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21.
John Stanley David N. Dowling William J. Broughton 《Molecular & general genetics : MGG》1988,215(1):32-37
Summary The hemA gene which encodes -aminolaevulinic acid synthase (ALAS), was cloned and characterized from the broad host-range Rhizobium strain NGR234. A cosmid, identified by hybridization with the cloned gene of R. meliloti and complementation of an R. meliloti hemA mutant, was subcloned to yield a 5.5 kb fragment containing the entire NGR234 gene. A physical-genetic map was made and the interposon was introduced into a single EcoRI site which bisects the gene. The mutated gene was homogenotized into NGR234 to generate a hemA mutant, with a view to evaluating the role of rhizobial bacteroid ALAS activity for a wide variety of legume symbioses. The mutant strain formed an ineffective (Fix–) symbiosis with all tested host plants. These included tropical legumes that produce either indeterminate (Leucaena) or determinate (Desmodium, Macroptilium, Lablab, Vigna) root nodules.Abbreviations ALA
-aminolaevulinic acid
- ALAS
aminolaevulinic acid synthase
- Lb
leghaemoglobin
- Lb-haem
haem moiety of leghaemoglobin 相似文献
22.
L. Sossountzov R. Maddiney B. Sotta I. Sabbagh Y. Habricot M. Bonnet E. Miginiac 《Planta》1988,175(3):291-304
Post-embedding immunocytochemical techniques using peroxidase-antiperoxidase or immunoglobulin G-gold as markers were used for the localization of cytokinins (CKs) in two isogenic lines, Craigella (C) and Craigella lateral suppressor (Cls), of tomato Lycopersicon esculentum Mill. Terminal buds, nodes, hypocotyl segments and root tips were submitted to a periodate-borohydride procedure, to obtain the coupling of isopentenyladeosine and zeatin riboside to cellular proteins, followed by a fixative step with a paraformaldehyde and glutaraldehyde mixture. Enzyme-linked immunosorbent assay tests performed on ovalbumin-coated microtitration plates have shown that this method was effective for CK riboside and base coupling to proteins. Paraffin-wax- or Spurr's-resin-embedded sections were cleared of wax or resin before incubation with anti-zeatin riboside or anti-isopentenyladenosine antibodies. The procedure was thoroughly investigated and many controls were done in order to eliminate artefacts. The immunostaining patterns observed along the plants showed a basipetally decreasing gradient of CKs along the stem and in the roots. Immunolabelling was higher in the actively growing regions of the stem bud and root apices. Terminal buds of Cls appeared to be less immunoreactive than C, whereas no differences were detected in root-tip immunolabelling. The staining patterns are consistent with the idea that root and bud apices have a different CK metabolism. The absence of axillary bud formation in Cls is correlated with low CK levels in the organogensis sites.Abbreviations C
Craigella, isogenic line
- CK
cytokinin
- Cls
Craigella lateral suppressor
- EDC
1-(3-dimethylaminopropyl)3-ethylcarbodiimide hydrochloride
- ELISA
enzyme-linked immunosorbent assay
- 2iP
isopentenyladenine
- 2iPA
isopentenyladenosine
- PAP
peroxidase-anti-peroxidase
- PFAG
paraformaldehyde/glutaraldehyde mixture
- Z
zeatin
- ZR
zeatin riboside 相似文献
23.
Effect of nitrogen stress and abscisic acid on nitrate absorption and transport in barley and tomato
F. Stuart Chapin III David T. Clarkson John R. Lenton Colin H. S. Walter 《Planta》1988,173(3):340-351
The potential of barley (Hordeum vulgare L.) and tomato (Lycopersicon esculentum Mill.) roots for net NO
3
-
absorption increased two-to five fold within 2 d of being deprived of NO
3
-
supply. Nitrogen-starved barley roots continued to maintain a high potential for NO
3
-
absorption, whereas NO
3
-
absorption by tomato roots declined below control levels after 10 d of N starvation. When placed in a 0.2 mM NO
3
-
solution, roots of both species transported more NO
3
-
and total solutes to the xylem after 2 d of N starvation than did N-sufficient controls. However, replenishment of root NO
3
-
stores took precedence over NO
3
-
transport to the xylem. Consequently, as N stress became more severe, transport of NO
3
-
and total solutes to the xylem declined, relative to controls. Nitrogen stress caused an increase in hydraulic conductance (L
p) and exudate volume (J
v) in barley but decrased these parameters in tomato. Nitrogen stress had no significant effect upon abscisic acid (ABA) levels in roots of barley or flacca (a low-ABA mutant) tomato, but prevented an agerelated decline in ABA in wild-type tomato roots. Applied ABA had the same effect upon barley and upon the wild type and flacca tomatoes: L
p and J
v were increased, but NO
3
-
absorption and NO
3
-
flux to the xylem were either unaffected or sometimes inhibited. We conclude that ABA is not directly involved in the normal changes in NO
3
-
absorption and transport that occur with N stress in barley and tomato, because (1) the root ABA level was either unaffected by N stress (barley and flacca tomato) or changed, after the greatest changes in NO
3
-
absorption and transport and L
p had been observed (wild-type tomato); (2) changes in NO
3
-
absorption/transport characteristics either did not respond to applied ABA, or, if they did, they changed in the direction opposite to that predicted from changes in root ABA with N stress; and (3) the flacca tomato (which produces very little ABA in response to N stress) responded to N stress with very similar changes in NO
3
-
transport to those observed in the wild type.Abbreviation and symbols ABA
abscisic acid
- Jv
exudate volume
- Lp
root hydraulic conductance 相似文献
24.
Production of gibberellins and indole-3-acetic acid by Rhizobium phaseoli in relation to nodulation of Phaseolus vulgaris roots 总被引:2,自引:0,他引:2
Similar ranges of gibberellins (GAs) were detected by high-performance liquid chromatography (HPLC)-immunoassay procedures in ten cultures of wild-type and mutant strains of Rhizobium phaseoli. The major GAs excreted into the culture medium were GA1 and GA4. These identifications were confirmed by combined gas chromatographymass spectrometry. The HPLC-immunoassays also detected smaller amounts of GA9- as well as GA20-like compounds, the latter being present in some but not all cultures. In addition to GAs, all strains excreted indole-3-acetic acid (IAA) but there was no obvious relationship between the amounts of GA and IAA that accumulated. The Rhizobium strains studied included nod
– and fix
– mutants, making it unlikely that the IAA- and GA-biosynthesis genes are closely linked to the genes for nodulation and nitrogen fixation.The HPLC-immunoassay analyses showed also that nodules and non-nodulated roots of Phaseolus vulgaris L. contained similar spectra of GAs to R. phaseoli culture media. The GA pools in roots and nodules were of similar size, indicating that Rhizobium does not make a major contribution to the GA content of the infected tissue.Abbreviations EIA
enzyme immunoassay
- GAn
gibberellin An
- GC-MS
gas chromatography-mass spectrometry
- HPLC
high-performance liquid chromatography
- IAA
indole-3-acetic acid
- Me
methyl ester
- RIA
radioimmunoassay
- TLC
thin-layer chromatography 相似文献
25.
Brian A. McGaw Roger Horgan Jim K. Heald George J. Wullems Rob A. Schilperoort 《Planta》1988,176(2):230-234
The levels of the major cytokinins, zeatin, zeatin riboside, zeatin riboside-5-monophosphate and zeatin-7-glucoside were measured in tobacco (Nicotiana tabacum L.) crown-gall tissues carrying insertion and deletion mutations in the T-DNA. Measurements were made by combined gas chromatography-mass spectrometry using selected ion monitoring with 15N- and 2H-labelled internal standards. The results demonstrate that, relative to wild-type tumour tissue, cytokinin levels are considerably elevated in tissues lacking functional T-DNA auxin-biosynthetic genes. From a detailed analysis of the major cytokinin metabolites it is concluded that a reduction in the extent of cytokinin degradation via N6-side-chain cleavage is an important factor leading to increased cytokinin levels in these tissues.Abbreviations IAA
indole-3-acetic acid
- SIM
selected ion monitoring
- Z
zeatin
- [7G]Z
zeatin-7-glucoside
- [9R]Z
zeatin-9-riboside
- [9R-5P]Z
zeatin riboside-5-monophosphate 相似文献
26.
We have isolated a series of monoclonal antibodies that react to antigens in flowers of Nicotiana tabacum L. (tobacco) displaying specificity or preferentiality in their cell and tissue distributions. We immunized mice with extracts from tobacco flowers and then screened the hybridomas by enzyme-linked immunosorbent assay (ELISA) against extracts from leaves, sepals, petals, stamens and pistils; twenty five were chosen from the total screened. The antigens detected by about half of the antibodies were periodate-sensitive, implying that the epitopes were carbohydrate. Competition ELISA assays were used to determine if any antibodies were reacting to the same epitopes. Western blot analysis showed that while some antibodies reacted to specific bands, the bulk either failed to react or reacted to multiple bands, consistent with a glyco-conjugate nature for many of the antigens. Analysis of the spatial pattern of antigen distribution within tobacco flowers by immunolocalization showed that some antibodies recognized epitopes that were limited to very specific cells and tissues. We used the immunolocalization technique to analyze a mutant with stigmoid anthers: an antibody recognizing a pistil transmitting-tract antigen also reacted to cells in stigmoid anthers. Our results with this antibody set imply that biochemical differentiation within the tobacco flower includes cell-and tissue-specific glyco-moeities, and also that similarities, at the biochemical level, exist between a normal floral organ and the abnormal organ in a phenotype with a developmental switch.Abbreviations ELISA
enzyme-linked immunosorbent assay
- Fg
immunoglobulin
- kDa
kilodalton 相似文献
27.
Masahito Natori 《Primates; journal of primatology》1988,29(2):263-276
The dental and cranial morphologies of all species ofSaguinus, S. oedipus, S. geoffroyi, S. leucopus, S. nigricollis, S. fuscicollis, S. labiatus, S. mystax, S. imperator, S.
bicolor, andS. midas are examined. The following hypotheses are developed by cladistic methodology, using only synapomorphic characters to assess
the interspecific relationships ofSaguinus.Saguinus are divided into two main groups; one consists ofS. oedipus, S. geoffroyi, andS. leucopus, and the other includesS. inustus, S. nigricollis, S. fuscicollis, S. labiatus, S. mystax, S. imperator, S. bicolor, andS. midas. In the former group,S. oedipus is more closely related toS. geoffroyi than either is toS. leucopus. In the latter group,S. labiatus, S. mystax, andS. imperator are classified into one group, andS. bicolor andS. midas form one monophyletic group. 相似文献
28.
29.
Nicole Benhamou Pierre J. Lafontaine Dominique Mazau Marie-Thérèse Esquerré-Tugayé 《Planta》1991,184(4):457-467
An antiserum raised against deglycosylated hydroxyproline-rich glycoproteins (HPGPs) from melon (Cucumis melo L.) was used to study the relationship between Rhizobium infection and induction of HRGPs in bean (Phaseolus vulgaris L.) root nodule cells infected with either the wild-type or a C4-dicarboxylic acid mutant strain of Rhizobium leguminosarum bv. phaseoli. In effective nodules, where fixation of atmospheric dinitrogen is taking place, HRGPs were found to accumulate mainly in the walls of infected cells and in peribacteroid membranes surrounding groups of bacteroids. Internal ramifications of the peribacteroid membrane were also enriched in HRGPs whereas the peribacteroid space as well as the bacteroids themselves were free of these glycoproteins. In mutant-induced root nodules, HRGPs were specifically associated with the electron-dense, laminated structures formed in plastids as a reaction to infection by this mutant. The presence of HRGPs was also detected in the host cytoplasm. The aberrant distribution of HRGPs in infected cells of mutant-induced nodules likely reflects one aspect of the altered host metabolism in relation to peribacteroid-membrane breakdown. The possibility that the antiserum used for HRGP localization may have cross-reacted with ENOD 2 gene products is discussed in relation to amino-acid sequences and sites of accumulation. 相似文献
30.
研究一系列有明确意义改性胰岛素的结构将有益于新型胰岛素分子的设计。经蛋白质工程制备的AspA21—人胰岛素(ADHI)基本保持了生物活力,而在酸性溶夜(pH3—4)中的稳定性比天然胰岛素在中性溶液中高5—10倍。本文报道ADHI的结晶、初步晶体学研究及中等分辨率衍射数据的收集。ADHI晶体属R3空间群:a_H=b_H=82.72埃,c_H=34.02埃。 相似文献