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A tomato waterlogging resistant gene rf and its application

A flood-resistant and tomato technology, applied in the field of agricultural biology, can solve problems such as plant growth, tomato seedling hypocotyl elongation, leaf yellowing, etc., and achieve the effects of improving absorption, improving flooding resistance, and maintaining biological yield.

Active Publication Date: 2022-03-11
FUJIAN AGRI & FORESTRY UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Tomato gene discovered in traditional mutant study au The mutation causes hypocotyl elongation, yellowing of leaves and leggy plants in tomato seedlings

Method used

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  • A tomato waterlogging resistant gene rf and its application
  • A tomato waterlogging resistant gene rf and its application
  • A tomato waterlogging resistant gene rf and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 rf Mutant and WT phenotypes and flooding resistance analysis;

[0024] obtained by comparing the Micro-Tom mutant library rf mutants and wild-type WT, where rf Mutants can autonomously develop adventitious root phenotypes ( figure 1 a, 1b). Our analysis of tomato WT and rf The mutants were subjected to water stress treatment respectively, and the number of adventitious roots of the plants in the 3d flooding period was counted and the growth of the plants in the 5th day of flooding was observed. See figure 1 , under normal growth conditions, tomato wild-type (WT) cannot produce the adventitious root phenotype (a), but rf The hypocotyls in the mutant can autonomously produce obvious aerial roots (b), when the plants are induced by waterlogging, rf The number of aerial roots on the mutants will increase significantly. When the flooding is induced for 3 days (+3 DPF, day post flooding), rf The most distal stem internodes of the mutant were also capable of ...

Embodiment 2

[0026] Example 2: rf Mapping of mutant candidate genes;

[0027] for positioning rf mutant candidate genes, will rf mutants were crossed with WT, and the F 1 Plant selfing produces F 2 group, and finally analyzed F using Chi-square test 2 Generation phenotype segregation ratio. Using the modified CTAB method from F 2 Genomic DNA was extracted from 55 mutant phenotype plants and 55 WT individual plants in the population. Equal amounts (ng) of DNA from single plants that passed the test were mixed, and mutant pools and WT pools were respectively constructed and sent to the company for BSA-seq. Finally, we performed linkage analysis on all SNP-indexes in the obtained clear-data, and selected SNP sites with SNP-index values ​​close to 100% for mutation identification.

[0028] Using BSA sequencing, the candidate gene linkage group RF was located on the short arm of tomato chromosome 1 (a), and the analysis of the sites with SNP-index = 100% showed that only one SNP site wa...

Embodiment 3

[0029] Example 3: Verification of Genetic Complementation RF gene function;

[0030] for further exploration RF In mediating the biological functions in the occurrence of AR, we use Gateway® technology to rf The mutant candidate genes were constructed by genetic complementation to confirm the biological functions of the mapped genes. Candidate genes AU The full-length cDNA (SEQ ID NO.1) was cloned into an entry intermediate vector pDONR221. Then the candidate gene was cloned into the expression vector pHellsgate 8 driven by CAMV35s promoter. Finally, using Agrobacterium C58 to transform rf mutants that acquire OE- AU / rf transgenic plants.

[0031] from image 3 Genetic complementation experiments proved that in the mutant rf Medium overexpression AU After the gene, the occurrence of aerial roots can be completely and completely suppressed. Under normal conditions, neither the WT (a) nor the transgenic lines could produce aerial roots (c), slice analysis found ...

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Abstract

A tomato flooding resistance gene of the present invention RF and its application, belonging to the field of biotechnology, in AU A SNP mutation in the gene protein domain caused increased HO‑1 enzyme activity in hypocotyls and aerial root production. Flooding test confirmed that this aerial root trait has a very significant effect on improving the waterlogging resistance of tomato germplasm materials. It was proved by grafting experiments and metabolic pathway analysis that when au The secondary metabolite heme in the mutant leaves may be transported to the hypocotyl through a long distance, and then decomposed by the activated HO-1 to further induce the occurrence of aerial roots of the hypocotyl. This method of inducing the autonomous aerial rooting of tomato through the long-distance transport and communication of the secondary metabolite heme in plant leaves has great application value for quickly solving the resistance problem of many tomato varieties that are not tolerant to flooding in current production.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and in particular relates to a tomato flooding resistance gene RF and its application. Background technique [0002] The frequent occurrence of floods and natural disasters caused by global warming is a serious challenge faced by the agricultural production in the world today. Compared with the research on the characteristics of plant flooding resistance in rice, the research on the flooding resistance of tomato and other vegetable crops has not been well studied. Tomato is one of the vegetable crops with the largest planting area in the world, but its own anti-flooding characteristics are very weak, and it may even lead to extinction due to flooding. Therefore, the identification of flood tolerance genes that do not affect tomato yield has important application value and research significance for tomato breeding. [0003] Plants have formed different flood tolerance mechanisms in the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/00A01H6/82
CPCC07K14/415C12N15/8271
Inventor 吴双吴俊清程洁许春苗孙文茹
Owner FUJIAN AGRI & FORESTRY UNIV
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