Application of paddy rice genes OsMADS25 to promoting tap root system plant lateral root growth

A gene and rice technology, applied in the application field of rice OsMADS25 gene in promoting the growth of lateral roots of taproot plants, can solve problems such as internal factors that have no profound explanation for changes, and achieve improved nitrogen use efficiency, important theoretical significance and potential applications Value, the effect of promoting early growth and development

Inactive Publication Date: 2014-11-12
ZHEJIANG UNIV
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  • Abstract
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  • Claims
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Problems solved by technology

[0005] Although the above research results have accumulated very valuable raw data for the study of nitrogen on the morphogenesis of rice roots and the uptake and utilization of nitrogen by roots, most of these studies have focused on the observation of phenomena and the determination of changes in substance content. However, there is no profound explanation of the internal cause of the change, so there is still a long way to go to understand the nature of the relationship between nitrogen and rice root growth.

Method used

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  • Application of paddy rice genes OsMADS25 to promoting tap root system plant lateral root growth
  • Application of paddy rice genes OsMADS25 to promoting tap root system plant lateral root growth
  • Application of paddy rice genes OsMADS25 to promoting tap root system plant lateral root growth

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Embodiment 1

[0027] Example 1 Preparation of transgenic Arabidopsis

[0028] (1) Take the roots of rice seedlings, grind them with liquid nitrogen, add TRIzol reagent (Invitrogen Company), and extract RNA. 0.5 μg RNA was taken for reverse transcription reaction to obtain cDNA.

[0029] (2) Design specific primers according to the OsMADS25 gene sequence on Genbank, add SalI and NotI restriction enzyme sites to the 5' ends of the primers respectively, and the sequences are as follows:

[0030] Upstream primer: 5'-GCGTCGACATGGGGAGAGGGAAGATTG-3' (SEQ ID No.2);

[0031] Downstream primer: 5'-TTGCGGCCGCTGTAGTAGGCGTGATGCTG-3' (SEQ ID No. 3).

[0032] (3) Perform PCR amplification using the full-length cDNA as a template.

[0033] high fidelity enzyme HS DNA Polymerase (TaKaRa Code: DR010A) system:

[0034] 5×PrimeSTAR Buffer (Mg 2+ plus) 10 μl,

[0035] dNTP Mixture (2.5mM each) 4μl,

[0036] Upstream primer (10 μM) 1 μl,

[0037] Downstream primer (10 μM) 1 μl,

[0038] Template cDNA ...

Embodiment 2

[0059] The lateral root growth observation statistics of embodiment 2 transgenic Arabidopsis

[0060] Arabidopsis is a taproot plant. The main root grows first, and the lateral roots grow after the main root grows for a period of time. To the obtained transgenic Arabidopsis (T of embodiment 1 0 T obtained after generation seed planting 3 Homozygous) in different concentrations of KNO 3 The growth of the root system on the B5 modified medium was counted.

[0061] Put the plate with transgenic Arabidopsis in the refrigerator at 4°C for two days, then put it vertically into the growth room, check the root system every day, mark the time when the first lateral root occurs and the time when the main root reaches 2cm, and calculate the transgenic Arabidopsis Growth promotion rate of first lateral root and main root relative to wild-type Arabidopsis.

[0062] The calculation formula of Advancement (growth promotion rate, %) is:

[0063] Advancement (growth promotion rate, %) = (...

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Abstract

The invention discloses application of paddy rice genes OsMADS25 to promoting tap root system plant lateral root growth. The base sequence of the genes OsMADS25 is shown in SEQ ID No.1. The application includes the steps that the paddy rice genes OsMADS25 are connected into a plant expression vector, and a recombinant expression vector is obtained through establishment; the recombinant expression vector is transformed into a receiver plant. According to the application, by cloning the paddy rice genes OsMADS25 and conducting genetic transformation on tap root system plants such as arabidopsis thaliana through the genes, external source paddy rice genes OsMADS25 are over-expressed through the plants, and therefore generation and growth of plant lateral roots can be promoted. The test result shows that an external source NO3- and the genes OsMADS25 are cooperative in promoting growth of the receiver plant. By the adoption of the application, nitrogen key controlling genes relevant to paddy rice lateral root growth can be researched, and the application has significance in screening and breeding plant varieties capable of efficiently absorbing nutrients.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to the application of rice OsMADS25 gene in promoting the growth of taproot system lateral roots. Background technique [0002] Rice is one of the most important food crops in the world, and my country is the largest producer and consumer of rice in the world. The successful use of heterosis has greatly increased the yield of rice and made a great contribution to solving the food crisis in the world. But hybrid rice production has stagnated since the 1980s. The input of nitrogen fertilizer in rice production is becoming one of the powerful measures to increase yield, but the input of a large amount of nitrogen fertilizer not only reduces the nitrogen use efficiency, but also brings a series of environmental problems such as waste of resources and eutrophication of water bodies. At present, people adjust in two aspects to improve nitrogen use efficiency, one is t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84C12N1/21A01H5/00C12R1/01
Inventor 甘银波于春燕苏莎黄琳丽徐一淳
Owner ZHEJIANG UNIV
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