Sisal aux / iaa gene and protein in plants

Abstract

The application belongs to the field of agricultural biotechnology, and particularly relates to a sisal Aux / IAA gene and application of the protein in plants. The sisal Aux / IAA gene provided by the application comprises a first SNP site or a second SNP site. Research finds that, Aux / IAA the gene can increase the number of plant vascular tissues, construct a plant model related to vascular tissues, and cultivate a plant with multiple vascular tissues; and the first SNP site or the second SNP site located on the gene is extremely significantly related to the number of vascular bundles of sisal leaves, and can be used for screening and cultivating a plant with multiple vascular tissues, and provides a theoretical basis and gene resources for genetic breeding.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of agricultural biotechnology, and particularly relates to a Aux / IAA gene and its protein in plants. BACKGROUND

[0002] Aux / IAA As one of the key gene families of auxin signaling pathway, the Aux / IAA gene family encodes proteins that specifically bind to ARF (Auxin response factor) and thus participate in the regulation of auxin signaling pathway. Whole genome analysis of the Aux / IAA gene family has been reported in more than 30 plants. Aux / IAA With the increasing number of published plant genomes year by year, it has become possible to study the evolutionary mechanism of the Aux / IAA gene family at the whole genome level. Aux / IAA With the increasing amount of biological information year by year, it is increasingly important to conduct in-depth molecular biology research on the Aux / IAA gene family and thus to analyze its regulation of plant development mechanism. Aux / IAA

[0003] The identification of the function of the Aux / IAA gene in Arabidopsis was carried out early, and the biological functions of multiple genes have been analyzed, such as the regulation of apical and root tropism development, hypocotyl elongation, leaf shape development, and lateral root development. However, most of the related research in Arabidopsis involves auxin-mediated developmental regulation, and research on specific agronomic traits is relatively less. In recent years, with the in-depth development of crop molecular biology research, Aux / IAA reports on the regulation of agronomic traits by the Aux / IAA gene have gradually increased. For example, Aux / IAA can improve drought tolerance and regulate tiller number in rice, OsIAA6 participate in the regulation of vascular development, IAA11 can improve plant type and yield in oilseed rape, corn, and peach, BnaA3.IAA7 the Aux / IAA gene participates in the regulation of pericarp development. In addition, research reports on eucalyptus and poplar have confirmed that Aux / IAA and PtoIAA9 play an important regulatory role in xylem development. EgrIAA4

[0004] Fibraurea efficiency (Fibraurea efficiency) is a hard leaf fiber crop and is widely planted in tropical and subtropical regions. It has a wide range of uses, mainly involving fisheries, navigation, aerospace, mining, transportation, oil fields, and textile industries. Fibraurea efficiency is a unique leaf fiber crop among hemp crops, which is different from seed fiber crops represented by cotton and bast fiber crops represented by ramie and flax. Fibraurea efficiency fiber is mainly developed from leaf vascular tissue, and the mechanism of fiber trait formation has not been clearly understood, so it is crucial to excavate functional genes that regulate vascular tissue traits for genetic improvement of fibraurea efficiency fiber traits. There is no report on the Aux / IAA gene family in fibraurea efficiency. Agave sisalana Perr. ex Engelm. Aux / IAA ​​​Research on gene regulation of vascular tissue traits and molecular markers. SUMMARY

[0005] The application aims to provide a sisal Aux / IAA Gene and its protein in plants, which can be used to increase the number of vascular tissues in plants, construct plant models related to vascular tissues, cultivate and screen plants with multiple vascular tissues.

[0006] The application provides a sisal Aux / IAA Gene, which comprises a first SNP site or a second SNP site. Aux / IAA

[0007] The nucleotide sequence of the sisal Aux / IAA Gene comprises any one of (a1)-(a3):

[0008] (a1). As shown in SEQ ID NO. 1;

[0009] (a2). The nucleotide sequence containing the first SNP site is shown in SEQ ID NO. 7;

[0010] (a3). The nucleotide sequence containing the second SNP site is shown in SEQ ID NO. 8.

[0011] Preferably, in the same plant germplasm, the number of leaf vascular bundles of the A genotype is significantly more than that of the G genotype;

[0012] The A genotype is that the nucleotide sequence of the sisal Aux / IAA Gene is shown in SEQ ID NO. 7 or SEQ ID NO. 8;

[0013] The G genotype is that the nucleotide sequence of the sisal Aux / IAA Gene is shown in SEQ ID NO. 1.

[0014] The application provides a sisal Aux / IAA Gene encoding a sisal Aux / IAA protein, and the amino acid sequence of the sisal Aux / IAA protein is shown in SEQ ID NO. 2.

[0015] The application provides a biological material, which comprises a recombinant expression vector or a recombinant microorganism containing a sisal Aux / IAA Gene.

[0016] The application provides a sisal Aux / IAA Gene, the sisal Aux / IAA protein or the biological material in plants.

[0017] ​Preferably, the plants include one or more of Agave sisalana, Zea mays, Oryza sativa, Nicotiana tabacum and Arabidopsis thaliana.

[0018] Preferably, the applications include one or more of increasing the number of vascular tissues of plants, constructing plant models related to vascular tissues, cultivating and screening plants with multiple vascular tissues.

[0019] Preferably, the vascular tissues of plants include vascular bundles in leaves and / or vascular columns in flower stalks.

[0020] The present application provides a method for cultivating plants with multiple vascular tissues, comprising the following steps:

[0021] Increasing the expression of the Agave sisalana Aux / IAA gene or increasing the content of Agave sisalana Aux / IAA protein in plants to obtain plants with multiple vascular tissues.

[0022] The present application provides a method for screening Agave sisalana with multiple vascular bundles, comprising the following steps:

[0023] Taking the DNA of the Agave sisalana to be tested as a template, sequentially performing PCR amplification and sequencing analysis;

[0024] When the nucleotide sequence of the PCR product is as shown in SEQ ID NO. 7 or SEQ ID NO. 8, the Agave sisalana to be tested is Agave sisalana with multiple vascular bundles.

[0025] When the nucleotide sequence of the PCR product is as shown in SEQ ID NO. 1, the plant to be tested is Agave sisalana with few vascular bundles.

[0026] Beneficial effects:

[0027] The present application provides an Agave sisalana Aux / IAA gene, the Agave sisalana Aux / IAA gene includes a first SNP site or a second SNP site; the nucleotide sequence of the Agave sisalana Aux / IAA gene includes any one of (a1) to (a3): (a1) as shown in SEQ ID NO. 1; (a2) the nucleotide sequence containing the first SNP site is as shown in SEQ ID NO. 7; (a3) the nucleotide sequence containing the second SNP site is as shown in SEQ ID NO. 8. It is found through the research of the present application that the Agave sisalana Aux / IAA gene can increase the number of vascular tissues of plants, construct plant models related to vascular tissues, cultivate plants with multiple vascular tissues; the first SNP site or the second SNP site located on the gene is extremely significantly related to the number of vascular bundles in the leaves of Agave sisalana, and can be used for screening plants with multiple vascular tissues. Therefore, the technical scheme provided by the present application can be applied to the genetic breeding of plant vascular tissue traits, and provides a theoretical basis and gene resources for cultivating new varieties or seedlings of plants with increased number of vascular columns or vascular bundles. Attached Figure Description

[0028] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the embodiments will be briefly described below.

[0029] Figure 1 The sisal provided by the present invention Aux / IAA Gene agarose gel electrophoresis verification results;

[0030] Figure 2 The sisal germplasm resources provided for this invention Aux / IAA The polymorphism at position 456 of the gene is associated with the number of vascular bundles, where ** indicates p <0.01.

[0031] Figure 3 The sisal germplasm resources provided for this invention Aux / IAA The polymorphism at gene position 602 is associated with the number of vascular bundles, where ** indicates p <0.01.

[0032] Figure 4 Comparison of wild-type Arabidopsis vascular tissue (A) provided for this invention with sisal Aux / IAA gene overexpression in Arabidopsis (B). Detailed Implementation

[0033] Unless otherwise specified, the reagents and methods used in this invention are all conventional reagents and methods.

[0034] This invention provides a sisal Aux / IAA The gene, obtained from the sisal transcriptome database, is from sisal. Aux / IAAThe nucleotide sequence of the gene is shown in SEQ ID NO. 1, and is as follows: 5'-ATGGGTGATGCATGTCCAAAGTTGCTTGATTTGATGGTCGCAAAGGAGAGAGATTGGATGGTGAGGGACTCAGGAGGAGGGAGTGGGCTTGGTGTTCAAGAGGAGAAGAAGCTGGAGCTGAAGCTGGGCCCACCAGGAGCTGAGGACTGGGGATCATCAATGGAGAGGAAAGCAGAAGACCCTTCTGCTCTCTCTCTTGGTTACTTCCCTAAAGCCTCTAAAACCACCAAAAGGGGATTTTTAGACACAATTACCTCAAAAAATGAAGGGTTTCAGCAGCAACAGCCTCCTCAGAGCCTTGAGAGGAAGGCATGTTGCCCACCACCATCACATGCTGCTGCTGTCTCTGTTAATGCAGCTAGAATCAATAGCAACAGCTCTCACTCTCAGCCAAGAACTGCTTCTGTTCCAGTAGTTGGGTGGCCTCCCATCCGATCTTTTAGGAAGAATCTCGCGAGCAGCTCCTCCAAGCTGCCACTGGTTGTATCACAGAATGGGGGATCTGAGACTGGAGCAAAGCCTCATATCTGCAAGAAAGGTCTTTTTGTGAAAATTAACATGGATGGGATCCCTATTGGAAGGAAAGTGGACCTCAAGGCCTGTGACAGCTATGAGAAGCTCTCTTCTGCTGTGGGAGAGCTCTTTCAAGGTCTTCTTGCAGCTCAAAAGGATCCAGCTGCTGCTGGAGTTCAGGCCACTGCAGAACAGAAAAAGGCATTTACAGGCTTATTGGATGGGACTGGTGAATACACTTTGGTTTATGACGACAATGAAGGAGACAGGATGCTAGTTGGGGATGTCCCATGGGAAATGTTTGTTTCAACTGCCAAGAGATTGAGGGTGCTGAAGAGCTCTGAACTCTCGACGCTAGCTTTGGGAGCAGTGAGCAGGAAGAGAACAGCAACCGAGTGCCGAGTGGAGTGA-3'.

[0035] The SNP site in the nucleotide sequence represented by SEQ ID NO. 1 in the application; the SNP site includes: a first SNP site or a second SNP site; the first SNP site is located at the 456th site from the 5' end of the nucleotide sequence represented by SEQ ID NO. 1, and G or A mutation exists at the site; when the first SNP site is A, it is recorded as A genotype, and the nucleotide sequence is as shown in SEQ ID NO.7, specifically as follows: 5'-ATGGGTGATGCATGTCCAAAGTTGCTTGATTTGATGGTCGCAAAGGAGAGAGATTGGATGGTGAGGGACTCAGGAGGAGGGAGTGGGCTTGGTGTTCAAGAGGAGAAGAAGCTGGAGCTGAAGCTGGGCCCACCAGGAGCTGAGGACTGGGGATCATCAATGGAGAGGAAAGCAGAAGACCCTTCTGCTCTCTCTCTTGGTTACTTCCCTAAAGCCTCTAAAACCACCAAAAGGGGATTTTTAGACACAATTACCTCAAAAAATGAAGGGTTTCAGCAGCAACAGCCTCCTCAGAGCCTTGAGAGGAAGGCATGTTGCCCACCACCATCACATGCTGCTGCTGTCTCTGTTAATGCAGCTAGAATCAATAGCAACAGCTCTCACTCTCAGCCAAGAACTGCTTCTGTTCCAGTAGTTGGGTGGCCTCCCATCCGATCTTTTAGGAAGAATCTCGCAAGCAGCTCCTCCAAGCTGCCACTGGTTGTATCACAGAATGGGGGATCTGAGACTGGAGCAAAGCCTCATATCTGCAAGAAAGGTCTTTTTGTGAAAATTAACATGGATGGGATCCCTATTGGAAGGAAAGTGGACCTCAAGGCCTGTGACAGCTATGAGAAGCTCTCTTCTGCTGTGGGAGAGCTCTTTCAAGGTCTTCTTGCAGCTCAAAAGGATCCAGCTGCTGCTGGAGTTCAGGCCACTGCAGAACAGAAAAAGGCATTTACAGGCTTATTGGATGGGACTGGTGAATACACTTTGGTTTATGACGACAATGAAGGAGACAGGATGCTAGTTGGGGATGTCCCATGGGAAATGTTTGTTTCAACTGCCAAGAGATTGAGGGTGCTGAAGAGCTCTGAACTCTCGACGCTAGCTTTGGGAGCAGTGAGCAGGAAGAGAACAGCAACCGAGTGCCGAGTGGAGTGA-3'; when the first SNP site is G, it is recorded as a G genotype, and the nucleotide sequence is shown in SEQ ID NO. 1.

[0036] The second SNP site in the application is located at the 602th site from the 5' end of the nucleotide sequence shown in SEQ ID NO. 1, and G or A exists at the site; when the second SNP site is A, it is recorded as A genotype, and the nucleotide sequence is shown in SEQ ID NO.8, specifically as follows: 5'-ATGGGTGATGCATGTCCAAAGTTGCTTGATTTGATGGTCGCAAAGGAGAGAGATTGGATGGTGAGGGACTCAGGAGGAGGGAGTGGGCTTGGTGTTCAAGAGGAGAAGAAGCTGGAGCTGAAGCTGGGCCCACCAGGAGCTGAGGACTGGGGATCATCAATGGAGAGGAAAGCAGAAGACCCTTCTGCTCTCTCTCTTGGTTACTTCCCTAAAGCCTCTAAAACCACCAAAAGGGGATTTTTAGACACAATTACCTCAAAAAATGAAGGGTTTCAGCAGCAACAGCCTCCTCAGAGCCTTGAGAGGAAGGCATGTTGCCCACCACCATCACATGCTGCTGCTGTCTCTGTTAATGCAGCTAGAATCAATAGCAACAGCTCTCACTCTCAGCCAAGAACTGCTTCTGTTCCAGTAGTTGGGTGGCCTCCCATCCGATCTTTTAGGAAGAATCTCGCGAGCAGCTCCTCCAAGCTGCCACTGGTTGTATCACAGAATGGGGGATCTGAGACTGGAGCAAAGCCTCATATCTGCAAGAAAGGTCTTTTTGTGAAAATTAACATGGATGGGATCCCTATTGGAAGGAAAGTGGACCTCAAGGCCTATGACAGCTATGAGAAGCTCTCTTCTGCTGTGGGAGAGCTCTTTCAAGGTCTTCTTGCAGCTCAAAAGGATCCAGCTGCTGCTGGAGTTCAGGCCACTGCAGAACAGAAAAAGGCATTTACAGGCTTATTGGATGGGACTGGTGAATACACTTTGGTTTATGACGACAATGAAGGAGACAGGATGCTAGTTGGGGATGTCCCATGGGAAATGTTTGTTTCAACTGCCAAGAGATTGAGGGTGCTGAAGAGCTCTGAACTCTCGACGCTAGCTTTGGGAGCAGTGAGCAGGAAGAGAACAGCAACCGAGTGCCGAGTGGAGTGA-3'; when the second SNP site is G, it is recorded as a G genotype, and its nucleotide sequence is as shown in SEQ ID NO.1. The SNP site is significantly related to the number of vascular bundles in the leaf of the sisal hemp plant, that is, the number of vascular bundles in the leaf of the sisal hemp seedling with the A genotype is significantly more than that of the sisal hemp seedling with the G genotype.

[0037] The application provides a sisal hemp plant Aux / IAA The sisal hemp Aux / IAA protein encoded by the sisal hemp gene has an amino acid sequence as shown in SEQ ID NO. 2, and specifically as follows: MGDACPKLLDLMVAKERDWMVRDSGGGSGLGVQEEKKLELKLGPPGAEDWGSSMERKAEDPSALSLGYFPKASKTTKRGFLDTITSKNEGFQQQQPPQSLERKACCPPPSHAAAVSVNAARINSNSSHSQPRTASVPVVGWPPIRSFRKNLASSSSKLPLVVSQNGGSETGAKPHICKKGLFVKINMDGIPIGRKVDLKACDSYEKLSSAVGELFQGLLAAQKDPAAAGVQATAEQKKAFTGLLDGTGEYTLVYDDNEGDRMLVGDVPWEMFVSTAKRLRVLKSSELSTLALGAVSRKRTATECRVE*. The SNP site is a synonymous mutation, and thus the nucleotide sequences as shown in SEQ ID NO. 1, SEQ ID NO. 7 and SEQ ID NO. 8 can all encode the sisal hemp Aux / IAA protein with the amino acid sequence as shown in SEQ ID NO. 2. The sisal hemp Aux / IAA gene, the sisal hemp Aux / IAA protein and the vascular tissue trait of the plant are related, and thus the above gene or protein can be used to increase the number of vascular tissues of the plant, construct a plant model related to the vascular tissue, and cultivate and screen a plant with multiple vascular tissues.

[0038] Example 1 Cloning of a sisal hemp Aux / IAA gene

[0039] The primer sequence is as follows:

[0040] The forward primer is 5'-ATGGGTGATGCATGTCCAA-3' (SEQ ID NO. 3);

[0041] The reverse primer is 5'-TCACTCCACTCGGCACTCGGT-3' (SEQ ID NO. 4).

[0042] The PCR amplification reaction system (20 μL) is specifically as follows:

[0043] 1 μL (10 pmol) of forward primer, 1 μL (10 pmol) of reverse primer, 10 μL of EasyTaq® PCR SuperMix, 1 μL of sisal cDNA (template) and 7 μL of ddH2O.

[0044] The PCR amplification reaction program was as follows: denaturation at 95℃ for 5 min, followed by 30 cycles of amplification at 95℃ for 15 s, 60℃ for 15 s, and 72℃ for 15 s, and then 72℃ for 2 min.

[0045] The amplified PCR products were subjected to Sanger sequencing, and the electrophoresis results are as follows: Figure 1 As shown (in) Figure 1 In the image, the left band represents the electrophoresis result of the PCR product, and the right band represents the marker. Sanger sequencing results show that the sequencing results of the PCR product are consistent with the sequence described in SEQ ID NO.1.

[0046] Example 2: Sisal Aux / IAA Validation of the correlation between gene SNP marker sites and seedling vascular bundle number

[0047] Primers were designed using the nucleotide sequence shown in SEQ ID NO.1 as a template, and the genome sequences of different sisal germplasm resources (all of which were derived from the Institute of Environment and Plant Protection, Chinese Academy of Tropical Agricultural Sciences) were amplified by PCR to analyze the sequence shown in SEQ ID NO.1. Aux / IAA Genetic diversity, primer sequences are as follows:

[0048] Forward primer: 5'-CCTCCCATCCGATCTTTTAGGAA-3' (SEQ ID NO.5);

[0049] Reverse primer: 5'-GCAAGAAGACCTTGAAAGAGC-3' (SEQ ID NO.6).

[0050] The PCR reaction system and procedure were the same as those in Example 1. The template for the PCR reaction was DNA from different sisal germplasm resources.

[0051] Genetic diversity analysis of the Aux / IAA genome sequences amplified from different sisal germplasm seedlings revealed that variations exist at positions 456 or 602, starting from the 5' end of the nucleotide sequence described in SEQ ID NO.1, with both mutation sites being G or A. Codon analysis showed that both sites were synonymous mutations.

[0052] The leaves of different sisal germplasm resource seedlings were made into sections and the number of vascular bundles was counted under a microscope, and correlation analysis was performed with the two SNP loci, and the results are shown in Table 2 Figure 2 and Figure 3 .

[0053] The results show that: the number of vascular bundles of the sisal germplasm resource seedlings with A genotype at the 456th locus is higher than that with G genotype, and there is a very significant difference between the two (P<0.01) Figure 2 ); the number of vascular bundles of the sisal germplasm resource seedlings with A genotype at the 602nd locus is higher than that with G genotype, and there is a very significant difference between the two (P<0.01) Figure 3 ).

[0054] The above results show that: Aux / IAA The two SNP loci of the gene are extremely significantly correlated with the number of vascular bundles of sisal seedlings, and can be used to identify the number of vascular bundles of sisal germplasm resources or breeding material seedlings, and have application value in the genetic breeding of sisal vascular bundle traits, and provide a theoretical basis for the cultivation of new sisal varieties.

[0055] Example 3 Sisal Aux / IAA Application of the gene in increasing the number of plant vascular bundles

[0056] The sisal Aux / IAA gene cloned in Example 1 was introduced into a pEGAD expression vector, and the expression vector was introduced into Agrobacterium GV3101 strain by electroporation method. The Agrobacterium containing the expression vector was cultured in LB liquid medium overnight (28°C), and cultured to the logarithmic phase (OD600 value was 0.8-1.0). The bacteria were collected by centrifugation at 4000 gp / min for 5 min at room temperature, resuspended in resuspension medium to OD600 value of 0.6, and 100 μmol / L of acetyl-syringone (AS) was added. Arabidopsis was transformed by dipping method. The fruit pods of the Arabidopsis plants that had already flowered were removed, and the flowers were immersed in the bacterial solution (60s) and kept moist overnight. One week later, the transformation was repeated.

[0057] The seeds collected from the transformed plants were disinfected with sodium hypochlorite and then inoculated on MS medium containing 50 mg / L kanamycin. After incubation at 22°C±2°C for one week, normal germination positive plants were selected and planted in soil. The T1 generation seeds were collected, planted, and the T1 generation plant leaves were cut for transgenic plant PCR detection to obtain transgenic positive plants. The transgenic positive plants were cultured and managed and T2 generation seeds were collected. Transgenic T2 generation seeds and wild type Arabidopsis seeds were planted in soil and incubated under the same conditions until flowering. The T2 plants and wild type plants were cut to prepare paraffin sections for histomorphological observation, and the results are shown in Table 3 Figure 4 ).

[0058] In combination with Figure 4The results show that the number of vascular columns of the transgenic Arabidopsis thaliana plants is increased by 33.33% compared with the wild type, indicating that the Agrobacterium tumefaciens HPT gene can regulate the development of vascular tissue. Aux / IAA The results show that the number of vascular columns of the transgenic Arabidopsis thaliana plants is increased by 33.33% compared with the wild type, indicating that the Agrobacterium tumefaciens HPT gene can regulate the development of vascular tissue. Aux / IAA The Agrobacterium tumefaciens HPT gene can regulate the development of vascular tissue, and can be applied to the genetic breeding of vascular tissue traits to cultivate new plant varieties or seedlings with increased number of vascular columns.

[0059] Although the above embodiment has made a detailed description of the present application, it is only a part of the embodiments of the present application, but not all the embodiments, and other embodiments can be obtained according to the present embodiment without creativity, which all belong to the protection scope of the present application.

Claims

1. Fibre hemp Aux / IAA Gene, characterized in that, The said plantain Aux / IAA the nucleotide sequence of the gene is any one of (a1) - (a3): (a1). As shown in SEQ ID NO. 1; (a2). As shown in SEQ ID NO. 7; (a3). As shown in SEQ ID NO.

8.

2. The sword of claim 1 Aux / IAA a gene encoding a sword hemp Aux / IAA protein, the amino acid sequence of the sword hemp Aux / IAA protein being set forth in SEQ ID NO.

2.

3. A biomaterial, characterized by, The biomaterial includes: a recombinant expression vector or a recombinant microorganism containing a gene of sisal Aux / IAA ​ The said plantain Aux / IAA The nucleotide sequence of the gene is as set forth in SEQ ID NO. 1, SEQ ID NO. 7 or SEQ ID NO.

8.

4. The Manila hemp of claim 1 Aux / IAA use of the gene, the Aux / IAA protein of claim 2 or the biological material of claim 3 in a plant. The application is: increasing the expression amount of the Agave sisalana L. Aux / IAA gene in the plant or increasing the content of the Agave sisalana L. Aux / IAA protein in the plant to increase the number of vascular tissues of the plant, or constructing a plant model related to the number of vascular tissues. The plant is: sisal or Arabidopsis.

5. Use according to claim 4, characterized in that, The plant vascular tissue includes: vascular bundle in leaf and / or vascular column in floral stalk.

6. A method of growing a multi-dimensional vascular tissue plant, comprising: The method comprises the following steps: Increasing sisal in plants Aux / IAA expression of the gene, or increasing the content of Aux / IAA protein in plants, to obtain plants with more vascular tissues The plant is: sisal or Arabidopsis. The said plantain Aux / IAA The nucleotide sequence of the gene is as set forth in SEQ ID NO. 1, SEQ ID NO. 7 or SEQ ID NO.

8.

7. A method of selecting multi-dimensional bundle raphia, characterized by, The method comprises the following steps: Taking the DNA of the sisal to be tested as a template, sequentially performing PCR amplification and sequencing analysis; When the nucleotide sequence of the PCR product is as shown in SEQ ID NO. 7 or SEQ ID NO. 8, the sisal to be tested is multi-vascular bundle sisal; When the nucleotide sequence of the PCR product is as shown in SEQ ID NO. 1, the plant to be tested is few-vascular bundle sisal; The primer for PCR amplification is: forward primer with the nucleotide sequence as shown in SEQ ID NO. 5 and reverse primer with the nucleotide sequence as shown in SEQ ID NO. 6.

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