Application of CaNAC100 gene in regulating carotenoid synthesis in plants

Abstract

The present invention discloses the use of the CaNAC100 gene in regulating carotenoid synthesis in plants, belonging to the field of plant genetic engineering technology. The nucleotide sequence of the CaNAC100 gene is shown in SEQ ID NO. 1; the protein sequence encoded by this gene is shown in SEQ ID NO. 2. The expression pattern of CaNAC100 is highly correlated with carotenoid synthesis in pepper fruit, and this gene can significantly promote plant carotenoid synthesis. Cloning this gene will not only facilitate the analysis of the regulatory mechanism of carotenoids in pepper fruit, but also help cultivate pepper varieties with higher carotenoid content and improve pepper fruit quality, thus possessing great application value.

Description

Technical Field

[0001] The present invention relates to the technical field of plant genetic engineering, and in particular to the application of the CaNAC100 gene in regulating the synthesis of plant carotenoids. Background Art

[0002] Peppers (Capsicum spp.) are a common spicy condiment and vegetable belonging to the Solanaceae family and an important agricultural and horticultural crop in my country. Fruit color, a key characteristic of peppers, has attracted attention from breeders and consumers. The pigments in pepper fruits primarily include chlorophyll, carotenoids, and anthocyanins. The presence and relative abundance of these pigments determine the fruit's color. Peppers of varying varieties and maturity exhibit a wide range of hues, from green, yellow, and orange to red and purple. This rich variety of colors offers diverse culinary and decorative applications.

[0003] Carotenoids are naturally occurring yellow, orange, and red lipophilic molecules found in the leaves, flowers, and fruits of higher plants. They belong to the terpenoid family of pigments. The concentration of carotenoids in pepper fruit ranges from 0.03 to 3 mg per 100 g fresh weight. These substances, primarily composed of β-carotene, capsanthin, lutein, and zeaxanthin, play a crucial role in plant physiology, development, ecology, and evolution. Carotenoids in pepper are synthesized into digeranyl geranyl diphosphate (GGPP) via the methylerythritol phosphate pathway (MEP). In the first step, phytoene synthase (PSY) condenses two GGPP molecules head-on to form phytoene; in the second step, dehydrogenase and isomerase form lycopene; in the third step, lycopene is cyclized to form α-carotene or β-carotene; in the fourth step, α-carotene and β-carotene are hydroxylated to form lutein and zeaxanthin, respectively; finally, under the catalysis of capsanthin / capsorubin synthase (CCS), 5,6-epoxyzeaxanthin and violaxanthin are converted into capsanthin and capsorubin, respectively. This step is a unique synthesis pathway in peppers.

[0004] The NAC family can participate in regulating the synthesis of plant carotenoids. However, there are few studies on the regulation of carotenoid synthesis in pepper fruit by NAC family transcription factors. Summary of the Invention

[0005] The present invention aims to provide a method for regulating carotenoid biosynthesis in plants using the CaNAC100 gene to address the aforementioned problems in the prior art. This method, through genetic verification, provides a target gene for breeding new pepper varieties and provides genetic material and theoretical basis for genetic improvement of pepper varieties.

[0006] To achieve the above object, the present invention provides the following solutions:

[0007] One of the technical solutions of the present invention is the use of the CaNAC100 gene in regulating the synthesis of carotenoids in plants. The nucleotide sequence of the CaNAC100 gene is shown in SEQ ID NO.1.

[0008] The second technical solution of the present invention is the use of a recombinant vector, expression cassette, transgenic cell line or recombinant bacteria containing the CaNAC100 gene in regulating carotenoid synthesis in pepper plants.

[0009] The third technical solution of the present invention is the use of the protein encoded by the CaNAC100 gene in regulating the synthesis of carotenoids in plants, and its amino acid sequence is shown in SEQ ID NO.2.

[0010] A fourth technical solution of the present invention is a method for improving the quality of pepper fruits, which promotes the synthesis of carotenoids in pepper fruits by regulating the expression level of the CaNAC100 gene in pepper plants.

[0011] The fifth technical solution of the present invention is the use of the CaNAC100 gene in cultivating transgenic pepper plants with high carotenoid content.

[0012] The sixth technical solution of the present invention is a method for cultivating transgenic pepper plants with high carotenoid content, comprising the following steps: introducing the CaNAC100 gene into the target plant.

[0013] Based on the above technical solution, the present invention has the following technical effects:

[0014] This study, published in Nature Communications, has cloned and validated CaCaNAC100, a key transcription factor that regulates carotenoid synthesis in pepper fruit. This transcription factor promotes the synthesis of carotenoid compounds in pepper fruit, impacting the development of pepper fruit quality. The study also provides recombinant plasmids, transgenic bacteria, and transgenic plants containing the CaCaNAC100 gene. This study enriches our understanding of pepper fruit secondary metabolism and the genetic mechanisms underlying fruit quality, particularly color development. This study provides a theoretical and practical basis for selective breeding of pepper fruit for agronomic traits. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0016] Figure 1This is a diagram showing the expression of CaNAC100 in different tissues of pepper fruit at different developmental stages and the expression differences of CaNAC100 and carotenoid synthesis genes.

[0017] Figure 2 This is a diagram of the subcellular localization analysis of the CaNAC100 gene in tobacco leaves.

[0018] Figure 3 This figure compares the effects of the CaNAC100 gene on the synthesis of carotenoid compounds in pepper fruit. Figure a shows fruits silenced by different silencing systems: CK represents uninoculated pepper fruit, pTRV2::00 represents fruit inoculated with the pTRV2 empty vector, and pTRV2::CaNAC100 represents CaNAC100-silenced fruit. Figure b shows the expression of CaNAC100 in CaNAC100-silenced fruit. Figure pTRV2::00 represents fruit inoculated with the pTRV2 empty vector, while pTRV2::CaNAC100 represents CaNAC100-silenced fruit. Figure c shows the expression of carotenoid biosynthesis-related genes in CaNAC100-silenced fruit tissues. Figure d shows the total carotenoid content. Figure pTRV2::00 represents fruit inoculated with the pTRV2 empty vector, while pTRV2::CaNAC100 represents CaNAC100-silenced fruit.

[0019] Figure 4 Figure 1 shows the accumulation of carotenoid compounds in pepper fruit regulated by the CaNAC100 gene. (a) GUS staining of fruits silenced by different overexpression systems. CK represents uninoculated pepper fruit tissue, 1301::00 represents the pCambia1301::00 empty vector-inoculated strain, and 1301::CaNAC100 represents CaNAC100-overexpressing fruit tissue. (b) CaNAC100 expression levels in CaNAC100-overexpressing fruit tissue. (Figure 1301::00 represents the pCambia1301::00 empty vector-inoculated strain, and 1301::CaNAC100 represents CaNAC100-overexpressing fruit tissue.) (c) Expression of carotenoid biosynthesis-related genes in CaNAC100-overexpressing fruit tissue. (d) Total carotenoid content. (Figure 1301::00 represents the pCambia1301::00 empty vector-inoculated strain, and 1301::CaNAC100 represents CaNAC100-overexpressing fruit tissue.) DETAILED DESCRIPTION

[0020] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0021] It should be understood that the terms described herein are intended only to describe particular embodiments and are not intended to limit the present invention. In addition, for numerical ranges herein, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Each smaller range between any intermediate value within a stated value or stated range and any other stated value or intermediate value within the stated range is also encompassed by the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded within the scope.

[0022] Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of any conflict with any incorporated document, the contents of this specification shall prevail.

[0023] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from the present invention. The present description and examples are intended to be illustrative only.

[0024] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0025] Unless otherwise specified, the technical solutions described in the present invention are all conventional solutions in the field, and the reagents or raw materials used, unless otherwise specified, are purchased from commercial channels or have been disclosed.

[0026] The embodiment of the present invention provides the use of the CaNAC100 gene in regulating carotenoid synthesis in plants. The nucleotide sequence of the CaNAC100 gene is shown in SEQ ID NO.1.

[0027] In some specific embodiments, overexpression of the CaNAC100 gene promotes the synthesis of carotenoids, while silencing or knocking out the CaNAC100 gene inhibits the synthesis of carotenoids.

[0028] In some specific embodiments, the plant comprises pepper.

[0029] The embodiments of the present invention also provide the use of a recombinant vector, an expression cassette, a transgenic cell line or a recombinant bacterium containing the CaNAC100 gene in regulating carotenoid synthesis in pepper plants.

[0030] The present invention also provides an embodiment of the invention that provides an application of a protein encoded by the CaNAC100 gene in regulating the synthesis of carotenoids in plants. The amino acid sequence of the protein is shown in SEQ ID NO.2.

[0031] In some specific embodiments, the plant comprises pepper.

[0032] The embodiment of the present invention also provides a method for improving the quality of pepper fruits, by regulating the expression level of the CaNAC100 gene in pepper plants to promote the synthesis of carotenoids in pepper fruits.

[0033] The embodiments of the present invention also provide the use of the CaNAC100 gene in cultivating transgenic pepper plants with high carotenoid content.

[0034] The embodiment of the present invention also provides a method for cultivating transgenic pepper plants with high carotenoid content, comprising the following steps: introducing the CaNAC100 gene into the target plant.

[0035] In some specific embodiments, the CaNAC100 gene is introduced into the target plant via the recombinant vector, expression cassette, transgenic cell line or recombinant bacteria.

[0036] Preferably, the carotenoids include β-carotene, zeaxanthin and capsanthin.

[0037] Example 1

[0038] 1. Cloning and sequence analysis of CaNAC100 gene

[0039] CaNAC100 gene, a NAC-type transcription factor gene in pepper, was cloned and sequenced.

[0040] Mature fruits of the Yunnan pepper variety Dianjiao No. 13 (registration number GPD(2022)530217) were grown in a greenhouse at the Institute of Sericulture and Beekeeping in Caoba Town, Mengzi City, Honghe Prefecture, Yunnan Province, and were used for RNA extraction. Total RNA was extracted using a Rapid Universal Plant RNA Extraction Kit (Beijing Huayueyang) according to the manufacturer's instructions, and RNA content and quality were determined using a spectrophotometer.

[0041] use Synthesize first-strand cDNA using the III 1st Strand cDNA Synthesis SuperMix Reverse Transcription Kit. After optimization, use an appropriate amount of the reverse transcription product for subsequent PCR. Amplify the CaNAC100 gene using conventional PCR using the first-strand cDNA as the RT-PCR template.

[0042] The upstream primer is: (5'-TCCTTAGAGCAATGGAAGAA-3');

[0043] Downstream primer: (5'-CCGAGTCAATCATTACAAGT-3').

[0044] 25μL PCR reaction system: MegaFi TM Fidelity 2X PCR Master Mix 12.5 μL, upstream and downstream primers 1 μL each, template 1 μL, ddH2O 9.5 μL.

[0045] The reaction program was as follows: 98°C for 2 min, 1 cycle, 98°C for 10 sec, 57°C for 30 sec, 72°C for 57 sec, 35 cycles.

[0046] The PCR product CaNAC100 gene was purified and recovered, and then ligated into the pClone007 Versatile SimpleVector vector (Qingke Biotechnology, China) to obtain the pMDTM19-T-CaNAC100 plasmid. The plasmid was transformed into Escherichia coli DH5a competent cells and sent to Shanghai Bioengineering Co., Ltd. for sequencing. The nucleotide sequence of the obtained CaNAC100 gene is shown in the sequence listing as SEQ ID NO. 1:

[0047] SEQ ID NO.1:ATGGAAGAAAATCTCCCTCCAGGGTTTAGGTTCCATCCAAGT。

[0048] The amino acid sequence encoded by the CaNAC100 gene is shown in SEQ ID NO.2:

[0049] SEQ ID NO.2:MEENLPPGRFHPSDEELITYYLSNKVSDFNFTCRAIADVDLNK SEPWDLPAKASMGEKEWYFFSQKDRKYPTGLRTNRATEAGYWKTTGKDKEVYRGGVHLVGMKKTLVFYRGRAPKGEKTNWVMHEYRLETNLGFKPSKEEWVVCRVFHKNSTAKKPQPTSSSQQSDHEDQSTCDTYHH TLPNEFGDMDLTNNFNLATPIGNGSLTNISLHNYSNIQENMNIMNLAALPSVNSLLFRALQLRGNFNNQPRDDIIPATTITSTSNYPFNIQPQGTISAFGVDFDPVLEPPSSSSHVVFNPLQPQGEQSYKLDSNIWE.

[0050] 2 Expression analysis of CaNAC100 gene in different tissues of pepper fruit at different parts and developmental stages

[0051] The improved Yunnan pepper variety Dianjiao No. 13 (registration number GPD(2022)530217) was grown in the greenhouse of the Sericulture and Bee Research Institute in Caoba Town, Mengzi City, Honghe Prefecture, Yunnan Province. Mature fruits were used for RNA extraction. Total RNA was extracted from three different tissues at six different stages of pepper fruit development. Flowers were used as controls. qRT-PCR was used to determine the relative expression levels of the CaNAC100 gene in different parts (seeds, placenta, and pericarp) at different developmental stages.

[0052] Figure 1 The following is a graph showing the expression differences of CaNAC100 in different tissues of pepper fruit at different developmental stages. Figure 1As shown, CaNAC100 is expressed in the seeds, placenta, and pericarp of pepper fruit, but expression levels vary significantly. Ten days after anthesis, CaNAC100 expression in the placenta was significantly higher than in the pericarp and seeds (P < 0.05). After 20 and 50 days of anthesis, expression levels in the placenta and seeds were significantly higher than in the pericarp (P < 0.05). After 30 and 60 days of anthesis, expression levels in the seeds were significantly higher than in the placenta and pericarp (P < 0.05). However, in samples 40 days after anthesis, expression levels in the pericarp were significantly higher than in the seeds and placenta (P < 0.05). Overall, CaNAC100 expression in pepper reached its peak 20 days after anthesis in the placenta and seeds, while peak expression in the pericarp occurred 30 days after anthesis.

[0053] 3. Subcellular localization of CaNAC100 gene

[0054] The PC1300S-EGFP plant transient fluorescence expression vector was treated with KpnI and BamHI for linearization, and CaNAC100 ORF cloning primers F and R (see Table 1) with PC1300S-EGFP vector homology arms were designed to amplify the CaNAC100 gene CDS region as an insert. TM HD Cloning kit was used to recombine the insert fragment and linearized vector into pCaNAC100-GFP recombinant vector according to the instructions.

[0055] Table 1

[0056]

[0057]

[0058] The pCaNAC100-GFP recombinant vector and the empty vector PC1300S-EGFP were transformed into Escherichia coli. After plating on resistance plates, positive bacteria were selected and cultured overnight at 37°C for plasmid extraction. pCaNAC100-GFP and the empty vector PC1300S-EGFP were co-transformed with a nuclear marker and injected into tobacco leaves. The leaves were cultured under low light for 8-10 hours, and the subcellular localization of CaNAC100 was observed under a laser confocal microscope.

[0059] Figure 2 The figure shows the subcellular localization analysis of CaNAC100 gene in tobacco leaves after injection. Figure 2 As shown, CaNAC100 has an obvious transcription factor nuclear localization signal in Arabidopsis protoplasts, which indicates that CaNAC100 may regulate the transcriptional expression of related genes in the cell nucleus.

[0060] Functional verification of the CaNAC100 gene in pepper fruit

[0061] (1) VIGS silencing of the CaNAC100 gene in pepper fruit

[0062] Based on the SGN VIGS Tool online gene silencing tool (https: / / vigs.solgenomics.net / ), the pepper genome database was selected to screen for silenced fragments. The purified amplification product of pepper CaRAP2-12 was used as a template, and the amplification primers were as follows:

[0063] Upstream primer: 5′-TAAGGTTACCGAATTCTTACTCCCAAATATTAGAGTC-3′;

[0064] Downstream primer: 5'-GCTCGGTACCGGATCCCAAGAGAACATGAATATCAT-3').

[0065] The insert fragment was cloned using primers CaNAC100-VIGS, and the procedure and system were the same as before (annealing temperature was 65°C, extension time was 10 sec). TM The purified CaNAC100 gene silencing fragment was recombined into the pTRV2 plasmid using the HD Cloning kit, and the recombinant plasmid was transformed into DH5α Escherichia coli. Finally, the positive colonies were screened and the plasmid was extracted and transformed into GV3101 Agrobacterium.

[0066] Prepare a solution containing 10mM MES, 10mM MgCl2, and 150μM AS, adjust the pH to 5.6, and use it as the infection solution after sterilization. Use centrifugation at 4000rpm / min for 10min to collect Agrobacterium, discard the supernatant, and resuspend it in the prepared infection solution. Use the infection solution as a control, and finally adjust the OD value of each strain suspension. 600nm The resuspension was incubated in a shaker at 28°C in the dark for 2-3 hours. A 1:1 ratio of GV3101 pTRV1 and pTRV2::CaNAC100 and pTRV2::00 inoculum was prepared. 0.5 ml of the inoculum was injected into the flesh of pepper fruits using a 1 ml sterile needleless syringe. The inoculated pepper fruits were incubated at 18°C ​​in a dark environment with 60% relative humidity for 48 hours, then placed in an incubator at 18°C ​​with a 16-hour light / 8-hour dark cycle. After 15 days of incubation, the silenced pepper fruits were sampled, and successful silencing was verified using qRT-PCR.

[0067] Table 1 qRT-PCR primers

[0068]

[0069]

[0070] The results show that ( Figure 3 ) After inoculation, pepper fruits injected with Agrobacterium containing the pTRV2::CaNAC100 plasmid showed obvious green color near the injection hole compared with the control. Using qRT-PCR to screen CaNAC100 silenced fruits, the CaNAC100 gene was downregulated ( Figure 3 b). Determination of carotenoid content in silent positive fruit tissues ( Figure 3 (d). The results showed that the total carotene content in the silenced samples was significantly reduced compared to pTRV2::00. When the CaNAC100 gene was silenced, the expression levels of CaZISO, CaZDS, CaCRTZISO02, CaLCYB, CaCHYB, and CaCRTY genes were significantly upregulated in pepper fruits compared to pTRV2::00 (P<0.05), while the expression levels of other carotenoid biosynthesis structural genes were significantly downregulated (P<0.05) ( Figure 3 (c)

[0071] (2) Overexpression of the CaNAC100 gene in pepper fruit

[0072] Using the In-Fusion Cloning Primer Design Tool, seamless cloning primers with 15 bp pCambia1301 plasmid homology arms were designed for both ends of the CaNAC100 gene ORF:

[0073] Upstream primer: 5′-GGACTCTTGACCATGGCCCTTCCGAGTCAATCATTACA-3′;

[0074] Downstream primer: 5'-CTCAGATCTACCATGGCCCTTAAATAATCTCCTTAGAGCAA-3'.

[0075] The CaNAC100 gene sequence was cloned and purified using the template to obtain the insert. The CaNAC100 gene insert was ligated to the NcoI site of the pCambia1301 vector using a homologous recombination kit to construct an overexpression vector. TMThe purified CaNAC100 gene overexpression fragment was recombined into the pCambia1301 plasmid using the HD Cloning Kit. The recombinant plasmid was then transformed into DH5α Escherichia coli. Positive colonies were screened, and the plasmid was extracted and transformed into GV3101 Agrobacterium tumefaciens. The inoculum was prepared using the same method as the silencing method, and injected into pepper fruits. After 10 days, the fruits were sampled, and positive samples for successful overexpression were verified using qRT-PCR.

[0076] The CaNAC100 gene was overexpressed in pepper detached fruits. GUS staining was performed on tissues near the injection hole 10 days after inoculation. The results showed that ( Figure 4 (a) Pepper fruit tissue injected with Agrobacterium containing pCambia1301::00 and pCambia1301::CaNAC100 plasmids showed a distinct blue color after staining compared to control fruit. qRT-PCR screening of CaNAC100-overexpressing fruit tissue revealed significantly higher CaNAC100 gene expression in overexpressing fruit tissue than in pCambia1301::00 fruit tissue, indicating successful CaNAC100 gene overexpression ( Figure 4 In the transient overexpression samples, the total carotenoid content of pepper fruit increased significantly (P<0.05) ( Figure 4 (d) Compared with the control, the expression levels of carotenoid biosynthesis structural genes, except for the CaLCYE gene, were upregulated to varying degrees, and the differences were significant (P<0.05).

[0077] In summary, the CaNAC100 protein and its encoding gene are related to the regulation of the synthesis and metabolism of carotenoid compounds in pepper fruit, and can significantly improve the quality formation of pepper fruit. The CaNAC100 gene can be used to regulate the synthesis of carotenoids in pepper fruit and the formation of pepper fruit quality.

[0078] The expression pattern of CaNAC100 is highly correlated with carotenoid synthesis in pepper fruit. Transient overexpression of this gene in pepper fruit promoted carotenoid synthesis and accumulation, significantly increasing the expression of carotenoid-related genes. Inhibiting CaNAC100 expression through VIGS significantly reduced the content of carotenoid-related compounds and the expression of carotenoid-related genes in pepper fruit. Cloning this gene will not only facilitate the deciphering of the regulatory mechanisms of carotenoid production in pepper fruit but also aid in the development of pepper varieties with higher carotenoid content and improved fruit quality, thus possessing significant application value.

[0079] This invention, for the first time, cloned and verified the key transcription factor CaNAC100, which regulates carotenoid synthesis in pepper fruit. This transcription factor promotes the synthesis of carotenoid compounds in pepper fruit, influencing the development of pepper fruit quality. The invention also provides a recombinant plasmid, transgenic bacteria, and transgenic fruit containing the CaNAC100 gene. This invention enriches our understanding of pepper fruit secondary metabolism and the genetic mechanisms underlying pepper quality development. It also provides a theoretical and practical basis for selective breeding of pepper varieties for agronomic traits.

[0080] Obviously, the above embodiments of the present invention are merely examples for the purpose of clearly illustrating the present invention and are not intended to limit the implementation methods of the present invention. A person skilled in the art will be able to make other variations or modifications based on the above description. It is not necessary and impossible to enumerate all implementation methods here. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention shall be included within the scope of protection of the claims of the present invention.

Claims

1. Application of the CaNAC100 gene in regulating carotenoid synthesis in pepper plants, characterized in that: The nucleotide sequence of the CaNAC100 gene is shown in SEQ ID NO.

1.

2. The use according to claim 1, characterized in that Overexpression of the CaNAC100 gene promoted the synthesis of carotenoids in pepper plants, while silencing the CaNAC100 gene inhibited the synthesis of carotenoids in pepper plants.

3. Use of a recombinant vector, expression cassette or recombinant bacterium containing the CaNAC100 gene in regulating carotenoid synthesis in pepper plants, characterized in that: The nucleotide sequence of the gene is shown in SEQ ID NO.

1.

4. Use of a protein encoded by the CaNAC100 gene in regulating carotenoid synthesis in pepper plants, characterized in that: Its amino acid sequence is shown in SEQ ID NO.

2.

5. A method for improving the quality of pepper fruit, characterized in that: By increasing the expression level of the CaNAC100 gene in pepper plants, the synthesis of carotenoids in pepper fruits is promoted. The nucleotide sequence of the CaNAC100 gene is shown in SEQ ID NO.

1.

6. The use of the CaNAC100 gene in cultivating transgenic pepper plants with high carotenoid content, characterized in that: The nucleotide sequence of the CaNAC100 gene is shown in SEQ ID NO.

1.

7. A method for cultivating transgenic pepper plants with high carotenoid content, characterized in that: The method comprises the following steps: introducing the CaNAC100 gene according to claim 1 into a target plant.

8. The method according to claim 7, characterized in that The CaNAC100 gene according to claim 1 is introduced into the target plant via the recombinant vector, expression cassette or recombinant bacteria according to claim 3.

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