Maize auxin transport gene ZmABCB15 as well as application thereof in resisting rough dwarf virus

A technology for transferring genes and auxin, which is applied in application, genetic engineering, plant genetic improvement, etc., can solve the problem that corn germplasm resources cannot meet the needs of corn production, achieve reliable materials and data support, reduce proliferation rate, and improve resistance. sexual effect

Active Publication Date: 2019-12-27
HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is to provide a kind of maize auxin transport gene ZmABCB15 and its application in resistan...

Method used

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  • Maize auxin transport gene ZmABCB15 as well as application thereof in resisting rough dwarf virus
  • Maize auxin transport gene ZmABCB15 as well as application thereof in resisting rough dwarf virus
  • Maize auxin transport gene ZmABCB15 as well as application thereof in resisting rough dwarf virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Maize auxin transport carrier encoding gene ZmABCB15

[0035] 1. Select 5 maize materials with different resistance to rough dwarf disease commonly used in current production, which are two rough dwarf resistant materials (Chang 72, P138), hybrids of Zheng 58 and Chang 7-2 ( Zheng Dan 958) and two rough dwarf disease-susceptible materials (478, Zheng 58) used corn rough dwarf virus SRBSDV to infect the above materials.

[0036] The specific method of virus inoculation test is as follows:

[0037] (1) Feeding experiment of SBPH

[0038] In spring, SBPH was captured from wheat seedlings and weeds in the field, and after removing the source of heterogeneous insects, it was brought back to the laboratory for artificial breeding and subculture. Large-scale artificial rearing of SBPH is carried out in a special insect culture room, using the beaker rearing method. Keep a constant temperature of 25°C in the insect rearing room, and adjust the indoor humidity to ...

Embodiment 2

[0047] Example 2: Maize auxin transport carrier gene after different resistant maize materials were infected by crude contraction virus ZmABCB15 Expression changes and virus particle content analysis

[0048] The virus content and virus proliferation rate of 5 different resistant corn materials were measured before and after corn infection with rough dwarf disease virus RBSDV (same as Example 1).

[0049] The concrete step method of virus proliferation detection experiment in corn plant is:

[0050] Total RNA from maize leaves was extracted using ZYMO RESEARCH (USA) Direct-zol™ RNA MiniPrep kit, and reverse transcribed into cDNA. According to the nucleotide sequence of the RBSDV S6 fragment, RT-PCR specific primers specially used for amplifying RBSDV were designed, namely:

[0051] F: 5'--TCA GCA AAA GGT AAA GGA ACG--3';

[0052] R: 5'--GCT CCT ACT GAG TTG CCT GTC--3'.

[0053] Applying the fluorescent quantitative PCR absolute quantitative method to detect unknown samples...

Embodiment 3

[0063] Example 3: ZmABCB15 protein subcellular localization

[0064] ZmABCB15 The cDNA clone of the gene was then constructed into the intermediate pMDT-20 vector. After sequencing, the 35S-GFP vector pH7FWG2.0 was constructed to obtain the recombinant vector 35S-ZmABCB15:GFP, and transformed into tobacco leaf protoplasts. Membrane protein marker protein 1008 (provided by the State Key Laboratory of Plant Physiology and Biochemistry, Zhejiang University) was used for co-infection, and the localization was observed with a LSM710 confocal microscope.

[0065] Such as Figure 7 As shown, the ZmABCB15 protein functions as an auxin transport carrier cell membrane localized.

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Abstract

The invention discloses application of a maize auxin transport gene ZmABCB15 as well as application thereof in resisting rough dwarf virus and aims to solve the technical problem that maize germplasmresources in the prior art cannot meet the current maize production requirement. The maize auxin transport gene ZmABCB15 and transport protein thereof are obtained by differential gene expression analysis, and the auxin transport gene ZmABCB15 can be applied to resistance of plants to rough dwarf virus and breeding of anti-rough dwarf virus plants. The invention confirms for the first time that the expression level of the maize auxin transport gene ZmABCB15 is in positive correlation with resistance of maize to the rough dwarf virus, and the ZmABCB15 gene is over-expressed in maize to improveresistance of maize to the rough dwarf virus; and the study of anti-rough dwarf virus genetic mechanism and disease-resistant molecular breeding of maize.

Description

technical field [0001] The invention relates to the technical field of plant molecular biology, in particular to a maize auxin transport gene ZmABCB15 And its application in anti-rough dwarf disease. Background technique [0002] Maize rough dwarf is a worldwide maize disease caused by a plant virus, which has seriously threatened my country's maize production for a long time. In recent years, with the adjustment of the planting structure of food crops and the change of the natural environment, the incidence of rough dwarf disease in my country has shown an obvious upward trend. The intensified outbreak of rough dwarf disease has brought very serious damage to local corn production and food security. Maize seedlings are infected with rough dwarf disease, the plants are severely dwarfed, the internodes become shorter and thicker, and abnormal heading occurs, which seriously leads to yield reduction or even infertility. Southern rice black-streaked dwarf virus (SRBSDV) is a...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00A01H6/46A01H6/82
CPCC07K14/415C12N15/8283
Inventor 岳润清铁双贵刘璐陈娜卢董华徐心志
Owner HENAN ACAD OF AGRI SCI
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