This presentation introduces the advances inbiological nitrogen fixation research abroad, in particular,describes the great progress and achievements on itsresearch in China as follows: collection of rhizobial resources and establishment of the largest database of Rhizobium inChina, correction and development of Rhizobium taxonomy in international; discovery of a couple of nif genes,identification and unification of linkage among the nif gene operons of Klebsiella pneumoniae, finding of regulative mechanism of positive regulation nif gene and its sensitivity to oxygen, temperature; finding of the activity of nodulation gene nodD3 product in Sinorhizobium meliloti which is notcontrolled by flavonoid produced from its host alfalfa;finding of the association between expression of genes coding the products for carbon utilization and nitrogen metabolism and their regulations; chemical synthesis of nodulationfactor of Sinorhizobium meliloti; constructions of engineered nitrogen fixers and utilization in practice based on theresearch of gene expression and regulation; chemicalsimulation of the structure and function of nitrogenase and bringing forward the model of nitrogenase active center for the first time in international and synthesis of modelcompounds which were paid attention by colleagues abroad. Finally, the development of nitrogen fixation research inChina in future has been put forward, suggesting that the nifgene regulation and its role in providing crops with nitrogen element, signal transduction and molecular interactions between Rhizobium and legume, coupling between carbonand nitrogen metabolisms, nitrogen fixation andphotosynthesis, and functional genomics of nitrogen-fixing nodule symbiosis, etc., would be actively worked on.
SHEN Shihua & JING Yuxiang Key Laboratory of Photosynthesis and Plant-Environment MolecularPhysiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China
Plasmid pVK1001 which carried the gfp gene of GFPmut2, a mutant of GFP, was introduced into Azospirillum brasilense Yu62 by electroporation. Maize seedlings were inoculated with the GFP-labelled baeteria and grown gnotobiotically in flask with semi-solid agar medium. Observations were performed with confocal laser scanning microscopy (CLSM) and electron microscopy, respectively, at 8 d and 12 d after inoculation. Confocal laser scanning microscopy showed that A. brasilense Yu62 could penetrate into the cortex tissue, colonizing in the intercellular spaces of the parenchyma cells of the cortex tissue. Transmission and scanning electron microscopy (TEM) showed that the majority of the bacteria colonized on the root surface and only a minority of them resided in the root interior.
