毛乌素沙地中间锦鸡儿根瘤菌的多样性及其抗逆性

从内蒙古毛乌素沙地优势物种中问锦鸡儿收集根瘤,分离根瘤菌15株,对其进行同工酶电泳,聚类分析证明中间锦坞儿的遗传多样性对其耐盐性、耐酸碱性、生长温度范围、产酸产碱性、过氧化氢酶活性以及唯一碳源利用能力进行比较研究,结果表明,所有中间锦鸡儿根瘤菌均分泌H^+,都具有过氧化氢酶活性,73.3%的菌株可以在含3％NaCl的YMA培养基上生长,80％的菌株可以抵抗50℃高温,除对乳糖和淀粉利用表现出差异外,中间锦鸡儿根瘤菌对其余碳源没有选择性,证明中间锦鸡儿根瘤菌具有较强的抗逆性,但仍存在菌株间差异,是对毛乌素沙地多种景观生态类型的适应作为一种新的种质资源,中间锦鸡儿根瘤菌可用于恶劣生境固氮。     

毛乌素沙地中间锦鸡儿根瘤菌遗传多样性及16S rDNA全序列分析

野生豆科植物中间锦鸡儿是毛乌素沙地的优势种。从蛋白质和DNA水平分析与其共生的根瘤茵的遗传多样性。在蛋白质水平上,24株中间锦鸡儿根瘤菌和9株参比菌株分为2组,A组包含95．8％供试中间锦鸡儿根瘤菌,参比菌株聚为B组,菌株GH72不与其余供试根瘤菌聚类。应用16S　rDNA　PCR—RFLP方法将供试菌株分为22种基因型,中间锦鸡儿根瘤菌组成12种,其余10种由参比菌株构成。表明中间锦鸡儿根瘤菌具高水平遗传多样性。选取代表菌株GH2001进行16S　rDNA全序列测定。与已知相关根瘤菌菌株16S　rDNA进行同源性比较,构建系统发育树状图。GH2001位于Rhizobium分支,与且Agrobacterium radiobacter,Ag.rubi,Rhizobium giardinii,R.mongolense,R.yanglingense,R.galegae和R.huautlense的序列同源性分别达到99％、98．3％、96．3％、95．5％、95．6％、95．27％和95．7％。     

沙冬青根瘤菌的抗逆性

沙冬青(Ammopiptanthus mongolicus)是西北荒漠地区珍稀常绿阔叶灌木,综合沙冬青分布区内气候、土壤和水分等因子,选取宁夏中卫县沙坡头、内蒙古磴口县、内蒙古阿拉善左旗、内蒙古乌拉特后旗作为样区,于2004年4月中下旬沙冬青的花期调查和收集根瘤,并结合地理环境对分离得到的根瘤菌进行抗逆性初步分析。调查发现根瘤的生成及其在宿主植物上的着生部位、大小、形状、颜色等因生境条件的不同而有所差异,水分是其主要的影响因子;分离得到根瘤菌17株,对其耐盐性、耐酸碱性、生长温度范围和抗生素抗性进行了研究。结果表明,64.7%的菌株可以在含3%NaCl的YMA培养基上生长;94.1%的菌株可以在pH 5~11的范围内生长;全部菌株在60 ℃处理10 min后仍能生长;不同菌株对不同抗生素表现出不同的抗性,ZW₄和WH₄¹对各抗生素表现出较强的抗性。分离自磴口县的根瘤菌普遍表现出较强的抗酸碱和抗高温的能力,这是对其环境的适应。本研究证明沙冬青根瘤菌具有较强的抗逆性,但菌株间存在差异,这可能是对沙冬青分布区多种景观生态类型的适应。同时测定了几种伴生植物中间锦鸡儿(Caragana intermedia)、猫头刺(Oxytropis aciphylla)和柠条(Caragana corshinskii)根瘤菌的抗逆性,与沙冬青根瘤菌表现出一定的相似性,具较强的抗逆性,说明生态环境对豆科植物-根瘤菌共生体有重要的影响。沙冬青的高抗性可能与沙冬青根瘤菌的高抗性有关。沙冬青与其根瘤菌的共生关系有利于其植物群落的发育和多样性的维系。     

沙埋对中间锦鸡儿幼苗生长发育的影响

在沙地环境中,沙埋是影响植物幼苗存活的一个重要因素。中间锦鸡儿(C arag ana interm ed ia)是浑善达克沙地和毛乌素沙地固定和半固定沙丘上常见的一种沙生灌木。为了研究沙埋对其幼苗存活以及生长的影响,对出土后生长了1个月的中间锦鸡儿实生苗进行沙埋实验,实验处理:TC为对照,T1为沙埋幼苗地上部的1/3,T2为沙埋幼苗地上部的1/2,T3为沙埋到幼苗的近顶端。实验持续4周,每周收获1次。结果表明:TC、T1和T2没有幼苗死亡,T3有20%的幼苗死亡;沙埋对中间锦鸡儿幼苗的生物量、根冠比、根生物量比和茎生物量比有显著影响,叶片生物量比各处理没有显著差异,沙埋对幼苗的叶面积没有显著影响,在第1周T2和T3的小叶片面积显著大于TC,在其余3周,TC与T1、T2的小叶片面积没有显著差异,与全埋则有显著差异;沙埋对幼苗的相对生长速率(RGR)和净同化速率(N AR)产生显著影响,实验结束时,T3处理幼苗的RGR和N AR显著地小于其它3个处理。上述结果表明,部分沙埋(T1、T2)对沙生灌木中间锦鸡儿幼苗的生长发育没有显著的影响,而全部沙埋(T3)对其幼苗的生长发育产生显著的抑制,中间锦鸡儿是一个比较耐沙埋的物种。     

新疆乌恰县豆科植物根瘤菌数值分类研究

新疆乌恰县具有独特的地理环境与气候条件,蕴藏着丰富的结瘤豆科植物资源.本研究从乌恰县豆科植物根瘤中分离到40株根瘤菌,并进行唯一碳源利用、唯一氮源利用、对抗生素和染料的抗性、耐盐性、初始pH生长、生长温度范围及石蕊牛奶反应、氧化酶、过氧化氢酶、脲酶共118项生理生化性状测定.结果表明:乌恰县豆科植物根瘤菌在碳氮源利用、抗生素敏感性、对染料抗性程度等方面存在着差异.供试根瘤菌能耐受较低温度,并具有较强的耐盐、碱能力,均能在pH 6~12的YMA培养基上生长,有60%的菌株能耐受0.86mol/L的NaCl.数值分类结果表明,在84.2%的相似水平上40株供试菌株构成了4个新的表观群.第Ⅰ、Ⅱ、Ⅲ和Ⅳ类群分别有8、9、3和3株菌,中心菌株分别为NWWQ73-2、NWWQ13-1、NWWQ1-2、NWWQ6-2.此外,乌恰县豆科植物根瘤菌与模式菌株的相似性较低,它们所形成的4个独立群可能有新种存在.     

新疆苦豆子根瘤菌的数值分类研究

苦豆子(Sophora alopecuroides)对于干旱荒漠地区的畜牧业发展有着非常重要的意义,其生长特性与根瘤菌有密切关系。我们对分离自新疆苦豆子根瘤的67株根瘤菌及36个模式菌株进行了118项表型性状的测定,包括唯一碳源利用、唯一氮源利用、对抗生素和染料的抗性、耐盐性、初始pH值生长范围、生长温度范围及石蕊牛奶反应、氧化酶、过氧化氢酶和脲酶。对测定结果用聚类分析方法进行了分析,获得数值分类树状图。结果表明：新疆苦豆子根瘤菌在碳氮源利用、抗生素敏感性以及对染料的抗性程度等方面存在着差异。新疆苦豆子根瘤菌能耐受低温,并具有较强的耐盐、碱能力,所有供试菌株均能在初始pH值为9－12的YMA培养基上生长,92.5％的菌株能耐受3.0％的NaCl,91.0％的菌株能耐受4.0％的NaCl,有18株菌甚至能耐受5.0％和6.0％的NaCl。聚类结果表明, 在84.8%的相似性水平上,67个供试菌株构成了4个新的表观群,第Ⅰ、Ⅱ、Ⅲ、Ⅳ类群分别有21、7、4、3个菌株,中心菌株分别为NWBC152、NWTKX101、NWYJS12、NWLP112。此外,数值分类结果还表明,苦豆子根瘤菌与模式菌株的相似性较低,它们所形成的4个独立群可能有新种出现。     

解糖热解纤维素菌F32降解未预处理秸秆及产纤维素酶特性分析

【目的】明确极端嗜热厌氧木质纤维素降解菌解糖热解纤维素菌F32代谢特征,并分析其产酶特性。【方法】使用细胞计数法绘制菌株的生长曲线,使用离子色谱及气相色谱进行产物和残糖量分析,以DNS法及对硝基苯酚法检测菌株胞外蛋白的酶活性。【结果】解糖热解纤维素菌F32在以葡萄糖、微晶纤维素和未经预处理小麦秸秆为碳源时生长状况优于解糖热解纤维素菌DSM 8903。在以葡萄糖为碳源进行培养时,与菌株DSM 8903相比,菌株F32具有产乳酸较多,而产氢气较少的特点。在以微晶纤维素和未经预处理小麦秸秆为碳源进行培养时,与菌株DSM 8903相比,菌株F32胞外蛋白具有较高的内切纤维素酶活性和木聚糖酶活性。【结论】解糖热解纤维素菌F32表现出较强的木质纤维素降解能力,其与DSM 8903的产物组成及胞外蛋白的酶活性具有明显差异。     

生态脆弱带不同区域近缘优势灌木的生理生态学特性

小叶锦鸡儿、中间锦鸡儿和柠条锦鸡儿分别为中国北方生态脆弱带典型区域——科尔沁沙地、毛乌素沙地和腾格里沙漠的优势灌木。为了探讨生长在地理位置相距很远的不同气候类型区、但亲缘关系较近的沙漠植物种之间特性的差异及近缘种间引种成功的可能性 ,对 3种灌木在原产地和引种地 (腾格里沙漠 )的气体交换、叶绿素荧光参数、β胡萝卜素和脯氨酸累积等特性进行了对比研究 ,结果显示 :各区优势灌木的气体交换特征不同 ,主要依照区域光照和温湿条件的不同组合而变化 ,各区域的环境条件组合最利于其建群种的生长 ;引进种对引种地环境变化的反应调节较乡土种敏感 ;地理位置相距较近的近缘种间生理特性相近 ,引种易于成功     

黄芪根瘤菌的多样性分析

选用23株分离自山西、内蒙古、宁夏、甘肃和吉林的黄芪根瘤菌菌株,进行了营养利用、抗生素抗性、耐逆性和酶活性测定,发现不同地区来源、甚至同一地区来源或同种寄主的不同菌株存在着较大的差异,这种不一致现象在来源于甘肃的7株菌中尤为明显,特别是G156、G130和GO13在碳源利用能力方面表现出诸多不同于本多数黄芪根瘤菌的特点。G156、G012和G022能在50℃生长。多数被试菌株在pH11的条件下生长,少数菌株耐4％～5％NaCl,这些结果均说明黄芪根瘤菌是一个极其多样化的类群,它们也明显不同于已知根瘤菌。     

沙埋和种子大小对柠条锦鸡儿种子萌发、出苗和幼苗生长的影响

研究了沙埋深度和种子大小对内蒙古毛乌素沙地植被群落中占优势的柠条锦鸡儿(Caragana korshinskii Kom.)种子萌发、出苗、幼苗存活和生长的影响.结果表明,沙埋深度显著影响柠条锦鸡儿的种子萌发率、休眠率、出苗率、幼苗存活率及生物量.在0.5-2cm的浅层沙埋下,种子萌发率、出苗率、幼苗存活率及生物量最高,休眠率最低;沙埋深度≥4 cm时,柠条锦鸡儿的种子萌发率、出苗率、幼苗存活率及生物量随着沙埋深度增加显著降低,而休眠率却显著升高;沙埋深度≥12 cm时,柠条锦鸡儿种子不能够出苗,幼苗也不能够存活.种子大小对柠条锦鸡儿种子萌发率没有显著影响,但对出苗率、幼苗存活率及生物量影响显著.在各个沙埋深度下,不同大小的柠条锦鸡儿种子间的萌发率没有显著差异.当沙埋深度≤6 cm时,不同大小的柠条锦鸡儿种子在同一沙埋深度下的出苗率间没有显著差异;但当沙埋深度≥8 cm时,在同一沙埋深度下,大种子的出苗率显著高于中种子和小种子的出苗率,而中种子和小种子出苗率间没有显著差异.0.5-10 cm的沙埋深度中,除6 cm和8 cm深度下中种子和小种子萌发幼苗的生物量间没有显著差异外,其余深度下,大种子萌发的幼苗的存活率及生物量显著高于同一沙埋深度下中种子萌发的幼苗的存活率及生物量,后者又显著高于小种子萌发的幼苗的存活率及生物量.可能正是种子萌发对沙埋环境的忍耐或响应能力以及种子的多态性提高了柠条锦鸡儿在毛乌素沙地的适合度,为其在流动或半流动沙丘环境中成功定居并形成优势群落奠定了基础.     

Genetic diversity of rhizobia isolated from Caragana intermedia in Maowusu sandland,north of China

AIMS: Thirty-three rhizobial strains isolated from nodules of Caragana intermedia in Maowusu sandland were examined for their genetic diversity and putative phylogenetic position. METHODS AND RESULTS: Isolates from Caragana intermedia were classified into 12 genotypes by 16S rDNA polymerase chain reaction-restriction fragment length polymorphism (RFLP), which showed no distinct relationships with those of the reference strains. The genotypes of rhizobia were not related to geographical location. Thr 16S rDNA sequence of representative strain GH2001 from dominant genotype 2 shared high homologuey with some Rhizobium species: Rh. giardinii (96.4%), Rh. huautlense (95.3%), Rh. galegae (95.7%), Rh. yanglingense (95.2%), Rh. mongolense (95.6%), Rh. radiobacter (99%) and Rh. rubi (98.3%). CONCLUSIONS: A high degree of genetic diversity existed among rhizobia nodulating Caragana intermedia in Maowusu sandland. Most of the new isolates might belong to Rhizobium. SIGNIFICANCE AND IMPACT OF THE STUDY: The results suggest that the rich diversity of rhizobia might have contributed to the adaptation of the arid region. These strains could be valuable at the economic and ecosystem level.     

卷荚相思根瘤菌的生物学特性

为探讨卷荚相思根瘤菌的生物学特性,从广西高峰林场卷荚相思林分采集分离到12株根瘤菌,对这些菌株进行了生理生化试验,并利用这些菌株对厚荚相思苗木进行接种结瘤试验。结果表明:(1)依据生长代时和单个菌落形成测定结果,分离获得的12个菌株可以分为9个快生菌株和3个慢生菌株。(2)大部分的供试根瘤菌菌株可以利用无机氮源作为其生长所需的氮源,但对供试用的糖源利用存在着较大的差别。(3)所有供试菌株在3一酮基乳糖反应、牛肉膏蛋白胨、淀粉水解试验中呈阴性,所有供试的12个菌株均能进行利用柠檬酸盐,在BTB试验中均产酸,在石蕊牛奶反应均产碱。(4)12个卷荚相思根瘤菌菌株接种到厚荚相思苗木后,均能使其根部结瘤,并进一步筛选出结瘤效果和固N效果比较好的供试菌株是快生型菌株(J003、J004、J005、J008)和慢生型菌株(J012)。这为从华南的酸性土壤中寻找和利用土著根瘤菌资源提供了科学依据。     

降雨对高寒沙地不同林龄中间锦鸡儿水分利用特征的影响

为明确不同林龄中间锦鸡儿水分来源对降雨的响应,利用稳定同位素技术测定青海共和盆地不同林龄的中间锦鸡儿(4a、9 a、17 a和31 a)在降雨前后土壤水、木质部水、地下水和雨水的δ2H、δ18O组成,运用Iso-Source模型计算植物对各潜在水源的利用比例.结果 表明:各林龄中间锦鸡儿的浅层(0～40 cm)土壤水δ...     

Increased metabolic potential of Rhizobium spp. is associated with bacterial competitiveness

Of 105 rhizobial isolates obtained from nodules of commonly cultivated legumes, we selected 19 strains on the basis of a high rate of symbiotic plant growth promotion. Individual strains within the species Rhizobium leguminosarum bv. trifolii, R. leguminosarum bv. viciae, and Rhizobium etli displayed variation not only in plasmid sizes and numbers but also in the chromosomal 16S-23S internal transcribed spacer. The strains were tagged with gusA gene and their competitiveness was examined in relation to an indigenous population of rhizobia under greenhouse conditions. A group of 9 strains was thus isolated that were competitive in relation to native rhizobia in pot experiments. Nineteen selected competitive and uncompetitive strains were examined with respect to their ability to utilize various carbon and energy sources by means of commercial Biolog GN2 microplate test. The ability of the selected strains to metabolize a wide range of nutrients differed markedly and the competitive strains were able to utilize more carbon and energy sources than uncompetitive ones. A major difference concerned the utilization of amino and organic acids, which were metabolized by most of the competitive and only a few uncompetitive strains, whereas sugars and their derivatives were commonly utilized by both groups of strains. A statistically significant correlation between the ability to metabolize a broad range of substrates and nodulation competitiveness was found, indicating that metabolic properties may be an essential trait in determining the competitiveness of rhizobia.     

腾格里沙漠东南缘不同植被类型土壤种子库多样性

为探究荒漠地区不同植被类型土壤种子库动态与分布规律,揭示不同植被类型群落的植被演替趋势与多样性特征,以腾格里沙漠东南缘胡杨群落、柠条群落、赖草群落和沙蓬群落为对象,采集0～2、2～5、5～10、10～20 cm土层土样,研究土壤种子库特征及其与地上植被的关系。结果表明: 4个植被类型的土壤种子库中共发现13种植物,隶属于3科12属,1、2年生草本植物种子占多数,藜科植物较多;土壤种子库密度较低,呈现柠条群落＞赖草群落＞胡杨群落＞沙蓬群落;种子库密度和植物种类整体随土壤剖面的加深而降低;综合Shannon指数、Simpson指数、Pielou指数和Patrick丰富度指数进行分析,土壤种子库物种多样性呈现胡杨群落＞赖草群落＞柠条群落＞沙蓬群落,柠条群落土壤种子库与地上植被相似性系数最高,沙蓬群落次之,胡杨群落和赖草群落最低。总体上,以胡杨群落为代表的乔木林较为稳定,以沙蓬群落为代表的沙地生态系统最为脆弱。     

Diversity and effectiveness of rhizobial strains from <Emphasis Type="Italic">Gliricidia sepium</Emphasis> native to Reunion Island,Kenya and New Caledonia

The aim of this study was to use several approaches to assess the diversity occuring in a collection of rhizobial strains that had been isolated from Gliricidia sepium (Jacq.) Walp. originating from Kenya, Reunion Island and New Caledonia. Results showed that G. sepium establishes an effective symbiosis only with fast-growing rhizobia. This suggests that this legume nodulates preferentially with this type of rhizobia, although ineffective nodules were observed on root systems of plants inoculated with slow-growing rhizobia. It was possible to distinguish several groups of strains which have the same behaviour in terms of utilization of amino acids and sources of carbon and intrinsic resistance to antibiotics and salinity. Six molecular groups were distinguished by profiles obtained using PCR/RFLP techniques. No relationship was demonstrated between phenotypic groups and molecular groups. Neither was it possible to find a relationship between the geographic origins of the strains and their distribution in the several phenotypic and/or molecular groups. In an experiment carried out under glasshouse conditions, we observed that all strains from the collection significantly improved the growth of G. sepium cultivated in a substrate totally devoid of nitrogen. The strain GsK5 induced the greatest stimulation of plant growth.     

Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis Protein Banding Patterns among Rhizobium leguminosarum biovar phaseoli Strains Isolated from the Mexican Bean Phaseolus coccineus

Several rhizobial strains were isolated from Phaseolus coccineus root nodules and were determined to be Rhizobium leguminosarum biovar phaseoli strains after reinfection of the same host plant. These strains were characterized by cultural procedures (growth on different carbon sources and intrinsic antibiotic resistance) and electrophoretic procedures (sodium dodecyl sulfate-polyacrylamide gel electrophoresis of total proteins). Our results showed that these rhizobia are very similar to each other, especially in their electrophoretic protein banding patterns, suggesting that they might belong to isolated populations.