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Applications of rice OsPCF7 gene in culturing high-tillering rice varieties

A technology for high tillering and rice, applied in the directions of application, genetic engineering, plant genetic improvement, etc., can solve problems such as no high tillering rice varieties, and achieve the effect of promoting stable rice yield and increasing the number of tillers.

Inactive Publication Date: 2017-02-22
CHONGQING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The nucleotide sequence of the OsPCF7 gene of the rice TCP protein family has been reported in the literature, but so far, there has been no report on the use of this gene in breeding high tillering rice varieties

Method used

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  • Applications of rice OsPCF7 gene in culturing high-tillering rice varieties
  • Applications of rice OsPCF7 gene in culturing high-tillering rice varieties
  • Applications of rice OsPCF7 gene in culturing high-tillering rice varieties

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Experimental program
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Effect test

Embodiment 1

[0030] Embodiment 1, the cloning of rice OsPCF7 gene reading frame fragment

[0031] According to the OsPCF7 gene candidate sequence (NM_001050819) found in GenBank, use the sequence processing online toolkit (SMS) biological software network http: / / www.bio-soft.net / sms to find its largest open reading frame sequence, using PrimerPremier 5.0 The software designed primers OsPCF7-f: 5'-catgccatgggcatgcgcaacgccaagg-3' (SEQ ID No.1, the underlined part is the NcoI restriction site) and OsPCF7-r: 5'-gggtaacctcacctgatcacctcacttcc-3' (SEQ ID No.2, the underlined part is the BstEII restriction site). Take different Zhonghua No. 11 rice seedlings, extract total RNA, and then synthesize the first strand of cDNA according to the instructions of the TaKaRa RNA PCR Kit (AMV) Ver.3.0 kit, and then use the first strand of cDNA as a template , using primers OsPCF7-f and OsPCF7-r to amplify the reading frame sequence of OsPCF7 gene. The PCR amplification system consists of: 2×Taq Mix 25μl, O...

Embodiment 2

[0033]Embodiment 2, the construction of rice OsPCF7 gene plant expression vector

[0034] The purified PCR amplified product was digested with NcoI and BstEII to obtain the open reading frame fragment of the OsPCF7 gene, which was then ligated with the backbone of the vector pCAMBIA1301 that had undergone the same double digestion to obtain the plant overexpression binary vector pCAMBIA1301-OsPCF7. The constructed overexpression binary vector pCAMBIA1301-OsPCF7 was amplified with primers OsPCF7-f and OsPCF7-r for PCR identification, the results were as follows figure 2 shown. The results showed that a DNA fragment with a length of 833bp was obtained, which was consistent with the size of the open reading frame fragment of OsPCF7 gene. At the same time, the constructed overexpression binary vector pCAMBIA1301-OsPCF7 was identified by double enzyme digestion with NcoI and BstEII, and the results were as follows image 3 shown. The lengths of the digested products were 9771bp...

Embodiment 3

[0035] Embodiment 3, the preparation of rice PCF7 gene Agrobacterium engineering strain

[0036] The overexpression binary vector pCAMBIA1301-OsPCF7 was transformed into Agrobacterium tumefaciens AGLO competent cells by liquid nitrogen cold shock method, and spread on YEB solid medium (containing 1.5% (w / w) agar, 50 mg / L kana (Kan), 500 mg / L streptomycin (Sm) and 50 mg / L rifampicin (Rif)), pH 7.0), cultured at 28±2°C in the dark for 2 days, picked single Bacterial colonies were cultured with YEB liquid medium (containing 50mg / L of Kan, 500mg / L of Sm and 50mg / L of Rif, pH7.0) for 2 days, and the bacterial liquid was taken for PCR identification. The positive recombinant was the rice OsPCF7 gene Engineering strains of Agrobacterium, frozen at -80°C, for later use.

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Abstract

The invention discloses applications of a rice OsPCF7 gene in culturing high-tillering rice varieties, wherein the nucleotide sequence of the rice OsPCF7 gene is shown as SEQ ID No.3. The experiment proves that the tiller number of the transgenic positive plants obtained by transforming the rice by adopting the rice OsPCF7 gene is obviously increased compared with that of the non-transgenic rice plants, and therefore, the rice OsPCF7 gene can be used for culturing the high-tillering rice varieties. The invention further discloses applications of plant expression vectors and microbial conversion bodies which contain the rice OsPCF7 gene in culturing the high-tillering rice varieties, therefore, a foundation is laid for improving the tillering capacity of rice and promoting the stable and high yield of rice, and the application prospects are very good.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of rice OsPCF7 gene in cultivating high tillering rice varieties. Background technique [0002] As a typical representative of monocotyledonous plants, rice is one of the three most important food crops in the world. More than half of the people in the world eat rice, and more than one-third of the population uses rice as a staple food. The rapid development of social economy has promoted the growth of the demand for food and increased our demand for food. Nowadays, food safety production has become a serious problem that must be solved, but the developmental morphogenesis of rice plants will directly affect the yield of plants. Rice is a tillering crop, and tillering is a physiological genetic characteristic of rice itself. Rice tillers are essentially the branches of rice stalks, which are an important part of rice plant architecture and also a determi...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82A01H5/00
CPCC07K14/415C12N15/8261
Inventor 胡廷章朱姗姗何帅陈国平胡宗利王贵学
Owner CHONGQING UNIV