Crisp caudex 2 gene family and correlation method and purpose

A genome and transgenic plant technology, applied in the field of plant molecular biology, can solve problems such as collapse and reduce mechanical strength

Inactive Publication Date: 2009-03-04
PIONEER HI BRED INT INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Mutations in three CesA genes from Arabidopsis thaliana lead to col

Method used

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  • Crisp caudex 2 gene family and correlation method and purpose
  • Crisp caudex 2 gene family and correlation method and purpose

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0223] Characterization of maize cDNA encoding Bk2-like protein

[0224]The corn crispy stem 2 (bk2) phenotype was first reported in 1940 (Langham, MNL 14:21-22 (1940)) by phenotype mapping chr9L between the markers umc95 and bn17.13 around the 100 centimorgan region (Howell et al., MNL 65:52-53 (1991 )). Previously, clone csc1c.pk005.k4:fis (SEQ ID NO: 1) was confirmed to encode the fragile stem 2 polypeptide (SEQ ID NO: 2) (International Application No. PCT / US2005 / 035450, which claims to be filed on October 6, 2004 priority of U.S. Provisional Application No. 60 / 615,868, the entire contents of which are incorporated herein by reference). Two additional members of the Bk2 gene family are also disclosed (SEQ ID NO: 7 and 8 and SEQ ID NO: 13 and 14). In this disclosure, these genes have been named Bk2-like (Bk2L).

[0225] Using the BLASTN or TBLASTN algorithm provided by the National Center for Biotechnology Information (NCBI), several databases were searched for other ma...

Embodiment 2

[0236] Gene expression analysis of Bk2-like proteins

[0237] Massively Parallel Signal Sequencing (MPSS) using Solexa TM ) technology (see Table 3) (Brenner et al., Nat.Biotechnol.18:630-634 (2000); Brenner et al., Proc.Natl.Acad.Sci.U.S.A.97:1665-1670 (2000)), checked Tissue specificity of expression of the Bk2-like gene family disclosed in Table 1. MPSS TM Contains a 17-base signaling flag established from an mRNA sample that has been reverse transcribed. The markers are sequenced simultaneously and assigned to genes or ESTs. Numerical values ​​normalized to parts per million (PPM) are given for the abundance of these markers and then compared for marker expression or marker abundance between different tissues. Therefore, MPSS TM The platform can be used to determine the expression pattern of a specific gene, and its expression level in different tissues. Numbers are averages of multiple libraries per tissue listed in the second column.

[0238] table 3

[0239]...

Embodiment 3

[0249] prophetic embodiment

[0250] By overexpressing a maize Bk2-like gene under the control of a strong stem-specific promoter, a Builds enhanced stem strength

[0251] Construct chimeric transgenes to directly overexpress Bk2 gene / polypeptide in a tissue-specific manner. The transgenic construct comprises a maize cDNA encoding Bk2L3 and / or Bk2L6 (e.g., SEQ ID NO:5 or SEQ ID NO:11) operably linked to the promoter of the stem-specific S2A gene from alfalfa (Abrahams et al. Human, Plant Mol. Biol. 27:513-528 (1995)). DNA containing the Bk2L3 or Bk2L6 ORF was then fused to the S2A promoter on the 5' end and the pinII terminator on the 3' end to generate image 3Expression cassettes indicated. The construct was then ligated to a selection marker cassette containing the CaMV 35S promoter driven bar gene and pinll terminator. Those skilled in the art will appreciate that different promoters, 5' end sequences and / or 3' end sequences can be used to achieve comparable expr...

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Abstract

This invention relates to isolated polynucleotides encoding BRITTLE STALK 2-like (Bk2L) family polypeptides. The invention also relates to the construction of a chimeric gene encoding all or a portion of a Bk2L polypeptide, in sense or antisense orientation, wherein expression of the chimeric gene results in production of altered levels of the Bk2L polypeptide in a transformed host cell.

Description

[0001] field of invention [0002] The field of invention relates to plant molecular biology, especially brittle stem 2-like (BRITTLESTALK·2-Like) gene, brittle stem 2-like polypeptide, and its use. Background of the invention [0003] Primary plant growth is primarily driven by cell enlargement, which is achieved through the irreversible generation of intracellular turgor by the primary cell wall. Although cell division is required to generate new cells, the growth resulting from the expansion of these cells does not result solely from their division. Cellulose microfibrils embedded in the hemicellulose matrix and lignin in the cell walls are the main determinants of tensile strength (Appenzeller et al., Cellulose 11:287-299 (2004)). Cells usually expand along an axis perpendicular to the direction of the microfibrils. For example, radial deposition of microfibrils facilitates cell expansion along the longitudinal axis. [0004] The secondary wall differs from the primary...

Claims

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Application Information

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IPC IPC(8): C12N15/82
CPCC12N15/8246
Inventor K·S·杜加
Owner PIONEER HI BRED INT INC
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