D-glufosinate as a male sterility agent for hybrid seed production.

A chemical male sterility system based on anther-localized conversion of the inactive D-enantiomer of the herbicide, glufosinate (2-amino-4-(methylphosphinyl)-butanoate) to the phytotoxic L is described. Highly pure D-glufosinate was isolated in >98% enantiomeric excess from the racemate via fermentation with a strain of Escherichia coli expressing the PAT (L-glufosinate N-acetyl transferase) gene and purification of the unreacted D-enantiomer from the broth by ion exchange. A modified (F58K, M213S) form of the D-amino acid oxidase (DAAO) (EC 1.4.3.3) from Rhodosporidium toruloides was designed, tested in vitro and found to efficiently oxidize D-glufosinate to its 2-oxo derivative [2-oxo-4-(methylphosphinyl)-butanoic acid]. Tobacco (Nicotiana tabacum) plants were transformed to express this modified oxidase under control of the TAP1 tapetum-specific promoter. A number of the resultant transgenic lines exhibited complete male sterility that persisted for two or more weeks immediately following foliar treatment with 75 or 200 g/ha of D-glufosinate without exhibiting obvious phytotoxic symptoms or any measurable decline in female fertility. Similarly, plants containing the same construct and, additionally, a PAT gene expressed from a plastocyanin promoter exhibited significantly reduced male fertility and no reduction in female fertility following foliar application of racemic glufosinate. Thus, foliar application of d-glufosinate either purified or as the commercial herbicide, combined with anther expression of a modified DAAO promises to provide a cost-effective conditional chemical male sterility system with the characteristics necessary for practical F1 hybrid seed production.

Technical performance of some commercial glyphosate-resistant crops.

Glyphosate-resistant (GR) crops have been sold commercially in the USA since 1996. The use of glyphosate alone or with conventional pre- and post-emergence herbicides with different modes of action gives growers many options for affordable, safe, easy, effective wide-spectrum weed control. Despite the overwhelming popularity of this technology, technical issues have surfaced from time to time as US growers adopt these crops for use on their farms. The types of concern raised by growers vary from year to year depending on the crop and the environment, but include perceptions of increased sensitivity to diseases, increased fruit abortion, reduced pollination efficiency, increased sensitivity to environmental stress, and differences in yield and agronomic characteristics between transgenic and sister conventional varieties. Although several glyphosate-resistant crops are commercially available, maize, soybean and cotton constitute the largest cultivated acreage and have likewise been associated with the highest number of technical concerns. Because glyphosate is rapidly translocated to and accumulates in metabolic sink tissues, reproductive tissues and roots are particularly vulnerable. Increased sensitivity to glyphosate in reproductive tissues has been documented in both glyphosate-resistant cotton and maize, and results in reduced pollen production and viability, or increased fruit abortion. Glyphosate treatments have the potential to affect relationships between the GR crop, plant pathogens, plant pests and symbiotic micro-organisms, although management practices can also have a large impact. Despite these potential technical concerns, this technology remains popular, and is a highly useful tool for weed control in modern crop production.