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Myrothamnus flabellifolia gene MfbHLH145 and application thereof

A kind of milo wood and genetic technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve problems such as unclear mechanism

Inactive Publication Date: 2019-01-11
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the past, researchers mainly studied the drought resistance mechanism of the plant through morphological, physiological, biochemical and other aspects, but the genes related to dehydration response and the molecular mechanism of drought resistance are still unclear, and the drought resistance bhLh transcription in Milo Reports about factors and their applications

Method used

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  • Myrothamnus flabellifolia gene MfbHLH145 and application thereof
  • Myrothamnus flabellifolia gene MfbHLH145 and application thereof
  • Myrothamnus flabellifolia gene MfbHLH145 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Cloning Miromu MfbHLH145 gene

[0034] 1. Take the leaves of A. milo, quick-frozen in liquid nitrogen, and store them in a -80°C refrigerator for extraction of total RNA; the total RNA was extracted using the Plant Total RNA Isolation Kit purchased by Chengdu Lambo Biological Company; the cDNA of A. milo The synthesis of the first chain was performed using Reverse Transcriptase M-MLV (RNaseH-) from Dalian Bao Biotechnology Co., Ltd., according to its product manual.

[0035] The first strand of cDNA synthesized by the above kit was used as the amplification template, and the designed F: 5'-CATGCCATGGGAAAGGACTGTGGATCC-3' (SEQ ID NO.3) and R: 5'-GCTAGTGAGAGAATCAAGTCCTAAAGCTTTG-3' (SEQ ID NO.4 ) as primers, for subsequent enzyme digestion and recombination connection, when designing primers, add NcoI and SpeI these two enzyme digestion sites, use RT-PCR to carry out cDNA amplification, the amplification system is shown in Table 1, and the amplification conditions...

Embodiment 2

[0039] Example 2 Sequence Homology and Homology Analysis of Miroma MfbHLH145

[0040] According to the gene homology comparison map, it can be seen that the amino acids with a high degree of overlap are not concentrated in the predicted functional domain of bHLH, and MfbHLH145 is divided into a separate category in the phylogenetic tree, indicating that the MfbHLH145 gene has high specificity. There is little research information on bHLH-like genes. It can be seen from the phylogenetic tree (such as figure 2 ), and the plants with higher affinity to MfbHLH145 are all woody plants, and belong to the phylum Angiosperms and the class Dicotyledons. Analysis of the growth environment of plants with high homology, such as cocoa (Theobroma cacao), found that most of the plants with high homology came from similar growth environments with Milo, and these plants prefer growth with low humidity and high temperature. environment, and most of them grow in subtropical and tropical regio...

Embodiment 3

[0041] Example 3 The expression level of MfbHLH145 in different dehydration stages of milo wood

[0042] Using the Trizol method to extract the total RNA of leaves in the dehydration stage of 0, 10%, and 25% of Milo, respectively, and using the extracted total RNA as a template, reverse transcription was performed using Reverse Transcriptase M-MLV (RNaseH-) from Dalian Bao Biotechnology Co., Ltd. Obtain cDNA.

[0043] Primers were designed by primer design software, the annealing temperature was set at 58°C, Actin gene was used as an internal reference, and each reaction was repeated three times. The real-time fluorescent quantitative PCR instrument was used for fluorescent quantitative PCR, and the results were analyzed using the software that comes with the program. The primer sequences used are shown in SEQ ID NO.5, SEQ ID NO.6, SEQ ID NO.7 and SEQ ID NO.8, and the results are as follows image 3 shown.

[0044] Actin primer sequence:

[0045] F: 5'-ATCACCATTGGGGCTGAACG-...

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Abstract

The invention discloses a Myrothamnus flabellifolia gene MfbHLH145 and an application thereof. The nucleotide sequence of the gene is represented by SEQ. ID. NO.1; and the amino acid sequence of the encoded protein of the gene is represented by SEQ. ID. NO.2. The drought-resistant bHLH transcription factor of Myrothamnus flabellifolia is studied to improve the drought resistance of plants in orderincrease the drought resistance of the plants.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and in particular relates to a milowood gene MfbHLH145 and its application. Background technique [0002] Drought has extremely adverse effects on plant growth and development. my country is a country seriously affected by drought, and the arid and semi-arid area accounts for about 1 / 2 of the country's land area. Population growth and the rapid development of urban construction not only lead to increasingly fierce competition for civil, industrial, and agricultural water use, but also objectively lead to increased pollution, ecological system damage, and water resource shortages. Therefore, discovering drought-resistant genes and cultivating new drought-resistant and water-saving plant varieties is of great importance for accelerating the construction of a resource-saving and environment-friendly society, vigorously developing the green economy, enhancing the ability of sustain...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/20
Inventor 黄卓蒋才忠靳斯涵朱培蕾郭涵度刘玲李西马均蔡仕珍陈佳邱嘉睿向香盈
Owner SICHUAN AGRI UNIV
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