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Mulberry powdery mildew related gene and application thereof

A technology for powdery mildew and mulberry, applied in the field of cloning and its expression under stress conditions

Active Publication Date: 2017-09-08
SERICULTURAL RES INST ANHUI ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as a defense protein, there are still few studies on the relationship between MLO genes and abiotic environmental stress

Method used

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  • Mulberry powdery mildew related gene and application thereof
  • Mulberry powdery mildew related gene and application thereof
  • Mulberry powdery mildew related gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Cloning of embodiment 1 Mulberry powdery mildew related gene MmMLO

[0022] 1.1 Design primers

[0023] The mulberry species tested was Yu 71-1, which was preserved in the National Germplasm Zhenjiang Mulberry Garden of the Institute of Sericulture, Chinese Academy of Agricultural Sciences. Utilize the obtained ESTs sequences containing MmMLO cDNA fragments, design primers, clone in segments, and verify, the primer sequences are as follows:

[0024] MmMLO-F1: 5'-ATGAGTGGCGGAGGGAGTG-3';

[0025] MmMLO-R1: 5′-CTCAGCAACTTCATGTGCCAA-3′

[0026] MmMLO-F2: 5'-AAATACATGATACGTGCCCTT-3';

[0027] MmMLO-R2: 5′-GACAACCGAAAACAATGATATG-3′

[0028] According to the results of the cloned and verified middle fragment of the gene, design the downstream specific primers for the target gene required for 5′RLM-RACE nested PCR amplification. The primer sequences are as follows:

[0029] GSP-outer: 5′-CAAAAGCATCAGTCCTCC-3′

[0030] GSP-inner: 5′-AGGCTTCTGGTTCTTCCTC-3′

[0031] Primers...

Embodiment 2

[0040] Example 2 Bioinformatics Analysis of Mulberry Powdery Mildew Related Gene MmMLO

[0041]The online tool PSORT was used to predict the nuclear localization sequence of the target gene and analyze the related conserved gene sequence according to the reported literature.

[0042] Utilize DNAStar software and online analysis software ExPASy (http: / / prosite.expasy.org / prosite.htm1 / ), SMART (http: / / smart.emb1-heidelberg.de / ) to carry out comparatively comprehensive analysis on mulberry powdery mildew related gene MmMLO According to the analysis of the amino acid sequence encoded by the gene, SEQ ID No. 2, the isoelectric point of the MmMLO protein encoded by the gene is 9.03, and the molecular mass is 62.48kDa.

[0043] Use NCBI's Blast tool to search and compare the homology of MmMLO proteins, and download some homologous amino acid sequences; use Clustal x software to identify the amino acid sequence encoded by MmMLO related gene MmMLO in mulberry with the amino acid sequen...

Embodiment 3

[0045] Example 3 Expression patterns of mulberry powdery mildew-related gene MmMLO under various stress conditions

[0046] When the new shoots of the tested mulberry seedlings grew to about 20 cm long, the mulberry seedlings were subjected to drought, high-salt, and low-temperature stress treatments. During the whole process of stress induction, the abiotic conditions such as temperature, light intensity, photoperiod and humidity of all experimental seedlings remained unchanged. When the mulberry seedlings showed obvious symptoms of damage under various stress treatments, the young mulberry leaves of the control and various stress conditions were taken respectively, wrapped in aluminum foil paper, quickly put into liquid nitrogen, and stored in a refrigerator at -80°C . Fluorescent quantitative PCR was used to determine the expression level of the target gene in mulberry under stress conditions, using the stably expressed actin gene (β-actin) as an internal reference. All s...

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Abstract

The invention discloses a mulberry powdery mildew related gene and application thereof. A gene sequence and a protein sequence are shown in SEQ ID NO.1 and SEQ ID NO. 2. By adoption of the technical scheme provided by the invention, whether plants are in a growing environment of drought, high salinity, low temperature and the like or not can be identified.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to the cloning of a mulberry powdery mildew related gene (MmMLO) and its expression under stress conditions. Background technique [0002] During the long-term evolution process, plants have formed a series of complex and rigorous defense mechanisms to protect themselves from pathogens, which involve the expression and regulation of a series of disease resistance-related genes. MLO (Mildew resistance locus0) gene is a kind of gene related to broad-spectrum and high-efficiency plant resistance. It is of great value in the theoretical and applied research of plant broad-spectrum resistance to powdery mildew, and plays a role in regulating plant resistance to biotic and abiotic stress. important role. [0003] The MLO gene was originally discovered in barley and has a complete set of seven transmembrane domains (TM1-TM7). MLO plays a negative regulatory role in the process of...

Claims

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Application Information

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IPC IPC(8): C12N15/31C07K14/37C12Q1/68
CPCC07K14/37C12Q1/6895C12Q2600/158
Inventor 李瑞雪赵卫国汪泰初王钰婷夏家凤王伟
Owner SERICULTURAL RES INST ANHUI ACADEMY OF AGRI SCI
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