Unlock instant, AI-driven research and patent intelligence for your innovation.

Method for producing transgene paddy with amount of spikelet being increased

A technology of transgenic rice and genes, applied in the field of genetic engineering, can solve the problem of not getting it

Inactive Publication Date: 2005-03-02
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI +1
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the results of in situ hybridization showed that the RFL gene is not a floral meristem attribute gene like FLO and LFY, and RFL may be related to the development of primary and secondary branches
Moreover, the rice RFL gene was overexpressed in Arabidopsis, and the results consistent with 35S LFY were not obtained.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for producing transgene paddy with amount of spikelet being increased
  • Method for producing transgene paddy with amount of spikelet being increased
  • Method for producing transgene paddy with amount of spikelet being increased

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Isolation of OsCen1 and OsCen2 Genes in Rice CENTRORADIALIS(CEN)-like

[0074] Using the conservative fragment of the fourth exon of the CEN gene of Snapdragon as a probe, the rice (Guangluai 4, Indica) genomic DNA library was screened by in situ hybridization. After three rounds of screening, six positive clones were obtained. Two positive clones with the strongest signals, FD229 and FD322, were selected for physical map construction and subcloning, and sequencing was completed. Two CEN-like genes OsCen1 (also called FDR1) and OsCen2 (also called FDR2) were obtained.

[0075] According to the genome sequence of OsCen1 and OsCen2, specific primers were designed, and the reverse transcription product of the mRNA in the rice inflorescence meristem was used as the template, and the corresponding OsCen1 and OsCen2 genes were isolated by 5'RACE and 3'RACE methods. CDNA, and completed sequencing. OsCen1 cDNA is 907bp long, and OsCen2cDNA is 956bp long. Comparing cDNA and genomic ...

Embodiment 2

[0078] Obtain the complete ORF of OsCen1 and OsCen2

[0079] (A) Obtain the complete ORF of OsCen1

[0080] For transgenic needs, a complete ORF must be obtained, so the 5′ primer is designed before the start codon of the OsCen1 gene, namely OsCen1-ORF-5′:

[0081] ttacctgtccctccaaggc (SEQ ID NO: 3)

[0082] And design a 3′ primer after the stop codon, namely OsCen1-ORF-3′:

[0083] cacaatctag tgatgtggc (SEQ ID NO: 4)

[0084] And using the reverse transcription product of the mRNA in the rice inflorescence meristem as a template, the ORF of OsCen1 plus a 3′UTR region was amplified by conventional RT-PCR, and the total length was 643bp, which was cloned into the pGEM-T vector , The number of the positive clone is pCSL0034.

[0085] The amplified 643bp cDNA of OsCen1 is shown in Figure 1 and SEQ ID NO:1. The amino acid sequence of the encoded OsCen1 protein is shown in SEQ ID NO:2.

[0086] (A) Obtain the complete ORF of OsCen2

[0087] Design a 5′ primer before the start codon of ...

Embodiment 3

[0094] Construction of sense expression vector for OsCen1 and OsCen2

[0095] (A) Construction of OsCen1 sense expression vector

[0096] After digestion with SpeI, fill in, then digest with NcoI to cut the 643bp cDNA of OsCen1 in pCSL0034 plasmid, and connect it to the intermediate vector pJIT163 (available from the British JOIN INNES Research Center), and then add Two 35S strong promoters plus the forward OsCen1 cDNA gene and the fragment of CaMV polyA were cloned into the plant expression vector pCAMBIA1301 (available from the CAMBIA Research Center) to obtain the sense expression vector of OsCen1, named pCSL0040, this vector The total length is 14000bp, the resistance in E. coli is kanamycin resistance, and the resistance in plants is hygromycin resistance. The vector also carries the GUS gene in the T-DNA region. The plasmid map of pCSL0040 is shown in Figure 3.

[0097] (B) Construction of OsCen2 sense expression vector

[0098] The construction process of OsCen2 sense expre...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

A process for preparing transgenic rice with increased number of spicula includes transferring the exogenous genes OsCen1 and / or OsCen2 into cell of rice to obtain the transgenic cells or tissue, and culturing rice plants. The carrier and host cell carrying genes OsCen1 and / or OsCen2 are also disclosed.

Description

Technical field [0001] The invention relates to the field of genetic engineering. More specifically, the present invention relates to a method for producing transgenic rice with an increased number of spikelets by using OsCen1 and OsCen2 genes, and related vectors and transgenic plants. Background technique [0002] Monocotyledonous rice is one of the main food crops in the world. The development status of young rice ears directly affects grain yield. Therefore, research on the relationship between the occurrence of rice ears and genes can provide a theoretical basis for the artificial regulation of rice reproductive growth and development . [0003] At present, the research on the flower development of dicotyledonous plants is more in-depth than the research on flower development of monocotyledonous plants. Especially in the study of the dicotyledonous model plants Arabidopsis and snapdragon, the "ABC model" of floral organogenesis was proposed. Generally speaking, the research ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): A01H1/00A01H4/00A01H5/00C12N5/10C12N15/29C12N15/63
Inventor 罗达张淑红孙崇荣
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI