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A gene related to eggplant verticillium wilt resistance, an acquiring method thereof and applications of the gene

A technology against verticillium wilt and eggplant, applied in the fields of molecular biology and agriculture, can solve problems that have not yet entered the level of molecular biology

Pending Publication Date: 2016-11-30
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research on eggplant polyphenol oxidase and Verticillium wilt resistance has not yet reached the level of molecular biology

Method used

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  • A gene related to eggplant verticillium wilt resistance, an acquiring method thereof and applications of the gene
  • A gene related to eggplant verticillium wilt resistance, an acquiring method thereof and applications of the gene
  • A gene related to eggplant verticillium wilt resistance, an acquiring method thereof and applications of the gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] In Example 1, the polyphenol oxidase gene was cloned using the RACE method using wild eggplant eggplant cDNA and genomic DNA as templates. The specific steps are as follows:

[0041] 1) Preparation of spore suspension:

[0042] Verticillium dahliae of eggplant was cultured on PDA medium plate for 7 days, and a mycelium block with an area of ​​about 1cm*1cm was picked from the edge with a dissecting needle, transferred into PL culture medium, and cultured on a shaker at 120rmp / min at 25°C for 7 days. -9 days, at this time, the PDA culture fluid (potato dextrose agar medium) changed from a transparent liquid to a milky white liquid. Filter the milky white liquid with double-layer gauze, keep the filtrate, observe the number of spores with a hemocytometer, calculate the number of spores per milliliter, dilute the bacterial spores with sterilized distilled water, and finally prepare a concentration of 1x10 7 spores / mL of the spore suspension, which was then used for inoculat...

Embodiment 2

[0074] In this example, the eggplant gene cloned in Example 1 was analyzed and detected, and it was confirmed by cluster analysis and amino acid comparison that the gene belongs to the polyphenol oxidase gene family, which is similar to the polyphenol oxidase gene in cultivated eggplant. The homology of the gene is high; and, semi-quantitative PCR analysis shows that its expression is related to the resistance of Verticillium wilt in watermelon, further confirming its function. details as follows:

[0075] 1) Bioinformatics analysis of StPPO1 gene and its encoded protein in Solanum mellifera

[0076] A cDNA sequence with a length of 2090bp was obtained by RACE technology and electronic cloning splicing, and it was named StPPO1. The 5'UTR of the gene contains 110bp, and the 3'UTR contains 180bp. The gene does not contain an intron structure, which is consistent with the structure of PPO genes reported in other dicotyledonous plants.

[0077] Through ORF Finder combined with ...

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Abstract

A gene related to eggplant verticillium wilt resistance, an acquiring method thereof and applications of the gene are provided. The gene has a nucleotide sequence shown as SEQ ID NO.1 in a sequence table. The method includes 1) extracting total RNA and genome DNA of a solanum torvum sample, 2) designing specific primers (a 3'-RACE primer GSP2 and a 5'-RACE primer GSP1) through adopting cDNA obtained by subjecting the RNA extracted in the step 1) to inverse transcription as templates, and performing RACE-PCR amplification, 3) cloning by adopting the genome DNA extracted in the step 1) as templates and by utilizing specific primers gP1F and gP1R designed based on the gene obtained in the step 2), and 4) subjecting a PCR product obtained in the step 3) to recovering, sequencing and sequence analyzing. The gene and the method are suitable for eggplant verticillium wilt research, plant molecular marker assisted breeding and inheritable character improvement in the genetic engineering field, and lay a primary base for verticillium wilt-resisting molecular breeding.

Description

technical field [0001] The invention relates to the fields of molecular biology and agriculture, in particular to a gene related to resistance to Verticillium wilt of eggplant, an acquisition method and application thereof. Background technique [0002] Eggplant (Solanum melongena L.) is one of the important vegetables in my country. It is rich in nutrition and has high economic benefits. However, most cultivated eggplants are susceptible to Verticillium wilt. Verticillium wilt of eggplant is a soil-borne vascular disease caused by Verticillium fungus. Its symptoms usually manifest as yellowing and wilting of half of the plant or half of the leaves. When the disease is severe, the stems are split and the vascular bundles turn brown. The disease has a long incubation period, strong transmission ability, and a wide range of damage. When the disease is severe, the output loss can reach more than 60%. The United States, Japan and India have all had outbreaks of eggplant Vertici...

Claims

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Application Information

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IPC IPC(8): C12N15/53C12N9/02C12N15/10
CPCC12N9/0059C12Q1/686C12Y110/03001C12Q2531/113
Inventor 杨旭张宇成玉富
Owner YANGZHOU UNIV
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