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

Huimin Cui,Piwu Wang,Abraham Lamboro,Shuo Qu,Yaolei Jiao

doi:10.32604/phyton.2022.020010

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.

Highlights

Soybean (Glycine max L., Merr) is an important oil and food crop in the world [1]
3.1 GmPR1 Gene Source The GmPR1 gene comes from high-throughput sequencing of the differential transcriptome to obtain differentially expressed genes
It can be seen that the characteristics of soybean gray spot infection are continuous, and the infection process is from work to pod to seed [17]

Summary

Introduction

Soybean (Glycine max L., Merr) is an important oil and food crop in the world [1]. It is one of the main sources of protein and fat for mankind. In China, the protein in soybeans accounts for about 40%, and the fat content accounts for about 20% [2,3]. In 2017, the total domestic consumption of soybeans reached 1.2 × 1011 Kg. In 2019, the total domestic consumption of soybeans reached 1.964 × 1011 Kg, and soybean consumption accounted for 5.4% of the world [4]. In 2019, the total domestic consumption of soybeans reached 1.964 × 1011 Kg, and soybean consumption accounted for 5.4% of the world [4] This shows that soybeans play an important role in people’s lives.

Methods

Results

Discussion

Conclusion