Maize (Zea mays L.) is a tropical crop, and low temperature has become one of the main abiotic stresses for maize growth and development, affecting many maize growth processes. The main area of maize production in China, Jilin province, often suffers from varying degrees of cold damage in spring, which seriously affects the quality and yield of maize. In the face of global climate change and food security concerns, discovering cold tolerance genes, developing cold tolerance molecular markers, and creating cold-tolerant germplasm have become urgent for improving maize resilience against these conditions and obtaining an increase in overall yield. In this study, whole-genome sequencing and genotyping by sequencing were used to perform genome-wide association analysis (GWAS) and quantitative trait locus (QTL) mapping of the two populations, respectively. Overall, four single-nucleotide polymorphisms (SNPs) and 12 QTLs were found to be significantly associated with cold tolerance. Through joint analysis, an intersection of GWAS and QTL mapping was found on chromosome 3, on which the Zm00001d002729 gene was identified as a potential factor in cold tolerance. We verified the function of this target gene through overexpression, suppression of expression, and genetic transformation into maize. We found that Zm00001d002729 overexpression resulted in better cold tolerance in this crop. The identification of genes associated with cold tolerance contributes to the clarification of the underlying mechanism of this trait in maize and provides a foundation for the adaptation of maize to colder environments in the future, to ensure food security.
PCR detection, quantitative real-time PCR (q-RTPCR), outdoor insect resistance, and disease resistance identification were carried out for the detection of genetic stability and disease resistance through generations (T 2 , T 3 , and T 4 ) in transgenic maize germplasms (S3002 and 349) containing the bivalent genes (insect resistance gene Cry1Ab13-1 and disease resistance gene NPR1) and their corresponding wild type. Results indicated that the target genes Cry1Ab13-1 and NPR1 were successfully transferred into both germplasms through tested generations; q-PCR confirmed the expression of Cry1Ab13-1 and NPR1 genes in roots, stems, and leaves of tested maize plants. In addition, S3002 and 349 bivalent gene-transformed lines exhibited resistance to large leaf spots and corn borer in the field evaluation compared to the wild type. Our study confirmed that Cry1Ab13-1 and NPR1 bivalent genes enhanced the resistance against maize borer and large leaf spot disease and can stably inherit. These findings could be exploited for improving other cultivated maize varieties.
Soybean oil is a traditional edible oil. Increasing the oleic acid content is an important direction of soybean breeding. The soybean FAD2 family consists of seven genes that regulate how oleic acid is converted into linoleic acid. Five genes of the soybean FAD2 gene family, GmFAD2-1B, GmFAD2-1A, GmFAD2-2B, GmFAD2-2C, and GmFAD2-2D, were taken as target genes in this study. Firstly, a multivalent CRISPR/Cas9 gene-editing vector was constructed to regulate FAD2 gene expression. Multiple knockout vectors were inserted into the soybean varieties JN38, T6098, and T7010 using Agrobacterium-mediated soybean cotyledon transformation. The functional analysis, agronomic character analysis, and comparison of the mutant lines of the offspring of different genotypes indicated that the JN38 mutant was significantly taller in terms of plant height than the receptor JN38. The fatty acid content of the three groups showed the same trend. The fatty acid contents of mutant plants were higher than those of recipient plants, and the linoleic acid contents of mutant plants were lower than those of recipient plants. The best-performing among the three groups was the JN38 mutant, whose oleic acid content increased from 18.58% to 54.07% and whose linoleic acid content decreased from 57.79% to 26.17%. In conclusion, the knockout expression of multiple FAD2 genes increased the soybean oleic acid content and decreased the linoleic acid content in different receptors.
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