A reproducible system for the generation of fertile, transgenic maize plants has been developed. Cells from embryogenic maize suspension cultures were transformed with the bacterial gene bar using microprojectile bombardment. Transformed calli were selected from the suspension cultures using the herbicide bialaphos. Integration of bar and activity of the enzyme phosphinothricin acetyltransferase (PAT) encoded by bar were confirmed in all bialaphos-resistant callus lines. Fertile transformed maize plants (R0) were regenerated, and of 53 progeny (R1) tested, 29 had PAT activity. All PAT-positive progeny analyzed contained bar. Localized application of herbicide to leaves of bar-transformed R0 and R1 plants resulted in no necrosis, confirming functional activity of PAT in the transgenic plants. Cotransformation experiments were performed using a mixture of two plasmids, one encoding PAT and one containing the nonselected gene encoding [beta]-glucuronidase. R0 plants regenerated from co-transformed callus expressed both genes. These results describe and confirm the development of a system for introduction of DNA into maize.
Glyphosate is the world's most widely used herbicide. It is nonselective and has been used to control a broad range of weed species for the past 20 yr, without the appearance of resistant weed biotypes. However, a biotype ofLolium rigidumfrom a field in Northern Victoria, Australia, in which glyphosate had been used for the past 15 yr, failed to be controlled by label recommended rates. Based on LD50values from pot dose-response experiments, this biotype exhibited resistance to glyphosate and was nearly 10-fold more resistant compared to the susceptible biotypes tested. The biotype was resistant to three different salts of glyphosate. The biotype was also nearly threefold more resistant to diclofop-methyl but was susceptible to other commonly used selective and broad-spectrum herbicides. Between the two-leaf and tillering stages of development, a susceptible biotype exhibited a small but significant decrease in tolerance to glyphosate, whereas tolerance of the resistant biotype remained unchanged with age. The resistant phenotype was verified in experiments in which seed was germinated in the presence of glyphosate. Observations on shoot and root growth of seedlings in these experiments suggested that the resistance mechanism might be associated more with the shoot than with the root.
SummaryThe maize mutation high chlorophyll¯uorescence 60-mutable 1 (hcf60-m1), generated through Activator (Ac) tagging, has insuf®cient photosynthetic electron transport. Here we show that the Hcf60 gene encodes a protein with substantial amino acid similarity to plant plastid and bacterial ribosomal small subunit protein 17 (RPS17) proteins. The lack of detectable HCF60 transcripts in mutant leaves, and insertion of the transposed Ac element 17 bp upstream of the start of translation in the mutated locus, suggest that little if any RPS17 is produced. The mutant phenotype is consistent with reduced plastid translation. Seedling lethal hcf60-m1 plants display temperature and light-dependent chlorophyll de®ciencies, a depletion of plastid rRNA pools, and few high-molecular-weight polysomal complexes. Growth under moderate light conditions (27°C, 100 mE m ±2 sec ±1
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