Cell wall acid invertase inhibitor gene lccif and its application

An acid invertase and inhibitor technology, used in applications, genetic engineering, plant genetic improvement, etc., can solve the problems of unclear molecular mechanism research, poor fruit setting, etc. early developmental effects

Active Publication Date: 2018-11-27
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The most important reason is that the early endosperm abortion of Nuomi Ci fruit leads to poor fruit setting, and the molecular mechanism of Nuomi Ci early endosperm abortion is not clear in the prior art.

Method used

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  • Cell wall acid invertase inhibitor gene lccif and its application
  • Cell wall acid invertase inhibitor gene lccif and its application
  • Cell wall acid invertase inhibitor gene lccif and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Cloning of litchi cell wall acid invertase inhibitor gene

[0023] 1. RNA extraction and quality detection of litchi samples

[0024] Huayueyang ultra-fast RNA extraction kit (refer to the instruction manual for the specific method) was used to analyze the leaves, flowers, pericarp, fruit pedicle, seed coat, seed kernel and other tissues of "feizixiao", "black leaf" and "nuomici" litchi The RNA was extracted, and the concentration of RNA was measured with an ultraviolet nucleic acid protein detector. Take 1 μl to measure the ratio of 260 nm to 280 nm. If it is between 1.80 and 2.00, the next experiment can be carried out. If the RNA concentration is high, it can be diluted to a certain number of times before detection.

[0025] The results of agarose gel electrophoresis figure 1 , it can be seen that RNA presents two clear bands of 28S and 18S, and the brightness of 28S is about twice that of 18S, there is no tailing phenomenon, and no DNA pollution. It sh...

Embodiment 2

[0028] Example 2 Functional verification of litchi cell wall acid invertase inhibitor gene

[0029] one, LCIF Cloning of gene fragments

[0030] Take lychees LCIF Design primers for fragments of about 500bp, and add pTRV2 before the primers Bam HI and Sma I. A 15 bp carrier sequence adapter homologous to the restriction site. The primer sequences are shown in Table 1 below. The PCR amplification conditions are 94°C, 2min; 98°C, 10s, 55°C, 30s, 72°C, 30s, 30 cycles ; 72°C, 10 min. Take 2 μl for detection by agarose gel electrophoresis, and purify and recover.

[0031]

[0032] 2. Digestion of virus silencing vector pTRV2 (pYL156)

[0033] The structure diagram of virus silencing vector pTRV1 (pYL192), pTRV2 (pYL156) is as follows image 3 , select pTRV2 (pYL156) on Bam HI and Sma Carry out double restriction endonucleases on the vector with two enzyme cutting sites, and the restriction endonuclease is selected from NEB (NewEngland Biolabs) company Bam H I HF a...

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Abstract

Belonging to the technical field of molecular biology, the invention in particular discloses a cell wall acid invertase inhibitor gene LcCIF and application thereof. The LcCIF nucleotide sequence is shown as SEQ ID No:1. The inhibitor can combine with cell wall acid invertase to inactivate it, through short-term silencing of the cell wall acid invertase inhibitor, the cell wall acid invertase activity can be enhanced, and liquid endosperm early development can be promoted. The cell wall acid invertase inhibitor gene LcCIF can be used for improving the fruit setting rate of Nuomici litchi, and has wide application prospect in production.

Description

technical field [0001] The invention relates to the field of molecular biology technology, more specifically, to the cell wall acid invertase inhibitor gene LCIF and its application. Background technique [0002] Endosperm is ubiquitous in angiosperm seeds. The development of endosperm is an important component of embryo development. The fertilized polar nucleus begins to undergo mitosis without dormancy, and a large number of endosperm cells are formed after multiple divisions. The main function of the endosperm is to provide for the developing embryo Nutrition . 3 days after flowering and pollination of litchi, the fusion of sperm cells can be seen in the section. 6 days after flowering, a small amount of endosperm (free nucleus) exists. 20-30 days after flowering, the endosperm in the embryo sac is already very rich, and by 50 days after flowering In about a day, the endosperm cells have all been absorbed. Some studies believe that the development of the endosperm d...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/11C12N15/84A01H5/00A01H6/00
Inventor 王惠聪赵杰堂张洁琼吴子辰刘恋黄旭明
Owner SOUTH CHINA AGRI UNIV
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