Rapid Enzyme-Linked Immunosorbent Assay for Isopentenyl adenosine Group Cytokinins

doi:10.3321/j.issn:1000-7091.1996.01.018

Abstract

Abstract: Abstract The rapid ELISA used specifically for the determination arf iPAs (iP,iPA,iPA-5'P) has been developed. The hapten iPA has been selected and conjugated,via its hydroxy groups of the ribose moiety,to free amino groups of bovine serum albumin(BSA) with NaIO₄ oxidation and the immuogen iPA-N₉-BSA has been synthesized. Anti-iPA antiserum has been raised in rabbit and its titer, determined by ELISA, reaches 1：34500. This antiserum basically recognized iPA group cytokinins. Because of higher quality of this antiserum direct ELISA (that is,poly antibodies as solid phase)has been set up and it has been observed that the immunoreaction almost reaches to equilibrium during 30 minutes at 4℃·The detection limit of this ELISA is 4.7 fmol/well and the linear detection range is 0.O1一50 pmol/well. The coefficient variation(CV) within plates is 3.7% and the CV between plates is 5.3% . The dilution curves of sample extracts parallel with standard curve in this ELISA. And the recovery is as high as 87. 1%.These results suggest that the simple sample preparation procedure(i. e. extraction with 80} methanol,then passing througn Sep-pak C18 column)used in this ELISA is effective. The iPA-group cytokinins contents in various parts of soybean plants have been determined with this ELISA. It is shown that iPAs levels in root nodules and root tips are greatly higher than those in young leaves and apical buds of soybean plants.

Key words: Cytokinins, Isopetenyladenosine, ELISA

CLC Number:

Chen Yifeng, Zhou Xie. Rapid Enzyme-Linked Immunosorbent Assay for Isopentenyl adenosine Group Cytokinins[J]. ACTA AGRICULTURAE BOREALI-SINICA, 1996, 11(1): 97-102. doi: 10.3321/j.issn:1000-7091.1996.01.018.

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