EFFECTS OF pH ON GROWTH AND NODULATION OF TWO FORAGE LEGUMES

A study was conducted to compare the responses to acid pH of Medicago sativa and Lotus glaber, two forage legumes with different environmental requirements, either supplied with inorganic nitrogen (N) or inoculated with different strains of their nodule bacteria. Medicago sativa showed, in both treatments, a significant reduction in total dry weight at pH below 6.0. In contrast, Lotus glaber grew equally well at all the pHs assayed in the presence of adequate N. Under inoculated conditions, in the absence of N supply, plant growth was dependent on the bacterial strain used. When the ability of each strain to multiply in culture medium was examined, it was observed that Sinorhizobium meliloti strains showed a pH-sensitive response that inhibited growth at pH 4.0, whereas Mesorhizobium loti strains showed normal growth at this pH. These results suggest that for the effectiveness of Mesorhizobium loti–Lotus glaber symbiosis in acid soils the major factor to be considered is the tolerance of the bacterial strain to acid conditions, while the limiting factors for the Sinorhizobium meliloti–Medicago sativa symbiosis are the sensitivity to low pH of both the plant and its bacteria.     

外源硝态氮对菊芋耐海水胁迫的影响

A hydroponic experiment with six treatments, i.e., 0% seawater （control）, 10% seawater, 25% seawater, 0% seawater ＋ N （7.5 mmol L-1 NaNO3）, 10% seawater ＋ N （7.5 mmol L-1 NaO3）, and 25% seawater ＋ N （7.5 mmol L-1 NaNO3）, was carried out to study the effect of nitrogen addition on the growth and physiological and biochemical characteristics of Jerusalem artichoke （Helianthus tuberosus） seedlings under seawater stress. The 10% seawater stress treatment had the least effect on plant growth while at 25% seawater growth was significantly inhibited. The malondialdehyde content and electrolyte leakage in leaves under 10% seawater were similar to those of the control, but significantly higher under the 25% seawater stress. The activities of superoxide dismutase, peroxidase and catalase in the leaves increased concomitantly with increasing seawater concentration and time. Proline and soluble-sugars in the leaves and Na^＋, K^＋, and Cl- contents in shoots and roots increased significantly with the concentration of seawater increasing. Nitrogen addition resulted in increasing fresh and dry weights of shoots and roots compared with seawater treatment without N. Nitrogen supplementation also significantly enhanced the activities of antioxidant enzymes in leaves. Addition of N to seawater enhanced the contents of proline and soluble-sugars in the leaves and K^＋ and total-N in the aerial parts and roots of H. tuberosus, but it resulted in declined concentrations of Na^＋ and Cl- in the aerial parts and roots. Nitrogen addition ameliorated the toxicity of seawater by improving the antioxidative enzymes, accumulating of proiine and soluble-sugars, and altering the distribution of inorganic ions in H. tuberosus.     

紫云英根瘤菌分子遗传学研究进展

总结了紫云英根瘤菌分子遗传学研究进展。这类根瘤菌的寄主范围较窄，早先根据互接种族概念将其定名为Rhizobium astragali。后来定名为Rhizobium huakuii。最近并入Mesorhizobium属，对大量菌株进行的16S和23SrDNAPCR-RFLP分析和代表菌株的部分16SrDNA碱基测序，揭示了7个不同基因型，表明这类菌具有遗传多样性。可以认为存在不同的种，它们都有质粒，数量1-5个，因菌株而异，可分为不同质粒型。共生基因常定位在最大的一个质粒上，即共生质粒，它们的结瘤基因具有独特结构。采用包括nodA,nodB。noC,nodD的苜蓿根瘤菌突变株进行基因互补，从紫云英根瘤蓖基因文库中获得了相应的转移接合子，这类结瘤基因的这种结构在7个基因型的菌株中都是一致的，说明结瘤基因的保守性。     

Enhancement of Nodulation and Yield of Chickpea by Co-inoculation of Indigenous Mesorhizobium spp. and Plant Growth–Promoting Rhizobacteria in Eastern Uttar Pradesh

We study the effect of plant growth–promoting rhizobacteria (PGPR) along with Mesorhizobium sp. BHURC02 on nodulation, plant growth, yield, and nutrient content of chickpea (Cicer arietinum L.) under field conditions. A similar study has been conducted for nodulation and plant growth of chickpea in pot experiment under glasshouse conditions. The treatment combination of Mesorhizobium sp. BHURC02 and Pseudomonas fluorescens BHUPSB06 statistically significantly increased nodule number plant^–1, dry weight of nodule plant^–1, and root and shoot dry weights plant^–1 over the control under a glasshouse experiment. The maximum significant increase in nodule number, dry matter, and nutrient content were recorded in co-inoculation of Mesorhizobium sp. BHURC02 and P. fluorescens BHUPSB06 followed by co-inoculation of Mesorhizobium sp., Azotobacter chroococcum, and Bacillus megatrium BHUPSB14 over uninoculated control in a 2-year field study. Hence, co-inoculation of Mesorhizobium sp. and P. fluorescens may be effective indigenous PGPR for chickpea production.     

发光酶基因标记的华癸根瘤菌JS5A16L在紫云英根圈的定殖动态

在土壤一盒栽、盆栽微宇宙系统（Ｍｉｃｒｏｃｏｓｍ）中和田间条件下，研究了发光酶基因（ｌｕｘＡＢ）标记的华癸根瘤菌ＪＳ５Ａ１６Ｌ在紫云英根圈的定殖动态、分布范围及结瘤情况。在盒栽系统中，ＪＳ５Ａ１６Ｌ的定殖密度在紫云英出苗２天后达到最高水平（７．８８ ｌｐｇ ｃｆｕ／克根）（ｃｆｕ为ｃｏｌｏｎｙ　ｆｏｒｍｉｎｇ　ｕｎｉｔ的缩写），然后开始下降，并保持在一个相对稳定的水平；５８天后又有所回升，且能散布至种子下方２２ｃｍ处的根段部位。盆栽条件下的定殖动态与盒栽系统中的相似，紫云英播种３天后，ＪＳ５Ａ１６Ｌ可达最高定殖水平（６．９２ ｌｏｇ ｃｆｕ／株根系），随后开始缓慢下降，直至３２天时仍维持在一个相对稳定水平。ＪＳ５Ａ１６Ｌ在田间条件的定殖动态则不同，播种后３０天时定殖密度达到最大值（７ ．０３ ｌｏｇ ｃｆｕ／克根），９０天后降到最低值（５． ２４ ｌｏｇ ｃｆｕ／克根），然后又开始上升，１６０天时为７．８７ ｌｏｇ ｃｆｕ／克根，甚至在地上部植株收割后２０天仍可维持在７．８９ ｌｏｇ ｃｆｕ／克根。接种该菌株可显著提高紫云英的生物学产量。     

紫云英根瘤菌种内不同菌株群体感应信号分子差异性和交互性分析

根瘤菌可以和豆科植物共生固氮,在很多根瘤菌中群体感应系统参与了结瘤固氮等相关生理过程的调节。本研究从野外不同生境紫云英宿主根部瘤体中分离了26株根瘤菌,体外定量检测了各菌株产生自体诱导物(AI)信号分子的能力,以及不同菌株AI激活中慢生华癸根瘤菌Mh菌株AHLs合成酶基因mrh I表达的情况,发现紫云英根瘤菌种内不同菌株间存在群体感应交互性。通过导入外源Att M水解酶观察菌株AI活性的变化,应用薄层层析法定性分析不同菌株的AI结构,初步揭示了不同紫云英根瘤菌产生AI特性的差异和多样性。     

华癸中生根瘤菌(Mesorhizobium huakuii)研究进展

综述了华癸中生根瘤菌(Mesorhizobium huakuii)遗传多样性、质粒多样性、质粒的功能、结瘤因子、结瘤基因、胞外多糖等方面的研究进展。这类菌只能在紫云英上结瘤,1997年并入Mesorhizobium。选取大量菌株进行REP-PCR、16S和23S rDNAPCR-RFLP等分析,揭示其分为16种16S rDNA基因型。选用不同16S rDNA基因型的部分代表菌株进行部分16S rDNA测序。能够在紫云英上结瘤的根瘤菌可以分为2个类群。表明这类菌具有遗传多样性,存在不同的种。它们多数含共生质粒,共生质粒定位于最大或次大质粒上,质粒数1~5个不等,其分子量范围在53~906 kb之间。质粒缺失菌株表现为不结瘤(Nod-)和结瘤而不固氮(Nod ,Nif-)。某些质粒缺失还影响LPS合成、抗酸性、竞争结瘤能力。其结瘤因子能诱导紫云英根毛变形,它们的结瘤因子结构类似于苜蓿根瘤菌的结瘤因子。它们的结瘤基因具独特性,nodA与nodBC分隔一定距离,且具有保守性。它们产生的胞外多糖在侵染结瘤过程中发挥重要作用。     

华癸中慢生根瘤菌7653R MCHK_8182基因的共生固氮功能

根据华癸中慢生根瘤菌7653R全局转录组分析结果,获得在共生固氮阶段显著上调表达的基因MCHK_8182;设计重叠延伸PCR引物,构建MCHK_8182双交换突变重组质粒载体;通过三亲本杂交,将突变载体导入7653R;在蔗糖压力培养下进行筛选,最终获得MCHK_8182双交换突变株。PCR及测序验证双交换突变株构建成功。植物盆栽试验结果显示,与7653R野生型接种的植株相比,MCHK_8182突变菌株接种的植株根瘤数减少,但固氮酶活未见明显差异。     

中慢生华癸根瘤菌nodD-突变株的筛选及其对紫云英的促生作用

通过二亲接合的方式,将含有sacB基因和nodD基因部分片段的质粒与中慢生华癸根瘤菌Mesorhizobium huakuii)93(Mh93)菌株进行同源重组,在含10％蔗糖的TY平板上成功地筛选到了nodD基因突变株Mh93-27.利用温室盆栽试验研究了紫云英单作及紫云英-小麦间作体系Mh93-27对紫云英的促生作用,结果表明,接种Mh93-27后,紫云英的地上部生物量和全氮含量以及根部生长得到了不同程度的提高,但只有在小麦孕穗期和收获期时,根瘤菌才表现出显著的促生作用.     

Compatibility of Mesorhizobium sp. Cicer with seed treatment of fungicide and insecticide in chickpea

This investigation was undertaken to study the compatibility of Mesorhizobium sp. Cicer with captan (fungicide) and chlorpyrifos (insecticide) for growth, symbiotic parameters and yield in chickpea. In an in vitro experiment, a significant reduction in the number of viable mesorhizobia was observed in Mesorhizobium sp. Cicer treated chickpea seeds at the recommended doses of captan (3 g kg^?1 seed) and chlorpyrifos (10 ml kg^?1 seed) after 4 h storage at 4°C, and further reduction was seen after 8–16 h contact with Mesorhizobium. The results showed that captan was more toxic than chlorpyrifos. In field experiments, improved growth and symbiotic parameters (plant height, dry weight of shoot, nodulation, leghaemoglobin content, chlorophyll content and nitrogen content) and a reduction in per cent damaged by termites and diseases were observed in the Mesorhizobium alone treatment compared with the uninoculated control. Grain yield was increased significantly in treatments with Mesorhizobium alone or in a mixture with fungicide and insecticide (captan and chlorpyrifos) compared with the control treatment. It is evident that chemically treated seeds should always be sown as soon as possible after inoculation. Recommended rates of captan and chlorpyrifos application with Mesorhizobium inoculant as a seed treatment was innocuous to chickpea–Mesorhizobium symbiosis.