KR20160032990A - Method for improving fast growth or biomass increase of plant using CBR1 in plants - Google Patents

Abstract

The present invention relates to a method for promoting plant growth and an increase in an amount of biomass. More specifically, the present invention relates to a method for promoting plant growth or an increase in an amount of biomass by over-expressing cytochrome b5 reductase1 (CBR1). According to the present invention, it has been found that effects of promoting growth and resistance to cold weather or salt stress are improved in a seed or a plant in which CBR1 is over-expressed. Thus, it is anticipated that the method can be useful for improvement of crops of which human can harness.

Description

[0001] The present invention relates to a method for promoting plant growth or biomass growth using CBR1,

The present invention relates to a method for promoting plant growth or biomass increase, and more particularly, to a method for promoting plant growth or biomass increase by regulating the expression of CBR1 (Cytochrome b5 reductase 1) and plant thereof.

As the era of high oil prices has arrived, the importance of biomass as a raw material for generating bioenergy has been emphasized. Production of first-generation biofuels using the saccharification process of starch present in grains has resulted in a rise in prices of crops, food and economic difficulties, and the development of related technologies has been delayed. In contrast, second-generation bioenergy research using wood-based cellulose, which is present in non-edible biomass crops such as Butchers, Switchgrass, and Poplar, is actively under way. However, in order to produce a substantial amount of energy that can replace practically fossil fuels, basically efforts will be needed to increase and sustain the production of biomass in plants.

On the other hand, cold weather and salt pollution are the main factors that reduce plant growth and crop productivity, with 1.4 billion hectares of farmland suffering from colds in the world, 45 million hectares of farmland damaged by salt damage, Is expected to be larger. In addition, Korea depends heavily on imports of major food crops such as corn, wheat, and soybeans from the US and China, and imports of agricultural products are several times higher than exports.

Thus, the development of plant genetic engineering provides crops or plants with improved economic, agricultural and horticultural traits, and the research and development of transgenic plants with increased growth and biomass is a way to reduce the supply of food resources . Therefore, it is possible to produce various transgenic plants with increased growth and biomass using genes derived from various plants (corn, soybean, pepper, Arabidopsis, tobacco or rice) or microorganisms (Synechocystis, Pseudomonas, Bacillus or Anabaena etc.) (Korean Patent No. 10-0814941), but it is still not enough.

DISCLOSURE OF THE INVENTION The present invention has been made in order to solve the above-mentioned problems, and the present inventors confirmed the effect of promoting growth, cold stress resistance and salt stress resistance of CBR1 over-expressing seeds or plants, .

Accordingly, an object of the present invention is to provide a method for producing a plant cell, which comprises transforming a plant cell with a recombinant vector comprising CBR1 (Cytochrome b5 reductase1) gene; And over-expressing CBR1 in the transformed plant. The present invention also provides a method for promoting plant growth or biomass growth.

It is another object of the present invention to provide a method for producing a plant cell, which comprises transforming a plant cell with a recombinant vector comprising the CBR1 gene; And regenerating the plant from the transgenic plant cell. The present invention also provides a method for producing a transgenic plant having increased growth or biomass.

It is still another object of the present invention to provide a plant in which an increase in growth or biomass produced by the above method is promoted.

However, the technical problem to be solved by the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.

In order to accomplish the above object, the present invention provides a method for producing a plant cell, comprising the steps of: transforming a plant cell with a recombinant vector comprising CBR1 (Cytochrome b5 reductase1) gene; And over-expressing CBR1 in the transformed plant. The present invention also provides a method for promoting plant growth or biomass increase.

In one embodiment of the present invention, the CBR1 gene may be composed of the nucleotide sequence of SEQ ID NO: 1.

The present invention relates to a method for transforming a plant cell with a recombinant vector comprising the CBR1 gene; And regenerating the plant from the transformed plant cell. The present invention also provides a method for producing a transgenic plant,

The present invention provides a plant in which an increase in growth or biomass produced by the above method is promoted.

In an embodiment of the present invention, the plant may be one in which the content of TAG (triacylglycerol) or minerals is increased.

In another embodiment of the present invention, the plant may be one which is improved in cold weather or salt stress resistance.

The present invention provides a method for screening a substance that promotes growth or biomass increase, comprising treating a candidate substance and then measuring CBR1 overexpression.

The method of promoting plant growth or biomass increase according to the present invention includes the step of overexpressing CBR1. As described above, by overexpressing CBR1 as described above, it has been confirmed that the effect of promoting plant growth, cold damage or stress resistance is improved , And CBR1 expression can be used to improve the crops available to human beings.

1 is a schematic diagram of a DNA construct for inducing overexpression of CBR1.
2 shows the appearance of the CBR1 overexpressed seed (CBR1 OX) and the wild type control (WT) under the electron microscope (A), (B) size, and (C) weight.
FIG. 3 shows electron microscopic observations of the inside of CBR1 over-expressing seeds and wild type control group.
FIG. 4 is a graph showing the amount (nmol) of TAG per seed (A) of CBR1 overexpressed seed and wild type control, (B) the content (nmol) of each fatty acid species contained in TAG per seed, and (C) the amount of triacylglycerol ) As a relative value (%).
FIG. 5 shows the contents of iron, manganese, phosphorus, and calcium in the CBR1 over-expressing seed and the wild type control group.
Fig. 6 shows the germination state (A) germination state and the germination rate (B) for 5 days of the CBR1 overexpressed and wild-type control groups.
FIG. 7 shows the results of visual observation of (A) growth state and (B) fresh weight of CBR1 overexpressing plants and wild type control at 23 ° C.
FIG. 8 shows the results of visual observation of (A) growth state and (B) fresh weight of CBR1 overexpressing plant and wild type control at 6 ° C.
FIG. 9 shows the survival rates of CBR1-overexpressing plants and wild-type control by temperature changes (0 ° C., -5 ° C., -7 ° C., and -9 ° C.).
Fig. 10 shows the results of visual observation of growth of (a) wild-type and (b) survival rate of CBR1 overexpressed plants and wild-type control by NaCl treatment.

The present inventors confirmed the effect of increasing weight, size, TGA (triacylglycerol), mineral content, and germination rate in seeds overexpressing CBR1 (Cytochrome b5 reductase1) gene. In addition, the present inventors completed the present invention on the basis of confirming the effects of promoting growth, cold stress resistance, and high salt stress resistance in a plant overexpressing the CBR1 gene.

Hereinafter, the present invention will be described in detail.

The present invention relates to a method for producing a plant cell, which comprises transforming a plant cell with a recombinant vector comprising CBR1 (Cytochrome b5 reductase1) gene; And over-expressing CBR1 in the transformed plant. The present invention also provides a method for promoting plant growth or biomass increase.

In the present invention, CTV1 (Cytochrome b5 reductase 1) overexpressing in the present invention has a strong reducing ability to transfer electrons of NADH to cytochrome b5 by using Flavin adenine dinucleotide (FAD) group in the electron transport system, thereby converting methemoglobin into hemoglobin in the blood In mice, myristoylation is known to regulate the migration of mitochondrial membrane and endoplasmic reticulum membrane. In the present invention, the relationship between the expression of CBR1 gene and plant growth, cold weather or salt stress has been clarified. Thus, the present invention is characterized by promoting plant growth or biomass growth by overexpressing CBR1. In addition, the CBR1 gene of the present invention preferably comprises the nucleotide sequence of SEQ ID NO: 1, and the CBR1 protein preferably comprises the amino acid sequence of SEQ ID NO: 2, but is not limited thereto.

As used herein, the term "growth" refers to the quantitative increase of various organs in plants. In particular, in the case of plant cells, the cell size only increases without cell division, And the cell becomes larger. The term "biomass " as used in the present invention refers collectively to crops, wastes, woods, organisms and the like which can be converted into energy or produce energy through a specific process. In accordance with the method of the present invention, Thereby promoting plant growth or biomass growth and producing crops with increased yields.

In one embodiment of the present invention, an increase in the size, weight, TAG, mineral content (see Examples 2 to 5) and germination rate of CBR1 over-expressing seeds were confirmed by preparing CBR1 overexpressing plants (see Example 1) See Example 6). In addition, it was confirmed that plants resistant to cold or salt stress of CBR1 overexpressing plants are excellent (see Examples 7 and 8), and plants capable of promoting growth or biomass increase can be produced.

Accordingly, the present invention provides a method for producing a plant cell, which comprises transforming a plant cell with a recombinant vector comprising the CBR1 gene; And regenerating the plant from the transgenic plant cell. The present invention also provides a method for producing a transgenic plant,

Further, in another aspect of the present invention, there is provided a plant in which an increase in growth or biomass produced by the above method is promoted.

The amount of triacylglycerol, iron (Fe), manganese (Mn), phosphorus (P) and calcium (Ca) is increased in the CBR1 overexpressing seeds or plants according to the present invention and the resistance to cold weather or salt stress is improved , Biomass, and improved crops that are promoted to grow.

The plant according to the present invention is a food crop including rice, wheat, barley, corn, soybean, potato, red bean, oats, sorghum; Vegetable crops including Arabidopsis, cabbage, radish, red pepper, strawberry, tomato, watermelon, cucumber, cabbage, melon, squash, onions, onions and carrots; Special crops including ginseng, tobacco, cotton, sesame seed, sugar cane, beet, perilla, peanut, and rapeseed; Fruit trees including apple trees, pears, jujube trees, peaches, sheep grapes, grapes, citrus fruits, persimmons, plums, apricots, and bananas; Roses, gladiolus, gerberas, carnations, chrysanthemums, lilies, tulips; And feed crops including ragras, red clover, orchardgrass, alpha-alpha, tall fescue, perenniallaigrus, and the like, preferably Arabidopsis , but are not limited thereto.

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the following examples.

Example  One. CBR1  Manufacture of overexpressed plants

To investigate the function of CBR1 (Cytochrome b5 reductase1), CBR1 overexpressed plants were prepared. First, CBR1 DNA was obtained by PCR in Arabidopsis cDNA library (leaf tissue) using primer Forward 5-CCGCTCGAGATGGATACCGAGTTTCTCCGA-3 (SEQ ID NO: 4) and primer Reverse 5-GGGGTACCGAACTGGAATTGCATCTCC-3 (SEQ ID NO: 5). After treatment with XhoI and KpnI restriction enzymes, ligation was performed with a pUC-GFP vector containing CaMV 35S promoter, sGFP, and Nos terminator treated with XhoI and BamHI. For subcloning with a binary vector, CBR1: sGFP DNA obtained by XhoI and EcoRI treatment was ligated with a restriction vector pBIB121 (hygromycin) binary vector. The DNA construct thus obtained was introduced into Agrobacterium tumefaciens by electroporation. Arabidopsis transformants were prepared by the vacuum-infiltration method and transformants were identified through Gamborg B5 medium (Duchefa; G0210.0050) containing 50 mg L1 hygromycin. Then, the homo-line was selected in the fourth-generation switchgear and used for the experiment.

Example  2. CBR1  Phenotypic analysis of overexpressed seeds

In order to confirm phenotypic changes due to overexpression of CBR1, the size and weight of wild type control and CBR1 overexpressed seeds were compared.

The size of the seeds was measured by measuring the lengths of the horizontal and vertical three sets of 14 seeds. Photographs were taken on an OLYMPUS SZ61 microscope and the length was measured using the iSolution Lite program. One seed weighed three sets of 500 seeds, and weighed the two weights by dividing the average of the weights by 500.

As shown in Fig. 2, the CBR1 overexpressed seed (CBR1 OX) showed a chunky morphology compared to the wild type control (WT). In addition, quantitative measurements confirmed that the size and weight of CBR1 overexpressed seeds increased by 9.5% and 17.5%, respectively, compared with the wild type control.

Example  3. CBR1  Electron microscopic analysis of overexpressed seeds

To investigate the reason for the increase in weight of CBR1 overgrown seed, the inside of the seed was examined under a microscope.

First, the seeds were cut in half with a razor blade to ensure a good pre-treatment. The seeds were fixed at 4 ° C for 20 hours using 2% paraformaldehyde, 2% glutaraldehyde and 0.05M sodium cacodylate buffer (pH 7.2). The sample was washed three times with 0.05M sodium cacodylate buffer (pH 7.2), and the second fixation was performed at 4 ° C for 2 hours using 1% osmium tetroxide and 0.05M sodium cacodylate buffer (pH 7.2). Washed twice with water, and stained with 0.5% uranyl acetate at 4 ° C for 30 minutes. 30, 50, 70, 80, 90, 100, 100, 100% ethanol for 6 hours each, and propylene oxide and Spurrs resin mixture were treated for 5 days. The sample was polymerized at 70 ° C for 24 hours and sectioned using Ultramicrotome (MT-X, RMC, Tucson, AZ, USA). 2% uranylacetate was stained for 7 minutes, Reynolds lead citrate was stained for 2 minutes, and the sample was observed using a TEM (JEM-1011, JEOL, Tokyo, Japan).

As shown in FIG. 3, it was confirmed that CBR1 overexpressed seeds had a form in which the oil body was filled more densely than the control group, thereby reducing the pore size in the cells.

Example  4. CBR1  Overexpressed seed TAG  Extraction and identification

The amount of TAG of the CBR1 over-expressing seeds was measured in consideration of the fact that the electron microscopic morphological characteristics of the CBR1 overexpressed seeds of Example 3 were also observed in the overexpressed oleosin and AtABCA9 transporter which increase the TAG reported above.

500 seeds were placed in a glass tube with 2 mL of isopropanol and boiled at 80 ° C for 10 minutes. After cooling with ice, 2 mL of chloroform was added and the seeds were ground with a Polytron homogenizer (Hitachi Koki). TAG spots were separated on a silica gel TLC plate and were subjected to transmethylation in methanol containing 2.5% sulfuric acid (3 mL) at 90 ° C for 45 min. The fatty acid composition of the whole TAG was analyzed by GC / MS (SHIMADZU GC-2010, HP-INOWAX capillary column, 30m, 0.25mm).

As shown in FIG. 4, the amount of TAG of the CBR1 over-expressing seeds was increased by 19.4% as compared with that of the control. In order to further elucidate the increase in the amount of TAG, it was confirmed that the amount of unsaturated fatty acids of 18: 1, 18: 2 and 18: 3 in CBR1 overexpressed material was significantly increased by checking the kinds of fatty acids of lipid.

Example  5. CBR1  Overexpressed seed Minute amount  Content analysis

To determine the mineral content of CBR1 overexpressed seeds, 50 seeds were dissolved in 60% nitric acid at 120 ° C for 6 hours. The amount of each mineral was identified through inductively coupled plasma spectrometry (ICP) (iCAP6300 DUO ICO-OES; Thermo Electron Corporation).

5, it was confirmed that iron (Fe), manganese (Mn), phosphorus (P) and calcium (Ca) were increased by 19.9%, 54.6%, 29.7% and 23.6% in the CBR1 overexpressed seeds, respectively. This can be an application point for promoting human health by increasing mineral content of seeds.

Example  6. CBR1  Analysis of germination rate of transgenic transgenic plants

To determine the germination rate of CBR1 over-expressing seeds, wild-type Arabidopsis ( Arabidopsis thaliana : (Colombia ecotype)) and CBR1-overgrown seeds were germinated in 1/2 MS medium supplemented with 1% sucrose in a chamber set at 40% humidity, 16 hours day / 8 hour night cycle (long day condition) .

As shown in FIG. 6, it was confirmed that the germination rate of the CBR1 over-expressing seed was increased by 12.5% as compared with the control. This means that since the plant uses TAG as an energy source at the time of germination, an increase in the amount of TAG of the overexpressed CBR1 can increase the germination rate.

Example  7. CBR1  Cold stress stress resistance of overexpressed plants

In order to investigate the enhancement of CBR1 overexpression resistance to cold stress, the growth state, fresh weight, and survival rate at subzero temperature of CBR1 overexpressed plant (OX1, OX2) Respectively.

First, for low-temperature treatment, the cells were grown in 1/2 MS, 1% sucrose medium for 2 weeks and then transferred to soil and observed at 6 ° C for 2 weeks. Observations at subzero temperatures were followed by cold treatment at 4 ° C for 2 days to reduce the soil temperature after the rock treatment, and helped the soil temperature to be lowered by placing the used pots on ice chips. Thereafter, the temperature was lowered by 1 [deg.] C for two hours to reach the desired subzero temperature. The water was evenly sprayed at the desired subzero temperature to cool the upper part of the plant evenly and to obtain a frost effect. After maintaining the state for 2 hours in this state, the temperature was restored by raising the temperature by 1 ° C in one hour to 6 ° C again. After one day incubation at 6 ° C, the cells were transferred to 22.5 ° C, long-day conditions, and the survival rate of the plants after 5 days was recorded for one week.

As shown in FIGS. 7 to 8, the CBR1 overexpressed plants were larger than the control group at normal temperature (23 ° C) and showed a fresh weight increase of 12.8%, and grown at normal growth conditions (23 ° C) When treated, the fresh weight was increased by 45% compared to the control.

As shown in FIG. 9, at -5 ° C, over-expressing plants of CBR1 showed a similar survival rate of 89%, whereas at lower temperatures of -7 ° C and -9 ° C, 22% and 11% Respectively. These results indicate that CBR1 overexpression affects not only the seed but also the plant growth, and that overexpression of CBR1 can induce resistance to low temperature and freezing stress.

Example  8. CBR1  Cold-stress Stress Resistance of Over-expressing Transgenic Plants

CBR1 overexpression plants (OX1, OX2) and wild-type controls were grown in normal growth inducing soil (14days) to gradually increase the salinity of the soil After exposure to high concentrations of salt (300 mM NaCl, 15 days), the degree of yellowing and the survival rate were determined

As shown in Fig. 10, overexpressed plants of CBR1 showed a low yellowing phenomenon and 22% higher survival rate compared to the control group. It was confirmed that overexpression of CBR1 could induce increased resistance to salt stress.

It will be understood by those skilled in the art that the foregoing description of the present invention is for illustrative purposes only and that those of ordinary skill in the art can readily understand that various changes and modifications may be made without departing from the spirit or essential characteristics of the present invention. will be. It is therefore to be understood that the above-described embodiments are illustrative in all aspects and not restrictive.

<110> POSTECH ACADEMY-INDUSTRY FOUNDATION <120> Method for improving fast growth or biomass increase of plant          using CBR1 in plants <130> P2014204-01-KR-PB14-12199 <160> 5 <170> Kopatentin 2.0 <210> 1 <211> 834 <212> DNA <213> Artificial Sequence <220> <223> Cytochrome b5 reductase DNA sequence <400> 1 atggataccg agtttctccg aaccctagat cgtcagattc ttttgggtgt cttcgttgct 60 ttcgtcgccg ttggtgctgg tgctgcttat tttcttacat cctccaagaa acgcagagtg 120 tgtttggtcc gagaatttca aggagttcaa gcttgttaag agacatcagc ttagtcacaa 180 tgtggccaag ttcgtttttg aactcccaac ttctacttct gtgttgggtc ttcccattgg 240 acaacacatc agtgcaggga aaggatggtc aaggagagga tgttattaag ccatacaccc 300 cgactacgtt agactctgac gttggacgtt tcgaacttgt cattaagatg tatccgcaag 360 gacggatgtc tcatcattta gggaatgcgt gttggagacc atcttgccgt aaagggacca 420 aagggtaggt tcaagtatca accaggtcag tttagggcat ttggaatgct tgctggaggt 480 tcaggcatca ctcccatgtt ccaagggcca ggcaattcta gaaaacccaa cagacaagac 540 aaaggtgcat ctcatttacg ccaacgtcac atacgacgac attctcttga aggaagaatt 600 ggagggtctt actaccaatt accctgaaca attaaaattt ctatgttttg aaccagcctc 660 cggaagtatg ggatggtggt gttggatttg tatcaaagga aatgattcag actcattgcc 720 ctgcacctgc atctgatatt cagatcctaa gatgcggcca ccgccatgaa caaggccatg 780 gctgcaaacc ttgaagctct gggatactct ccggagatgc aattccagtt ctga 834 <210> 2 <211> 279 <212> PRT <213> Artificial Sequence <220> <223> Cytochrome b5 reductase protein sequence <400> 2 Met Asp Thr Glu Phe Leu Arg Thr Leu Asp Arg Gln Ile Leu Leu Gly   1 5 10 15 Val Phe Val Ala Phe Val Ala Val Gly Ala Gly Ala Ala Tyr Phe Leu              20 25 30 Thr Ser Ser Lys Lys Arg Arg Val Cys Leu Asp Pro Glu Asn Phe Lys          35 40 45 Glu Phe Lys Leu Val Lys Arg His Gln Leu Ser His Asn Val Ala Lys      50 55 60 Phe Val Phe Glu Leu Pro Thr Ser Thr Ser Val Leu Gly Leu Pro Ile  65 70 75 80 Gly Gln His Ile Ser Cys Arg Gly Lys Asp Gly Gln Gly Glu Asp Val                  85 90 95 Ile Lys Pro Tyr Thr Pro Thr Thr Leu Asp Ser Asp Val Gly Arg Phe             100 105 110 Glu Leu Val Ile Lys Met Tyr Pro Gln Gly Arg Met Met Ser His His Glu         115 120 125 Met Arg Val Gly Asp His Leu Ala Val Lys Gly Pro Lys Gly Arg Phe     130 135 140 Lys Tyr Gln Pro Gly Gln Phe Arg Ala Phe Gly Met Leu Ala Gly Gly 145 150 155 160 Ser Gly Ile Thr Pro Met Phe Gln Val Ala Arg Ala Ile Leu Glu Asn                 165 170 175 Pro Thr Asp Lys Thr Lys Val His Leu Ile Tyr Ala Asn Val Thr Tyr             180 185 190 Asp Asp Ile Leu Leu Lys Glu Glu Leu Glu Gly Leu Thr Thr Asn Tyr         195 200 205 Pro Glu Gln Phe Lys Ile Phe Tyr Val Leu Asn Gln Pro Pro Glu Val     210 215 220 Trp Asp Gly Gly Val Gly Phe Val Ser Lys Glu Met Ile Gln Thr His 225 230 235 240 Cys Pro Ala Pro Ala Ser Asp Ile Gln Ile Leu Arg Cys Gly Pro Pro                 245 250 255 Pro Met Asn Lys Ala Met Ala Ala Asn Leu Glu Ala Leu Gly Tyr Ser             260 265 270 Pro Glu Met Gln Phe Gln Phe         275 <210> 3 <211> 2692 <212> DNA <213> Artificial Sequence <220> &Lt; 223 > CaMV 35S promoter: CBR1: sGFP: NOS terminator DNA sequence <400> 3 agattagcct tttcaatttc agaaagaatg ctaacccaca gatggttaga gaggcttacg 60 cagcaggtct catcaagacg atctacccga gcaataatct ccaggaaatc aaataccttc 120 ccaagaatta aagatgcagt caaaagattc aggactaact gcatcaagaa cacagagaaa 180 gatatatttc tcaagatcag aagtactatt ccagtatgga cgattcaagg cttgcttcac 240 aaaccaaggc aagatagaga ttggagtctc taaaaaggta gttcccactg aatcaaaggc 300 catggagtca aagattcaaa tagaggacct aacagaactc gccgtaaaga ctggcgaaca 360 gttcatacag agtctcttag atcaatgaca agaagaaaat cttcgtcaac atggtggagc 420 acgacacact tgtctactcc aaaaatatca aagatacagt ctcagaagac caaagggcaa 480 ttgagacttt tcaacaaagg gtaattccga aacctcctcg gattccattg cccagctatc 540 tgtcacttta ttgtgaagat agtggaaaag gaaggtggct cctacaaatg ccatcattgc 600 gataaaggaa aggccatcgt tgaagatgcc ttgccacagt ggtcccaaag atggaccccc 660 acccacgagg agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga 720 ttgatgtgat atctccactg acgtaaggga tgacgcaaat ccactatcct tcgcaagacc 780 cttcctctat ataaggaagt tcatttcatt tggagagaac acgggggact ctagaggatc 840 tcgagatgga taccgagttt ctccgaaccc tagatcgtca gatcttttgg gtgtcttcgt 900 tgctttcgtc gccgttggtg ctggtgctgc ttattttctt acatcctcca agaaacgcag 960 agtgtgtttg gaccagagaa tttcaaggag ttcaagcttg ttaagagact cagcttagtc 1020 acaatgtggc caagttcgtt tttgaactcc caacttctac ttctgtgttg ggtcttccca 1080 ttggacaaca catcagttga ggggaaagga tggtcaagga gaggatgtta ttaagcatac 1140 accccgacta cgttagactc tgacgttgga cgtttcgaac ttgtcattaa gatgtatccg 1200 caaggacgga tgtctcatca tttcaggaga tgcgtgttgg agaccatctt gccgtaaagg 1260 gccaaagggt aggttcaagt atcaaccagg tcagtttagg gcatttggaa tgcttgctgg 1320 aggttcaggc atcactccca tgttccaagt ggcagagcaa ttctagaaaa cccaacagac 1380 aagacaaggt gcatctcatt tacgccaacg tcacatacga cgacattctc ttgaaggaag 1440 aattggaggg tcttactacc aattaccctg aacaatttaa atcttctatg ttttgaacca 1500 gcctccggaa gtagggatgg tggtgttgga tttgtatcaa aggaaatgat tcagactcat 1560 tgccctgcac ctgcatctga tattcagatc ctaagatgcg gaccacccca atgaacaagg 1620 ccatggctgc aaaccttgag ctctgggata ctctccggag atgcaattcc agttcggatc 1680 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga 1740 cggcgacgta aacggccaca agttcacgtg tccggcgagg gcgagggcga tgccacctac 1800 ggcaagctac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc 1860 tcgtgaccac cttcacctac ggcgtgcagt gctcagccgc taccccgacc acatgaagca 1920 gcacgacttc ttcaatccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc 1980 aaggacgacg gcaactacaa gacccgcgcc gaggtgaatt cgagggcgac accctggtga 2040 accgcatcga gctgaagggc atgacttcaa ggaggacggc aacatcctgg ggcacaagct 2100 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacagcagaa gaacggcatc 2160 aaggtgaact tcaagatccg ccacaacatg aggacggcag cgtgcagctc gccgaccact 2220 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac actacctgag 2280 cacccagtcc gccctgagca aagaccccaa cgagaacgcg atcacatggt cctgctggag 2340 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggcgcc 2400 cggctgcaga tcgttcaaac atttggcaat aaagtttctt aagattgaat cctgttgccg 2460 gtcttgcgat gattatcata taatttctgt tgaattacgt taagcatgta ataattaaca 2520 tgtatgcatg acgttattat gagatgggtt tttatgatta gagtcccgca attatacatt 2580 taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc gcgcgcggtg 2640 tcatctatgt actagatccg atgtaagctg tcaaacatga actagtgaat tc 2692 <210> 4 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> CBR1 primer Forward <400> 4 ccgctcgaga tggataccga gtttctccga 30 <210> 5 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> CBR1 primer reverse <400> 5 ggggtaccga actggaattg catctcc 27

Claims (6)

A method for promoting plant growth or an increase in biomass comprising the steps of:
(a) transforming a plant cell with a recombinant vector comprising CBR1 (Cytochrome b5 reductase1) gene; And
(b) overexpressing CBR1 in the transformed plant.
The method according to claim 1,
Wherein the CBR1 gene comprises the nucleotide sequence of SEQ ID NO: 1.
A method for producing a transgenic plant which promotes an increase in growth or biomass comprising the steps of:
(a) transforming a plant cell with a recombinant vector comprising the CBR1 gene; And
(b) regenerating the plant from the transformed plant cell.
A plant produced by the method of claim 3, wherein the increase of growth or biomass is promoted.
5. The method of claim 4,
Wherein said plant is characterized by increased TAG (triacylglycerol) or mineral content.
5. The method of claim 4,
Wherein the plant is improved in cold-tolerance or salt-stress resistance.

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