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Tomato yellowing-to-green gene SlRHBDD2 and application thereof

A tomato yellow and gene technology, applied in the field of genetic engineering, can solve problems such as limitations, and achieve the effect of simplifying the screening steps

Active Publication Date: 2021-04-16
ZHEJIANG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the currently cloned genes controlling leaf color mutants are limited to model plants such as Arabidopsis or rice. For example, multiple genes controlling chlorophyll synthesis and chloroplast development were isolated and identified from leaf color mutants lacking pigment

Method used

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  • Tomato yellowing-to-green gene SlRHBDD2 and application thereof
  • Tomato yellowing-to-green gene SlRHBDD2 and application thereof
  • Tomato yellowing-to-green gene SlRHBDD2 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1. Acquisition and Character Identification of Tomato Yellow-to-Green Leaf Color Mutant "cely403"

[0024] 1. Acquisition of tomato yellow-turned-green leaf color mutant "cely403"

[0025] The tomato yellow-to-green leaf color mutant was derived from the high-generation inbred line "T12404" of the American tomato germplasm resource bank (TGRC) "LA3320". A mutant plant with natural leaf color was found during field planting. The cotyledons turn yellow, the heart leaves are yellow-green at the seedling stage, and the leaves gradually turn green and return to normal at the later stage. Mutant plants were self-seed to obtain mutant lines. After multiple generations of self-propagation and selection, the mutant traits have been stabilized. The mutant was named "cely403" (Cotyledon and EarlyLeaf Yellowing). The Tomato Research Group of the Technical College saves and makes available to the outside world.

[0026] 2. Character identification of tomato yellow-to-green...

Embodiment 2

[0028] Embodiment 2, comparison of chlorophyll content and photosynthetic characteristics of tomato yellowing to green leaf color mutant

[0029] At different periods after sowing, randomly select 12 individual plants with the same growth from the mutant and the wild type, select the second true leaf that is fully expanded under the growth point, cut it into 2-3cm, weigh 0.2g and soak it in 25mL A mixture of acetone and ethanol (volume ratio 95% acetone: absolute ethanol = 2:1), sealed and protected from light, and cultured at 26° C. in the dark for 24 hours. Measure the OD value of the extract solution at three wavelengths of 470nm, 645nm and 663nm with an ultraviolet spectrophotometer, and repeat 3 times. According to the Arnon calculation method improved by Livchtenthaler, the contents of chlorophyll a, chlorophyll b and carotenoids were calculated respectively, and the total chlorophyll content was the sum of chlorophyll a and chlorophyll b. The results showed that, compa...

Embodiment 3

[0031] Example 3, Genetic Analysis and Gene Mapping of Tomato Yellowing to Green Leaf Color Mutation Character

[0032] 1. Genetic analysis of the mutation trait of tomato from yellow to green leaf color

[0033] Crossing mutant cely403 with wild-type 404 yielded F 1 Generation, self-crossing to get F 2 Generation, using F 2 Genetic analysis of segregating populations. The results showed that F 1 The leaf color of the generation plant is green, which is the wild-type phenotype; sowing F 2 In this generation, the leaf color of 483 plants was counted, and the number of plants with normal leaf color and yellowed leaf color was 378 and 105, and the chi-square test results (X 2 =2.572 0.05,1 =3.84) in line with the segregation ratio of 3:1, the above results were repeated three times, and the experimental results all showed that the mutant trait was controlled by a recessive single gene.

[0034] 2. Gene mapping of tomato yellow to green leaf mutation trait

[0035] Use F ...

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Abstract

The invention discloses a tomato yellowing-to-green gene SlRHBDD2 coding regionc DNA sequence SEQ ID No.1 and a coding proteinamino acid sequence SEQ ID No. 2. The gene SlRHBDD2 for controlling the tomato leaf color is cloned from an early tomato leaf color yellowing mutant "cel y 403" by a map-based cloning technology, and a function complementation test proves that the SlRHBDD2 is a gene for controlling the tomato leaf color yellowing to green. According to the leaf color yellowing-to-green character controlled by the gene, early cotyledons and core leaves are yellow green, the chlorophyll content is reduced, and the net photosynthetic rate is reduced; and the leaves in the later stage are turned into green, the chlorophyll content and the net photosynthetic rate are recovered, so the gene can be used as a seedling stage marker for early selection of filial generation in the breeding process of new tomato varieties, such as the seedling stage identification of a nuclear male sterile line, can also be applied to the aspects of seed production, purity identification and the like, and has good application prospect.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to a tomato yellowing-transforming gene S1RHBDD2 and its application. Background technique [0002] Tomato (Solanum lycopersicum L.) belongs to Solanaceae tomato genus, is one of the most widely cultivated vegetables in the world, and plays an important role in the production and consumption of vegetables in my country. Leaf photosynthetic efficiency is a key factor determining tomato yield and quality. The high efficiency of photosynthesis depends on the synthesis of chlorophyll and the normal development of chloroplasts, so it is of great significance to study the metabolism of chlorophyll and the development of chloroplasts. [0003] Leaf color is an important agronomic trait, and under normal conditions, plant leaves are green. When plants are affected by the external environment or their own genetics, resulting in defects in chloroplast development and blockage ...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/82C12N15/84A01H5/12A01H6/82C12Q1/6895
Inventor 卢钢刘悦邹滔李孟卓许华超宋建伟余小林
Owner ZHEJIANG UNIV
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