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Modification of lignin biosynthesis via sense suppression

A biosynthesis and lignin technology, applied in the fields of plant genetic improvement, sugar derivatives, biochemical equipment and methods, etc., can solve the problems that the nutritional value cannot meet the metabolic requirements of lactating dairy cows, and the feeding value decreases.

Pending Publication Date: 2016-11-16
DAIRY AUSTRALIA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Significant declines in the feeding value of grasses observed in Australian temperate pastures are in late spring and early summer when the nutritional value of perennial ryegrass based pastures cannot meet the metabolic demands of lactating cows

Method used

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  • Modification of lignin biosynthesis via sense suppression
  • Modification of lignin biosynthesis via sense suppression
  • Modification of lignin biosynthesis via sense suppression

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0241] Three 4-coumaric acid-CoA ligases (4CL) from perennial ryegrass (Lolium perenne)

[0242] Isolation and characterization of cDNAs

[0243] Materials and methods

[0244] plant material

[0245] Plant and germ cell suspensions of perennial ryegrass (Lolium perenne L.) cv Ellet and tall fescue (Festuca arundinacea Schreb.) cv Triumph were established and maintained as previously described (Heath et al., 1998). Wounding experiments were performed using 10 day old perennial ryegrass (cv Ellet) as previously described (Heath et al., 1998).

[0246] Screening of cDNA library

[0247] A cDNA library prepared from RNA isolated from perennial ryegrass seedlings (Heath et al., 1998) was used in [ 32 P] dCTP-labeled rice part 4CL probe screening. The rice 4CL probe and the 4CL specific sequence consisting of 844bp were inserted into PUC 119. This insertion sequence has 93% sequence identity with the cDNA sequence of rice 4CL (Genbank, L43362, base 453-1300). The cDNA insert w...

Embodiment 2

[0269] Isolation and Characterization of Cinnamoyl-CoA Reductase (CCR) cDNA from Perennial Ryegrass (Lolium perenne)

[0270] A total of 500,000 phages were screened using a maize CCR probe from a cDNA library constructed from 10 day old etiolated perennial ryegrass (L. perenne) shoots. Ninety-three positive plaques were found in the initial screen and 5 were subsequently analyzed by restriction digestion. 4 out of 5 are the same. One of the 4 identical cDNAs, LpCCR1, was selected for further analysis ( Figure 9 ).

[0271] Nucleic acid sequence analysis of CCR cDNA of perennial ryegrass

[0272] The full-length nucleotide sequence of LpCCR1 was obtained, and the amino acid sequence was predicted ( Figure 10). The cDNA of LpCCR1 is 1395bp, with 149bp 5' non-coding region and 160bp 3' non-coding region. The 1086 bp open reading frame encodes a protein of 362 amino acids. The composition of the coding region was found to be 68% G+C rich. Codon usage was also examined a...

Embodiment 3

[0278] Isolation and Characterization of Cinnamyl Alcohol Dehydrogenase (CAD) cDNAs from Perennial Ryegrass (Lolium perenne)

[0279] A 558 bp cinnamoyl dehydrogenase (CAD) fragment was amplified from cDNA synthesized from total RNA prepared from perennial ryegrass seedlings. Conserved amino acid regions among CADs of radiata, alfalfa, Aralia cordata, eucalyptus, and Arabidopsis were used to design oligonucleotides to amplify CAD of perennial ryegrass. The forward oligonucleotide was designed in the conserved amino acid region CAGVTVYS, and the reverse oligonucleotide was designed in the conserved region DVRYRFV. A 551 bp PCR fragment was cloned and sequenced to confirm that it corresponds to the PCR fragment of perennial ryegrass CAD. An eDNA library prepared from RNA extracted from perennial ryegrass seedlings was screened with a 551 bp PCR fragment specific to perennial ryegrass CAD. Eight cDNAs were isolated and divided into 6 groups by restriction analysis. One represe...

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Abstract

The present invention relates to the modification of lignin biosynthesis in plants, to enzymes involved in the lignin biosynthetic pathway and nucleic acids encoding such enzymes and, more particularly, to methods of modifying lignin biosynthesis via sense suppression and to related nucleic acids and constructs.

Description

[0001] This application is a divisional application, the filing date of the original application is July 17, 2008, the application number is 200880107482.3, and the name is "Improvement of lignin biosynthesis through justice inhibition". technical field [0002] The present invention relates to the improvement of lignin biosynthesis in plants, enzymes comprising lignin biosynthetic pathways and nucleic acids encoding these enzymes, and more particularly, methods of improving lignin biosynthesis by sense inhibition, related nucleic acids and structures. [0003] The present invention also relates to regulatory elements, and more specifically, promoters capable of causing expression of exogenous genes in plant cells, such as genes encoding enzymes involved in the lignin biosynthetic pathway in plants. [0004] The invention also relates to vectors comprising the nucleic acids and regulatory elements of the invention, plant cells, plants, seeds and other parts of plants transforme...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12N15/54C12N15/60C12N15/52C12N15/82A01H5/00
CPCC12N9/0006C12N9/0008C12N9/0073C12N9/0077C12N9/1007C12N9/88C12N9/93C12N15/8255C07H21/00C12N15/8251
Inventor 杰曼·斯潘根贝格安杰拉·简·利杰特罗宾·路易丝·希思拉塞尔·莉·麦金尼斯达米安·保罗·林奇乌尔里克·彼得·约翰艾登·莫拉多夫梅甘·格里菲思
Owner DAIRY AUSTRALIA
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