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A method for producing regenerated rice tillers in advance to improve the yield of regenerated rice

A technology for ratooning rice and yield, applied in the field of agricultural biology, can solve the problems of short growth period, lack of application value, and less time for photosynthesis, and achieve the effects of increasing yield, good grain filling, and prolonging the growth period

Active Publication Date: 2019-03-12
HUNAN AGRICULTURAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because the number of tillers in multi-tiller transgenic rice materials obtained through transgenic technology is often too large, which is unfavorable to the rice yield in the first season of rice, therefore, even if it can increase the tillering ability of rice or the number of spikes of regenerated rice, it is still lacking in actual production. Value
[0006] So far, the rice varieties (materials) with multi-tiller characteristics selected by traditional breeding methods have not been ideal for increasing yield
Even if a variety with multiple regenerated rice spikes is obtained, the yield increase effect is not good due to the short growth period resulting in less photosynthesis time, the length of the spike and the filling degree of the grain.

Method used

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  • A method for producing regenerated rice tillers in advance to improve the yield of regenerated rice
  • A method for producing regenerated rice tillers in advance to improve the yield of regenerated rice
  • A method for producing regenerated rice tillers in advance to improve the yield of regenerated rice

Examples

Experimental program
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Effect test

Embodiment 1

[0017] Example 1 Cloning of 12 different rice genes in OsMIR156a-l

[0018] Using the online Primer-BLAST software, the upstream and downstream primers were designed for 12 different OsMIR156a~l genes. The main setting or modification parameters during primer design include: PCR product size is 150-1000bp; Primermelting temperatures (Tm) is 55-65°C; Organism is set to Oryza sativa (japonicacultivar-group) (taxid: 39947); Database selection It is Refseq representative genomes, and the rest of the parameters are the default values ​​of the system. After the primers were set, they were sent to Huada Genomics or Wuhan Jinkairui Biological Company for primer synthesis.

[0019] The CTAB method was used to extract DNA from Nipponbare, Zhonghua 11 or other rice materials, and the synthetic primers corresponding to different OsMIR156a~l genes designed by the above method were used for PCR amplification. The PCR reaction system was 50 μL: DNA template 1.0 μL, normal Direct primer 1.0...

Embodiment 2

[0032] Example 2 Cloning of Tissue and Organ-Specific Expression Promoter Sequences

[0033] The stem-specific expression promoter is preferably the promoter of OsGA3ox2 gene. The about 2.1Kb promoter of the OsGA3ox2 gene (referred to as the D18 promoter), wherein the relevant information of the D18 promoter sequence is reported in the literature (SakamotoT, Morinaka Y, Ishiyama K, Kobayashi M, Itoh H, Kayano T, Iwahori S, MatsToka M, Tanaka H. Genetic manipulation of gibberellin metabolism in transgenic rice. Nat Biotechnology. 2003, 21:909-913). Utilize the PCR method described in this reference to obtain the fragment and connect it to the pBluescriptSK+(pBS) carrier fragment obtained by EcoR V enzyme digestion to obtain the intermediate vector pBS-D18p, and send it to Huada Gene Biotechnology Company for sequencing analysis, and the sequencing result shows that it is consistent with the obtained Knowing that the sequences were completely consistent, the pBS-D18p plasmid wa...

Embodiment 3

[0034] Example 3 Construction of 12 stem-specific expression vectors pWM-D18p-OsMIR156a~l (abbreviated as D18Pro-OsMIR156a~l)

[0035] After digesting the vector pWM101 and BamH I / sal I with HindⅢ / sal I and the intermediate vector pBS-OsMIR156a~l respectively, the required fragments were recovered, and the three fragments were ligated with T4ligase and D18p promoter. The system is: HindⅢ / sal I digestion vector pWM101 product 1.0 μl, HindⅢ / BamH I digestion pBS-D18p plasmid 1.0 μl, BamH I / sal I digestion intermediate vector pBS-OsMIR156a~l 1.5 μl, 5×T4ligaseBuffer 1.0 μl and T4ligase 0.5 μl, the above reaction ligation system was kept at 16°C for 30min or at 4°C overnight. Then adopt conventional molecular test methods to transform the ligation product into strains of competent Escherichia coli, and use Kan + Through resistance screening, 12 different vectors pWM-D18p-OsMIR156a~l were obtained.

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Abstract

The invention relates to a method for producing ratooning rice tillering in advance and further increasing the yield of ratooning rice. The method comprises the following steps: specifically expressing nucleotide sequences capable of influencing the tillering development of the rice such as OsMIR156a-1 gene sequences shown as SEQ ID No.1 to SEQ ID NO. 12 in a sequence table in a rice stem by utilizing a rice stem specific expression promoter. An effect of a transgenic plant obtained by implementing the technology shows that compared with a wild type, when the specific promoter is used for overexpressing the OsmiR156 transgenic plant, the transgenic strain specifically expressing D18pro-OsMIR156 in the tissue and organ can form the tillering at the mature period of a first rice crop, but the tillering growing in advance has no obvious influence on characters related to the yield such as thousand seed weight of the first rice crop; however, the height and the rice ear length of the ratooning rice can be increased, the grain plumpness can be increased, and the yield of the ratooning rice is further increased.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and specifically relates to a method for producing ratooned rice tillers in advance to increase the yield of ratooned rice, more specifically, using a specific promoter to specifically express the OsmiR156 family gene in stems to generate ratooned rice tillers in advance and further A method for increasing the yield of ratooning rice. Background technique [0002] Since the beginning of the 20th century, the production of ratooning rice in my country has entered a period of rapid development, and has successively experienced development stages such as the combined utilization of high-stalk ratooning rice, the combined utilization of dwarf-stalk ratooning rice, and the regeneration and utilization of hybrid rice. The research progressed rapidly, and a batch of new varieties (combinations) with good rice quality and strong regeneration ability were successively obtained, such as Pei 64 / E32,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82A01H6/46
CPCC12N15/8205C12N15/8226C12N15/8261C12N2800/60C12N2840/007
Inventor 刘清王若仲肖浪涛
Owner HUNAN AGRICULTURAL UNIV
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