Effects of Nitrogen Level during Seed Production on Wheat Seed Vigor and Seedling Establishment at the Transcriptome Level

Nitrogen fertilizer is a critical determinant of grain yield and seed quality in wheat. However, the mechanism of nitrogen level during seed production affecting wheat seed vigor and seedling establishment at the transcriptome level remains unknown. Here, we report that wheat seeds produced under different nitrogen levels (N0, N168, N240, and N300) showed significant differences in seed vigor and seedling establishment. In grain yield and seed vigor, N0 and N240 treatments showed the minimum and maximum, respectively. Subsequently, we used RNA-seq to analyze the transcriptomes of seeds and seedlings under N0 and N240 at the early stage of seedling establishment. Gene Ontology (GO) term enrichment analysis revealed that dioxygenase-activity-related genes were dramatically upregulated in faster growing seedlings. Among these genes, the top three involved linoleate 9S-lipoxygenase (Traes_2DL_D4BCDAA76, Traes_2DL_CE85DC5C0, and Traes_2DL_B5B62EE11). Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis revealed that pathways involved in nutrient mobilization and the antioxidant system showed enhanced expression under N240. Moreover, seeds with faster growing seedlings had a higher gene expression level of α-amylase, which was consistent with α-amylase activity. Taken together, we propose a model for seedling establishment and seed vigor in response to nitrogen level during seed production

Universal Multiplex PCR: a novel method of simultaneous amplification of multiple DNA fragments

<p>Abstract</p> <p>Background</p> <p>Multiplex PCR has been successfully applied in many areas since it was first reported in 1988; however, it suffers from poor universality.</p> <p>Results</p> <p>A novel method called Universal Multiplex PCR (UM-PCR) was created, which simultaneously amplifies multiple target fragments from genomic DNA. The method has two steps. First, the universal adapter-F and universal adapter-R are connected to the forward primers and the reverse primers, respectively. Hairpin structures and cross dimers of five pairs of adapter-primers are detected. Second, UM-PCR amplification is implemented using a novel PCR procedure termed “Two Rounds Mode” (three and 28–32 cycles). The first round (the first three cycles) is named the “One by One Annealing Round”. The second round (28–32 cycles) combines annealing with extension. In the first two cycles of the first round, primers only amplify the specific templates; there are no templates for the universal adapters. The templates of universal adapters begin to be synthesized from the second cycle of the first round, and universal adapters and primers commence full amplification from the third cycle of the first round.</p> <p>Conclusions</p> <p>UM-PCR greatly improves the universality of multiplex PCR. UM-PCR could rapidly detect the genetic purity of maize seeds. In addition, it could be applied in other areas, such as analysis of polymorphisms, quantitative assays and identifications of species.</p

Dynamic Changes in Seed Germination under Low-Temperature Stress in Maize

Low-temperature stress delays seed germination in maize. Different maize inbred lines display various low-temperature resistance, but the dynamic changes in seed germination under low-temperature stress in maize remain unknown, especially at the transcriptome level. In this study, low-temperature-resistant maize (RM) inbred line 04Qun0522-1-1 had a significantly faster germination speed than low-temperature-sensitive maize (SM) line B283-1 under low-temperature stress. Moreover, the total antioxidant capacity, superoxide dismutase, and peroxidase activities were notably higher in the RM line than in the SM line from 3 to 6 d. In contrast, the SM line showed significantly higher malondialdehyde (MDA) content than the RM line at 6 d. Gene ontology (GO) enrichment analysis showed that in 2dvs0d, both SM and RM lines displayed the downregulation of ribosome-related genes. Moreover, photosystem II and heat shock protein binding-related genes were also downregulated in the SM line. In 4dvs2d, the RM line showed a higher degree of upregulation of the ribosome and peroxidase (POD)-related genes than the SM line. In 6dvs4d, POD-related genes were continuously upregulated in both SM and RM lines, but the degree of upregulation of the genes was higher in the SM line than in the RM line. Moreover, vitamin B6-related genes were specifically upregulated in the RM line. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that in 6dvs4d, phenylpropanoid biosynthesis was the most significantly enriched pathway in both SM and RM lines. Moreover, phenylpropanoid biosynthesis was also enriched in the RM line in 4dvs2d. More than half of the differentially expressed genes (DEGs) in phenylpropanoid biosynthesis were peroxidase, and the DEGs were similar to the GO enrichment analysis. The results provide new insights into maize seed germination in response to low-temperature stress

Photosynthesis and antioxidant metabolism modulate the low-temperature resistance of seed germination in maize

AbstractSpring maize is usually subjected to low-temperature stress during seed germination, which retards seedling growth even if under a suitable temperature. However, the mechanism underlying maize seed germination under low-temperature stress modulating seedling growth after being transferred to normal temperature is still ambiguous. In this study, we used two maize inbred lines with different low-temperature resistance (SM and RM) to investigate the mechanism. The results showed that the SM line had higher lipid peroxidation and lower total antioxidant capacity and germination percentage than the RM line under low-temperature stress, which indicated that the SM line was more vulnerable to low-temperature stress. Further transcriptome analysis revealed that seed germination under low-temperature stress caused down-regulation of photosynthesis related gene ontology (GO) terms in two lines. Moreover, the SM line displayed down-regulation of ribosome and superoxide dismutase (SOD) related genes, whereas genes involved in SOD and vitamin B6 were up-regulated in the RM line. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis revealed that photosynthesis and antioxidant metabolism related pathways played important roles in seed germination in response to low-temperature stress, and the photosynthetic system displayed a higher damage degree in the SM line. Both qRT-PCR and physiological characteristics experiments showed similar results with transcriptome data. Taken together, we propose a model for maize seed germination in response to low-temperature stress.One sentence summaryDamage degree of photosynthesis and total antioxidant capacity (especially SOD activity) determine diverse low-temperature resistance among maize inbred lines at the germination stage.</jats:sec

Maize Seed Germination Under Low-Temperature Stress Impacts Seedling Growth Under Normal Temperature by Modulating Photosynthesis and Antioxidant Metabolism

Spring maize is usually subjected to low-temperature stress during seed germination, which retards seedling growth later even under a suitable temperature. However, the mechanism underlying maize seed germination under low-temperature stress impacting seedling growth is still ambiguous. In this study, we used one low-temperature sensitive maize (SM) and one low-temperature resistance maize (RM) to investigate the mechanism. The results showed that the SM line had higher malondialdehyde content and lower total antioxidant capacity (TAC) and germination percentage than the RM line under low-temperature stress, indicating the vulnerability of SM line to low-temperature stress. Further transcriptome analysis revealed that seed germination under low-temperature stress caused the down-regulation of photosynthesis-related gene ontology terms in two lines. Moreover, the SM line displayed down-regulation of ribosome and superoxide dismutase (SOD) related genes, whereas genes involved in SOD and vitamin B6 were up-regulated in the RM line. Kyoto Encyclopedia of Genes and Genomes enrichment analysis revealed that photosynthesis and antioxidant metabolism-related pathways played essential roles in response to low-temperature stress during seed germination. The photosynthetic system displayed a higher degree of damage in the SM line. Both qRT-PCR and physiological characteristics experiments showed similar results with transcriptome data. Taken together, we propose a model for maize seed germination in response to low-temperature stress.</jats:p

Dynamic Changes in Seed Germination under Low-Temperature Stress in Maize

Low-temperature stress delays seed germination in maize. Different maize inbred lines display various low-temperature resistance, but the dynamic changes in seed germination under low-temperature stress in maize remain unknown, especially at the transcriptome level. In this study, low-temperature-resistant maize (RM) inbred line 04Qun0522-1-1 had a significantly faster germination speed than low-temperature-sensitive maize (SM) line B283-1 under low-temperature stress. Moreover, the total antioxidant capacity, superoxide dismutase, and peroxidase activities were notably higher in the RM line than in the SM line from 3 to 6 d. In contrast, the SM line showed significantly higher malondialdehyde (MDA) content than the RM line at 6 d. Gene ontology (GO) enrichment analysis showed that in 2dvs0d, both SM and RM lines displayed the downregulation of ribosome-related genes. Moreover, photosystem II and heat shock protein binding-related genes were also downregulated in the SM line. In 4dvs2d, the RM line showed a higher degree of upregulation of the ribosome and peroxidase (POD)-related genes than the SM line. In 6dvs4d, POD-related genes were continuously upregulated in both SM and RM lines, but the degree of upregulation of the genes was higher in the SM line than in the RM line. Moreover, vitamin B6-related genes were specifically upregulated in the RM line. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that in 6dvs4d, phenylpropanoid biosynthesis was the most significantly enriched pathway in both SM and RM lines. Moreover, phenylpropanoid biosynthesis was also enriched in the RM line in 4dvs2d. More than half of the differentially expressed genes (DEGs) in phenylpropanoid biosynthesis were peroxidase, and the DEGs were similar to the GO enrichment analysis. The results provide new insights into maize seed germination in response to low-temperature stress.</jats:p