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Shine clade of transcription factors and their use

a transcription factor and clade technology, applied in the field of new transcription factor clade, can solve the problems of high cost, high labor intensity, and inability to meet the needs of a large number of people, and achieve the effect of reducing labor intensity, reducing labor intensity, and reducing labor intensity

Inactive Publication Date: 2012-11-01
STICHTING DIENST LANBOUWKUNDIG ONDERZOEK
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0030]Based on the finding that the SHN1 expression resulted in an increase in cuticular water loss, it was even more surprising to find that 35S::SHN1 transformants showed enhanced drought tolerance and recovery. Thus apparently the increased non-stomatal water loss through the altered cuticle was outweighed by the effect of the reduced stomatal index.
[0147]The method comprises in one embodiment the steps of mutagenizing plant seeds (e.g., EMS mutagenesis), pooling of plant individuals or DNA, PCR amplification of a region of interest, heteroduplex formation and high-throughput detection, identification of the mutant plant, sequencing of the mutant PCR product. It is understood that other mutagenesis and selection methods may equally be used to generate such mutant plants. Seeds may for example be radiated or chemically treated and the plants screened for a modified SHN phenotype, such as enhanced drought tolerance.

Problems solved by technology

Without the protective cuticle, transpiration of most land plants would be so rapid that death would result.
Apart from that, also plants grown in regions with relatively high precipitation may suffer spells of drought throughout the growing season.
Although breeding (e.g., marker assisted) for drought tolerance is possible and is being pursued for a range of crop species (mainly cereals, such as maize, upland rice, wheat, sorghum, pearl millet, but also in other species such as cowpea, pigeon pea and Phaseolus bean), it is extremely difficult and tedious because drought tolerance or resistance is a complex trait, determined by the interaction of many loci and gene-environment interactions.
Premature shattering causes significant yield losses in Brassica species, such as Brassica napus (oilseed rape or “canola” if erucic acid and glucosinolate levels are below a certain threshold value).
One of the difficulties in transgenic podshatter approaches is that on the one hand it is desired to prevent easy separation of the two pod valves, on the other hand it must still remain possible to separate the valves in order to harvest the seeds.

Method used

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  • Shine clade of transcription factors and their use
  • Shine clade of transcription factors and their use
  • Shine clade of transcription factors and their use

Examples

Experimental program
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Effect test

example 1

Material and Methods

1.1 Plant Material and Drought Tolerance Experiment

[0181]All plants, including the activation tag population (Marsch-Martinez et al., 2002, Plant Physiol. 129:1544-1556) and transgenic lines were grown in the greenhouse at around 22° C. and were in the Arabidopsis ecotype Wassilewskija (Ws). For the drought tolerance experiments, soil mixture comprised 1 part of sand and perlite and 2 parts of compost. Seeds were sown (after 3 nights at 4° C.) at density of six plants per 4 cm pot in a tray with 51 pots (Aracon containers, BetaTech, Belgium). Mineral nutrients were supplied 10 days after germination and at two weeks after germination the plants were subjected to drought (for 9, 10, 11 or 12 days) by transferring the pots to dry trays (after drying each pot from outside). Every 2 days in drought, the plants were moved within the tray to nullify pot position effects. Subsequently, plants were rehydrated and observed for recovery after one week. The drought experime...

example 2

Identification of the Shine Mutant

[0193]By screening a collection of 2000 Arabidopsis transposon activation tag lines (Marsch-Martinez et al., 2002) a mutant plant was identified which showed leaf surface alterations (not shown). Both rosette and cauline leaves of the mutant (termed shine, shn) had a more brilliant, shiny green color when compared to wild type plants and often had curved-down edges (not shown). The stem of mature plants was often bowed-down, siliques were slightly smaller than wild type and also showed a more brilliant surface. Structure of other floral organs and plant fertility did not seem to be affected in shn. Progeny analysis of the self-pollinated shn mutant line suggested a dominant mutation (three quarters of the plants exhibited the shn phenotype).

example 3

Alterations to Wax Load in the shn Mutant

[0194]Scanning electron microscopy (SEM) was utilized for a detailed comparison between the surfaces of wild type plant organs and those of shn. The surfaces of stems and siliques of Arabidopsis are covered by a dense mixture of different types of wax crystals while leaf surfaces normally exhibit only small numbers of epicuticular wax crystals. In contrast to wild type we detected more wax crystals on both adaxial and abaxial sides of rosette and cauline leaves of shn (data not shown). The leaf surface was not entirely covered by crystals, as in the case of wild type siliques and stems, but rather had irregular patches of plate-like wax crystals. An additional characteristic of the shn mutant was the presence of cuticular ridges on the surface of both cauline leaves and siliques, which were not detected in the wild type (data not shown). Such cuticular ornamentation was not visible on either the adaxial or abaxial surfaces of shn rosette leav...

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Abstract

The present invention relates to the field of transgenic plants with novel phenotypes, especially plants with enhanced drought tolerance. Provided are SHINE proteins and nucleic acid sequences encoding these, which are useful in conferring novel phenotypes to plants.

Description

FIELD OF THE INVENTION[0001]The present invention relates to a novel class of transcription factors, referred herein to as the SHINE clade of transcription factors, and their use to confer various novel phenotypes onto plants, such as drought tolerance and other water use related phenotypes, indehiscence of plant dehiscence zones (conferring for example male sterility or pod shatter resistance) and modification of other cell layers involved in cell separation or in the cell—environment interface. The invention provides nucleic acid sequences encoding SHINE (SHN) proteins, or functional fragments thereof, which are useful for modifying, or newly conferring, one or more novel plant phenotypes. Further provided are isolated SHINE proteins, chimeric genes, nucleic acid vectors, recombinant microorganisms and plants, as well as methods and means for using SHINE (SHN) nucleic acid sequence to confer novel plant phenotypes.BACKGROUND OF THE INVENTION[0002]The interface between plants and t...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N15/29C12N15/82A01H5/08A01H5/00A01H5/10C07K14/415
CPCC12N15/8273C07K14/415
Inventor AHARONI, ASAPHDIXIT, CHITALPEREIRA, ANDY
Owner STICHTING DIENST LANBOUWKUNDIG ONDERZOEK
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