While transformation of the major monocot crops is currently possible, the process typically remains confined to one or two genotypes per species, often with poor agronomics, and efficiencies that place these methods beyond the reach of most academic laboratories. Here, we report a transformation approach involving overexpression of the maize (Zea mays) Baby boom (Bbm) and maize Wuschel2 (Wus2) genes, which produced high transformation frequencies in numerous previously nontransformable maize inbred lines. For example, the Pioneer inbred PHH5G is recalcitrant to biolistic and Agrobacterium tumefaciens transformation. However, when Bbm and Wus2 were expressed, transgenic calli were recovered from over 40% of the starting explants, with most producing healthy, fertile plants. Another limitation for many monocots is the intensive labor and greenhouse space required to supply immature embryos for transformation. This problem could be alleviated using alternative target tissues that could be supplied consistently with automated preparation. As a major step toward this objective, we transformed Bbm and Wus2 directly into either embryo slices from mature seed or leaf segments from seedlings in a variety of Pioneer inbred lines, routinely recovering healthy, fertile T0 plants. Finally, we demonstrated that the maize Bbm and Wus2 genes stimulate transformation in sorghum (Sorghum bicolor) immature embryos, sugarcane (Saccharum officinarum) callus, and indica rice (Oryza sativa ssp indica) callus.
Light and fungal elicitor induce mRNA encoding 3-deoxy-Darabino-heptulosonate 7-phosphate (DAHP) synthase in suspension cultured cells of parsley (Petroselinum crispum L.). The kinetics and dose response of mRNA accumulation were similar for DAHP synthase and phenylalanine ammonia-lyase (PAL). Six micrograms of elicitor from Phytophthora megasperma f. glycinia gave a detectable induction within 1 hour. Induction of DAHP synthase and PAL mRNAs by light was transient, reaching maximal levels at 4 hours and retuming to pretreatment levels after 24 hours. Our data suggest that either light or fungal elicitor transcriptionally activate DAHP synthase. A coordinate regulation for key enzymes in the synthesis of primary and secondary metabolites is indicated.
Soybean [Glycine max (L.) Merr.] oil is one of the most consumed and highest produced vegetable oils. However, the high percentage of polyunsaturated fatty acids in soybean oil limits its stability and shelf life. Here we report the generation and characterization of a high oleic acid soybean event (305423 soybean) that has an elevated content of monounsaturated oleic acid and reduced presence of polyunsaturated linoleic acid and linolenic acid. This transgenic event was generated by the insertion of a soybean ω‐6 desaturase (FAD2) gene fragment (gm‐fad2‐1), resulting in the suppression of endogenous FAD2‐1 expression, and a modified version of the soybean acetolactate synthase (ALS) gene (gm‐hra), used as a selectable marker. Molecular characterization revealed that 305423 soybean harbors four DNA inserts containing multiple copies of complete and partial gm‐fad2‐1 gene cassettes and a single copy of the intact gm‐hra gene cassette. However, these DNA inserts seemed to be inherited together without segregation. The suppression of endogenous FAD2‐1 led to an increase of oleic acid (18:1) from 211 to 765 g kg–1 along with concurrent reduction of both linoleic acid (18:2) from 525 to 36.2 g kg–1 and linolenic acid (18:3) from 93.5 to 53.9 g kg–1 out of total fatty acyl groups. The 305423 soybean was also subjected to nutrient composition analyses and phenotypic and agronomic performance evaluations and found to be comparable to the control soybean except for the expected traits.
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