Cloning and application of key gene Wo for controlling tomato hair generation

A gene, tomato technology, applied in the field of cloning of the key gene Wo

Inactive Publication Date: 2011-03-16
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But there is no gene that has the characteristics of the tomato hair gene Wo: it not only regulates the differentiation and development of the hair on the body surface, but also participates in the growth regulation of the embryo

Method used

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  • Cloning and application of key gene Wo for controlling tomato hair generation
  • Cloning and application of key gene Wo for controlling tomato hair generation
  • Cloning and application of key gene Wo for controlling tomato hair generation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: the precise location of Wo gene of the present invention

[0038] Through the analysis of the existing classic tomato genetic map Tomato-EXPEN 2000, Tomato-ILs and integrated map (see: Tanksley, 1992, http: / / sgn.cornell.edu / ), it is known that the tomato fuzz regulation gene (Woolly, Wo, the gene not cloned) in the interval where the introgressed fragments of IL2-3 and IL2-5 (cited from the Tomato Genetic Resource Center (TGRC), http: / / tgrc.ucdavis.edu / ) were located. In the present invention, the applicant first uses IL2-5 as the female parent and the hairy mutant LA3186 (quoted from the Tomato Genetic Resource Center of the United States, http: / / tgrc.ucdavis.edu / ) as the male parent for hybridization, and pulls out the F 1 Medium to less hairy plants, selfed more hairy plants and harvested F 2 Generation seeds, followed by sowing F 2 106 of the less-haired plants were randomly selected as the mapping population, using the reported molecular markers U23...

Embodiment 2

[0048] Embodiment 2: Cloning and sequence analysis of Wo gene

[0049] According to the information of the candidate Wo gene, PCR primers capable of amplifying the full length of the gene were designed: forward primer: 5'-TTCAAGATGTTTAATAACCACCAGC-3', reverse primer: 5'-ACCTTTCATGCATTTGCGGAAGTTAC-3'. Genomic RNA was extracted from tomato leaves, and the RNA was reverse-transcribed into cDNA using reverse transcriptase (purchased from TOYOBO Company) (refer to J. Society, 2002 version), the full-length gene product of Wo was obtained by PCR amplification, detected by 0.8% agarose gel, and the target fragment was recovered with a recovery kit (purchased from Axgen Company), and the pMD18-T vector (purchased from Baobio Engineering (Dalian) Co., Ltd.) cloning, take 5 μ L of the ligation product and carry out heat shock treatment, the specific method refers to J. Sambrook et al., translated by Huang Peitang et al., Molecular Cloning Experiment Guide (Third Edition), Science Press,...

Embodiment 3

[0052] Example 3: Determination of the stage of embryo abortion in hairy mutant plants

[0053] The fuzzy mutant LA3186 has the characteristic of homozygous embryonic lethality, and the applicant used LA3186 as a material to observe its embryonic lethality period microscopically. The seeds in the tomato fruit from the 4th day to the 40th day after pollination were collected (tomato plants grow uniformly, and the third sedge was adopted) and were fixed, stained, dehydrated and other steps were taken to make paraffin sections (refer to Wang Zaoan, 1992 ), observed and counted the development of tomato embryos in each period after flower pollination, and found that the development of the embryos on the 7th day after pollination was not much different. , the embryonic stalk began to degrade, and the embryos showed obvious differentiation, and then gradually matured through globular embryos, torpedo-shaped embryos, and cotyledon-shaped embryos. However, the aborted embryos stagnat...

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Abstract

The invention belongs to the technical field of plant genetic engineering, and specifically relates to cloning and an application of a Wo gene which is related to tomato hair generation. The cDNA (complementary deoxyribonucleic acid) sequence of the cloned Wo gene is as shown in SEQ ID NO.1(sequence identity number 1) in a sequence table, the corresponding amino acid sequence is as shown in SEQ ID NO.2, and the Wo gene encodes 731 amino acids. The invention further relates to a promoter of the Wo gene and cloning of two allelic genes of the Wo gene. The Wo gene is over-expressed in tomatoes and can improve the amount of epidermal hair of the tomatoes, significantly enhance the insect resistance of the tomatoes and enhance the anti-virus capability.

Description

technical field [0001] This patent belongs to the technical field of plant genetic engineering. It specifically relates to the cloning, functional identification and application of a key gene Wo controlling the occurrence of tomato fuzz. The gene has significant functions in regulating the quantity and type of tomato hairs, and can be applied in tomato resistance to insects, viruses and the like. Homozygous for this gene can lead to embryonic lethality, and it is also a key gene for tomato embryo development. technical background [0002] It is one of the defense mechanisms formed in the process of plant evolution to use the structure of the hairy tissue structure of the plant body as a plant to resist and expel herbivorous pests. Plant epidermis is a specialized unicellular or multicellular epidermal structure, including glandular and non-glandular hairs, which is a model material for studying the regulation of plant cell differentiation. Epidermal hairs, as the first ba...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/113C07K14/415A01H5/00
Inventor 李汉霞杨长宪叶志彪王涛涛张俊红张余洋卢永恩
Owner HUAZHONG AGRI UNIV
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