水稻对耐氨固氮工程菌株的响应

耐氨固氮工程菌（下称工程菌）催娩克氏杆菌（Ｋｌｅｂｓｉｅｌｌａ）ＮＧ１３９０和阴沟肠杆菌（Ｅｎｔｅｒｏｂａｃｔｅｒｃｌｏａｃａｅ）Ｅ２６１３的基因组带有组成型ｎｉｆＡ,其固氮酶的生成不受氨的阻遏,因此在有氨条件下具有固氮能力。接种固氮菌分别在两个水稻品系,即籼稻品种Ｃ５４４４和粳稻品种秀水－０４进行试验。水稻种植在灭菌土壤的塑料桶,生长至扬花后期取样分析表明,接种固氮菌能促进水稻的生长和植株含氮量的增加。接种工程菌ＮＧ１３９０的植株其生物量、总氮和固氮率（Ｎｄｆａ％）超过接种野生型ＮＧ１３的水稻。Ｅ２６１３接种Ｃ５４４４也获得类似的结果。应用１５Ｎ同位素稀释法证明了固氮菌接种水稻形成的联合固氮体系,存在细菌的固氮作用,而植株中氮素成分的增加可能来自细菌固定的氮。     

固氮阴沟肠细菌(Enterobacter cloacae)某些生理特性的研究

从广东省水稻根际分离出的阴沟肠细菌是一种兼性厌氧细菌。它具有较高的固氮能力。在培养过程中产生乳酸和氢、二氧化碳、氨等大量气体,并能排出一些含肽物质。在好气条件下,介质中硝酸盐能诱导形成硝酸还原酶,其活性随硝酸盐浓度增加而升高,硝酸盐浓度过高时抑制其活性。固氮酶活性则相反,随硝酸盐浓度增加而急剧下降,直至全部丧失。在厌氧条件下E.cloacae具有需硝酸的固氮活性。     

阴沟肠杆菌Tn5－nifA重组质粒的构建及其nifA的表达分析

构建的重组质粒ＰＢＦ１０１带有阴沟肠杆菌（Ｅｎｔｅｒｏｂａｃｔｅｒｃｌｏａｃａｅ）Ｅ２６固氮基因ｎｉｆＡ片段,该质粒具有广泛宿主范围接合转移特性和转座子Ｔｎ５的转座作用。验证了Ｅ２６ｎｉｆＡ产物能激活肺炎克氏杆菌（Ｋｌｅｂｓｉｅｌｌａｐｎｅｕｍｏｎｉａｅ）ｎｉｆＨ启动子的表达,解除氨对固氮酶合成的阻遏作用。发现当ＰＢＦ１０１引入自生固氮催娩克氏杆菌（Ｋ．ｏｘｙｔｏｃａ）ＮＧ１３后,可观察到固氮酶的组成型生成,同时在３９℃高温条件下的细菌仍能检测到部分固氮活性。     

稻根联合固氮细菌的研究I．菌种的分离和鉴定

从水稻根内分离到两株固氮能力较强的细菌,A-15和 E-26。经鉴定,A-15的表现性状与粪产碱菌的性状相符,但其DNA中GC含量为62．95—63 93克分子％,略高于文献报道的58．9克分子％。 由于缺乏粪产碱菌的模式株,暂将A—15归属于粪产碱菌(Alcaligenesfaecalis)。E-26的表观性状与阴沟肠杆菌属相符,因此将E一26归属于阴沟肠杆菌(Entcroba-cter cloacae)。据报道,在玉米根际曾分离到固氮的阴沟肠杆菌,却未见到有分离出固氮的粪产碱菌者。     

水稻内生固氮细菌的分离及鹑鸡肠球菌在水稻根中的分布

从福建省推广的谷杆两用水稻201中分离出内牛细菌12株,鉴定为阴沟肠杆菌（Enterobacter cloacae）、鹑鸡肠球菌（Enterococcus gallinarum）、短小芽孢杆菌（Bacillus pumilus）、地衣芽孢杆菌（Bacillus licheniformis）、巨大芽孢杆菌（Bacillus megaterium）、球形芽孢杆菌（Bacillus sphaericus）、蜡状芽孢杆菌（Bacillus cereus）、脂肪杆菌（Pimelobacter）和节杆菌（Arthrobacter）。选择固氮酶活性较大的3个菌株,对其抗性、革蓝氏染色特性进行了研究。采用三亲交配法将标记基因nifH-lacZ导入到鹑鸡肠球菌中,用该固氮细菌回接水稻,对感染鹑鸡肠球菌的水稻根进行β-半乳糖苷酶组织化学染色、光学显微镜和电子显微镜观察。结果表明,固氮细菌鹑鸡肠球菌在水稻根表皮细胞、内皮层细胞、维管组织细胞和细胞间隙中存在。     

阴沟肠杆菌nifL对nifA蛋白激活固氮基因表达的影响

通过基因定位突变的方法 ,构建了阴沟肠杆菌E2 6染色体上基因nifL突变菌E11和E12 ,并对其固氮特性进行分析。结果显示 ,当细菌在有NH 4 的条件下 ,nifL作为固氮基因的负调节因子 ,可能是通过nifL蛋白与nifA蛋白之间的相互作用 ,形成某种复合物 ,导致NifA的失活 ,从而关闭nif基因的表达。高温条件下 (37℃ ) ,nifL并不参与对nif基因表达的阻遏作用 ,但是nifL的突变影响NifA激活nif基因表达对热的稳定性     

用遗传工程技术强化水稻结合性固氮的研究

本文介绍甘蔗、玉米、水稻等粮食作物和经济作物根际固氮微生物及固氮活性的存在,阐明水稻可以根据这一存在,运用工程技术的成就,将固氮基因引入水稻根面细菌,强化水稻形成结合性固氮,以便为水稻提供氮素营养的可能性。试验证明,带有固氮基因质核粒(PRD_1)转入根面受体菌或采用结合转移,均能获得成功。在获得转化体的同时,探讨培育抗氨、泌氨高效菌株的途径、提出水稻种子催芽拌种,或蘸根头肥时加入高效的固氮菌株以强化形成结合性固氮的可能性。     

粪产碱菌（Alcaligenes faecalis）对稻根氧化还原特性及水稻多元酚和内根际激素水平的影响

联合固氮粪产碱菌结合于水稻根表时能增强水稻根部还原力和稻根超氧化物歧化酶（ＳＯＤ）活性。实验室和田间试验证明粪产碱菌能提高水稻幼苗对高、低温不良环境的抗逆性。经浸种处理的水稻幼苗植株内多元酚含量增加了１２．５％。粪产以菌对接种水稻多元酚抽提物有强烈的趋化性,而该抽提物对粪产碱菌的固氮活性有明显的刺激作用。多元酚抽提物经双向纸层析和薄层层析表明接种诱导了至少一个特征组分含量提高。采用粪产减菌野生型Ａ１５０１（ｎｉｆ＋）和固氮缺陷型Ａ１５０６（Ｎｉｆ－）浸种能改变宿主水稻内源激素水平,提高内根际的ＩＡＡ和Ｚ的含量,促进植株及根系的生长发育,使其侧根和根毛数目明显增多。在根际联合体系形成过程中,多元酚或激素可能充当植物与细菌间相互作用的一类特异信号分子。     

水稻根际固氮菌与水稻根愈伤组织联合固氮体系的研究初报

组织培养法应用在豆科和非豆科共生体系的研究中已有报道,但应用于水稻与根际固氮菌联合共生的研究中至今还未见。我们在探讨水稻根际联合固氮体系的植物细胞与细菌的关系中,采田了水稻根组织培养的方法,建立了愈伤组织水平上的联合固氮体系。初步结果表明这种方法用在水稻的联合固氮研究中是可行的,并取得了有意义的结果,现报道如下。     

水稻内生联合固氮细菌的筛选,鉴定及其分布特性

利用乙炔还原法和固定１５Ｎ２ 活性测定法对分离自水稻（ Ｏｒｙｚａ ｓａｔｉｖａ Ｌ．）“越富”种子、根、茎和叶的内生细菌进行了筛选,获得２９ 株具有体外固氮能力的水稻内生联合固氮细菌。鉴定结果表明它们分属于根癌土壤杆菌（ Ａｇｒｏｂａｃｔｅｒｉｕｍ ｔｕｍｅｆａｃｉｅｎｓ （Ｓｍｉｔｈ ｅｔ Ｔｏｗｎｓｅｎｄ） Ｃｏｎｎ） ,放射土壤杆菌（ Ａ． ｒａｄｉｏｂａｃｔｅｒ （Ｂｅｉｊｅｒｉｎｃｋ ｅｔ ｖａｎ Ｄｅｌｄｅｎ） Ｃｏｎｎ） ;阴沟肠杆菌（ Ｅｎｔｅｒｏｂａｃｔｅｒ ｃｌｏａｃａｅ （Ｊｏｒｄａｎ） Ｈｏｒｍａｅｃｈｅ ｅｔ Ｅｄｗａｒｄｓ） ,成团肠杆菌（ Ｅ． ａｇｇｌｏｍｅｒａｎｓ （Ｂｅｉｊｅｒｉｎｃｋ） Ｅｗｉｎｇ ｅｔ Ｆｉｆｅ） ,坂崎肠杆菌（ Ｅ． ｓａｋａｚａｋｉｉ Ｆａｍｅｒ ｅｔ ａｌ．） ;皮氏产碱菌（ Ａｌｃａｌｉｇｅｎｅｓ ｐｉｅｃｈａｕｄｉｉ Ｋｉｒｅｄｊｉａｎ ｅｔ ａｌ．） ,反硝化产碱菌（ Ａ． ｄｅｎｉｔｒｉｆｉｃａｎｓ （Ｌｅｉｆｓｏｎ ｅｔ Ｈｕｇｈ） Ｒｕｇｅｒ ｅｔ Ｔａｎ） ;类产碱假单胞菌（ Ｐｓｅｕｄｏｍｏｎａｓ ｐｓｅｕｄｏａｌｃａｌｉｇｅｎｅｓ Ｓｔａｎｉｅｒ） ,产碱假单胞菌（ Ｐ． ａｌｃａｌ     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

Time of detection of recessive genes: effects of system of mating and number of examined individuals

Summary Anthers were cultured from two sets of seven lines of hexaploid wheat (Triticum aestivum L.) with different cytoplasms, the euplasmic nucleus donors, Siete Cerros 66 and Penjamo 62, as well as their six alloplasmic lines derived from wild relative species of the genera Triticum and Aegilops. Significant cytoplasmic and nuclear effects but no cytoplasmic-nuclear interaction were found for embryogenic anther response, with the best performance of Penjamo 62 in Ae. kotschyi cytoplasm. Plant regeneration was not affected significantly by the cytoplasmic background of the lines cultured. The possible genetic implications of the observed cytoplasmic and nuclear influences on the in vitro haploid induction of wheat are discussed.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

Scales of spatial patterns of distribution of intertidal invertebrates

Few comparative studies of spatial patterns at different scales have examined several species in the same habitat or the same species over a range of habitats. Therefore, variability in patterns among species or among habitats has seldom been documented. This study quantifies spatial patterns of a suite of intertidal snails and a species of barnacle using a range of statistical techniques. Variability in densities was quantified from the scale of adjacent quadrats (over a distance of centimeters) to tens of kilometers. Significant differences in abundances occurred primarily at two spatial scales. Small-scale differences were found at the scales of centimeters or 1–2 m and, for many species on many shores, these accounted for most of the variability in abundances from place to place. These are likely to be determined by behavioural responses to small-scale patches of microhabitat. Large-scale differences in abundance were also found in most species at the scale of hundreds of meters alongshore. These are likely to be due to variation in recruitment (and/or mortality) because of limited dispersal by adults of these species. There was little or no additional variation among shores, separated by tens of kilometers, than was shown among patches of shore separated by hundreds of meters. Identification of the scale(s) at which significant differences in abundance are found focus attention on the processes (and the scales at which these processes operate) that influence patterns of distribution and abundance. Some of the advantages and disadvantages of various procedures are discussed.     

Identification of the site of glycation of human insulin

This study evaluates the nature of glycated human insulin formed following exposure to hyperglycemic conditions in vitro. Glycated insulin was purified by RP-HPLC and its molecular mass (5971.3 Da) determined by plasma desorption mass spectrometry (MS). The difference in mass (163.7 Da) from nonglycated insulin (5807.6 Da) corresponds to a single reduced glucose (glucitol) residue. Following reduction of insulin disulfide bridges, MS confirmed that the B-chain was glycated. Enzymatic digestions with trypsin, endoproteinase Glu-C, and thermolysin, followed by RP-HPLC and identification of fragments by MS, localized glycation to the B-chain (1–5) region. Electrospray tandem MS identified the site of glycation as the B-chain NH₂-terminal Phe¹ residue. This was confirmed by automated Edman degradation with glycated human insulin.