Plants having improved growth characteristics and method for making the same

A characteristic, plant technology, applied in the field of molecular biology, to achieve the effect of improving growth characteristics and increasing yield

Inactive Publication Date: 2007-09-19
CROPDESIGN NV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, such breeding techniques have several disadvantages, namely that these techniques are often labor-intensive a

Method used

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  • Plants having improved growth characteristics and method for making the same
  • Plants having improved growth characteristics and method for making the same
  • Plants having improved growth characteristics and method for making the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0144] Embodiment 1: Gene cloning-gene coding for tobacco RNA binding protein

[0145] Genes encoding RNA-binding proteins were initially identified as expressed sequence tags from tobacco BY2 cells and isolated as partial sequences in a CDNA-AFLP experiment using samples from synchronized tobacco BY2 cell cultures (Nicotiana tabacum L. cv.BrightYellow-2) prepared cDNA execution. Based on this CDNA-AFLP experiment, cell cycle-regulated BY2 signatures were identified and selected for further clones. Screening of tobacco cDNA libraries and isolation of full-sequence cDNA using expressed sequence tags.

[0146] Synchronization of BY2 cells

[0147] Tobacco BY2 (Nicotiana tabacum L. cv. Bright Yellow-2) culture cell suspensions were synchronized in early S phase by arresting the cells with aphidicolin as follows. Cultured cell suspensions of tobacco Bright Yellow-2 were maintained as described (Nagata et al. Int. Rev. Cytol. 132, 1-30, 1992). For synchronization, 7-day-old sta...

Embodiment 2

[0160] Example 2: Rice Transformation

[0161] Mature dry seeds of Japanese cultivar Nipponbare (NB) were dehulled. by incubating in 70% ethanol for one minute, followed by 0.2% HgCl 2 30 minutes, followed by 6 washes with distilled water, 15 minutes each time for disinfection. The sterile seeds are then germinated on medium containing 2,4-D (callus induction medium). After four weeks of incubation in the dark, embryogenic scutellum-derived calli were excised and propagated in the same medium. After two weeks, the calli were propagated or propagated by subculturing in the same medium for a further 2 weeks. Three days prior to co-cultivation, embryogenic callus fragments were subcultured on fresh medium (in order to promote cell division activity). Agrobacterium strain LBA4404 containing the binary T-DNA vector was used for co-cultivation. Agrobacterium was inoculated in AB medium containing appropriate antibiotics and cultured at 28°C for 3 days. Bacteria are then harves...

Embodiment 3

[0162] Example 3: Evaluation and Results

[0163] Approximately 15 to 20 independent TO rice transformants were generated. Primary transformants were transferred from the tissue culture room to the greenhouse for growth and T1 seeds were harvested. Five events were retained in which the T1 progeny segregated 3:1 for the presence / absence of the transgene. Approximately 10 T1 seedlings containing the transgene (heterozygous and homozygous) and an equal number of approximately 10 T1 seedlings lacking the transgene (nullzygote) were selected in each event by monitoring the expression of the visible markers. The 4 T1 events were further evaluated in the T2 generation following the same evaluation method as the T1 generation, but with more individuals for each event that was further evaluated.

[0164] Statistical analysis: F-test

[0165] A two-way ANOVA (Analysis of Variation) was used as a statistical model for the overall assessment of plant phenotypic properties. F-tests were...

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Abstract

The present invention concerns a method for improving the growth characteristics of plants by increasing activity in a plant of an RNA-binding protein or a homologue thereof, wherein said RNA-binding protein or homologue thereof is either: (i) a polypeptide having RNA-binding activity and comprising either 2 or 3 RNA 10 recognition motifs (RRMs) and a motif having at least 75% sequence identity to motif I: PIYEAAVVALPVVVKERLVRILRLGIATRYD and/or a motif having at least 50% sequence identity to motif II: RFDPFTGEPYKFDP; or (ii) an RBP1 polypeptide or homologue thereof having (a) RNA-binding activity; (b) two RRM domains, (c) the following two motifs: (i) KIFVGGL; and (ii) 15 RPRGFGF, allowing for up to three amino acid substitutions and any conservative change in the motifs; and (d) having at least 20% sequence identity to the amino acid represented by SEQ ID NO: 15. The invention also concerns to transgenic plants having introduced therein an RNA-binding protein-encoding nucleic acid or variant thereof, which plants have improved growth characteristics relative to corresponding wild type plants. The present invention also concerns constructs useful in the methods of the invention.

Description

technical field [0001] The present invention relates generally to the field of molecular biology, and to methods of improving plant growth characteristics. More specifically, the present invention relates to methods for improving plant growth characteristics, especially yield, by increasing the activity of RNA binding proteins or homologues thereof in plants. The invention also relates to plants having increased activity of RNA-binding proteins or homologues thereof, which plants have improved growth characteristics relative to corresponding wild-type plants. The RNA binding protein or its homologue useful in the method of the present invention has RNA binding activity, and comprises 2 or 3 RNA recognition motifs (RRM), and comprises the base that has at least 75% sequence identity with motif I: PYEAAVVALPVVVKERLVRILRLGIATRYD sequence and / or a motif having at least 50% sequence identity to Motif II: RFDPFTGEPYKFDP. The RNA binding protein or homolog thereof useful in the met...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/00
CPCC12N15/8241
Inventor V·弗兰卡德C·鲁兹A·I·桑兹莫林纳罗
Owner CROPDESIGN NV
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