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Phyllostachys edulis senescence related NAP (neutrophil alkaline phosphatase) transcription factor as well as encoding gene and application thereof

A gene and bamboo technology, applied in the senescence-related NAP transcription factor of moso bamboo and its encoded gene and application field, can solve the problem of lack of regulatory function analysis, specificity and complexity increasing the difficulty of research on the senescence mechanism of moso bamboo, and unclear senescence regulation mechanism, etc. problem, to promote the aging process, the effect is significant

Active Publication Date: 2020-04-28
INT CENT FOR BAMBOO & RATTAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, the specificity and complexity of biological characteristics and senescence types of moso bamboo increase the difficulty of studying the senescence mechanism of moso bamboo
Therefore, there are very few studies on the aging phenomenon and aging mechanism of moso bamboo, and the aging regulation mechanism is even less clear, and the research on NAP members in moso bamboo aging is limited to evolutionary analysis and gene PeNAC1 (NAP subfamily member) cloning, lack of regulation Function Analysis

Method used

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  • Phyllostachys edulis senescence related NAP (neutrophil alkaline phosphatase) transcription factor as well as encoding gene and application thereof
  • Phyllostachys edulis senescence related NAP (neutrophil alkaline phosphatase) transcription factor as well as encoding gene and application thereof
  • Phyllostachys edulis senescence related NAP (neutrophil alkaline phosphatase) transcription factor as well as encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Analysis and identification of the senescence-related gene PheNAP8 in Phyllostachys pubescens

[0054] 1) According to Li (2019) results of postharvest senescence transcriptome sequencing, the number of up-regulated expression among NAC family members was the largest, among which there were 11 PheNAC genes whose expression was basically not detected at 0h postharvest (RPKM<0.3). The expression was significantly upregulated during storage (Table 1). Among them, PheNAP8 has a similar expression pattern, all of which are rapidly up-regulated at 12-24 hours after harvest, and the up-regulation fold (greater than 120) is higher than that of other PheNAC genes, which has an extremely significant or even unexpected up-regulation effect.

[0055] Table 1. Expression analysis of 11 PheNAC genes in Phyllostachys pubescens after harvest (transcriptome sequencing)

[0056]

[0057]

[0058] 2) Expression analysis of PheNAP8 in seedling leaves and flowering leaves o...

Embodiment 2

[0067] Example 2 PheNAP8 gene evolution analysis.

[0068] The amino acid sequences of the reported NAP subfamily members were downloaded from the NCBI database, respectively. Through ClustalX1.83 (http: / / www.clustal.org / ), the protein sequence encoded by the PheNAP8 gene of Phyllostachys pubescens was compared with the NAP protein sequences of other species (Thompson et al., 1997), and MEGA 7.0 A rootless phylogenetic tree was constructed using the neighbor-joining method (bootstrap value was set to 1000). The results showed that PheNAP8 of Phyllostachys pubescens had high homology with other reported NAP transcription factors, and PheNAP8 had the highest sequence similarity with BeNAC1 of Bamboo chinensis ( figure 2 ), but there is no 100% homologous gene, which proves that PheNAP8 is a brand-new gene that has not been reported in the prior art, and is a kind of NAC transcription factor. Members of the NAP subfamily mainly come from Arabidopsis (ANAC018, NP_175696; ANAC02...

Embodiment 3

[0069]Example 3 Acquisition of the gene encoding the PheNAP8 transcription factor of Phyllostachys pubescens

[0070] The material used in this example is moso bamboo leaves, which are stored at -80°C after quick-freezing in liquid nitrogen.

[0071] 1) Extraction of total RNA from leaves of Phyllostachys pubescens

[0072] Carry out the extraction of total RNA and the synthesis of first-strand cDNA according to the method of embodiment 1

[0073] 3) RT-PCR amplification of the PheNAP8 gene sequence

[0074] The full-length primers were designed according to the Phyllostachys pubescens genome database, which are: NAP1-F: 5'-ATGATGATGTCGAACCCGG-3' (SEQ ID NO.7) and NAP1-R: 5'-TCAGTTCATCCCCATGTTTGAAT-3' (SEQ ID NO.8). The reverse transcription product was used as a template, and NAP1-F and NAP1-R were used as primers for PCR amplification. PCR products were subjected to agarose gel electrophoresis ( image 3 ), recover the target product, connect with pGEMT-easy, transform E...

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Abstract

The invention discloses a phyllostachys edulis senescence related NAP (neutrophil alkaline phosphatase) transcription factor as well as an encoding gene and application thereof. The nucleotide sequence of the factor is shown in SEQ ID NO.1 in the description, and the protein sequence of the factor is shown in SEQ ID NO.2 in the description. Through cloning and expression of a PheNAP8 gene of phyllostachys edulis and allogenic transformation of arabidopsis thaliana, it finds that a transgenic arabidopsis thaliana plant with over-expressed PheNAP8 has remarkable phenotypes of growth retardationand pro-senescence, and it means that the gene has significant functions in senescence regulation and breeding of the phyllostachys edulis, can be applied to screening and identification on phyllostachys edulis varieties prone to senescence, and is applicable to industrial application.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and in particular relates to an aging-related NAP transcription factor of moso bamboo, its encoding gene and its application. Background technique [0002] Flowering senescence, shoot senescence, and post-harvest rapid senescence of bamboo shoots can all cause serious economic losses and ecological problems. At present, there are very few studies on the aging phenomenon and aging mechanism of moso bamboo, and the aging regulation mechanism is even less clear. Moso bamboo is a perennial, lignified herbaceous plant. In addition to the common individual aging, organ aging and gradual aging, its aging types also include group aging, stand aging and shoot senescence. The various aging processes of moso bamboo overlap with each other. The specificity and complexity of the biological characteristics and senescence types of Moso bamboo increase the difficulty of studying the senescence ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/11C12N15/82C12Q1/6895A01H5/12A01H6/20A01H6/46
CPCC07K14/415C12N15/8266C12Q1/6895C12Q2600/13C12Q2600/158
Inventor 高健谢丽华
Owner INT CENT FOR BAMBOO & RATTAN
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