Nucleic acid molecule for detecting corn plant ND6603 and detection method thereof
A nucleic acid molecule, corn technology, applied in the field of plant biology
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0097] Obtaining and screening of embodiment 1 transformation events
[0098] The cDNA sequence of the ZmNRT1.1B gene was cloned from maize, and inserted into the intermediate vector pCAMBIA1301-Ubi by restriction enzyme digestion. The intermediate vector already contained the bar gene, and finally constructed into a pCAMBIA1301-ZmNRT1.1B expression vector. For the physical map of the vector, see figure 1 . Using Agrobacterium infection technology, the recombinant plasmids were efficiently transformed into young embryos of the self-selected line y822 of the Institute of Agricultural Biotechnology, Jilin Academy of Agricultural Sciences, and 56 transformation events of ZmNRT1.1B and bar genes were obtained. Among them, the transgenic maize event ND6603 contains exogenous genes ZmNRT1.1B and bar, exhibits stable hereditary generation and outstanding nitrogen deficiency tolerance, and has a good application prospect.
[0099] The detection method of the exogenous ZmNRT1.1B and b...
Embodiment 2
[0102] Example 2 Nitrogen efficient utilization and nitrogen deficiency tolerance identification of transformation event ND6603
[0103] Maize ZmNRT1.1B protein has the function of promoting nitrate absorption. The invention expresses the ZmNRT1.1B protein in corn by adopting a constitutive expression method, improves the nitrogen utilization efficiency of the corn by improving the nitrate absorption capacity of the corn, and finally improves the tolerance under the condition of nitrogen deficiency. The present invention identifies the field growth morphology and physiological traits of ND6603 plants under different nitrogen fertilizer gradients.
[0104] The test site is at the transgenic test base of Jilin Academy of Agricultural Sciences, Gongzhuling City, Siping City, Jilin Province. The test soil is sandy loam with flat terrain. The plow layer (20cm) contains 125.94mg / kg of alkaline hydrolyzed nitrogen, 23.96mg / kg of available phosphorus, 149.24mg / kg of available potassi...
Embodiment 3
[0113] Example 3 The flanking sequence of the transformation event ND6603 exogenous sequence and the insertion position of the maize genome
[0114] In order to clarify the identity characteristics of the transformation event ND6603, the present invention further identified the insertion site of the exogenous sequence of ND6603 on the maize genome.
[0115] The invention adopts the third-generation sequencing method to locate the T-DNA insertion position of the transgenic maize ND6603. The third-generation sequencing technology used the nanopore sequencing technology of Oxford Nanopore Technologies (ONT) to re-sequence the transformant ND6603. The sequencing depth was 10 times. The average read length of the data was 20kb, and the longest read length was 60kb. The accuracy rate is 85%.
[0116] Using the T-DNA exogenous sequence to perform BLAST search and comparison on the third-generation sequencing results, it was found that a contg with a read length of about 33 kb contai...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More