Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Gene for regulating and controlling sugar content of tomato fruits and application thereof

A technology for sugar content, tomato, applied in the fields of biotechnology and genetic breeding

Active Publication Date: 2021-05-07
SHENYANG AGRI UNIV
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few studies reported in tomato, the mechanism of action of cytoplasmic invertase on the sugar metabolism of tomato fruit

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Gene for regulating and controlling sugar content of tomato fruits and application thereof
  • Gene for regulating and controlling sugar content of tomato fruits and application thereof
  • Gene for regulating and controlling sugar content of tomato fruits and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Construction of RNAi vector and recombinant Agrobacterium

[0029] The SlCIN2-RNAi vector was constructed using gateway technology.

[0030] 1. Extract tomato RNA, and reverse transcribe it into cDNA, use cDNA as template to amplify the target fragment SlCIN2 gene, the target fragment size is 170bp ( figure 1 A), its nucleotide sequence is shown in SEQ ID NO.1;

[0031] The primer sequences are as follows:

[0032] SlCIN2-F: 5'-CGCACCTGGTCCATTGTGTCGTCTT-3', shown in SEQ ID NO.2.

[0033] SlCIN2-R: 5'-GGTGTGTTCTTAGGGTCGCT-3', shown in SEQ ID NO.3.

[0034] The amplification reaction system is as follows:

[0035] High-fidelity DNA polymerase 10 μL, SlCIN2-F 1 μL, SlCIN2-R 1 μL, cDNA 2 μL, RNase-FreeddH2O 6 μL.

[0036] The amplification reaction procedure is as follows:

[0037] 98°C for 30s, 1 cycle; 98°C for 10s, 55°C for 20s, 72°C for 10s, 35 cycles; 72°C for 7min.

[0038] 2. Through TOPO reaction, take 4 μL of the amplification product of the target fragment,...

Embodiment 2

[0040] Transformation of tomato with recombinant Agrobacterium

[0041] Using Micro-Tom tomato as a test material, the recombinant vector plasmid was transformed into tomato by the leaf disc method to obtain stable genetic transformation materials.

[0042] When the tomato seedlings in the medium grow until the cotyledons are fully stretched ( figure 2 A), cut the 0.5cm long cotyledons and place them in the pre-medium for dark culture for 2 days, use OD 600 Agrobacteria reaching 0.6-0.8 infect the tomato cotyledons, and after infection, the back of the leaf is placed in the co-culture medium ( figure 2 B), after dark culture for 2 days, the explants were transferred to antibacterial medium for cultivation. When the explants were grown therein for two weeks, they were transferred to the germination medium and cultivated ( figure 2 C), after which the culture medium was changed every two weeks. When the adventitious buds grow to 2-3cm, cut the adventitious buds from the b...

Embodiment 3

[0044] Identification of Transgenic Tomato Plants Silencing SlCIN2

[0045] The DNA of the transgenic tomato leaves was extracted, and the screened silent SlCIN2 transgenic plants were identified by PCR. SlCIN2 silenced plants were detected using the Bar gene site on the pB7GWIWG2 vector plasmid to design specific primers.

[0046] The specific primer sequences are as follows:

[0047] Bar-F: 5'-GAAGTCCAGCTGCCAGAAA-3', as shown in SEQ ID NO.4.

[0048] Bar-R: 5'-CACCATCGTCAACCACTACA-3'T, as shown in SEQ ID NO.5.

[0049] Using DNA as a template for PCR amplification and electrophoresis detection, the detected specific fragment length is 439bp. As a result, it was found that a specific fragment of 439bp was detected in the 10 transgenic lines ( image 3 ), while negative controls (ddH2O and untransformed wild-type Micro-Tom) did not amplify any bands. These 10 lines are all transgenic positive plants and can be used for subsequent test analysis.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to the field of biotechnology and genetic breeding, and particularly relates to a gene for regulating and controlling the sugar content of tomato fruits and application thereof. The gene for regulating and controlling the sugar content of the tomato fruits is SlCIN2, and the nucleotide sequence of the gene is shown in SEQ ID NO. 1. The gene is prepared by the following steps of: firstly, extracting tomato RNA, performing reverse transcription on cDNA, designing an amplification primer, and amplifying the SlCIN2 gene by taking the cDNA as a template; and constructing the SlCIN2 gene into a pB7GWIWG2 vector by utilizing a gateway technology, screening correct positive clones, transforming the obtained positive clones into LBA4404 agrobacterium competence to obtain recombinant agrobacterium, and transforming the recombinant agrobacterium into tomatoes to obtain tomato plants with improved sugar content of the fruits.

Description

technical field [0001] The invention relates to the fields of biotechnology and genetic breeding, in particular to a gene regulating the sugar content of tomato fruit and its application. Background technique [0002] The fruit quality of tomato consists of appearance quality, flavor quality, nutritional quality and processing and storage quality. Among them, flavor quality includes sweetness, sourness, aroma, etc., which are related to soluble sugars, organic acids and volatile aromatic compounds in fruits. Loss of flavor can seriously affect the sensory quality of the fruit, which in turn affects its economic value. Improving the sweetness of tomato fruit is an important part of improving tomato quality. The sucrose content in the fruit of most tomato varieties is very low, mainly glucose and fructose. The content of glucose and fructose in tomato fruit is positively correlated with the activity of invertase. During tomato fruit development, sucrose metabolism is particu...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/56C12N9/24C12N15/84A01H5/08A01H6/82
CPCC12N9/2402C12N15/8245
Inventor 姜晶张琼琼刘欣
Owner SHENYANG AGRI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com