With the aim of improving the nutritive value of an important grain legume crop, a chimeric gene specifying seed-specific expression of a sulfur-rich, sunf lower seed albumin was stably transformed into narrow-leafed lupin (Lupinus angustifolius L.). Sunf lower seed albumin accounted for 5% of extractable seed protein in a line containing a single tandem insertion of the transferred DNA. The transgenic seeds contained less sulfate and more total amino acid sulfur than the nontransgenic parent line. This was associated with a 94% increase in methionine content and a 12% reduction in cysteine content. There was no statistically significant change in other amino acids or in total nitrogen or total sulfur contents of the seeds. In feeding trials with rats, the transgenic seeds gave statistically significant increases in live weight gain, true protein digestibility, biological value, and net protein utilization, compared with wild-type seeds. These findings demonstrate the feasibility of using genetic engineering to improve the nutritive value of grain crops.

Thalamic inputs strongly drive neurons in the primary visual cortex, even though these neurons constitute only approximately 5% of the synapses on layer 4 spiny stellate simple cells. We modeled the feedforward excitatory and inhibitory inputs to these cells based on in vivo recordings in cats, and we found that the reliability of spike transmission increased steeply between 20 and 40 synchronous thalamic inputs in a time window of 5 milliseconds, when the reliability per spike was most energetically efficient. The optimal range of synchronous inputs was influenced by the balance of background excitation and inhibition in the cortex, which could gate the flow of information into the cortex. Ensuring reliable transmission by spike synchrony in small populations of neurons may be a general principle of cortical function.

The pea (Pisum sativum) is an important grain legume crop plant that has gained worldwide economic importance as a source of protein for animal and human nutrition. In addition, it has well-defined genetics, and it has been commonly used as a model plant for research in plant physiology and biochemistry. The productivity and value of peas could be greatly increased by the introduction of stably inherited traits such as pest and disease resistance, herbicide resistance, and improved protein quality. These traits are not available in natural populations of near relatives of cultivated peas, but current advances in plant genetic engineering provide a potentially powerful tool for achieving these goals by another means.The prerequisites for the transfer of foreign genes into any plant species by genetic engineering are an efficient gene delivery system, such as Agrobacterium-mediated DNA transfer, an effective selectable marker for transformed cells, and the ability to regenerate mature, fertile, transgenic plants from transformed tissue in culture.Regeneration via embryogenesis or organogenesis has been described for a variety of pea explants, e.g. from immature leaflets (Mroginski and Kartha, 1981;Rubluo et al., 1984), from cotyledonary node (Jackson and Hobbs, 1990), from hypocotyls (Nielsen et al., 1991), from embryos (Kysely et * Corresponding author; fax 61-6-246-5000. 75 1 Natali and Cavallini, 1987;Tetu et al., 1990), from various organs of seedlings (Malmberg, 1979;Hussey and Gunn, 1984;Ezhova et al., 1985), and from protoplast cultures (Jacobsen and Kysely, 1984; Puonti-Kaerlas and Eriksson, 1988; Lehminger-Mertens and Jacobsen, 1989). Agrobacterium-mediated transfonnation of various pea explants has also been reported, e.g. stem explants (Lulsdorf et al., 199 l), embryonic axis and epicotyl segments (Filippone and Lurquin, 1989;Puonti-Kaerlas et al., 1989), nodus explants (De Kathen and Jacobsen, 1990; Nauerby et al., 1991), and root explants and protoplasts (Schaerer and Pilet, 1991). Tumors were induced in young pea plants by wild-type Agrobacterium . However, no mature transgenic pea plants were regenerated from any of the above transformation systems. The only report to date of stable transformation of peas and the production of mature, flowering, transgenic pea plants is by Puonti-Kaerlas et al. (1990), who achieved regeneration by organogenesis via callus formation using a gene encoding hygromycin phosphotransferase as a selectable marker.In this paper we report the development of a routine, reliable transformation and regeneration system for peas. The procedure has been used to introduce herbicide resistance and the expression of an antibiotic resistance gene into two cultivars of peas using an Agrobacterium tumefaciens-mediated delivery system. Integration of the two traits was stable, and their frequency in the first generation progeny followed the Mendelian pattem. MATERIALS AND METHODS Plant Material and Transformation ProcedurePea (Pisum sativum L.) cv Greenfeast and cv Rondo were grown in t...

Bruchid larvae cause major losses of grain legume crops throughout the world. Some bruchid species, such as the cowpea weevil and the azuki bean weevil, are pests that damage stored seeds. Others, such as the pea weevil (Bruchus pisorum), attack the crop growing i n the field. We transferred the cDNA encoding the a-amylase inhibitor (a-AI) found in the seeds of the common bean (Phaseolus vulgaris) into pea (Pisum sativum) using Agrobacferium-mediated transformation. Expression was driven by the promoter of phytohemagglutinin, another bean seed protein. The a-amylase inhibitor gene was stably expressed in the transgenic pea seeds at least to the T, seed generation, and a-AI accumulated i n the seeds up to 3% of soluble protein. This level is somewhat higher than that normally found in beans, which contain 1 to 2 % a-AI. In the 1 , seed generation the development of pea weevil larvae was blocked at an early stage. Seed damage was minimal and seed yield was not significantly reduced in the transgenic plants. These results confirm the feasibility of protecting other grain legumes such as lentils, mungbean, groundnuts, and chickpeas against a variety of bruchids using the same approach. Although a-AI also inhibits human a-amylase, cooked peas should not have a negative impact on human energy metabolism.The common bean (Phaseolus vulgaris L.) contains a family of structurally related seed proteins: PHA-E and -L, arcelin, and a-AI. PHA-E and PHA-L are strong agglutinins, i.e. classical lectins that bind carbohydrate, and arcelin, which is found only in certain wild accessions of the common bean, may be a weak agglutinin (Hartweck et al., 1991). The bean a-AI has 65 to 70% amino acid sequence identity with the other three but lacks at least one of the conserved residues needed for lectin activity. Its biochemical mode of action is to form a one-to-one complex with certain amylases (for reviews, see Chrispeels and Raikhel, 1991;Rouge et al., 1993).