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Application of wheat TaMADS6 gene in regulation of plant spike and grain development as well as flowering time

A flowering time and plant technology, applied in the field of plant molecular biology, can solve problems such as few reports on wheat function research and production application research, and achieve the effects of increasing the number of cob ears, delaying flowering time, and high application value.

Inactive Publication Date: 2020-02-18
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, there are few reports on the functional research and production application of TaMADS6 in wheat.

Method used

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  • Application of wheat TaMADS6 gene in regulation of plant spike and grain development as well as flowering time
  • Application of wheat TaMADS6 gene in regulation of plant spike and grain development as well as flowering time
  • Application of wheat TaMADS6 gene in regulation of plant spike and grain development as well as flowering time

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Construction of pAHC25-6*MYC-TaMADS6-D intermediate vector

[0048] To construct the overexpression vector of the wheat TaMADS6-D gene, first connect the TaMADS6-D gene to the pAHC25-6*MYC vector (pAHC25-6*MYC is a vector transformed by pAHC25, replace the GUS sequence in pAHC25 with 6*MYC Sequence, the plasmid map of pAHC25 as figure 1 shown), construct the intermediate vector pAHC25-6*MYC-TaMADS6-D, the specific method is as follows:

[0049] (1) Use primers with sequences such as SEQ ID NO.7 and SEQ ID NO.8 to amplify the TaMADS6-D fragment with Spe I and Sac I restriction sites using the cDNA of wheat tissue as a template, and use gel to recover The kit recovers the target fragment;

[0050] (2) double-enzyme digestion of pAHC25-6*MYC and the purified product in step (1), and recovery of the linearized vector and target fragment using a gel recovery kit;

[0051] (3) Ligation: The reaction system is: 2 μL of target fragment, 2 μL of linearized vector, 1...

Embodiment 2

[0053] Example 2 Construction of pCAMBIA3300-Ubi-MCS-6*MYC-TaMADS6-D overexpression vector

[0054] The overexpression vector used to overexpress the TaMADS6-D gene in wheat is pCAMBIA3300-UbiMCS (transformed with the pCAMBIA3300 vector as the backbone, and the Ubiquition promoter of corn is added before the multiple cloning site, and the herbicide-resistant Bar gene is driven by the 35s promoter , graph such as figure 2 shown).

[0055] Use the primer shown in SEQ ID NO.9 and SEQ ID NO.10 (the primer shown in SEQ ID NO.9 has a BamH I restriction site, and the primer shown in SEQ ID NO.10 has a Sma I restriction site;) from the pAHC25-6*MYC-TaMADS6-D intermediate vector constructed in Example 1, amplify the fusion gene fragment with 6*MYC and TaMADS6-D gene fragment, i.e. 6*MYC-TaMADS6 -D.

[0056] Digestion, ligation, transformation, and positive clone sequencing are the same as steps (2) to (4) of Example 1. The sequencing primers are SEQ ID NO.11 and SEQ ID NO.12. 6*MY...

Embodiment 3

[0057] Example 3 Construction of transgenic wheat overexpressing TaMADS6-D gene

[0058] 1. Transformation of TaMADS6-D gene overexpression vector into Agrobacterium

[0059] (1) Take out Agrobacterium competent C58C1 from -80°C refrigerator and melt on ice;

[0060] (2) Divide each competent tube into two tubes, add 2 μL pCAMBIA3300-Ubi-MCS-6*MYC-TaMADS6-D plasmid, flick and mix well, and place on ice for 30 minutes;

[0061] (3) Liquid nitrogen for 5 minutes;

[0062] (4) 37°C water bath for 5 minutes;

[0063] (5) 5 minutes on ice;

[0064] (6) In a clean bench, add 400 μL of anti-antibiotic-free LB medium to each tube, flick and mix evenly, 28°C, 200 r / min, and shake for 3-4 hours;

[0065] (7) The bacterial solution was applied to the solid plate containing Rif and Kan, sealed with a parafilm, and cultured upside down in an incubator at 28°C for 2 days;

[0066] (8) pick single clones in LB medium (containing Rif and carrier-resistant antibiotics), shake culture at 2...

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Abstract

The invention relates to the technical field of plant molecular biology, and specifically relates to application of a wheat TaMADS6 gene in regulation of plant spike and grain development as well as flowering time. It is discovered by the invention that a TaMADS6 protein has the functions of regulating plant flowering as well as spike and grain developing; and moreover, number of spikelets, numberof spikelet stalks and grain number per spike are significantly increased with flowering time obviously delayed by increasing expression of the TaMADS6 protein in plants. The TaMADS6 has relatively high application values, and can be applied to genetic breeding and germplasm resource improvement of the wheat; and thus, gene resources and new ideas are provided for cultivation of high-yield plants. Therefore, the TaMADS6 is of great significance to agricultural production.

Description

technical field [0001] The invention relates to the technical field of plant molecular biology, in particular to the application of wheat TaMADS6 gene in regulating the development of plant ears and grains and flowering time. Background technique [0002] Wheat is an important food crop, and about 40% of people in the world take wheat as their main food. At the same time, wheat, as one of the three major grains, produces almost all of it for food, and is the food crop with the world's total output second only to corn. With the increase of consumption level, higher requirements are put forward for the yield and quality of wheat. The three elements of wheat yield are the number of spikes per unit area, the number of grains per spike, and the weight of grains. The coordination of the relationship between the three is of great significance in improving wheat yield. Among them, the number of grains per spike depends on the number of spikelets and the development of florets, so ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C07K14/415C12N15/29C12N15/11A01H5/00A01H6/46
CPCC07K14/415C12N15/8261C12N15/827
Inventor 李爱丽孔星辰陶姝邓中印耿帅锋毛龙
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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