Open Access

ARTICLE

Bioinformatics Analysis of Disease Resistance Gene PR1 and Its Genetic Transformation in Soybeans and Cultivation of Multi-resistant Materials

by Huimin Cui, Shuo Qu, Abraham Lamboro, Yaolei Jiao, Piwu Wang^*

College of Agronomy, Jilin Agricultural University, Changchun, 1300118, China

* Corresponding Author: Piwu Wang. Email:

Phyton-International Journal of Experimental Botany 2022, 91(7), 1445-1464. https://doi.org/10.32604/phyton.2022.020010

Received 29 October 2021; Accepted 17 December 2021; Issue published 14 March 2022

Abstract

In agricultural production, a single insect-resistant and disease-resistant variety can no longer meet the demand. In this study, the expression vector pCAMBIA-3301-PR1 containing the disease-resistant gene PR1 was constructed by means of genetic engineering, and the PR1 gene was genetically transformed to contain the PR1 gene through the pollen tube method. In CryAb-8Like transgenic high-generation T₇ receptor soybean, a new material that is resistant to insects and diseases is obtained. For T₂ transformed plants, routine PCR detection, Southern Blot hybridization, fluorescence quantitative PCR detection, indoor and outdoor pest resistance identification and indoor disease resistance identification were performed. The results showed that there were 9 positive plants in the routine PCR test of T₂ generation. In Southern Blot hybridization, both PR1 and CryAb-8Like genes are integrated in soybeans in the form of single copies. Fluorescence quantitative PCR showed that the expression levels of PR1 and CryAb-8Like genes are different in different tissues. The average expression levels of PR1 gene in plant roots, stems, and leaves are 2.88, 1.54, and 5.26, respectively. CryAb-8Like genes are found in roots, stems, and leaves. The average expression levels were 1.36, 1.39, and 4.25, respectively. The insectivorous rate of the CryAb-8Like gene in outdoor plants with positive insect resistance identification was 3.78%. The disc partition method was used indoors for pest resistance identification, and the bud length of transformed plants increased significantly. The average mortality rate of untransformed plants in indoor disease resistance identification was as high as 56.66%, and the average mortality rate of plants transformed with PR1 gene was 10.00%, and disease resistance was significantly improved. Therefore, a new material with resistance to diseases and insects is obtained.

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