Liriodendron hybrids LhMKK2 gene and expression protein and application thereof

A technology of hybridizing Liriodendron tulipifera and genes, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problems of slow speed, shortage of good seedlings, and restriction of good hybrid plants, so as to improve stress resistance, stress response, The effect of good industrial application prospects

Active Publication Date: 2015-01-21
NANJING FORESTRY UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, hybrid Liriodendron is mainly propagated through traditional asexual propagation methods such as cuttings and grafting, and sexual propagation methods of artificial hybridization. However, due to the influence of seasons and hybrid seed productio

Method used

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  • Liriodendron hybrids LhMKK2 gene and expression protein and application thereof
  • Liriodendron hybrids LhMKK2 gene and expression protein and application thereof
  • Liriodendron hybrids LhMKK2 gene and expression protein and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Using hybrid Liriodendron tulipifera plants as materials, total RNA was extracted and reverse-transcribed into cDNA, corresponding primers were designed for PCR, and after detection by agarose gel electrophoresis, the target band was recovered and compared with pMD TM The 19-T Vector (Takara D102A) vector was ligated, transformed into Escherichia coli, sequenced and analyzed. Pick positive clones for plasmid extraction, add appropriate restriction sites, and simultaneously double-enzyme digest with the expression vector PBI121, connect them under the action of T4 ligase, and transfer them into Agrobacterium strain GV3101, and wait for Arabidopsis to age , transformed by flower organ immersion transformation method, observed T1, T2 phenotypes, carried out statistical analysis, and carried out stress resistance treatment to analyze its function.

[0029] (1) Extraction of total RNA

[0030] Using the young leaves of Liriodendron chinensis as material, according to the NO...

Embodiment 2

[0050] (1) Gene function analysis

[0051] First construct the hybrid Liriodendron 35S: LhMKK2 Overexpress the vector and transfer it into Agrobacterium strains, and transform two wild-type Arabidopsis Col (Columbia) and Landsberg Ler (Landsberg) by inflorescence soaking method to obtain overexpression LhMKK2 Gene-positive transgenic plants, and stress-treated T2 generation positive plants, research and analysis of hybrid Liriodendron LhMKK2 function.

[0052] (2) Construction of the carrier

[0053] The Escherichia coli strain used in the present invention is E. coli JM109 (purchased from Biovector (Dalian) Co., Ltd.); the expression vector is pBI121 (purchased from Biovector Co., LTD).

[0054] The specific process is as follows:

[0055] 1. By PCR in LhMKK2 Xba I and Sac I double restriction sites were added to the upstream and downstream of the gene fragment respectively. The PCR system and reaction conditions were the same as those for full-length amplification. The ...

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Abstract

The invention discloses a Liriodendron hybrids LhMKK2 gene, expression protein of the Liriodendron hybrids LhMKK2 gene and application of the liriodendron hybrids LhMKK2 gene. The nucleotide sequence is showed in SEQID NO.1 and the amino acid sequence is showed in SEQID NO.2. On the basis of the completely established somatic embryogenesis system of the Liriodendron hybrids and the fact that part of the transcriptome data is already obtained, the full length of the Liriodendron hybrids LhMKK2 gene, through the adoption of homology-based cloning, is obtained and is named as LhMKK2. Through gene expression analysis, that the Liriodendron hybrids LhMKK2 gene is widely expressed in plants and is involved in the growth and development of plants, and the adversity stress response process are proved; through functional analysis of the gene, it is discovered that the overexpression LhMKK2 gene is capable of enhancing resistance of plants to salt stress response, influencing the growth and development of plants and providing theoretical basis for stress resistance molecular breeding of Liriodendron hybrids and can be further widely used in adversity resistance stress of plants.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, in particular to a hybrid tulip tree LhMKK2 Genes and their expressed proteins and applications. Background technique [0002] Hybrid Liriodendron is an interspecies obtained from the hybrid experiment between Chinese mandarin tree (L.chinese (HEMSL.) Sarg) and North American Liriodendron (L.tulipifera Linn.) by Professor Ye Peizhong, a famous tree genetic breeder at Nanjing Forestry University. Hybrids with obvious heterosis. Because of its fast growth speed, strong resistance to diseases and insect pests, fine material texture, peculiar leaf shape and fast growth speed, hybrid tulipwood is an excellent tree species for afforestation on barren hills, industrial timber, and ornamental gardens. It is also used for garden greening, afforestation and timber Tree species with great market potential. At present, hybrid Liriodendron is mainly propagated through traditional asexual ...

Claims

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Application Information

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IPC IPC(8): C12N15/54C12N9/12C12N15/84A01H5/00
Inventor 施季森赵芳芳陈金慧王鹏凯郝兆东闫珊施帅男孟岩陆叶
Owner NANJING FORESTRY UNIV
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