五种主要土壤类型土著大豆根瘤菌的生态学特点及接种效应

本研究历时4年,对吉林省主要土类土著大豆根瘤菌的生态学特点及接种效果进行了调查与研究。结果表明:土著大豆根瘤菌的类型:黑土、草甸黑土以慢生型为主,淡黑钙土、白浆土全部为慢生型,轻碱土全部是快生型。血清型:黑土、草甸黑土和白浆土中以477—28、477—694为主,淡黑钙土中以35为主,而轻碱土中以113和516快生型为主。固氮酶活性:淡     

鲁黄1号大豆与根瘤菌的共生匹配性

大豆可与中华根瘤菌属及慢生根瘤菌属的多种根瘤菌共生固氮.研究大豆品种与不同种根瘤菌之间的共生匹配性,对获得高效根瘤菌用于接种,提高大豆的产量及品质有重要的理论和实践意义.本研究使用黄淮海地区的优质高蛋白大豆品种鲁黄1号从当地土壤内捕捉并分离纯化到27株根瘤菌.经持家基因recA的序列分析,发现其中18株属于中华根瘤菌属,9株属于慢生根瘤菌属.选用两个属的代表菌株各一株(Sinorhizobium fredii S6和Bradyrhizobium sp. S10),分别在蛭石、土壤盆栽及大田试验条件下,研究这两株菌单独及混合接种对鲁黄1号大豆的生长、结瘤、固氮活力、产量、种子蛋白含量及含油量的影响.结果表明： 与S10菌株相比,S6菌株对大豆的促生能力更强,对提高产量和品质的效果更好,从而确定S6为与鲁黄1号大豆相匹配的高效根瘤菌,可作为黄淮海地区推广种植鲁黄1号大豆时接种高效根瘤菌的菌种资源.
     

双拷贝固氮结瘤基因的大豆根瘤菌R.fredii基因工程菌的耐…

用热处理消除了快生型大豆根瘤菌Ｒ．ｆｒｅｄｉｉＵＳＤＡ１９４的非共生质粒,获得了突变株Ｈｏ４,以Ｈｃ４为受体菌,以Ｅ．ｃｏｌｉＨＢ１０１Ｅｃ８３（ＰＬＡＦＲＩ．ｎｏｄｎｉｆ）为供体菌,Ｅ．ＣｏｌｉＰＲＫ２０１３为助体,进行三亲本杂交,得到双重ｎｏｄｎｉｆ基因拷贝的杂合子Ｈｃ４－８。用它们完成共生固氮结瘤试验,结果表明,在初花期Ｈｃ４的固氮酶活性比出发株ＵＳＤＡ１９搞７５．７％Ｈｃ４－８比ＵＳＤＡ１     

快生型大豆根瘤菌在山东嘉祥获得了良好的田间应用效果

中国农业菌种保藏中心从我国九省、市、自治区的11个不同生态环境中收集到57株快生型大豆根瘤菌,通过水培、沙培试验,对其生物学特性进行了鉴定,通过固氮酶活性、植株干重、茎叶含氮量的测定,明确了快生型大豆根瘤菌总的共生特点.快生型大豆根瘤菌大多能与相适应的中国栽培大豆有效共生,固氮酶活性一般在200～300nMC_2H_4/克鲜瘤.分单株干重和茎叶含氮量比不接种对照提高1倍左右,比慢     

河西走廊豆科植物结瘤固氮特性初步研究

对甘肃河西走产野生豆科植物结瘤及固氮状况进行了调查。栽培豆科植物均能自然结瘤,野生豆科植物结瘤率仅５０％。该地区豆科植物根瘤大多数圆形、棒状或指状,形状较为规则。栽培植物根瘤多为粉红色,而野生揿为白色、黄色或棕色。ＡＲＡ测定表明,２５．６％根瘤为无效根瘤。不同种不同地区根瘤固氮活性相差较大,一般活力都较低,乙炔还原活性小１μｍｏｌＣ２Ｈ４／ｇＦＷ．ｈ的占检测样品数的２９．９％,最高者信为２８．１９     

耐盐高效大豆根瘤菌株的构建

快生型大豆根瘤菌RT19是一株耐盐而固氮能力差的菌株,在含有0．3 mol／L Nacl的YM培养液中生长良好,代时为3．5h,甚至在0．6mol／L NaCl条件下仍可生长。慢生型大豆根瘤菌USDA110对盐敏感,在含有0.1 mol／L Nacl的YM培养液中不生长,但结瘤固氮能力强。     

中国东北地区野大豆及其共生根瘤菌的生态分布与特性研究

中国东北地区野大豆及其共生根瘤菌共生体系资源的生态分布甚广。从5853份的资源中分离获得了快生型大豆根瘤菌331株。从野大豆及其共生根瘤菌共生体系的生物学特性和微生物生理生化特性来分,可以归并成五大类群,它们在质量性状基本是一致的,在数量性状则各不相同,而反映了遗传多样性和进化的规律,为今后共生固氮的深入研究奠定了重要的基础。     

类黄酮激活根瘤菌在油菜上结瘤和固氮的研究初报

１引言自然界大多数植物不具备共生固氮系统．在非豆科中除Ｐａｒａｓｐｏｎｉａ［８］外,至今还未发现其它非豆科植物能像豆科植物一样与根瘤菌形成共生．聂延富报道了２．４Ｄ能使小麦根系诱导结瘤的作用［２］．ＡｌＭａｌｅｎ等研究了根瘤菌在水稻和油菜幼苗根系...     

食物中牛磺酸含量

本研究测定了食品包括水果、蔬菜、种子、坚果、谷类、肉类、海产品和奶制品的牛磺酸含量。发现蚶和章鱼的牛磺酸;含量最高（４１．４μｍｏｌ／ｇ和３１．２μｍｏｌ／ｇ）,其次是小虾和鱼类（１２．４μｍｏｌ／ｇ和９．１μｍｏｌ／ｇ）。牛肉、猪肉和羊肉含有的牛磺酸范围为３．５～４．０μｍｏｌ／ｇ,鸡腿中的牛磺酸含量为６．６μｍｏｌ／ｇ,鸡胸肉为１．４μｍｏｌ／ｇ。鸡蛋（蛋黄和蛋白）、奶酪产品或蜂蜜中未发现含有牛磺酸。水果和蔬菜未检出牛磺酸。种子和谷类的检出结果为：稻米、玉米、麦片、裸麦、小麦、大麦、芝麻、咖啡豆、巧克力豆不含牛磺酸,南瓜子含１３．５ｎｍｏｌ／ｇ,黑豆含９．２ｎｍｏｌ／ｇ,蚕豆含１２．９ｎｍｏｌ／ｇ,嫩豌豆含１８．７ｎｍｏｌ／ｇ,花生未检出牛磺酸。胡桃、杏仁、如树坚果、榛子和松子中牛磺酸的含量范围为１５～４６ｎｍｏｌ／ｇ,阿月浑子含很少量的牛磺酸（４．９ｎｍｏｌ／ｇ）。所有分析是以不经烹调的样品完成的。这些结果的重要性可以认为是人体和动物牛磺酸缺乏有害结果的依据。     

八株弗氏中华根瘤菌的血清学和氮代谢研究

本文对从中国东北地区土壤中分离到的８株弗氏中华根瘤菌（Ｓｉｎｏｒｈｉｚｏｂｉｕｍｆｒｅｄｉｔ）进行了血清学和氢代谢研究。交叉凝集试验结果表明其中存在３种血清型,而Ｓｊ５与国内外目前发现的１４种Ｓ．ｆｒｅｄｉｉ接种的大豆依赖共生固氮作用,在其株木质部汁液中,含有大量的酰脲（尿囊酸＋尿囊素）,它是共生固氮氮素贮存和运输的主要形式,与接种Ｂ．ｊａｐｏｎｉｃｕｍ的值株木质部汁液中的氮运输特征基本相同。而施以无机氮源的大豆植株,其木质部汁液中酰脲含量相对较低,但却含有相对多的氮基酸［１］。     

外源硒对大豆产量、植株氮磷含量及土壤酶活性的影响

利用盆栽试验,研究了不同浓度、不同价态外源se对大豆产量、N、P含量及与N、P代谢密切联系的土壤脲酶和磷酸酶活性的影响．结果表明,施用浓度为0．25和0．5μg·g^-1土的Se^4+和Se^6+后,大豆产量、含N量及土壤脲酶活性增加,大豆含P量和土壤磷酸酶活性降低．当Se^6+浓度为0．5μg·g^-1土时,大豆植株含N量与土壤脲酶活性呈极显著的抛物线相关关系;Se^6+浓度为0．25μg·g^-1土时,大豆植株含P量与土壤磷酸酶活性呈极显著的抛物线相关关系。Se^4+处理的大豆N、P含量和土壤脲酶和磷酸酶活性均无显著相关关系。     

Acetylene reduction (nitrogen fixation) associated with corn inoculated with Spirillum.

Sorghum and corn breeding lines were grown in soil in field and greenhouse experiments with and without an inoculum of N2-fixing in Spirillum strains from Brazil. Estimated rates of N2 fixation associated with field-grown corn and sorghum plants were less than 4 g of N2/ha per day. The mean estimated N2-fixation rates determined on segments of roots from corn inoculated with Spirillum and grown in the greenhouse at 24 to 27 degrees C were 15 g of N2/ha per day (16 inbreds), 25 g of N2/ha per day (six hybrids), and 165 g of N2/ha per day for one hybird which was heavily inoculated. The corresponding mean rates determined from measurements of in situ cultures of the same series of corn plants (i.e., 16 inbreds, six hybrids, and one heavily inoculated hybrid) were 0.4, 2.3, and 1.1 g of N2/ha per day, respectively. Lower rates of C2H2 reduction were associated with control corn cultures which had been treated with autoclaved Spirillum than with cultures inoculated with live Spirillum. No C2H2 reduction was detected in plant cultures treated with ammonium nitrate. Numbers of nitrogen-fixing bacteria on excised roots of corn plants increased an average of about 30-fold during an overnight preincubation period, and as a result acetylene reduction assays of root samples after preincubation failed to serve as a valid basis for estimating N2 fixation by corn in pot cultures. Plants grown without added nitrogen either with or without inoculum exhibited severe symptoms of nitrogen deficiency and in most cases produced significantly less dry weight than those supplied with fixed nitrogen. Although substantial rates of C2H2 reduction by excised corn roots were observed after preincubation under limited oxygen, the yield and nitrogen content of inoculated plants and the C2H2-reduction rates by inoculated pot cultures of corn, in situ, provided no evidence of appreciable N2 fixation.     

Novel endophytic nitrogen-fixing clostridia from the grass Miscanthus sinensis as revealed by terminal restriction fragment length polymorphism analysis

Anaerobic nitrogen-fixing consortia consisting of N2-fixing clostridia and diverse nondiazotrophic bacteria were previously isolated from various gramineous plants (K. Minamisawa, K. Nishioka, T. Miyaki, B. Ye, T. Miyamoto, M. You, A. Saito, M. Saito, W. Barraquio, N. Teaumroong, T. Sein, and T. Tadashi, Appl. Environ. Microbiol. 70:3096-3102, 2004). For this work, clostridial populations and their phylogenetic structures in a stand of the grass Miscanthus sinensis in Japan were assessed by a 16S rRNA gene-targeted terminal restriction fragment length polymorphism (TRFLP) analysis combined with most-probable-number (MPN) counts. PCR primers and restriction enzymes were optimized for analyses of the plant clostridia. Clostridia were detected in strongly surface-sterilized leaves, stems, and roots of the plants at approximately 10(4) to 10(5) cells/g of fresh weight; they made up a large proportion of N2-fixing bacterial populations, as determined by MPN counts associated with an acetylene reduction assay. Phylogenetic grouping by MPN-TRFLP analysis revealed that the clostridial populations belonged to group II of cluster XIVa and groups IV and V of cluster I; this result was supported by a culture-independent TRFLP analysis using direct DNA extraction from plants. When phylogenetic populations from M. sinensis and the soil around the plants were compared, group II clostridia were found to exist exclusively in M. sinensis.     

酸性土壤上缺磷和铝毒对大豆生长的交互作用

以7个磷效率不同的大豆基因型为材料,通过土壤盆栽试验进行石灰和磷肥处理,研究酸性土壤上缺磷和铝毒对大豆生长的交互影响及其基因型差异.结果表明：缺磷和铝毒是酸性土壤上同时存在的影响大豆生长的主要障碍因子,其中铝毒对大豆生长的限制更为严重;缺磷和铝毒对酸性土壤上大豆生长的影响具有显著的交互作用.同时施用石灰（降低铝毒）和磷肥（提高磷有效性）比单施石灰或单施磷肥处理对大豆生长的促进效果更显著;缺磷和铝毒对大豆磷吸收的影响远大于对氮、钾吸收的影响.合理种植大豆对酸性土壤具有较好的改良作用.同时施用石灰［1.103 g Ca(OH)₂·kg^-1土］和磷肥（2.018 g KH₂PO₄·kg^-1土）可使酸性土壤pH值平均提高38.4%,交换性铝含量降低96.3%,有效磷含量提高3223.8％.种植磷高效大豆基因型比磷低效大豆基因型对酸性土壤的改良效果更好.     

四种除草剂对根瘤菌、AMF等土壤微生物的影响

本文采用盆栽试验, 研究4种除草剂在大田常规剂量下, 对根际土壤微生物和土著根瘤菌、AMF与苗期大豆共生的影响。结果表明, 除草剂对V2期大豆根际土壤中真菌数量和V3期细菌数量的抑制作用显著, 对V2和V3期AMF的侵染率、V3期根瘤数量影响显著。除草剂对土壤中真菌数量的负面影响可能是抑制菌根形成的重要因素, 对菌根形成产生的抑制可能会进一步导致根瘤形成受到抑制。     

粉煤灰改良砂姜黑土对麦田生态因子及重金属残留的影响

采用盆栽试验,研究了粉煤灰改良砂姜黑土对麦田生态因子及Cd、Cr、Pb、Hg和As残留的影响.结果表明,粉煤灰改良砂姜黑土可以降低土壤容重和土壤比重,减少土壤粘粒含量,增加土壤孔隙度和土壤渗透系数,提高耕层地温,促进土壤微生物活动和养分转化;土壤含水量高时具有散湿作用,土壤含水量低于10%后,具有保墒作用;在粉煤灰用量为6×10⁴～18×10⁴kg·hm^-2的条件下,土壤及小麦子粒中重金属元素的积累量远低于国际污染标准值,即在此范围内,施用粉煤灰改良砂姜黑土是安全可靠的.     

Characterization of a Mannitol-Utilizing, Nitrogen-Fixing Bradyrhizobium japonicum USDA 110 Derivative

We have isolated a colonial derivative of Bradyrhizobium japonicum USDA 110 (designated MN-110) that is both mannitol utilizing and N(2) fixing. Derivative MN-110 showed growth on mannitol and glucose similar to that of non-N(2)-fixing, mannitol-utilizing L2-110. Derivative MN-110 showed high constitutive and induced d-mannitol dehydrogenase activity (similar to L2-110) relative to N(2)-fixing, non-mannitol-utilizing I-110. Hybridization to EcoRI and HindIII total DNA digests with cloned USDA 110 nif DK and nif H genes revealed similar patterns for non-N(2)-fixing mannitol-utilizing derivative L1-110 and derivative MN-110. Symbiotic tests with soybean cultivars Ransom and Lee indicate MN-110 to be a superior N(2)-fixing derivative compared with derivative I-110 and the parent strain USDA 110. However, these differences were not revealed when comparing 28-day-old soybean-B. japonicum associations but were apparent in 49-day-old associations. It was apparent from this work that mannitol utilization was not necessarily correlated to symbiotic effectiveness in B. japonicum and that gene rearrangements were not responsible for differences in N(2) fixation between L1-110 or L2-110 and MN-110.     

根瘤菌菌剂的研究与开发现状

根瘤菌与豆科植物共生成为豆科植物固氮的重要方式,它可以为豆科植物提供所需氮量的1/2～1/3。因此,土壤中有效根瘤菌的数量是决定豆科植物产量的重要因素,而根瘤菌菌剂的使用可以有效地提高土壤中根瘤菌数量。本文从根瘤菌菌剂制备中高效菌种的选育及匹配、高密度菌剂的制备、菌剂保存方法等方面进行综述。比较了自然选育、杂交选育和诱变选育等各类选育方法及琼脂试管配对法和水培配对法的优缺点;总结了菌剂制备的一般过程和方法;论述了菌剂保藏过程中冷冻干燥法和各种保护剂的使用对菌剂保藏效果的影响。本文阐述了根瘤菌菌剂的制备工艺和发展方向,为根瘤菌剂的研制提供重要参考。     

Sensitivity of the common bean (Phaseolus vulgaris L.) symbiosis to high soil temperature

Summary Experiments were done to test whether N fixation is more sensitive to high soil temperatures in common bean than in cowpea or soybean. Greenhouse experiments compared nodulation, nitrogenase activity, growth and nitrogen accumulation of several host/strain combinations of common bean with the other grain legumes and with N-fertilization, at various root temperatures. Field experiments compared relative N-accumulation (in symbiotic relative to N-fertilized plants) of common bean with cowpea under different soil thermal regimes. N-fertilized beans were unaffected by the higher temperatures, but nitrogen accumulation by symbiotic beans was always more sensitive to high root temperatures (33°C, 33/28°C, 34/28°C compared with 28°C) than were cowpea and soybean symbiosis. Healthy bean nodules that had developed at low temperatures functioned normally in acetylene reduction tests done at 35°C. High temperatures caused little or no suppression of nodule number. However, bean nodules produced at high temperatures were small and had low specific activity. ForP. vulgaris some tolerance to high temperature was observed among rhizobium strains (e.g., CIAT 899 was tolerant) but not among host cultivars. Heat tolerance ofP. acutifolius andP. lunatus symbioses was similar to that of cowpea and soybean. In the field, high surface soil temperatures did not reduce N accumulation in symbiotic beans more than in cowpea, probably because of compensatory nodulation in the deeper and cooler parts of the soil.     

Nitrogen transfer from nodulating soybean to maize or to nonnodulating soybean in intercrops: the 15N dilution method

In 1985, 1986 and 1988, maize (Zea mays L.) was monocropped or intercropped with nodulating or nonnodulating soybean (Glycine max [L.] Merr.). In addition, nodulating soybean and nonnodulating soybean were each monocropped and grown as a mixture. In 1985 and 1986, treatments were grown at 0 and 60 kg N ha^–1 and in 1988, the treatments were grown without N fertilizer, on N-depeted soil and on non-N-depleted soil. ¹⁵N enriched N was applied to soil in all the aforementioned treatments to test for N transfer from nodulating soybean to non-N₂-fixing crops by the ¹⁵N dilution method.The ¹⁵N dilution method did not show the occurrence of N transfer in 1985 and 1986, but the N sparing effect was evident from the total N uptake of nonnodulating soybean, dwarf maize and tall maize, in 1986. In 1988, maize and nonnodulating soybean seed yields and seed N yields were higher on non-N-depleted soil than on N-depleted soil. On N-depleted soil, the ¹⁵N dilution method indicated N transfer from nodulating soybean to maize and to nonndulating soybean. At a population ratio of 67% nodulating soybean to 33% nonnodulating soybean, N transfer was also seen on non-N-depleted soil in 1988.