Application of PH8 protein or coding gene thereof in regulating plant height, ear length or lodging resistance of plant

Abstract

The invention relates to the technical field of plant molecular breeding, in particular to application of PH8 protein or a coding gene thereof in regulation of plant height, ear length or lodging resistance of plants. The PH8 protein is cytochrome P450 CYP78A15, and the plant is an oryza plant. The PH8 protein comprises an amino acid sequence as shown in SEQ ID NO.1. The application comprises the following steps: reducing the expression level of the PH8 protein in the plant, reducing the plant height and spike length of the plant, and improving the lodging resistance of the plant. The PH8 protein related to the plant height of the oryza plant is obtained through screening, and the plant height and the ear length of the oryza plant can be regulated and controlled by regulating and controlling the expression level of the PH8 protein in the oryza plant. The PH8 protein provided by the invention can be used for cultivating oryza plants with strong lodging resistance, does not influence the yield of the oryza plants, and has important application value.

Description

Technical Field

[0001] The present invention relates to the technical field of plant molecular breeding, and in particular to the application of PH8 protein or its encoding gene in regulating plant height, ear length or lodging resistance of plants. Background Art

[0002] The key to the green revolution in agriculture is the cultivation and promotion of semi-dwarf crop varieties. It was later discovered that mutations in genes related to gibberellin (GA) metabolism and signaling pathways in these varieties led to reduced plant height, increased harvest index, enhanced lodging resistance and nitrogen fertilizer tolerance, and ultimately a significant increase in crop yield per unit area. semi-dwarf 1 ( sd1 ) is a key gene of the rice green revolution, and is currently carried in almost all major rice varieties. sd1 Non-functional or weakly functional alleles. However, over-reliance on this single gene for semi-dwarf rice breeding often reduces the genetic diversity of cultivated rice, aggravates the trend of homogenization of rice varieties, and reduces the ability of rice to resist the risk of pests and diseases. Although a large number of semi-dwarf genes have been cloned in rice, most of these genes reduce stalk length while accompanied by severe yield losses. Therefore, further discovery of new and excellent rice semi-dwarf genes is of great significance to rice breeding and improvement.

[0003] Rice plant height is regulated by a variety of hormones, which interact with each other to varying degrees, forming a complex plant height regulation network. GA is one of the important hormones that regulate rice plant height. Blockage of the biosynthesis or signal transduction of active GA will lead to a decrease in rice plant height. sd1 The functional mutation of the GA20 oxidase it encodes leads to a reduction in the biosynthesis of active GA, which ultimately causes rice to become semi-dwarf. Other hormones such as brassinosteroid (BR), strigolactone (SL), cytokinin (CK), and indole-3-acetic acid (IAA) also play an important role in regulating rice plant height, but very few of these hormone biosynthesis or signal transduction pathway-related genes are widely used in rice semi-dwarf breeding. HTD1 / D17 Encoding carotenoid cleavage dioxygenase OsCCD7, which is involved in the first step of SL biosynthesis, partial functional loss of this gene can increase the number of tillers and improve rice yield; some studies have found that HTD1 The haplotype of Huanghua is sd1 Co-selection is widely used in modern rice variety improvement.

[0004] The premise of crop molecular breeding is to discover and clone excellent gene resources related to the traits to be improved, especially excellent alleles obtained through map-based cloning, which have more direct application value. For rice, a large number of major effect genes related to its important agronomic traits have been discovered. However, some genes with rare allelic variations are still worth discovering and utilizing. Summary of the invention

[0005] In order to solve the problems existing in the prior art, the present invention provides the use of PH8 protein or its encoding gene in regulating the plant height, ear length or lodging resistance of plants.

[0006] The present invention uses a chromosome segment replacement line constructed by Haobuka (HBK) and Chuan 7 (C7) to identify a major QTL for plant height PH8 , through the NIL-F2 population PH8 The gene was localized to the 26.8 kb region of chromosome 8 and confirmed by transgenic knockout and genetic complementation experiments. PH8 Control rice plant height. PH8 The C7 allele type ( PH8 C7 ) Only in a few aus It is distributed in rice and is a rare allele. It can significantly reduce plant height in both indica and japonica rice backgrounds, and has important application value in conventional rice semi-dwarf breeding. PH8 C7 Available in SD1 The weak functional allele (GR haplotype) background can reduce rice plant height without sacrificing yield; in addition, PH8 It has no dominant effect, and its heterozygous type can appropriately reduce plant height, so it can be used to improve the plant height of hybrid rice.

[0007] In a first aspect, the present invention provides the use of PH8 protein, or a gene encoding it, or a biological material comprising a gene encoding it, in regulating plant height, ear length or lodging resistance of a plant; The PH8 protein is cytochrome P450 CYP78A15, which is included in the National Rice Data Center. The present invention further provides the use of the PH8 protein or its encoding gene as an inhibition target in cultivating dwarf plants, short panicle length plants or plants with strong lodging resistance.

[0008] Those skilled in the art know that, based on the PH8 protein or its encoding gene as the inhibition target, the expression of PH8 protein can be inhibited, or the transcription of its encoding gene can be inhibited by various existing technical means, such as using the CRISPR-Cas9 gene editing system to knock out its encoding gene as a target, so as to achieve the inhibition of PH8 protein expression.

[0009] The present invention further provides the use of PH8 protein, or a gene encoding it, or a biological material comprising a gene encoding it in improving lodging resistance-related germplasm resources of rice plants.

[0010] Furthermore, by reducing the expression level of the PH8 protein in the plant, the plant height and ear length of the plant are reduced, and the lodging resistance of the plant is improved; and / or, by introducing the nucleotide sequence shown in SEQ ID NO.3 into the plant, the plant height of the plant is reduced and the lodging resistance of the plant is improved.

[0011] Further, the expression level of the PH8 protein in the plant is regulated (e.g., reduced) by any of the following methods: Transgenic, hybrid, backcross, selfing or asexual reproduction; The transgenic method preferably includes one or more methods selected from the group consisting of Ti plasmid, plant virus vector, direct DNA transformation, microinjection, gene gun, electroporation or Agrobacterium-mediated.

[0012] Furthermore, the PH8 protein comprises any one of the following amino acid sequences: (1) the amino acid sequence shown in SEQ ID NO.1; (2) The amino acid sequence of the amino acid sequence shown in SEQ ID NO.1 obtained by replacing, inserting or deleting one or more amino acids, and having the same functional protein.

[0013] SEQ ID NO.1: .

[0014] Furthermore, the coding gene of the PH8 protein includes any one of the following nucleotide sequences: (1) the nucleotide sequence shown in SEQ ID NO.2; (2) A nucleotide sequence encoding a protein with the same function obtained by replacing, deleting or inserting one or more nucleotides of the nucleotide sequence shown in SEQ ID NO.2; (3) A nucleotide sequence that can hybridize with the nucleotide sequence shown in SEQ ID NO. 2 under stringent conditions.

[0015] SEQ ID NO.2:

[0016] Furthermore, the plant is a rice plant, preferably rice.

[0017] Furthermore, the biological material is an expression cassette, a vector or a transgenic cell.

[0018] The transgenic cells described in the present invention do not include transgenic cells that have the ability to independently develop into a complete individual, that is, do not include plant varieties.

[0019] In a second aspect, the present invention provides a nucleic acid comprising: a nucleotide sequence as shown in SEQ ID NO.3.

[0020] The present invention provides a PH8 Rare alleles PH8 C7 ,exist PH8 HBK allele type ( PH8 HBK ) Based on the G to A mutation at the splicing junction of the first exon and the first intron, PH8 The alternative splicing of mRNA ultimately leads to PH8 C7 The mRNA produced a new stop codon and caused the translated peptide chain to terminate prematurely. Finally, the nucleotide sequence shown in SEQ ID NO.3 was obtained.

[0021] SEQ ID NO.3:

[0022] In a third aspect, the present invention provides a method for reducing plant height, comprising: introducing the aforementioned nucleic acid into the plant; the plant is a rice plant.

[0023] In a fourth aspect, the present invention provides a KASP primer combination, comprising: Kasp-PH8-F1: 5'-TTAATTACTGGAGTACTGTACATAC-3', Kasp-PH8-F2: 5'-TTAATTACTGGAGTACTGTACATAT-3', Kasp-PH8-R: 5'-GACTTCACCGACGTCCTTCTCTCC-3'.

[0024] The present invention has the following beneficial effects: The present invention screens and obtains a PH8 protein related to plant height of rice plants and its encoding gene. By gene editing the PH8 protein in rice plants, the plant height and panicle traits of rice plants can be effectively regulated. The PH8 protein provided by the present invention can be used to cultivate rice varieties with strong lodging resistance and improve rice germplasm resources, and has important application value in the field of cultivating high-quality rice varieties. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0026] Figure 1 The C7 genotype provided in Example 1 of the present invention PH8 Comparison of chromosome segment replacement line CSSL82 and parent HBK in terms of plant height, yield per plant and lodging index; A shows the plant morphology of parents HBK and C7, and B shows the plant morphology of HBK and CSSL82; the scale of A and B is 20 cm; C is the identification of the donor segment introduced into CSSL82, PH8 Located at the terminal region of the long arm of chromosome 8; D is the comparison of plant height between HBK and CSSL82; E is the comparison of single plant yield between HBK and CSSL82; F is the comparison of lodging index between HBK and CSSL82, and the lodging index of CSSL82 is significantly lower than that of HBK; ” indicates significant difference. P ≤0.01, the same below.

[0027] Figure 2 Embodiment 2 of the present invention provides SD1 Separate groups and PH8 Analysis of genetic effects of plant height in segregating populations; A and B are SD1 Separate groups and PH8 The high frequency distribution histogram of the isolated population; C shows SD1 and PH8 Genetic effect of plant height at the locus, where "CC" represents the C7 homozygous genotype, "HH" represents the HBK homozygous genotype, "CH" represents the heterozygous genotype, "A" represents the additive effect, "D" represents the dominant effect, and |D / A| represents the dominance.

[0028] Figure 3 is the plant height gene provided in Example 3 of the present invention PH8 Schematic diagram of the positional cloning process; the black, white and gray columns represent the C7 homozygous genotype (CC), HBK homozygous genotype (HH) and heterozygous type (CH), respectively; the different letters on the bar graph of the progeny test represent P The difference was significant at the ≤0.05 level.

[0029] Figure 4 The parent C7 and HBK provided in Example 3 of the present invention PH8 Schematic diagram of the functional variation sites and transcription products of candidate genes.

[0030] Figure 5 Embodiment 4 of the present invention provides PH8 Verification of transgenic functional mutation of genes; the upper part of A shows the schematic diagram of the target location of gene editing, and the lower part shows the different homozygous mutation types obtained in ZH11 and HBK varieties using the gene editing system; B shows the PH8 Comparison of plant morphology (left) and plant height (right) between homozygous mutants and transgenic negative controls; C shows the HBK background PH8 Plant morphology (left) and panicle length morphology (right) of homozygous mutants and transgenic negative controls; D is under the HBK background PH8 Comparison of plant height (left) and ear length (right) between the homozygous mutant and the transgenic negative control; the scale bars in B and C are both 10 cm.

[0031] Figure 6 Embodiment 4 of the present invention provides PH8 Verification of transgenic gene function complementation; A shows HBK-PH8 C7 Complementary HBK alleles in the background PH8 The plant height phenotype of the family; B is the comparison of plant height between the positive family of transgenic complementation and the negative control; C shows HBK-PH8C7 Complementary HBK alleles in the background PH8 The ear length phenotype of the family; D is the comparison of ear length between the transgenic complementation positive family and the negative control; the scale bars in A and C are both 10 cm.

[0032] Figure 7 This is the haplotype analysis result provided in Example 5 of the present invention.

[0033] Figure 8 Embodiment 6 of the present invention provides PH8 Application of the C7 allele in reducing the high yield of indica rice 9311 and japonica rice Koshihikari strain; A shows the 9311 wild type and the 9311 background with PH8 C7 Near isogenic line (9311-PH8 C7 ) Plant height; B shows the Koshihikari (KOS) wild type and the PH8 introduced into the KOS background C7 Near-isogenic line (KOS-PH8 C7 ) Plant height; C and E are 9311 and 9311-PH8, respectively C7 Comparison of plant height and yield per plant; D and F are KOS and KOS-PH8 respectively. C7 Comparison of plant height and single plant yield; the scale bars in A and B are both 20 cm.

[0034] Fig. 9 Embodiment 7 of the present invention provides PH8 Application of C7 allele in reducing the height of hybrid rice plants; A shows the hybrid rice Liangyoupeijiu (LYP9) and Peiai 64S (PA64S) and 9311-PH8 C7 Comparison of plant height of hybrid rice (LYP9-M) in the paired group; B shows the plant height of hybrid rice Yangliangyou 6 (YLY6) and Guangzhan 63-4S (GZ63-4S) and 9311-PH8 C7 Comparison of plant heights of hybrid rice (YLY6-M) in pairs; the scale bars in A and B are both 20 cm. DETAILED DESCRIPTION

[0035] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be clearly and completely described below in conjunction with the drawings of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0036] The experimental methods involved in the following examples, unless otherwise mentioned, are all conventional methods in the art, for example, reference may be made to experimental manuals in the art, or the conditions recommended in the manufacturer's instructions.

[0037] Unless otherwise specified, the experimental materials and reagents involved in the following examples can be obtained from commercial sources, for example:

[0038] Example 1: Plant height phenotypes of chromosome segment replacement line CSSL82 and parent HBK The parents HBK and C7 used in this example are local variety resources collected by our team independently, of which HBK is a local temperate japonica rice variety in Yunnan Province, and C7 is an aus rice from the Ganges River Basin. HBK and C7 have great differences in plant type, mainly manifested in that HBK is upright and compact with few tillers, while C7 has a larger tiller angle and many tillers (see Figure 1 A in the figure). The present invention constructed a chromosome segment replacement line population (BC4F3 generation) with HBK as the recipient parent and C7 as the donor parent, and identified two major QTLs for plant height, one of which was located on chromosome 1 and was related to the green revolution gene sd1 Co-localization, another QTL is located at the end of the long arm of chromosome 8 and is named PH8 ; One family, CSSL82, carries the C7 genotype PH8 (See Figure 1 B and Figure 1 The present invention investigated the plant height, yield per plant and lodging index of CSSL82 and HBK. Compared with the parent HBK, the plant height and lodging index of CSSL82 were significantly reduced (see Figure 1 B, D and F in Figure 2), but the yield per plant did not decrease significantly (see Figure 1 E in ), indicating PH8 The C7 genotype has certain breeding value because it can reduce the number of culms without reducing yield in the HBK background.

[0039] Embodiment 2: PH8 and SD1 Analysis of genetic effects of plant height The present invention obtains the genotype of the chromosome segment replacement line BC4F2 generation single plant by identifying the genotype of the chromosome segment replacement line BC4F2 generation single plant. PH8 and SD1 The individual plants with heterozygous genotype at the locus were planted and their offspring (BC4F3) were obtained respectively. PH8 and SD1 The F2 segregation population with loci segregation was used to examine the plant height of individual plants in these two populations and analyze PH8 and SD1 Genetic effects of plant height.

[0040] Identification using molecular markers RM431 and InDel1-32N SD1 The genotypes of the individual plants in the segregation population were analyzed by making a frequency distribution histogram based on the genotypes and plant height phenotypes of the individual plants in the segregation population and analyzing the genetic effect of plant height. The results showed that SD1 The segregating population showed a typical 3:1 (tall plant: short plant) segregation ratio (see Figure 2 A in the figure), where the plant height of the C7 homozygous genotype individual plant is the same as that of the heterozygous genotype individual plant and both are tall plants; SD1 The additive effect and dominant effect of are 16.7cm and 15.2cm respectively, and the dominance is 0.91. SD1 Inherited in a dominant manner (see Figure 2 Similarly, molecular markers RM267 and RM477 were used to identify PH8 The genotypes of individual plants in the population were separated and their genetic effects on plant height were analyzed; the results showed that PH8 The plant height of C7 homozygous genotype in the segregating population was significantly lower than that of HBK homozygous genotype; PH8 The plant height of heterozygous plants was between the two homozygous genotypes and close to the mid-parent value; PH8 The additive effect and dominant effect of are -13.6cm and -1.0cm respectively, and the dominance is 0.07. PH8 Inherited in an additive manner (see Figure 2 B and C in ). The molecular marker primer sequences are shown in Table 1.

[0041] Example 3: Plant height gene PH8 Fine mapping and functional variation analysis of candidate genes (1) PH8 Fine positioning The present invention obtains the PH8 The seeds of the locus heterozygous single plant (BC4F4) were used to plant a segregating population of 2100 plants, and the PH8 The molecular markers were located in the P14-M15 interval, which is approximately 95.1 kb (see Figure 3 ). Further expansion of the planting separation population (BC4F5), through the screening of recombinant plants, identification of recombinant plant genotypes by encrypted molecular markers and progeny testing, will eventually PH8 Localized to the 26.8 kb interval (ID38-S40) (see Figure 3 ); This interval contains only one complete annotated gene, LOC_Os08g43390, encoding a P450 cytochrome oxidase; therefore, the present invention uses this gene as PH8 The molecular marker primer sequences are shown in Table 1.

[0042]

[0043] (2) PH8 Functional variation analysis of candidate genes To determine PH8The variation of candidate genes between parents. The present invention compared the sequence differences of LOC_Os08g43390 between parents by Sanger sequencing (sequencing primer sequences are shown in Table 1). The sequencing results showed that most of the sequence differences of the promoter of this gene between parents were the same as those between typical indica and japonica rice (Minghui 63 and Nipponbare); but it is worth noting that the base at the splicing site of the first exon and the first intron of LOC_Os08g43390 in parent C7 mutated from G to A, resulting in alternative splicing of the mRNA of LOC_Os08g43390 in parent C7, generating a new stop codon in advance, and ultimately resulting in truncation of the encoded protein (see Figure 4 ).

[0044] Example 4: Plant height gene PH8 Transgenic knockout and complementation validation To further verify whether LOC_Os08g43390 regulates plant height, the present invention used the CRISPR-Cas9 gene editing system to knock out the gene in the HBK and ZH11 (the genotype of LOC_Os08g43390 is the same as that of HBK) backgrounds, in order to obtain plants with functional mutations of the gene (see Figure 5 The sequences of the gene editing targets are shown in Table 1. The gene was knocked out using dual targets. The present invention obtained two homozygous mutants with different mutation types in the ZH11 background, wherein the mutation type of ZH11-CR1 was a 2 bp deletion in the first target and a 1 bp insertion in the second target; the mutation type of ZH11-CR2 was a 233 bp deletion between the first and second target (see Figure 5 Compared with the transgenic negative control, the plant heights of ZH11-CR1 and ZH11-CR2 were reduced by 10.1 cm and 8.4 cm, respectively (see Figure 5 In the HBK background, the present invention also obtained two homozygous mutants of different mutation types, in which HBK-CR1 lacked 2bp and 3bp at the first and second target sites, respectively, and HBK-CR2 lacked 237bp between the first and second target sites (see Figure 5 Compared with the negative control in HBK background, the mutant had significantly shorter plant height and panicle length (see Figure 5 C in the figure); the plant height of HBK-CR1 and HBK-CR2 decreased by 17.4 cm and 24.5 cm, respectively, and the ear length decreased by 2.8 cm and 1.7 cm, respectively (see Figure 5 These results confirm that LOC_Os08g43390 is PH8 .

[0045] Further, the present invention is in NIL-PH8 C7 HBK allele PH8(3kb promoter + 1.8kb coding region + 1kb 3'UTR), transgenic complementary positive plants were obtained, and then self-pollinated twice to obtain genetically transformed stable families (see Figure 6 Compared with the negative family complemented by transgenic, the positive family had significantly higher plant height and longer panicle length; C7 -COM1 and HBK-PH8 C7 -COM2 plant height increased by 25.7cm and 32.5cm respectively (see Figure 6 B), the spike length increased by 2.4cm and 2.7cm respectively (see Figure 6 C and D in Figure ). It further confirmed that LOC_Os08g43390 is PH8 .

[0046] Embodiment 5: PH8 The C7 allele is a rare allele. Using 529 rice micro-core germplasm resource populations collected by this group, the present invention PH8 Haplotype analysis was performed (see Figure 7 , Different letters in the column of plant height effect comparison indicate significant differences at the P ≤ 0.01 level. ). According to the SNP variation information in its coding region, PH8 There are 8 haplotypes (each haplotype has N ≥ 3); HAP1 and HAP2 are mainly distributed in temperate japonica and tropical japonica subspecies, while HAP3, HAP4 and HAP6 are mainly distributed in indica I and indica II subgroups, HAP5 is distributed in indica I and Aus rice subgroups, and HAP8 is mainly distributed in Aus rice subgroup (see Figure 7 ); C7 type alternative splicing variant (HAP9) is only distributed in the Aus rice subpopulation, and only 4 Aus rice samples carry this variant; this variant accounts for only 0.8% of the entire micro-core germplasm population, indicating that PH8 The C7 allele is a rare allele (see Figure 7 ).

[0047] Embodiment 6: PH8 The C7 allele can be used to reduce the plant height of conventional indica and japonica rice PH8 The C7 allele (PH8 C7 ) is a rare allele and has hardly been used in modern cultivated rice. The present invention introduces PH8 into the background of the high-quality indica rice restorer line 9311 and the high-quality japonica rice Koshihikari (KOS). C7 Alleles were selected by using Kasp-PH8 molecular marker to examine the plant height and yield per plant of the near-isogenic lines (see Figure 8The results showed that compared with the wild type 9311, the PH8 C7 Allele 9311 (9311-PH8 C7 ) Plant height decreased by 20.5 cm (see Figure 8 A and Figure 8 C), but the yield per plant did not decrease significantly (see Figure 8 E); compared with the KOS wild type, the introduction of PH8 C7 Allele of KOS (KOS-PH8 C7 ) Plant height decreased by 10.7 cm (see Figure 8 B and D in Figure ), in terms of single plant yield, KOS-PH8 C7 There was no significant difference with the wild type (see Figure 8 These results suggest that PH8 C7 The allele can reduce plant height in both indica and japonica rice without significantly reducing yield.

[0048] Embodiment 7: PH8 The C7 allele can be used to reduce the height of hybrid rice because PH8 There is no dominant effect, and the plant height of the heterozygous type is close to the mid-parent value, so PH8 The C7 allele (PH8 C7 ) can be used to reduce the plant height of hybrid rice. The present invention utilizes 9311 wild type and 9311-PH8 C7 The hybrid rice LYP9 and LYP9-M were obtained by mating with the two-line sterile line PA64S. C7 The hybrid rice PA64S and GZ63-4S were mated with the two-line sterile line Guangzhan 63-4S (GZ63-4S) to obtain the hybrid rice Yangliangyou 6 (YLY6) and the improved Yangliangyou 6 (YLY6-M). PA64S and GZ63-4S have been widely circulated in the field. Their yield-related traits were investigated, and the results showed that the plant height of LYP9-M was significantly reduced by 5.7 cm compared with LYP9 (see Fig. 9 A in Figure 2 and Table 2), the number of effective ears per plant increased significantly, the thousand-grain weight decreased slightly, while other agronomic traits showed no significant differences, and the final yield per plant showed no significant difference (see Table 2); compared with YLY6, the plant height of YLY6-M was significantly reduced by 8.9 cm (see Fig. 9 B and Table 2), the number of effective ears per plant increased significantly, the fruit set rate and 1000-grain weight decreased slightly, other agronomic traits showed no significant differences, and the final yield per plant did not decrease significantly (see Table 2). These results show that PH8 C7The heterozygous allele can also reduce the height of rice plants without affecting the yield per plant, and can be used to reduce the height of hybrid rice plants.

[0049] Table 2 Yield performance of LYP9, YLY6 and their improved hybrid rice

[0050] and Respectively expressed in P ≤0.05 and P The difference was significant at the ≤0.01 level.

[0051] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. Application of PH8 protein, or its encoding gene, or biological materials containing its encoding gene in regulating plant height, panicle length or lodging resistance of plants; the PH8 protein is cytochrome P450 CYP78A15, and the plant is a rice plant.

2. Application of PH8 protein or its encoding gene as an inhibition target in cultivating dwarf plants, short panicle length plants or plants with strong lodging resistance; the PH8 protein is cytochrome P450 CYP78A15, and the plant is a rice plant.

3. Application of PH8 protein, or its encoding gene, or biological materials containing its encoding gene in improving plant germplasm resources related to lodging resistance; the PH8 protein is cytochrome P450 CYP78A15, and the plant is a rice plant.

4. The use according to any one of claims 1 to 3, characterized in that: By reducing the expression level of the PH8 protein in the plant, the plant height and ear length of the plant are reduced, and the lodging resistance of the plant is improved; and / or, by introducing the nucleotide sequence shown in SEQ ID NO.3 into the plant, the plant height of the plant is reduced and the lodging resistance of the plant is improved.

5. The use according to any one of claims 1 to 3, characterized in that: The PH8 protein comprises any one of the following amino acid sequences: (1) the amino acid sequence shown in SEQ ID NO.1; (2) The amino acid sequence of the amino acid sequence shown in SEQ ID NO.1 obtained by replacing, inserting or deleting one or more amino acids, and having the same functional protein.

6. The use according to any one of claims 1 to 3, characterized in that: The coding gene of the PH8 protein includes any one of the following nucleotide sequences: (1) the nucleotide sequence shown in SEQ ID NO.2; (2) A nucleotide sequence encoding a protein with the same function obtained by replacing, deleting or inserting one or more nucleotides of the nucleotide sequence shown in SEQ ID NO.2; (3) A nucleotide sequence that can hybridize with the nucleotide sequence shown in SEQ ID NO. 2 under stringent conditions.

7. The use according to any one of claims 1 to 3, characterized in that: The biological material is an expression cassette, a vector or a transgenic cell.

8. A nucleic acid, characterized in that The nucleic acid comprises: a nucleotide sequence as shown in SEQ ID NO.

3.

9. A KASP primer combination, characterized in that: These include: Kasp-PH8-F1: 5'-TTAATTACTGGAGTACTGTACATAC-3', Kasp-PH8-F2: 5'-TTAATTACTGGAGTACTGTACATAT-3', Kasp-PH8-R: 5'-GACTTCACCGACGTCCTTCTCTCC-3'.

10. A method for reducing plant height, characterized in that: include: The nucleic acid according to claim 8 is introduced into the plant; the plant is a rice plant.

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