Isolated DNA molecule, encoded protein molecule and application thereof

A DNA molecule and an isolated technology, applied in the field of genetic engineering, can solve the problems of reduction of adventitious roots, reduction of root-shoot ratio, etc., and achieve the effects of increasing yield, improving ability, and improving resistance to drought and salt stress.

Inactive Publication Date: 2019-08-09
SHANGHAI NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Mutation of the OsCOW1 gene results in fewer adventitious roots and a lower root-to-shoot ratio

Method used

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  • Isolated DNA molecule, encoded protein molecule and application thereof
  • Isolated DNA molecule, encoded protein molecule and application thereof
  • Isolated DNA molecule, encoded protein molecule and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Cloning of Example 1 Rice Gene OsARF25

[0044] 1. The rice variety Zhonghua11 was cultured in an incubator (SPX-250-GB, Shanghai, China): the growth conditions were photoperiod 16h / 8h (L / D), 28°C.

[0045] 2. DNA extraction. Take about 500mg of fresh rice plant tissue material, add 80μl Lysis Buffer, grind the plant tissue with a grinding rod, add 120μl ddH 2 O. After centrifugation at 12000rpm for 15min, the supernatant was transferred to a new centrifuge tube, the OD value was measured, and detected by electrophoresis.

[0046] 3. Cloning of genes. Using the extracted rice DNA as a template, the forward primer and the reverse primer are used to carry out PCR to obtain the full length of the gene. For specific sequence information, see SEQ ID NO.1.

Embodiment 2

[0047] Example 2 Analysis of expression pattern of rice OsARF25 gene organ

[0048] Extract total RNA from rice roots, stems, and leaves respectively, use a reverse transcription kit to reverse transcribe the total RNA into cDNA, and use primers SEQ ID NO.5 and SEQ ID NO.6 to perform Real-time PCR detection, such as figure 1 shown. The results showed that the gene was constitutively expressed, with the lowest expression level in leaves and the highest expression level in roots.

Embodiment 3

[0049] Example 3 Analysis of the expression pattern of rice gene OsARF25 under drought, high-salt stress and plant hormone treatment

[0050] Two-week-old rice seedlings were subjected to 150mM NaCl and 20% PEG drought treatment for 12 hours, and total RNA was extracted from the roots after 0, 2, 4, 8, and 12 hours of treatment; , 0.25, 0.5, 1, 1.5, 2, 4, 8, 12h after the total RNA in the leaves; 50μM IAA, 6-BA, KT, ABA, GA soaking treatment for 8h, extract the total RNA in the root. The total RNA was reverse-transcribed into cDNA using a reverse transcription kit, and Real-time PCR detection was performed using primers SEQ ID NO.5 and SEQ ID NO.6. The results showed that the expression level of OsARF25 increased rapidly after NaCl treatment, reached the maximum at 8h, and the expression level increased significantly, as shown in figure 2 As shown in a, after being treated with 20% PEG8000 for 4 hours, the expression level of OsARF25 was significantly increased and reached t...

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Abstract

The invention belongs to the technical field of genetic engineering and specifically relates to an encoding sequence of an auxin response factor gene which is involved in rice root growth and expressed in rice and application of the encoding sequence. The gene cloned from rice is recorded as OsARF25, and the total length is 6057 bp, wherein the open reading frame is 2700 bp, and the nucleotide sequence of the gene is shown as SEQ ID NO.1. The application specifically includes cloning of the gene OsARF25, spatial expression patterns of different organs and tissue which are endogenous to the rice gene, and changes of the expression amount of OsARF25 after different hormone treatments and different stress treatments. Sequencing identification of a Tilling mutant of the gene OsARF25 is disclosed. Phenotypic identification of the mutant shows that the OsARF25 plays an important role in plant root growth. The gene OsARF25 can be used for the study of a rice root growth mechanism and has a certain guiding effect on plant improvement.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and specifically relates to isolated DNA molecules and encoded protein molecules and applications thereof, namely the OsARF25 gene coding sequence expressed in rice and applications thereof. Background technique [0002] Root system is the general term for all roots of plants. It is a bridge for material exchange and information exchange between plants and soil, and plays a vital role in plant growth and development (Hochholdinger et al., 2004). It is also a source of plant hormones, organic acids and An important place for the synthesis and transformation of amino acids and other substances. Rice belongs to monocotyledonous plants, and its root system consists of seed roots, adventitious roots and lateral roots (Shi Qinghua., 1988). The growth of the rice root system directly affects the growth and development of the aboveground part and the formation of yield, and is the key factor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/11C07K14/415C12Q1/686A01H5/10A01H6/46
CPCC07K14/415C12N15/8261C12N15/8267C12Q1/686
Inventor 明凤盖若男王玉霞毛婵娟周子硕
Owner SHANGHAI NORMAL UNIVERSITY
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