Transgenic plants with modified sterol biosynthetic pathways

A plant, sterol technology, applied in the field of plant genetic engineering

Inactive Publication Date: 2000-03-15
MONSANTO CO (MONSANTO CY)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Unfortunately, such methods are only effective against specific endotoxin-sensitive insects

Method used

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  • Transgenic plants with modified sterol biosynthetic pathways
  • Transgenic plants with modified sterol biosynthetic pathways
  • Transgenic plants with modified sterol biosynthetic pathways

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Embodiment 1

[0094] EXAMPLES Example 1. Phytosterols

[0095] Sterol isomers were extracted from cotton and separated into homogeneous forms by chromatography. Identification of novel phytosterols with side chains not available to insects.

[0096] by mass spectrometry and 1 H and 13 C nuclear magnetic resonance (NMR) experiments, found that sterols have structural specificity (Table 1) (Guo et al., 1995).

[0097] Initial studies showed that 4-day-old maize produced mono- or di-alkylated sterols at C-24. Maize can produce those sterols because the isolated 24(28)-methylene and 24(28)-ethylene sterols were obtained from maize seedling tissue and their structures were determined by mass and proton NMR Resonance spectroscopy was identified.

[0098] Table 1

[0099] Sterol Components of Corn

[0100] MS a TLC a Sterol bd plant source c

[0101] ...

Embodiment 2

[0121] Figure 4 summarizes the kinetically favorable end-products Δ produced during seedling development into leaves and sheaths under dark-grown conditions. 5 -24-Alkylsterol pathway. Expression of SMT enzyme activity and data on sterol specificity early in the process of leaf and sheath production suggest that maize synthesizes at least two distinct SMT enzymes: SMT I Catalyzes the subsequent methyl conversion to generate Δ 24(28) -methylene sterol and Δ 24(28) - Ethylene sterol, while SMT II then catalyzes the conversion of the methyl group to Δ 23(24) -24-Methylsterol. Identification of sterols required for embodiment 2 plant growth

[0122] The phytosterols identified in Example 1 were individually tested for their ability to support growth. Due to the lack of phytosterol mutants available for this study, the yeast sterol auxotroph GL-7 was grown in the presence of the sterols identified in Example 1, see above (Li, 1996). This yeast mutant was used as a model syst...

Embodiment 3

[0123] Sterols can be classified according to their effects on growth processes. Growth-activating sterols include ergosterol. Sterols including cholesterol and sitosterol that are migrated to membrane and cellular structural components without affecting cell growth rate (Nes et al., 1993). The enzymatic research of embodiment 3 sterol converting enzyme

[0124] To illustrate the enzymatic basis for the sterols identified in Example 1, the sterol specificity of microsome-bound soluble SMTase from 4-day-old maize seedlings was determined. When using a microsome-bound enzyme system we found that cycloartenol was the preferred sterol acceptor and that 24(28)-methylene-4-methyl-7-encholestanol was methylated to generate 24 (28)-Ethylidene-4-methyl-7-encholestanol. Table 2 summarizes the specificity of soluble SMTases from maize seedlings for various sterol substrates.

[0125] Table 2

[0126] Sterol specificity of (S)-adenosyl-L-methionine: D 24 -Ste...

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Abstract

Plant phytosterol compositions are modulated in transgenic plants to confer resistance to insects, nematodes, fungi, and / or other environmental stresses, and / or to improve the nutritional value of the plants. Recombinant DNA molecules of the invention encode RNA or protein sequences capable of altering plant sterol profiles by affecting the expression or activity of sterol biosynthetic enzymes. The DNA molecules are transformed into plant cells and plants having altered sterol compositions are regenerated therefrom.

Description

[0001] The present invention relates broadly to plant genetic engineering. More specifically, it relates to manipulating the level and / or activity of endogenous phytosterol components as a strategy to minimize crop damage from plant insects or other pests and / or to enhance plant nutrition value. Background of the invention [0002] Harsh environmental factors such as drought, severe cold, weeds and crop-eating organisms can cause harm to agricultural production. Traditional methods of killing weeds and parasites rely almost entirely on chemical herbicides, pesticides and fungicides. However, the widespread use of these agricultural chemicals has led to the development of resistance. In fact, insects have been reported to be resistant to most major classes of insecticides, including organophosphates, chlorinated hydrocarbons, and carbamates. [0003] Sterols comprise a class of essentially natural compounds that are required to some extent by all eukar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/10C12N15/82C12P33/00
CPCC12P33/00C12N9/1007C12N15/8243C12N15/8286C12N15/8279Y02A40/146
Inventor W·D·内斯
Owner MONSANTO CO (MONSANTO CY)
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