CN110527688A - 84K poplar ARK1 gene and its utilization in Hybrid Poplar - Google Patents
84K poplar ARK1 gene and its utilization in Hybrid Poplar Download PDFInfo
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Abstract
The invention belongs to forestry breeding technical fields, specially 84K poplar ARK1 gene and its utilization in Hybrid Poplar.In the present invention, the ARK1 gene of 84K poplar, constructs new carrier, and has cultivated ' 717 ' Hybrid Poplars for turning ARK1 gene, and auxin related gene up-regulated expression can promote plant strain growth；Lignin synthesis related gene lowers expression simultaneously, will lead to content of lignin decline, improves its industrial utility value.
Description
Technical field
The invention belongs to forestry breeding technical fields, the specially method of hybrid poplar transgenic breeding.
Background technique
Poplar belongs to Angiospermae (Angiosperms) Dicotyledoneae (Dicots) Salicaceae (Salicaceae)
The fallen leaves xylophyta of Populus (Populus), growth of poplar is rapid, and wide adaptability and flexibility are strong, and it is anti-to be widely used in ecology
Shield, urban afforestation, industry and fiber production.Poplar is in addition to widely distributed, growth is rapid, yield is high, adaptive capacity to environment
Outside the various features such as strong and easy improvement, genome sequencing has been completed, and is that the idealized model of Forest Tree Genetic Breeding improvement is planted
Object.All poplar tree species all have the ability of vegetative propagation.Poplar is the natural host of Agrobacterium tumefaciems, convenient for utilizing crown gall
Agrobacterium-mediated transformation carry out plant genetic transformation, therefore poplar be considered as Forest-tree Gene Engineering research model plant it
One, furthermore poplar and its hybrid are considered as the preferred perennial plant of bioenergy raw material production.Poplar is that Temperate Region in China is raw
Long fastest tree species, have a variety of economical characters and a species advantage, and current biomass growth rate is close to 10-15kg/hm2
Every year；Biomass can be stored in root by irrigating rudiment regeneration, can reduce the investment to storage facilities to the maximum extent；
Poplar period of farming and harvesting compared with other fibre crops is shorter, highly beneficial to the net effect of greenhouse gas emission.
' 717 ' Hybrid Poplars (Populus tremula × P.alba ' INRA 717-1B4 ') are from trembling poplar and silvery white
The excellent variety selected in the hybrid Population of poplar, have stronger resistance of reverse, to revegetation, prevent erosion with it is saline and alkaline
Ground reparation has very big application value, it may also be used for biomass energy and fiber energy development.' 717 ' Hybrid Poplars are perennial
Plant exists compared to the annual herb plants such as rice (Oryza sativa) and arabidopsis (Arabidopsis thaliana)
Formation and the seasonality suspend mode etc. for studying timber have unique advantage, and poplar generates a large amount of weights during evolution
Its regulatory mechanism of multiple sequence is increasingly complex, therefore seems outstanding by research object progress gene function analysis of ' 717 ' Hybrid Poplars
It is important.
Summary of the invention
The invention discloses the ARK1 genes of 84K poplar, it is characterised in that the sequence of ARK1 gene such as nucleotide sequence is such as
Shown in SEQ.ID.NO.1.
' 717 ' Hybrid Poplars for turning ARK1 gene are cultivated in the utilization of the ARK1 gene of 84K poplar.
The utilization of the ARK1 gene of 84K poplar is to construct the transgene carrier of ARK1 gene (from 84K poplar) simultaneously
' 717 ' the poplar plant for turning ARK1 gene are obtained by transgenosis.Transgenic hybrid poplar of the invention internode away from, stem thickness,
There were significant differences on petiole length, leaf width, leaf length and height of seedling, has on the related gene that the growth and development of plant and lignin regulate and control
Difference.The ARK1 gene of 84K poplar takes part in the growth course of plant, is an important gene relevant to growth, is making
It has broad application prospects in terms of the improvement of object germ plasm resource, genetic breeding.
In the present invention, the ARK1 gene of 84K poplar, constructs new carrier, and has cultivated and turned ' the 717 ' of ARK1 gene
Hybrid Poplar, auxin related gene up-regulated expression, can promote plant strain growth；Lignin synthesis related gene lowers expression simultaneously,
It will lead to content of lignin decline, improve its industrial utility value.
Detailed description of the invention
Fig. 1 is PCR detection figure, and M is D2000 maker in figure；CK is that the DNA of ' 717 ' Hybrid Poplar plant of non-transgenic is
The PCR product of template；1-11 is using the DNA of ' 717 ' Hybrid Poplar different plants of transgenosis as the PCR product of template respectively；
Fig. 2 is ' 717 ' Hybrid Poplar of non-transgenic and turns ' 717 ' Hybrid Poplar aspect graph of ARK1 gene, A: non-transgenic ' 717 '
Hybrid Poplar；B and C: expression turns ' 717 ' Hybrid Poplar of ARK1 gene；
Fig. 3 microexamination cytological map A, B are ' 717 ' Hybrid Poplar of non-transgenic；C, D turn ' 717 ' Hybrid Poplar of ARK1 gene；
Fig. 4 shows gene expression difference.
Specific embodiment
Step 1, design of primers and synthesis
1) target gene amplimer
Primer ARBF, sequence 5 '-GGAGAGGACACGCTCGAGATGGAGGGTGGTGATGGTG-3 ', length 37bp, sequence
Column are as shown in SEQ.ID.NO.2.
Primer ARBR, sequence 5 '-ATCCTTGTAGTCGAATTCAAGCAGTGTGGGAGAGATGTC-3 ', length 39bp,
Sequence is as shown in SEQ.ID.NO.3.
2) target gene amplification and glue recycling, recycle target gene band using hundred Tyke plastic recovery kits.
3) target gene is connect with PART-CAM-FLAG, carrier construction.
PART-CAM-FLAG-gene constructing plan:
1) XhoI and EcoRI double digestion plasmid PART-CAM-FLAG is used, large fragment is recycled；
Double enzyme digestion reaction system:
| Reaction system | Usage amount |
| 10xTango buffer | 4.0μl |
| XhoI | 1μl |
| EcoRI | 1μl |
| Plasmid | 1μg |
| dH2O | up to 20μl |
37 DEG C of digestion 2.5h；65 DEG C: 20min inactivation.
2) target gene is cloned with upstream and downstream primer, with XhoI and EcoRI double digestion PCR product, recycles small fragment；
3) by 1,2 gained segments recombination ligase recombination connection, conversion to DH5 α competent cell；Bacterium colony PCR detection；
4) picking positive bacterium colony sequence verification obtains and constructs successful carrier and related strain.
Because Component Vectors can connect together with foreign gene ARK1 gene, will not partially connect together with foreign gene, portion
Divide also in linear condition, partially can be in cyclic annular state, the carrier only containing foreign gene and in cyclic annular state has been only
The carrier imitated and can expanded in Escherichia coli, thus need to select positive colony.The carrier of building compares in Escherichia coli
It is easy amplification, and is easy to screen the gene of ARK1 containing foreign gene and carrier annular in shape.But it can not be infected with great Yang bacillus
Poplar, only Agrobacterium tumefaciems can just infect poplar.So to screen the Escherichia coli of the genophore containing ARK1, and expands and arrive
Enough amounts, and separate and be transformed into Agrobacterium tumefaciems, it just can be carried out infecting for next step.
Step 2, carrier convert Agrobacterium tumefaciems
The upgrading grain from the DH5 α of the genophore containing ARK1, and Agrobacterium tumefaciens strain LBA4404 is converted, PCR amplification is chosen
Positive strain is selected, for disseminating in next step.
Step 3, Hybrid Poplar transgenosis and detection
1) foundation of ' 717 ' Hybrid Poplar tissue culture systems
With 75% ethanol postincubation 15s, 0.1% mercuric chloride handles 10min and carries out explant disinfection；In evoked callus,
Used medium is MS+1.0mg/L6-BA+1.2mg/l NAA.Adventitious buds differentiation formula is MS+1.0mg/l6-BA+0.4mg/l
ZT.The formula that seedling takes root is 1/2MS+0.02mg/L NAA+0.6mg/LIBA.
2) ' 717 ' Hybrid Poplar blade genetic conversion systems are constructed
On the basis of ' 717 ' Hybrid Poplar blade tissue culture system of foundation, the choosing of ' 717 ' Hybrid Poplar blade Kan concentration is determined
Selecting pressure is 100mg/L, and carbenicillin optimum allowable concentrations are 200mg/L.Bacterial concentration OD600 value is 0.3, and immerged time is
When 10min, the conversion Hybrid Poplar blade effect of Agrobacterium is best, and method for transformation is leaf disk method.Include the following steps:
A) it infects
In on superclean bench, ' 717 ' the Hybrid Poplar blades through preculture are taken out from culture bottle, are put into early period through making a living
It is soaked for a period of time in the bacterium solution of change.It then takes out ' 717 ' Hybrid Poplar explants and is placed on aseptic paper the bacterium solution for sucking attachment.
B) it co-cultures
By ' 717 ' the Hybrid Poplar blade inoculations infected with Agrobacterium in callus inducing medium (MS+
NAA1.0mg/L+6-BA 1.0mg/L) on, 2~4d is co-cultured under the conditions of 28 DEG C of dark cultures, it is seen that agriculture is generated around blade
Bacillus bacterial plaque.
C) selection culture
By the callus by co-culturing with sterile water wash 3 times or so, with aseptic paper suck dry moisture, it is then transferred into
On de- bacterium differential medium (MS+1.0mg/L 6-BA+0.4mg/L ZT) added with selection pressure (carbenicillin+Kan), in light
According to carrying out selection culture under the conditions of period 16/8h, 28 DEG C.
D) subculture selection culture
28d or so replaces a Selective agar medium and induces its differentiation, induces new callus and budding.
E) culture of rootage
When adventitious bud grows to 2~3cm, the root media (1/2MS containing selection pressure (carbenicillin+Kan) is moved on to
+ 0.02mg/L NAA+0.6mgL IBA) in carry out culture of rootage, after plant grows adventitious root, be subsequently moved to greenhouse practice seedling.
3) Molecular Detection of transgenic plant
According to the expression vector of building, corresponding specific detection primer is designed, upstream primer is then ARK1 mesh gene internal
Sequence, downstream primer use carrier NPT II in sequence, the sequence of primer are as follows:
Forward 5 '-AAGATCCAGCCCTTGACCAA-3 ' sequence is as shown in SEQ.ID.NO.4；
Reverse 5 '-CATTGCCATCACCACAACCA-3 ' sequence is as shown in SEQ.ID.NO.5
Using above-mentioned primer, PCR reaction is carried out according to following system.
After being separately added into above-mentioned each ingredient, of short duration centrifugation is carried out by liquid and is collected into tube bottom, uses Takara company
Prime STAR Max Premix archaeal dna polymerase carries out PCR reaction.
PCR amplification program is as follows:
98 DEG C, 3min；35Cycles；94 DEG C of 30s, 55 DEG C, 30s；72℃5min；72℃10min.
Then PCR reaction is carried out, PCR reaction step is consistent with bacterium colony PCR.Then with 1.2% Ago-Gel (0.5 μ g/
Mol ethidium bromide) electrophoresis detects pcr amplification product.
The DNA for turning ' 717 ' Hybrid Poplar blades of ARK1 gene is extracted, PCR detection is carried out, discovery positive rate is 31.43%,
' 717 ' Hybrid Poplar of preliminary proof ARK1 over-express vector genetic transformation success (as shown in Figure 1).
4) transgenic plant morphological change detects
The morphological difference of the tissue-cultured seedling of ' 717 ' Hybrid Poplar transgenic plants and nontransgenic plants is compared, is measured
Stem section thickness, internode number and the panel length of identical growth cycle, every group is done 3 repetitions.
The result shows that: ' 717 ' Hybrid Poplar seedling of transgenosis compared to non-transgenic seedling internode away from, stem thickness, petiole be long, leaf
There were significant differences on wide, leaf length and height of seedling.' 717 ' Hybrid Poplar seedling stem section of transgenosis is elongated, is in usually pencil and multi-branched, does not have
Elongated petiole, leaf-shaped is elongated, and some blades do not develop complete (as shown in Figure 2).
5) paraffin section and displaing microstructure observing
A) it draws materials: cutting the stem section under the 5th leaf of ' 717 ' Hybrid Poplars rapidly, be about 3~5mm, it is solid to immerse FAA immediately
Determine liquid (50.0% alcohol, 5.0% acetic acid, 3.7% formaldehyde, 41.3% water).
B) fixed: to be fixed at room temperature in FAA fixer more than for 24 hours.
C) it is dehydrated: being first washed with distilled water；Serial dehydration then is carried out with the mixed liquor of the tert-butyl alcohol and alcohol, is sequentially
70%-85%-95%-100%-100%, every step 2h.45min × 8 are evacuated, are dehydrated step by step to 100% tert-butyl alcohol, 100% uncle
Butanol is dehydrated 3h, is repeated 3 times.
D) waxdip: being added the paraffin of thawing in ' 717 ' Hybrid Poplar samples, is placed more than for 24 hours in 65 DEG C of incubator.
E) it embeds: the paraffin that 65 DEG C melt being poured into the carton of 3cm × 3cm, waits paraffin slightly to solidify, bottom is seen white
When, ' 717 ' Hybrid Poplar materials are erected and are inserted into paraffin, bubble is chosen with dissecting needle and is removed, it is quiet to put after wax stone solidification, put 5 DEG C of ice
Case saves stand-by.
F) it is sliced: repairing wax stone, stick on wooden unit, with slice 8 μm of slabs of machine-cut, slice is sticked to load glass with being stained with tablet
On piece, glass slide overnight, can also dry glass slide in 45 DEG C of degree incubators with room temperature.
G) dewax: after slice is dry through dimethylbenzene → dimethylbenzene → 100% alcohol of+1/2 alcohol → 2 time of 1/2 dimethylbenzene →
95% alcohol → 95% alcohol → 70% alcohol → 50% alcohol → distilled water 1min.
H) dye: hematoxylin dyes 10min, 50% alcohol → 70% alcohol → 85% alcohol → 95% alcohol → 70% wine
Essence → 2+50% alcohol → 2 time dimethylbenzene of 100% alcohol → 1/2 dimethylbenzene
I) microscopy: mounting, micro- sem observation slice.
The result shows that: paraffin section and ' 717 ' Hybrid Poplar seedling of displaing microstructure observing transgenosis are compared with non-transgenic seedling stem wood
Matter portion and phloem cell quantity reduce and cell dia becomes smaller (as shown in Figure 3).
4, transcript profile is sequenced
The mRNA of ' 717 ' Hybrid Poplar of transgenosis and ' 717 ' Hybrid Poplar sample of non-transgenic is extracted, reverse transcription is at cDNA, benefit
High pass measurement is carried out with cDNA of the microarray dataset HiSeq to ' 717 ' Hybrid Poplar of transgenosis and ' 717 ' Hybrid Poplar sample of non-transgenic
Sequence analysis, including sequencing data filtering, difference expression gene GO functional analysis and Pathway functional analysis, carry out relevant difference
The screening and analysis of expressing gene.
The result shows that: initial data is filtered by Quality Control, after the reads for removing redundant sequence and low quality value, is obtained altogether
To the Clean reads of 45.8GB, wherein sequencing quality Q30 base percentage is average in 91.85% or more, four sample
G/C content is 47.69%, illustrates that sequencing quality is good, meets requirement for construction data base.Simultaneously by Clean reads be sequenced and
After poplar carries out sequence alignment with reference to genome, comparison efficiency is differed from 55.61% to 60.61%, and there are about 57.53% sequences
Genome annotation can be referenced.Wherein, the Clean reads comparison rate that position is uniquely compared with reference sequences is 56.61%.In
In difference expression gene collection number of genes, 641 difference expression genes are filtered out, wherein up-regulation number of genes there are 389, are lowered
Gene has 252 (as shown in Figure 4).
By to the adjusting and the transduction of access Plant hormone signal that are grown with cell tip-growth and separate living tissue and phenylpropyl alcohol
The gene that significant difference is expressed in alkanes biosynthesis pathway is analyzed, and after differential gene functional annotation, we are screened
Difference expression gene to during 27 participation growth of poplar, including protide, transcription factor class and protein kinases, these
Gene is related to the growth and development of plant and lignin regulation.In transgenic plant, the relevant gene expression of plant growth is big
It is in up-regulation state more, means to raise in turning ' 717 ' Hybrid Poplar of ARK1 with growth related gene in this way, illustrate ARK1 gene pairs
The growth of plant has positive regulation effect；And lignin synthesis relevant enzyme such as hydroxyphenyl lignin (P- in phenylpropyl alcohol alkane approach
Hydroxyphenyl Lignin), guaiacyl lignin (Guaiacyl lignin), 5- hydroxyl guaiacyl lignin (5-
Hydroxyguaiacyl lignin), syringyl lignin (Syringyl lignin), p-Coumaric Acid (p-Coumaric
Acid it) and to the expression of coumaric acyl coacetylase (p-Coumaroyl-CoA) etc. lowers.And there is height in the phenotype of plant
The misgrowth of degree, diameter etc..
Lignin is one of three big main chemical compositions of timber (lignin, cellulose and hemicellulose), in plant
With important biological function.Lignin limits the development of paper industry, this is because can generation environment in paper-making process
Caused by polluting and needing a large amount of wood producing energy, the reduction of content of lignin not only can be improved paper pulp and make in trees
The economy and environmental benefit of paper industry, can also promote the decomposition of lignocellulosic, improve the transformation efficiency of sugar.
To sum up, the growth rate of poplar and its value in paper industry can be improved in this transgenic research.
<110>Southwest Forestry University
<120>84K poplar ARK1 gene and its utilization in Hybrid Poplar
<160> 5
<210> 1
<211> 1352bp
<212> DNA
<213>84K poplar
<400> 1
TATCATGCGATCATAGGCGTCTCGCATATCTCATTAAAGCAGGACTCTAGATTAGGGACGTCTCTTATCGTCA
TCGTCCTTATAGTCTTGCTTGTCATCGTCATCTTTATAATCGCCCTTGTCATCGTCATCCTTGTAGTCGAATTCAAG
CAGTGTGGGAGAGATGTCCATTGGGAAAGGATTGCCCAAAACATTATCCATGTAGTAATGAGGATGGCCGGCATCCA
TCACCACAAACTGCATGTCTTCCGATGGTTTCCAGTGTCGTTTCCTTTGGTTAATGAACCAGTTGTTTATTTGCTTC
TGATCCAGACCAGTGGATTCAGCAAGGGCCAGCTTCTGCGACTCCGATGGATATGGCCATTTGTAATGCCTGCTCCA
CCAATCTAGCAGCTGTTGCCTGGCTTCTTTCGGTAACTTCCCTTTCTTTCTCTTTTTCATGAATTCCTGCTTGAGAC
TCCCTAAATATCCACTGTACCTGCGAAGTAGCTGACCCTTCAGTTCTTGATCTTCAGCTTGAGGATCTATGAAGTTG
TTATTCACATCAACCTCTTCTTCAGATGACACATTCCTATCATTGCCATCACCACAACCAGAAATTGGAGAGGAAAC
AGTGAGAGCTTTGAATTGACACTCAACCCTCTGAAGAAAAAGCATGGCTTCCTTTAAGGGTTTAGAGAGTTCTTGCT
CATACTTGGTCAGCATCTCACAGTAAGCCTCCATGAATTGGTCAAGGGCTGGATCTTCGCCGATGCAGCCTGTGTTA
GCGGGGGCCATAGAACCAGCCGATGCGCAAGCTTCTTCTAGCCTAGCCACCACTTCAGGTGGTGCTCCAACCTTCTG
ACAATTAGCATAGGCAGCAAGGAGACGATGGTAGTGAGGATGAGCCATAATTTTGGCCTTCACAGAAGAACTACTAC
CATCATTGTTATCCATGAAATAACACCCAGTAGCAGTGACAGTGTTGTTGTGGTTGTGATCGTCAATAATCATAGAA
GAAGAGCCACTAGCATTACTATGGATACGGTTATGGCCTTGATTGTTTGTAGGAGGTAGAGGAAGGAATAAGGTATT
TGATATTGACGAATCGCCCTCATTAGCCCGATGATGATGATGATGATGATGAGCAGAAGACGAAGAAGACATGAGAG
GCGTCATCATCATCATAGGGCAAAGTCCATTGCTGTTGTCTCCAAAAGCCATCATACATGAAGTGGTACTTGAACCA
CCATCACCACCCTCCATCTCGAGCGTGTCCTCTCCAAATGAAATGAACTTCCTTATATAGAGGAAGGGTCTTGCGAA
GGATAGTGGGATTGTGCGTCATCCCTTACGTCAGTGGAGATGTCACA
<110>Southwest Forestry University
<120>84K poplar ARK1 gene and its utilization in Hybrid Poplar
<160> 5
<210> 2
<211> 37bp
<212> DNA
<213>84K poplar
<400> 2
5′GGAGAGGACACGCTCGAGATGGAGGGTGGTGATGGTG-3′
<110>Southwest Forestry University
<120>84K poplar ARK1 gene and its utilization in Hybrid Poplar
<160> 5
<210> 3
<211> 39bp
<212> DNA
<213>84K poplar
<400> 3
5′ATCCTTGTAGTCGAATTCAAGCAGTGTGGGAGAGATGTC -3′
<110>Southwest Forestry University
<120>84K poplar ARK1 gene and its utilization in Hybrid Poplar
<160> 5
<210> 4
<211> 20bp
<212> DNA
<213>84K poplar
<400> 4
5’ -AAGATCCAGCCCTTGACCAA-3′
<110>Southwest Forestry University
<120>84K poplar ARK1 gene and its utilization in Hybrid Poplar
<160> 5
<210> 5
<211> 20bp
<212> DNA
<213>II gene of NPT of carrier PART-CAM
<400> 5
5’ -CATTGCCATCACCACAACCA-3′
Claims (2)
- The ARK1 gene of 1.84K poplar, it is characterised in that the sequence of ARK1 gene such as nucleotide sequence such as SEQ.ID.NO.1 institute Show.
- 2. the utilization of the ARK1 gene of 84K poplar as described in claim 1, it is characterised in that cultivation turns ARK1 gene ' 717 ' Hybrid Poplars.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003000898A1 (en) * | 2001-06-22 | 2003-01-03 | Syngenta Participations Ag | Plant genes involved in defense against pathogens |
| US20110061133A1 (en) * | 2007-09-14 | 2011-03-10 | Basf Plant Science Gmbh | Plants having increased yield-related traits and a method for making the same |
| US20150007364A1 (en) * | 2012-02-06 | 2015-01-01 | A.B. Seeds Ltd. a corporation | Isolated polynucleotides expressing or modulating micrornas or targets of same, transgenic plants comprising same and uses thereof |
| CN109161553A (en) * | 2018-09-29 | 2019-01-08 | 安徽农业大学 | A kind of pears transcription factor PbBP and its application |
| CN109679965A (en) * | 2018-09-28 | 2019-04-26 | 中国林业科学研究院林业研究所 | A kind of poplar adjusted and controlled blade profile development gene and its application |
-
2019
- 2019-09-10 CN CN201910855290.0A patent/CN110527688A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003000898A1 (en) * | 2001-06-22 | 2003-01-03 | Syngenta Participations Ag | Plant genes involved in defense against pathogens |
| US20110061133A1 (en) * | 2007-09-14 | 2011-03-10 | Basf Plant Science Gmbh | Plants having increased yield-related traits and a method for making the same |
| US20150007364A1 (en) * | 2012-02-06 | 2015-01-01 | A.B. Seeds Ltd. a corporation | Isolated polynucleotides expressing or modulating micrornas or targets of same, transgenic plants comprising same and uses thereof |
| CN109679965A (en) * | 2018-09-28 | 2019-04-26 | 中国林业科学研究院林业研究所 | A kind of poplar adjusted and controlled blade profile development gene and its application |
| CN109161553A (en) * | 2018-09-29 | 2019-01-08 | 安徽农业大学 | A kind of pears transcription factor PbBP and its application |
Non-Patent Citations (5)
| Title |
|---|
| GROOVER,A.T.等: ""Populus alba x Populus tremula homeodomain protein ARBORKNOX1 (ARBORKNOX1) mRNA, complete cds"", 《GENBANK》 * |
| GROOVER,A.T.等: ""The Populus homeobox gene ARBORKNOX1 reveals overlapping mechanisms regulating the shoot apical meristem and the vascular cambium"", 《PLANT MOLECULAR BIOLOGY》 * |
| YE, QX等: ""Over-expression of transcription factor ARK1 gene leads to down-regulation of lignin synthesis related genes in hybrid poplar "717""", 《SCIENTIFIC REPORTS》 * |
| 张汉尧等: ""滇杨抗蛀干害虫转基因表达载体的构建"", 《西南林学院学报》 * |
| 赵岩秋等: ""杨树中 Ⅰ 类 KNOX 基因结构, 表达与功能分析"", 《林业科学研究》 * |
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