Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition.

We have systematically made a set of precisely defined, single-gene deletions of all nonessential genes in Escherichia coli K-12. Open-reading frame coding regions were replaced with a kanamycin cassette flanked by FLP recognition target sites by using a one-step method for inactivation of chromosomal genes and primers designed to create in-frame deletions upon excision of the resistance cassette. Of 4288 genes targeted, mutants were obtained for 3985. To alleviate problems encountered in high-throughput studies, two independent mutants were saved for every deleted gene. These mutants-the 'Keio collection'-provide a new resource not only for systematic analyses of unknown gene functions and gene regulatory networks but also for genome-wide testing of mutational effects in a common strain background, E. coli K-12 BW25113. We were unable to disrupt 303 genes, including 37 of unknown function, which are candidates for essential genes. Distribution is being handled via GenoBase (http://ecoli.aist-nara.ac.jp/).

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Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection.

Standardization of procedures for nitrogen fractionation of ruminant feeds

Rates of herbivory and defensive characteristics of young and mature leaves were measured for saplings of 46 canopy tree species in a lowland tropical rain forest (Barro Colorado Island, Panama). Grazing rates were determined in the field for sample periods in the early wet, late wet, and dry seasons. Leaf properties such as pubescence, toughness, water, protein, fiber, and phenolic contents explained over 70W% of the variation among plant species in the rates of herbivory on mature leaves. Leaf toughness was most highly correlated with levels of herbivory, followed by fiber content and nutritive value. Phenol content and phenol: protein ratios were not significantly correlated with damage. Mature leaves of gap-colonizing species were grazed six times more rapidly than leaves of shade- tolerant species. Gap-colonizers have less tough leaves, lower concentrations of fiber and phenolics, higher levels of nitrogen and water, shorter leaf lifetimes, and faster growth rates than do shade- tolerant species. Gap-colonizers did not escape discovery by herbivores to any greater extent than shade-tolerant species, as measured by the spatial distribution of plants or by the intraspecific dis- tribution of herbivore damage under natural or experimentally manipulated conditions. In 70W% of the species, young leaves suffered higher damage levels than mature leaves. Although young leaves are more nutritious and less tough and fibrous, they have two to three times the con- centrations of phenols. The temporal appearance of young leaves was not correlated with the distri- bution of herbivory among individuals of a species. Interspecific patterns of defense mechanisms are discussed in terms of current theories of plant apparency, and an alternative model for the evolution of plant defenses is presented.

Hk Goering

Papers

Forage Fiber Analysis (Apparatus, reagents, procedures and some applications)