Application of larch miR166a in plant development regulation and control

A plant and gymnosperm technology is applied in the field of plant molecular biology to achieve the effects of increasing plant growth rate and plant survival rate

Inactive Publication Date: 2015-03-25
INST OF FORESTRY CHINESE ACAD OF FORESTRY
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  • Abstract
  • Description
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  • Application Information

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  • Application of larch miR166a in plant development regulation and control
  • Application of larch miR166a in plant development regulation and control
  • Application of larch miR166a in plant development regulation and control

Examples

Experimental program
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Effect test

Embodiment 1

[0054] Example 1. Acquisition and identification of miR166a overexpression transgenic plants

[0055] The miR166a involved in this example is miR166a of Japanese larch (Larix leptolepis). Among them, the sequence of the mature miR166a sequence is sequence 1 in the sequence listing; the sequence of the precursor miR166a sequence is sequence 2 in the sequence listing; the full-length cDNA of the precursor miR166a sequence is sequence 3 in the sequence listing, named LaMIR166a gene. Among them, the RNA secondary structure of precursor miR166a is as follows figure 1 shown.

[0056] 1. Construction of recombinant expression vector pSuper::MIR166a

[0057] 1. Primer design

[0058] According to the full-length cDNA (LaMIR166a gene) of the precursor miR166a sequence shown in Sequence 3 in the sequence listing, primers were designed at about 200 bp at both ends of the miR166a stem-loop structure sequence on the cDNA, as follows:

[0059] Primer 1: 5'-CCC AAGCTT GCAACACCATACTTACG...

Embodiment 2

[0111] Embodiment 2, functional identification of transgenic larch

[0112] The transgenic larch a4 transformed into the recombinant expression vector pSuper::MIR166a obtained in Example 1, the transgenic larch transformed into pSuper1300+empty vector, and the non-transgenic larch were used as experimental materials. The mature somatic embryos of each experimental material were placed on the rooting medium a17 (1 / 2MS+IBA1.0mg / L+sucrose 20g / L+agar 5.5g / L), treated at 4°C in a dark environment for 7 days, and then placed in Cultivate under the conditions of 25°C and light (16h / 8h), observe and record the plant morphology, plant growth rate, and root system development of each experimental material. After the plants of each experimental material took root and germinated, the well-grown tissue culture seedlings were transplanted to the greenhouse for further cultivation. Experiments were repeated three times.

[0113] The results showed that when cultured on the rooting medium a...

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Abstract

The invention discloses an application of larch miR166a in plant development regulation and control. The application provided by the invention specifically is the application of nucleic acid molecules shown in any of (1)-(4) in plant development regulation and control, namely (1) RNA molecules shown in sequence 1 in a sequence table; (2) RNA molecules shown in sequence 2 in the sequence table; (3) RNA molecules shown in sequence 3 in the sequence table; (4) DNA molecules shown in 27th-539th of the sequence 3 in the sequence table. Experiments prove that the transgenic plants with overexpressed LaMIR166a genes generate lots of lateral roots and grow well when cultured on a rooting culture medium a17 for 4 months; non-transgenic control plants only elongate main roots, do not generate lateral roots and relatively grow slowly. The result indicates that the overexpressed miR166a is capable of promoting the growth speed of larch tissue culture seedling and the generations of the lateral roots, thus increasing the transplanting survival rate.

Description

technical field [0001] The invention belongs to the technical field of plant molecular biology, and relates to the application of larch miR166a in regulating plant development, in particular to the application of larix miR166a in promoting the development of plant lateral roots. Background technique [0002] Larch (Larix spp.) is an important fast-growing coniferous tree species for afforestation in northern my country, which can be used as raw materials for wood and papermaking. From a global perspective, larch is naturally distributed in temperate mountainous areas, cold temperate plains, and alpine climate zones. The first two zones are roughly between 48° and 60° north latitude, and have the characteristics of high latitude and high altitude. There are 10 species and 5 varieties of larch naturally distributed in my country. They are distributed in the mountainous and alpine areas of Northwest, North, Central, Northeast and Southwest respectively. Vigorously carrying ou...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/00
Inventor 张守攻李水根齐力旺张立峰韩素英朱彩虹
Owner INST OF FORESTRY CHINESE ACAD OF FORESTRY
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