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TaPPH (pheophytin pheophorbide hydrolase) wheat gene and encoded protein and cloning method of gene

A technology of pheophytin and pheophorbide is applied in the field of wheat genes, the key enzyme genes in the chlorophyll degradation process of wheat leaves, and can solve the problems of yield decline, inconsistent conclusions, and photosynthetic capacity decline, and achieve the effect of delaying senescence.

Pending Publication Date: 2016-12-07
SHANXI AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

4. Even for the same plant species, the conclusions of different researchers are inconsistent
[0005] The degradation of chlorophyll in wheat leaves will lead to a decrease in its photosynthetic capacity, which in turn affects the growth and development of wheat plants, and ultimately leads to a decrease in yield

Method used

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  • TaPPH (pheophytin pheophorbide hydrolase) wheat gene and encoded protein and cloning method of gene
  • TaPPH (pheophytin pheophorbide hydrolase) wheat gene and encoded protein and cloning method of gene
  • TaPPH (pheophytin pheophorbide hydrolase) wheat gene and encoded protein and cloning method of gene

Examples

Experimental program
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Effect test

Embodiment 1

[0024] Example 1: PPPHmolecular cloning.

[0025] Select full-grained Chang 4738 wheat seeds, place them in a petri dish, add deionized water, plant them in flower pots after germination, and place them in an artificial climate box for cultivation. The photoperiod is 12 hours of light, 12 hours of darkness, humidity 80%, and temperature 25 ℃. Wheat leaves were taken at the three-leaf stage, frozen in liquid nitrogen, and stored in a -80°C ultra-low temperature freezer for later use.

[0026] Total RNA from wheat seedling leaves was extracted using the Plant RNA Kit provided by Omega.

[0027] Using good quality wheat leaf total RNA as a template, using Trans Script ® All-in-One SuperMix for PCR synthesized cDNA by reverse transcription, and stored the product at -20°C for future use.

[0028] Arabidopsis AtPPH The gene sequence (GenBank registration number: NM_121383) was used as the query sequence, and the blastn program was used to search the Expressed sequence tags ...

Embodiment 2

[0045] Example 2: PPPH Sequence information and characterization of genes.

[0046] PPPH The full-length cDNA of the gene is 2490bp, including an open reading frame (open reading frame, ORF) of 1032bp, a 5' untranslated region (untranslated region, UTR) of 698bp and a 3'UTR of 760bp. The gene encodes a protein TaPPH consisting of 343 amino acids, and its molecular weight is predicted to be about 38.65kDa. TaPPH has the conserved domains possessed by PPH proteins: esterase-lipase superfamily domain and α / β hydrolase catalytic domain. Protein sequence comparison found that TaPPH has high similarity with PPH proteins of other plants, among which the similarity with OsPPH of rice is the highest, reaching 91%.

Embodiment 3

[0047] Example 3: PPPH Gene expression studies.

[0048] Using real-time quantitative PCR (quantitative real-time PCR, qPCR) method to analyze different wheat tissues, wheat leaves after dark induction and exogenous hormone treatment PPPH gene expression.

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Abstract

The invention discloses a TaPPH (pheophytin pheophorbide hydrolase) wheat gene and encoded protein and cloning method of the gene. A gene sequence is shown as SEQ ID No. 2 while an amino acid sequence of the protein is shown as SEQ ID No. 3. Expression quantities of different tissues of wheat are different, however, the expression quantities are the highest in leaves; expression quantities of the TaPPH are increased through dark treatment and exogenous application of plant hormones of abscisic acid (ABA) and ethephon (Eth). The gene is beneficial to discovery of degradation mechanism of chlorophyll of wheat leaves during aging, and a foundation is provided for breeding stay-green wheat varieties by means of biological engineering.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering and relates to a wheat gene, in particular to a key enzyme gene in the degradation process of wheat leaf chlorophyll. The present invention also relates to the cloning of the gene and its encoded protein. Background technique [0002] Wheat is the main food crop in our country. Two-thirds of wheat grain yield comes from assimilated products synthesized after anthesis. The senescence of wheat flag leaves is synchronized with grain formation and grain filling maturity. Premature senescence of flag leaves will seriously affect wheat yield, delay the occurrence of flag leaf senescence, and delay its Senescence can significantly increase wheat yield and improve grain quality. According to theoretical calculations, in the mature stage of crops, prolonging the life of functional leaves by one day can increase the yield by about 2%. Therefore, delaying wheat leaf senescence (keeping green) ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/55C12N9/18
CPCC12N9/18C12Y301/01082
Inventor 王曙光孙黛珍王绘艳马昊翔王玉斌周福平
Owner SHANXI AGRI UNIV