Immunoaffinity co-purification of cytokinins and analysis by high-performance liquid chromatography with ultraviolet-spectrum detection

Nicander, Björn; Ståhl, Ulf; Björkman, Per‐Olof; Tillberg, Elisabeth

doi:10.1007/bf00194427

Cited by 61 publications

(47 citation statements)

References 22 publications

(25 reference statements)

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“…The extract was filtered and taken through a precolumn containing 1 ml of Sepharose 6B (Pharmacia). The samples were chromatographed through an IAC column, essentially as described by Nicander et al (32) except that the column and solution temperatures were 30-35ЊC and for the comparison of extraction methods, ethanol was used as the eluent. The eluted fraction was evaporated under vacuum to about 300 l, diluted by the addition of PBS, and taken through the process a second time.…”

Section: ϫ2mentioning

confidence: 99%

Expression of early nodulin genes in alfalfa mycorrhizae indicates that signal transduction pathways used in forming arbuscular mycorrhizae andRhizobium-induced nodules may be conserved

Rhijn

Fang²,

Galili³

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

180

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Transcripts for two genes expressed early in alfalfa nodule development (MsENOD40 and MsENOD2) are found in mycorrhizal roots, but not in noncolonized roots or in roots infected with the fungal pathogen Rhizoctonia solani. These same two early nodulin genes are expressed in uninoculated roots upon application of the cytokinin 6-benzylaminopurine. Correlated with the expression of the two early nodulin genes, we found that mycorrhizal roots contain higher levels of trans-zeatin riboside than nonmycorrhizal roots. These data suggest that there may be conservation of signal transduction pathways between the two symbioses-nitrogen-fixing nodules and phosphate-acquiring mycorrhizae.Of the two commonly described symbioses of roots-nitrogenfixing nodules in response to members of Rhizobiaceae or Frankiaceae and phosphate-acquiring arbuscular mycorrhizae (AM) between roots and endophytic fungi-the more ancient of the two is AM. Structures identified as arbuscules have been found in Aglaophyton major, an early Devonian land plant, providing evidence for at least a Ͼ400-million-year-old association between terrestrial plants and fungi (1). In contrast, the Rhizobium-legume symbiosis is much younger, having been established no more than 65 to 136 million years ago, when the angiosperms evolved. However, unlike the AM symbiosis, which is found in almost all angiosperm families, Rhizobiuminduced nitrogen-fixing symbioses appear to be restricted to a single subclade of a larger nitrogen-fixing clade (2).LaRue and Weeden (3) proposed that nodulation evolved from the more ancient AM association because of certain similarities between the two symbioses. For example, some legumes are Nod Ϫ (absence of nodule formation) as well as Myc Ϫ

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Section: ϫ2mentioning

confidence: 99%

Expression of early nodulin genes in alfalfa mycorrhizae indicates that signal transduction pathways used in forming arbuscular mycorrhizae andRhizobium-induced nodules may be conserved

Rhijn

Fang²,

Galili³

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

180

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“…An unknown compound was isolated from potato tuber sprouts using immunoaffinity chromatography and HPLC (Nicander et al, 1993). The UV spectrum is shown in Figure 1, together with UV spectra of some cytokinin standards.…”

Section: Resultsmentioning

confidence: 99%

“…The details of this method have been published (Nicander et al, 1993). Sprouts from potato tubers were harvested, extracted, and immunopurified against anti-cytokinin antibodies.…”

Section: Kolation Of Cytokininsmentioning

confidence: 99%

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Identification of an N-Glucoside of cis-Zeatin from Potato Tuber Sprouts

1995

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A compound was isolated from potato (Solanum tuberosum 1. cv Bintje) tuber sprouts by immunoaffinity chromatography with antibodies against the cytokinins zeatin riboside and isopentenyladenosine. Analysis by ultraviolet spectroscopy and gas chromatography-mass spectrometry of derivatives identified the compound as a 9-glucoside of 6-[(Z)-4-hydroxy-3-methyl-2-butenylaminolpurine (cis-zeatin). Kglucosides have often been reported as metabolites of other cytokinins, but to our knowledge, they have never before been found for cis-zeatin. The finding gives proof that cis-zeatin, a modified base in tRNA, also exists as a free substance in plants, since the glucoside, unlike other tRNA-free cis-zeatins described earlier by others, cannot arise by enzymatic degradation of tRNA during plant extraction.

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“…Therefore plant hormones are usually conjugated to macromolecules to form antigens. Ulvskov and Nicander developed an IAC method in which antibodies are raised against IAA and cytokinins conjugated to bovine serum albumin (BSA) [51,52]. The antibody preparation step is the key factor for establishment of the IAC method because the antibody directly affects the specific affinity of the plant hormones.…”

Section: Purificationmentioning

confidence: 99%

Progress in quantitative analysis of plant hormones

Sun

Wang

et al. 2011

Chin. Sci. Bull.

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Plant hormones are small molecular natural products that regulate all plant developmental processes at low concentrations. Quantitative analysis of plant hormones is increasingly important for in-depth study of their biosynthesis, transport, metabolism and molecular regulatory mechanisms. Although plant hormone analysis remains a bottleneck in plant scientific research owing to the trace concentrations and complex components in plant crude extracts, much progress has been achieved in the development of extraction, purification and detection techniques in recent years. Solid phase extraction and chromatography/mass spectrometry have been applied widely for purification and quantitative analysis of plant hormones owing to their high selectivity and sensitivity. Purification methods such as liquid partition and immunoaffinity chromatography, and detection methods including immunoassay and electrochemical analysis, are employed. The advantages and disadvantages of these methods are discussed. In situ, real-time and multi-plant hormone profiling will comprise mainstream techniques for quantitative analyses in future studies on the regulatory mechanisms and crosstalk of plant hormones. defense responses are all triggered by plant hormones. A plant hormone, acting as a plant-development controller or a plant-environment mediator, can influence crop yield directly or indirectly, therefore research into plant hormone physiology has become an important target for agriculture development. However, it is noteworthy that endogenous level of each plant hormone is very low [1], and quantitative analysis of these compounds is still extremely difficult for both biologists and analytical chemists. It is a challenge for researchers to enrich these low-level natural compounds from plant extracts in which large amounts of interfering substances are present. In addition, the limited quantities of plant tissue available for hormone quantification is another challenge. Consequently, the analytic methods must be extremely selective and sensitive. The quantitative analysis of plant hormones typically includes the following key points: (i) the plant materials must

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Immunoaffinity co-purification of cytokinins and analysis by high-performance liquid chromatography with ultraviolet-spectrum detection

Cited by 61 publications

References 22 publications

Expression of early nodulin genes in alfalfa mycorrhizae indicates that signal transduction pathways used in forming arbuscular mycorrhizae andRhizobium-induced nodules may be conserved

Expression of early nodulin genes in alfalfa mycorrhizae indicates that signal transduction pathways used in forming arbuscular mycorrhizae andRhizobium-induced nodules may be conserved

Identification of an N-Glucoside of cis-Zeatin from Potato Tuber Sprouts

Progress in quantitative analysis of plant hormones

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