CN105925605A - Application of lotus cellulose synthase gene NnuCESA8 - Google Patents
Application of lotus cellulose synthase gene NnuCESA8 Download PDFInfo
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Abstract
The invention discloses application of lotus cellulose synthase gene NnuCESA8, and belongs to the technical field of plant genetic engineering. According to the application, a positive transgenic plant of a lotus gene NnuCESA8 trans-arabidopsis cesa8 homozygous mutant is obtained. Then, the NnuCESA8 gene in a lotus is transferred into a corresponding arabidopsis mutant through a transgenic technology to detect a functional complementation situation of the NnuCESA8 gene and the arabidopsis mutant and deduce the function expression situation of the gene so as to improve the rate of forest cellulose subsequently and purposefully through transgenosis and improve the availability. According to the application of the lotus cellulose synthase gene NnuCESA8, the situation that the phenotype of the arabidopsis cesa8 can be partially compensated by the gene is found through the comparison of a transgenetic plant, a homozygous mutant plant and a wild type plant; the gene has the function of a cellulose synthase and promotes the synthesis of the cellulose; the application determines that the gene participates in the synthesis of the cellulose in secondary cell walls of a plant.
Description
Technical field
The invention belongs to field of plant genetic, relate to the application of a kind of Cellulose-synthase gene, special
Do not relate to the application of a kind of lotus Cellulose-synthase gene NnuCESA8.
Background technology
Cellulose be cell wall mainly comprise one of composition, be also evaluate that paper making raw material performance is good and bad one
Important indicator.Plant cell wall main cellulose, hemicellulose, lignin three great achievement are grouped into, and this is three big
Component is Renewable resource the abundantest on the earth, and cellulose is the first big component of cell wall, by homogeneous
Pyrans formula D-Glucose be formed by connecting with β-Isosorbide-5-Nitrae glycosidic bond, its glucose residue about 2000～25000
(Pauly M,Keegstra K(2010)Plant cell wall polymers as precursors for biofuels.
Current Opinion in Plant Biology,95:305-312；Pauly M,Keegstra K(2008)
Cell-wall carbohydrates and their modification as a resource for biofuels.The Plant
Journal,54(4):559-568).And this β-1,4 glycosidic linkage is catalyzed and synthesized by cellulose synthase.
Richmond and Somerville has been found that the encoding gene of 10 cellulose synthases in arabidopsis
(existing named AtCESA) (Richmond T A, Somerville C R. (2000) The
cellulosesynthase superfamily1.Plant Physiology,124:496-498).AtCESA1,
AtCESA2, AtCESA3, AtCESA5, AtCESA6 and AtCESA9 close with the biological of plant primary wall
Become relevant (Arioli T, Peng L, Betzner A, et al. (1998) Molecularanalysis of cellulose
biosynthesis in Arabidopsis.Science,279:717-720；Burn J E,et al.Functional
analysis of the cellulose synthase genes CESA1,CESA2,and CESA3 in arabidopsis.
Plant Physiology,129(2):797-807；Pagant S,et al.(2002)KOBITO1 encodes a
novel plasma membrane protein necessary for normal synthesis of cellulose during
Cell expansion in arabidopsis.The Plant Cell Online, 14 (9): 2001-2013), secondary wall is fine
Dimension element synthase gene complex is made up of AtCESA4, AtCESA7 and AtCESA8, the sudden change of these genes
Body all shows as xylem (irregular xylem, irx) (Turner S R, the Somerville C R. caved in
(1997)Collapsed xylem phenotype of Arabidopsis identifies mutants deficient in
cellulose deposition in the secondary cell wall.Plant Cell,9:689-701；Szyjanowicz P
M J,Mckinnon I,Taylor N G,et al.(2004)The irregular xylem 2 mutant is an allele
of korrigan that affects the secondary cell wall of Arabidopsis thaliana.The Plant
Journal,37(5):730-740).Fibre in arabidopsis cesa4 mutant and cesa8 mutant secondary cell wall
Dimension element deposition occurs that major defect causes tracheid (Turner S R, the Somerville that caving in occurs in plant
C R(1997)Collapsed xylem phenotype of Arabidopsis identifies mutants deficient in
cellulose deposition in the secondary cell wall.Plant Cell,9:689-701；Taylor N G,
Howells R M,Huttly A K,et al.(2002)Interactions among three distinct CesA
proteins essential for cellulose synthesis.100(3):1450-1455)。
At present, the possible function of CESA mostly come from gene expression, UDP-G Binding experiment (Pear J R,
Kawagoe Y,Schreckengost W E,et al.(1996)Higher plants contain homologs of
the bacterial celA genes encoding the catalytic subunit of cellulose synthase.Proc
Natl Acad Sci U S A, 93 (22): 12637-12642), mutation analysis (Fagard M, Desnos T,
Desprez T,et al.(2000)PROCUSTE1 encodes a cellulose synthase required for
normal cell elongation specifically in roots and dark-grown hypocotyls of
Arabidopsis.The Plant Cell, 12 (12): 2409-2424) and gene silencing experiments (Burton et al,
2000)。
It is perennial that lotus (Nelumbo nucifera) belongs to Nymphaeceae (Nymphaeaceae) Nelumbo (Nelumbo)
Aquatic persistent root herbs.Containing substantial amounts of cellulose in Folium Nelumbinis handle, fractureing from centre, it is the most elongated to pull out
And the lotus fiber of flexible.The reports such as the performance of lotus fiber is the most special, Pan, between two sections of petioles broken into two with one's hands
Cellosilk can be elongated at least ten centimetres and not turn off, every cellosilk is by the fine fibre silk of about 20
Composition, (Pan Y, Han G, Mao Z in the parallel and vessel element being distributed in Folium Nelumbinis handle in a spiral form
(2011)The anatomy of lotus fibers found in petioles of Nelumbo nucifera.Aquatic
Botany,95:167-171).The content difference of cellulose, hemicellulose, lignin in the filametntary composition of lotus
It is 41.4 ± 0.29%, 25.87 ± 0.64%, 19.56 ± 0.32% (Pan Y, Han G, Mao Z (2011)
Structural characteristics and physical properties of lotus fibers obtained from
Nelumbo nucifera petioles.Carbohyd Polym,85:188-195).Clone the cellulose synthase of lotus
Gene, and utilize transgenic technology to study its functional expression, it is beneficial to further investigate lotus cellosilk and fiber
The synthesis mechanism of element, so on purpose utilize lotus Cellulose-synthase gene go to improve some forest tree specieses for
Increase its content of cellulose, improve utilizability.
Lotus and the filametntary pharmaceutical applications of lotus, chemical property and physical arrangement are the most studied and reported, also
Have the research of lotus gene level, the genome of lotus in 2013 announce (Ray M, Robert V, Yanling L,
et al.(2013)Genome of the long-living sacred lotus(Nelumbo nucifera Gaertn.).
Genome Biology, 14 (5): R41), this is that the clone of lotus related gene provides convenience with functional study.
Summary of the invention
In order to overcome the shortcoming of prior art with not enough, it is an object of the invention to provide a kind of lotus cellulose and close
The application of enzyme gene NnuCESA8.In view of internal, outside containing a large amount of cellosilks and petiole in the petiole of lotus
The phenomenon that content of cellulose is inconsistent, carries out furtheing investigate its cellulose for starting with in terms of degree of gene expression
Inside and outside for Folium Nelumbinis handle and three positions of blade are done transcript profile order-checking, and then are studied its cellulose synthase by synthesis
The expression of gene.The main object of the present invention is at petiole position expression ratio by transcript profile one section of discovery
The expression of higher Cellulose-synthase gene.Check order the primers obtained with lotus according to transcript profile
The total cDNA of petiole is that this gene is cloned out and checks order by template, to sequencing result row BLAST, sends out
It is existing that it is the highest with the AtCESA8 gene order similarity of arabidopsis, therefore we by named
NnuCESA8.Then it is subscribed to intend south from ABRC (Arabidopsis Biological Resource Center)
Mustard cesa8 mutant (Salk_046685c), and by transgenic technology by the NnuCESA8 gene in lotus
It is transferred in corresponding Arabidopsis Mutants detect its situation that has complementary functions, infers the functional expression feelings of this gene
Condition, in order to the follow-up purposive ratio by transgene improvement forest cellulose, improves utilizability.
The present invention, by building over-express vector and studying its functional expression by transgenic technology, determines this gene
There is cellulose synthase function and promote the cellulosic electrode of Plant Secondary Materials cell wall.
The purpose of the present invention is achieved through the following technical solutions:
The present invention provides a kind of lotus Cellulose-synthase gene NnuCESA8 application in promoting cellulosic electrode.
The present invention provides a kind of lotus Cellulose-synthase gene NnuCESA8 cellulose in promoting secondary cell wall
Application in synthesis.
The aminoacid sequence of described lotus Cellulose-synthase gene NnuCESA8 is as shown in SEQ ID NO:1.
The nucleotide sequence of described lotus Cellulose-synthase gene NnuCESA8 is as shown in SEQ ID NO:2.
The detailed process of above-mentioned application is as follows:
1. the acquisition of gene
According to transcript profile data, obtain the CESA gene order of 15 lotuses, to these 15 CESA genes
Carried out RT-qPCR, find wherein sequence expression the highest and inside and outside petiole position expression
It is significantly larger than blade, according to transcript profile sequencing result design primer with the total cDNA of Folium Nelumbinis handle for template by this base
Because clone out and checks order, to sequencing result row BLAST, find the AtCESA8 of this sequence and arabidopsis
Sequence similarity is the highest, and therefore we guess that it is closely related with the synthesis of secondary cell wall cellulose, and will
This unnamed gene is NnuCESA8, and then the lotus genome sequence announced of search, find this gene order with
NNU_08388 sequence is identical.
Described primer sequence is as follows:
NnuCESA8-F:5 '-GGGGACAAGTTTGTACAAAAAAGCAGGCTTCATGAT
GGAGTCCGGGGTCC-3′；
NnuCESA8-R:5 '-GGGGACCACTTTGTACAAGAAAGCTGGGTCTCAACA
ATCAATAGAAATGCAGC-3′。
2. the structure of carrier
The cDNA of the NnuCESA8 of amplification is built to over-express vector pEarlyGate100 composition double
Unit's expression vector, obtains process LAN recombiant plasmid；This carrier carries cauliflower mosaic virus (CaMV)
35S promoter, and there is Herbicid resistant, it is available for the screening of transgenic plant.
3. convert Agrobacterium
Process LAN recombiant plasmid obtained above use freeze-thaw method convert to Agrobacterium tumefaciems GV3101,
Choose positive bacteria and preserve, preparing transfection arabidopsis.
4. carry out plant transgene operation with arabidopsis cesa8 Heterozygous mutants:
Owing to arabidopsis cesa8 Mutants homozygous plant is short and small, really pod deformity, bears seeds hardly, therefore selects
Cesa8 Heterozygous mutants without phenotype carries out transgenic and collects seed in case positive plant of future generation screens, will
Positive Agrobacterium containing genes of interest obtained above converts the cesa8 heterozygosis of 45 days full-bloom stages with dip method
Mutant plants, really collects seed, with Herbicid resistant screening transgenic positive plant, and leads to after pod maturation
Cross gene identification and choose the positive plant that genetic background is cesa8 Mutants homozygous, be transfer-gen plant.With
Arabidopsis cesa8 Mutants homozygous is as negative control..
5. transfer-gen plant Phenotypic Observation
Arabidopsis cesa8 Mutants homozygous plant is short and small, and lotus throne leaf is narrow short dark green, and really pod deformity is short and small,
Almost without seed, content of cellulose is relatively low, and has the xylem structure seriously caved in.By turning Lian Jiyin
NnuCESA8 enters arabidopsis cesa8 Mutants homozygous, to turning of growth 6 to 7 week planted under the same terms
Gene plant, WT lines and the stem height of cesa8 Mutants homozygous plant, lotus throne leaf size and fruit pod
State compares observation record.Transfer-gen plant has grown shorter and smaller than wild type but can normally produce seeds
The fruit pod of son, lotus throne leaf length increases 56% than cesa8 Mutants homozygous, and stem stalk height also increases
43%.
In order to observe xylem change, the stem transverse section of transfer-gen plant is carried out Toluidine blue staining,
Optical microphotograph Microscopic observation, cesa8 Mutants homozygous caves in than more serious tracheid originally in lotus
Obtain part after the proceeding to of NnuCESA8 gene to recover.
Measure the content of cellulose of arabidopsis stem by nitric acid Ethanol Method, measurement result proves transfer-gen plant
Content of cellulose increases 38.6% than cesa8 Mutants homozygous, but not yet recovers to wild type degree.
Result shows: determine Lian Jiyin by phylogenetic analysis and transgenic technology and complementary plant phenotype analysis
NnuCESA8 has portion of cellulose synthase function, can participate in the synthesis of cellulose in secondary cell wall.
The present invention, relative to prior art, has such advantages as and effect:
(1) present invention amplification obtains Lian Jiyin NnuCESA8, has obtained containing Lian Jiyin NnuCESA8's
Over-express vector.
(2) present invention has obtained Lian Jiyin NnuCESA8 and turns the positive of arabidopsis cesa8 Mutants homozygous and turn
Gene plant.
(3) present invention is contrasted with WT lines by transfer-gen plant, Mutants homozygous plant, and finding should
The phenotype of gene energy partial complementarity arabidopsis cesa8 Mutants homozygous, has the function of cellulose synthase, promotes
The synthesis of cellulose, and determine the synthesis of cellulose in this gene involved in plant secondary cell wall.
Accompanying drawing explanation
Fig. 1 be in transcript profile order-checking 15 NnuCESA genes at the expression of different parts；Wherein, T1,
T2, T3 are inside blade, petiole outside and petiole respectively.
Fig. 2 is the RT-qPCR result at different parts of 15 NnuCESA genes；Wherein, T1, T2,
T3 is inside blade, petiole outside and petiole respectively.
Fig. 3 is expression vector establishment schematic diagram.
Fig. 4 is cesa8 Mutants homozygous, transfer-gen plant and WT lines phenotype comparison diagram；Wherein, figure
A is respectively cesa8 Mutants homozygous, transfer-gen plant and WT lines from left to right；Figure B in under
Supreme three rows are respectively the lotus throne blade of cesa8 Mutants homozygous, transfer-gen plant and WT lines；C schemes
Three rows are respectively the fruit pod of cesa8 Mutants homozygous, transfer-gen plant and WT lines from bottom to up.
Fig. 5 is cesa8 Mutants homozygous, transfer-gen plant and WT lines stem traverse section comparison diagram；
Wherein, A, B, C are respectively stem's horizontal stroke of cesa8 Mutants homozygous, transfer-gen plant and WT lines
Tangent plane Toluidine blue staining figure, the position of arrow indication is tracheid.
Detailed description of the invention
Below in conjunction with embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention
It is not limited to this.
The experimental technique of unreceipted specific experiment condition in the following example, generally according to conventional laboratory conditions or
According to the experiment condition proposed by manufacturer.
Embodiment 1: the acquisition of lotus NnuCESA8 gene order
First inside and outside for Flos Nelumbinis petiole and three positions of blade are done transcript profile order-checking and obtains each genetic fragment in difference
The expression at position, finds out the expression (result is shown in Fig. 1) of 15 CESA genetic fragments, and is RT-qPCR
Checking (result is shown in Fig. 2), each gene is in the result of relative expression quantity result and the transcript profile order-checking of different parts
Trend is basically identical, and wherein one section of CESA gene order expression is the highest and inside and outside at stem stalk
Expression is above blade position, according to transcript profile sequencing result design primer with the total cDNA of petiole as template
Clone out by this gene and check order, to sequencing result row BLAST, finding in this gene and arabidopsis
The Gene A tCESA8 similarity participating in secondary cell wall cellulosic electrode is high, therefore it is presumed that this gene
The synthesis of cellulose in secondary cell wall should be participated in, and by its named NnuCESA8, pass through sequence alignment
Find, this gene order and the gene in lotus genome website (http://lotus-db.wbgcas.cn/)
NNU_08388 sequence is identical.
Embodiment 2: the extraction of the total serum IgE of Folium Nelumbinis handle
Extract test kit (OMEGA) with RNA and extract the total serum IgE of Folium Nelumbinis handle respectively.Detailed step reference
Test kit description.
Embodiment 3: the acquisition of the total cDNA of lotus
Use OMEGA Reverse Transcription box to obtain the total cDNA of lotus with Folium Nelumbinis handle total serum IgE for template, walk in detail
Suddenly operate according to test kit operation instructions.
Embodiment 4: the amplification of purpose fragment
With the mRNA of Lian Jiyin NnuCESA8 for stencil design two ends primer, and at primer two ends plus using
Gateway adapter-primer in carrier construction.Upstream and downstream primer is respectively as follows:
NnuCESA8-F:5 '-GGGGACAAGTTTGTACAAAAAAGCAGGCTTCATGAT
GGAGTCCGGGGTCC-3′；
NnuCESA8-R:5 '-GGGGACCACTTTGTACAAGAAAGCTGGGTCTCAACA
ATCAATAGAAATGCAGC-3′。
With the total cDNA of Folium Nelumbinis handle as template, under annealing temperature is 52 DEG C and program that elongating temperature is 72 DEG C
Carry out PCR amplification, amplified production purification preserving after electroresis appraisal.
Embodiment 5: vector construction connects
Use Gateway binary vector con-struction method, amplified production and the carrier matter that will obtain in embodiment 4
Grain pDONR207 (buying in ABRC, http://abrc.osu.edu/) carries out BP recombining reaction, and restructuring is produced
Thing converts DH5 α competent cell (commercially available) and also obtains positive bacteria, and amplification culture is also carried out after extracting plasmid
Plasmid order-checking is identified.By the correct recombiant plasmid of order-checking and pEarlyGate100 (buy in ABRC,
Http:// abrc.osu.edu/) (this carrier carries the 35S promoter of cauliflower mosaic virus (CaMV), and has
Herbicid resistant, is available for the screening of transgenic plant) carry out LR recombining reaction, recombinant products converts large intestine bar
Bacterium DH5 α also obtains positive bacteria, extracts plasmid and checks order, by recombiant plasmid correct for order-checking after amplification culture
Save backup.Expression vector establishment schematic diagram is as shown in Figure 3.Sequencing result as shown in SEQ ID NO:2,
Identical with sequence in gene NnuCESA8 (gene I/D is NNU_08388).
Embodiment 6: convert Agrobacterium tumefaciems GV3101
1. by Agrobacterium tumefaciems (Agrobacterium tumefaciens) GV3101 competent cell (commercially available)
Thawed on ice, adds the recombiant plasmid in 1 to 5 μ L (< 300 μ g) embodiment 5, flicks mixing, ice bath
15min。
2. it is transferred to said mixture in liquid nitrogen suddenly freeze 5min, then 37 DEG C of heat shock 5min, is transferred quickly to
5min on ice.
3. adding 800 μ L nonresistant LB fluid medium, 28 DEG C, 200rpm shakes 4h.
4.5000rpm, centrifugal 1min, abandon 700 μ L of supernatant, and remaining is resuspended, takes 200 μ L and coats LB
Solid medium (Rif+: 50 μ g/mL, Gen+: 15 μ g/mL), it is inverted for 28 DEG C and cultivates 2 days.
5. choose positive bacterium colony and little shaking protects bacterium.
Embodiment 7: Agrobacterium infection arabidopsis
1. select bolting arabidopsis (Arabidopsis thaliana) cesa8 Heterozygous mutants plant after ten days,
Keep higher levels of humidity, prepare to convert.
2. recover in LB culture medium the above-mentioned Agrobacterium with recombiant plasmid respectively, and 28 DEG C, 220rpm is little
Distant 24h.
3.5000rpm centrifugal 15min collects thalline.Simultaneously preparation 1L re-suspension liquid (5% (w/v) Sucrose,
0.02～0.05%Silwet-77).
4. abandoning supernatant, residual bacterium solution 5% (w/v) sucrose further cleans, 8000rpm, centrifugal 10min
Collecting thalline, this step can more once.
5., with the resuspended thalline of re-suspension liquid to OD600 position 0.8, re-suspension liquid is transferred in 15cm culture dish.
6. the inflorescence of arabidopsis to be transformed is invaded 1min in re-suspension liquid.
7. being inverted by transformed plant and keep flat, 22 DEG C, relative humidity 60～70% light culture, after one day, transfer to
Normal cultivation, collected transformed the seed after one month.
Embodiment 8: screening transgenic positive plant
The planting seed that will obtain in embodiment 7, about 10 days herbicide sprayings of rudiment, obtain the plant that survives
Identified by DNA afterwards and determine whether genes of interest proceeds to and plant genotype background, carry out labelling, transgenic
After cesa8 Mutants homozygous can tie a small amount of seed, individual plant sowing preserving after fruit pod maturation.
Embodiment 9: arabidopsis observes phenotype
Choose growth wild type, cesa8 Mutants homozygous and the transfer-gen plant of 6～7 weeks to plant to observe contrast
Strain character mutation, each type of plant is chosen more than ten strains and measures its stem stalk height, lotus throne with scale
Leaf length, observes and Taking Pictures recording fruit pod size.As shown in Figure 4 and Table 1, result shows result, cesa8
The malformed fruit pod of Mutants homozygous is more, almost without seed；Transfer-gen plant has grown shorter than wild type
Little but can normal seed bearing fruit pod, lotus throne leaf length increases 56%, and stem than cesa8 Mutants homozygous
Stalk height also increases 43%.
Table 1 WT lines, Mutants homozygous and transfer-gen plant content of cellulose comparative result
| Wild type | Cesa8 Mutants homozygous | Transfer-gen plant | |
| Stem stalk fiber cellulose content (μ g/mg) | 300±8.97A | 159.225±6.89C | 220.6125±5.76B |
| Lotus throne leaf length (cm) | 4.41±0.13A | 1.29±0.08C | 2.02±0.09B |
| Stem stalk height (cm) | 33.4±2.37A | 11.15±1.53C | 15.96±1.33B |
Note: numerical value represents meansigma methods+standard deviation (n=4), different letters represents significant difference (P < 0.05；T checks).
Embodiment 10: arabidopsis stem content of cellulose compares
Cellulose extracts and uses nitric acid Ethanol Method.Concretely comprise the following steps:
First the arabidopsis stem collected is pulverized, through Benzene-ethanol extraction after drying, weigh 2g through Benzene-ethanol extraction
Sample, is placed in sample in the conical flask of 250mL clean dried, adds 25mL nitric acid alcohol mixeding liquid,
Load onto reflux, be placed on boiling water bath heating 1h.In heating process, conical flask should be swayed at any time, with
Prevent sample from beating.After boiling 1h, remove condensing tube, conical flask is taken off in water-bath, stand a moment.
After at the bottom of residue deposition bottle, with decantation filter through the 1G2 glass filter of constant weight, do not make sample flow out as far as possible.
With vacuum pump, the filtrate in filter is blotted, then with Glass rod, the residue flowing into filter is moved into weight in conical flask
Implement above-mentioned steps for several times again, till fiber bleaches.Finally conical flask content is all moved into filter,
Washing residue with 10mL nitric acid alcohol mixeding liquid, reusable heat water washs, and meets methyl orange not in acid to cleaning mixture
Property reaction till.Finally use washing with alcohol twice, blot washing liquid, filter is moved into baking oven, in 105 ± 2 DEG C
Dry to constant weight.Content of cellulose calculates: content of cellulose=(m1-m2)/m0 (1-w) × 100%, formula
Middle m1 is the quality of bone dry fiber element and glass filter, and m2 is over dry sky glass filter quality, and m0 is air-dried
The quality of sample, w is the water content of sample.Content of cellulose the results are shown in Table 1.As can be known from Table 1, survey
Determine result and prove that the content of cellulose of transfer-gen plant increases 38.6% than cesa8 Mutants homozygous, but not yet
Recover to wild type degree.
Embodiment 11: the making of arabidopsis stem section transverse section and observation
Choose growth wild type, cesa8 Mutants homozygous and the transfer-gen plant stem of 6～7 weeks respectively,
The above 3cm of upper soll layer takes 1cm stem section, by the Agarose embedding stem section of 3% (w/v), at Leica
Cut into slices on VT1000S vibrations microtome, thickness 40 μm, amino toluene indigo plant dyeing 1～2min, it is placed in microscope slide
On relatively and take pictures in optical microphotograph Microscopic observation.Result is as it is shown in figure 5, A, and B, C are respectively cesa8
Stem's cross section Toluidine blue staining figure of Mutants homozygous, transfer-gen plant and WT lines, arrow institute
The position referred to is tracheid, and the tracheid's Collapse Deformation ratio in figure A is more serious, the wood in figure B
Matter portion cell has obtained part and has recovered, but the cell still subsided with the presence of part deformity, it is not fully returned to open country
Raw type.
Above-described embodiment is the present invention preferably embodiment, but embodiments of the present invention are not by above-mentioned enforcement
The restriction of example, the change made, modifies, replaces under other any spirit without departing from the present invention and principle
In generation, combine, simplify, all should be the substitute mode of equivalence, within being included in protection scope of the present invention.
Claims (7)
1. kind of lotus Cellulose-synthase gene NnuCESA8 application in promoting cellulosic electrode.
Application the most according to claim 1, it is characterised in that: described lotus Cellulose-synthase gene
NnuCESA8 is the application in cellulosic electrode in promoting secondary cell wall.
Application the most according to claim 1 and 2, it is characterised in that: described lotus cellulose synthase base
Because the aminoacid sequence of NnuCESA8 is as shown in SEQ ID NO:1.
Application the most according to claim 1 and 2, it is characterised in that: described lotus cellulose synthase base
Because the nucleotide sequence of NnuCESA8 is as shown in SEQ ID NO:2.
5. the over-express vector containing lotus Cellulose-synthase gene NnuCESA8.
6. a transgenic plant, it is characterised in that: containing the over-express vector described in claim 5.
Transgenic plant the most according to claim 6, it is characterised in that: described plant is arabidopsis.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1344325A (en) * | 1998-10-13 | 2002-04-10 | 吉尼西斯研究及发展有限公司 | Materials and methods for modification of plant cell wall polysaccharides |
| WO2007019245A2 (en) * | 2005-08-03 | 2007-02-15 | Board Of Control Of Michigan Technological University | Methods for enhancing expression of secondary cell wall cellulose synthases in plants |
| WO2009049373A1 (en) * | 2007-10-19 | 2009-04-23 | Australian Centre For Plant Functional Genomics Pty Ltd | Modulation of plant cell wall deposition |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1344325A (en) * | 1998-10-13 | 2002-04-10 | 吉尼西斯研究及发展有限公司 | Materials and methods for modification of plant cell wall polysaccharides |
| WO2007019245A2 (en) * | 2005-08-03 | 2007-02-15 | Board Of Control Of Michigan Technological University | Methods for enhancing expression of secondary cell wall cellulose synthases in plants |
| WO2009049373A1 (en) * | 2007-10-19 | 2009-04-23 | Australian Centre For Plant Functional Genomics Pty Ltd | Modulation of plant cell wall deposition |
Non-Patent Citations (11)
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