxins (ranging from 1.1 to 12.1 mg l-1) produced by the rhizobacteria in vitro and amendment of the culture media with l-tryptophan (l-TRP), further stimulated auxin biosynthesis (ranging from 1.8 to 24.8 mg l-1). HPLC analysis confirmed the presence of indole acetic acid (IAA) and indole acetamide (IAM) as the major auxins in the culture filtrates of these rhizobacteria. A series of laboratory experiments conducted on two cv. of wheat under gnotobiotic (axenic) conditions demonstrated increases in root elongation (up to 17.3%), root dry weight (up to 13.5%), shoot elongation (up to 37.7%) and shoot dry weight (up to 36.3%) of inoculated wheat seedlings. Linear positive correlation (r = 0.99) between in vitro auxin production and increase in growth parameters of inoculated seeds was found. Based upon auxin biosynthesis and growth-promoting activity, four isolates were selected and designated as plant growth-promoting rhizobacteria (PGPR). Auxin biosynthesis in sterilized vs nonsterilized soil inoculated with selected PGPR was also monitored that revealed superiority of the selected PGPR over indigenous microflora. Peat-based seed inoculation with selected PGPR isolates exhibited stimulatory effects on grain yields of tested wheat cv. in pot (up to 14.7% increase over control) and field experiments (up to 27.5% increase over control); however, the response varied with cv. and PGPR strains. CONCLUSIONS: It was concluded that the strain, which produced the highest amount of auxins in nonsterilized soil, also caused maximum increase in growth and yield of both the wheat cv. SIGNIFICANCE AND IMPACT OF STUDY: This study suggested that potential for auxin biosynthesis by rhizobacteria could be used as a tool for the screening of effective PGPR strains.     

Fertilization regulates the response of wheat yield to interannual temperature variation in North China

Aims Understanding the effect of long-term fertilization on the sensitivity of grain yield to temperature changes is critical for accurately assessing the impact of global warming on crop production. In this study, we aim to assess the impacts of temperature changes on grain yields of winter wheat (Triticum aestivum L.) under different fertilization treatments in a long-term manipulative experiment in North China.Methods We measured grain yields of winter wheat under four fertilization treatments at the Yucheng Comprehensive Experimental Station each year from 1993 to 2012. We also measured air temperature at 0200, 0800, 1400 and 2000h each day since 1 January 1980. We then used the first-difference method and simple linear regression models to examine the relationship of crop yield changes to mean air temperature, mean daytime and nighttime air temperature in crop growing seasons.Important findings We found that increases in mean daily temperature, mean daytime temperature and mean nighttime temperature each had a positive impact on the grain yield of winter wheat. Grain yield increased by 16.7–85.6% for winter wheat in response to a 1°C increase in growing season mean daily temperature. Winter wheat yield was more sensitive to variations of nighttime temperature than to that of daytime temperature. The observed temperature impacts also varied across different fertilization treatments. Balanced fertilization significantly enhanced grain yields for winter wheat under a warming climate. Wheat plots treated with nitrogen and phosphorous balanced fertilization (NPK- and NP-treated plots) were more responsive to temperature changes than those without. This report provides direct evidence of how temperature change impacts grain yields under different fertilization treatments, which is useful for crop management in a changing global climate.