Method for improving disease resistance by transforming arethusa with longan ferredoxin gene

A technology of gene transformation and disease resistance, applied in the field of genetic engineering

Inactive Publication Date: 2014-07-02
FUJIAN AGRI & FORESTRY UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Previously, only the ferredoxin-like ferredoxin derived from sweet pepper was reported in terms of improving plant disease resistance. Whether different types of fd genes derived from other species have improved Disease resistance function remains unknown

Method used

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  • Method for improving disease resistance by transforming arethusa with longan ferredoxin gene
  • Method for improving disease resistance by transforming arethusa with longan ferredoxin gene
  • Method for improving disease resistance by transforming arethusa with longan ferredoxin gene

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Embodiment 1

[0021] Longan ferredoxin gene Dlfd3 , Dlfdc1 Design with restriction endonuclease sites Bgl II. Speech The corresponding primers upstream and downstream of I Dlfd3 F: 5'-GA AGATC TATGGCAACT GTGACCCTTAG-3', Dlfd3 R: 5'-GG ACTAGT GTAAAGTTCG CCTTCCTTGTG-3'; Dlfdc1 F: 5'-GA AGATCT ATGG CAACACTTCACTTCAG-3', Dlfdc1 R: 5'-GG ACTAGT ATCATCAGCA GTTGCCAGTTG-3'. 25μL PCR reaction system: 10 X PCR buffer ( plus Mg 2+ ) 2.5 μL, dNTPs ( 2.5 mmol L -1 ) 0.5 μL, forward primer (10 μmol L -1 ) 1 μL, reverse primer ( 10 μmol L -1 ) 1 μL, Taq ( 5 U μL -1 ) 0.2 μL, ddH 2 O 18.8 μL. PCR reaction program: pre-denaturation at 94°C for 4 min, denaturation at 94°C for 45 s, annealing at 54°C for 45 s, extension at 72°C for 40 s, 35 cycles, and finally extension at 72°C for 10 min. The PCR product was detected by 1.0% (W / V) agarose electrophoresis, and a band of about 500 bp was recovered, which was introduced into PMD18-T (TAKARA) to construct the PMD18-T plasmid. Using Fermen...

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Abstract

The invention provides a method of improving disease resistance by transforming arethusa with longan ferredoxin gene and belongs to the technical field of genetic engineering. The longan ferredoxin gene (Dlfd3) (Register number: JF733784) is guided to arethusa oncidium by means of a transgenic technology, and the obtained transformed plant represents higher transplanting survival rate and disease resisting capability. Compared with wild plants, the soft rot resistant capability of the transgenic plant is improved from high touch soft rot (Di=100) to intermediate soft rot resistance (Di=27.5). The transplanting survival rate is improved from 79.3% to 95.7%, thereby realizing extremely significant difference.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and relates to a method for transforming orchid plants with a longan ferredoxin gene to improve disease resistance, in particular to the application of the longan ferredoxin gene and orchid breeding. Background technique [0002] Oncidium ( Oncidium ), also known as Dancing Girl Orchid, King Phalaenopsis, Tumor Petal Orchid, etc., is an important economic cultivar of pot-cut flowers. Oncidium is susceptible to soft rot, and soft rot can bring devastating blows during the transplanting stage of tissue culture seedlings. At present, there is no particularly effective control method for this disease, so breeding oncidium varieties resistant to soft rot is an important topic in oncidium breeding. Oncidium can carry out a wide range of intra-genus and inter-genus hybrids, but both intra-genus and inter-genus hybrids have poor affinity, and the results are relatively rare under natural co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84A01H5/00
Inventor 赖钟雄叶炜林玉玲匡云波江金兰李永清雷伏贵周建金许旭明陈裕坤刘生财张梓浩
Owner FUJIAN AGRI & FORESTRY UNIV
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