Genome-wide association study of agronomic traits related to nitrogen use efficiency in Henan wheat.

BACKGROUND: Nitrogen use efficiency (NUE) is closely related to crop yield and nitrogen fertilizer application rate. Although NUE is susceptible to environments, quantitative trait nucleotides (QTNs) for NUE in wheat germplasm populations have been rarely reported in genome-wide associated study. RESULTS: In this study, 244 wheat accessions were phenotyped by three NUE-related traits in three environments and genotyped by 203,224 SNPs. All the phenotypes for each trait were used to associate with all the genotypes of these SNP markers for identifying QTNs and QTN-by-environment interactions via 3VmrMLM. Among 279 QTNs and one QTN-by-environment interaction for low nitrogen tolerance, 33 were stably identified, especially, one large QTN (r2 > 10%), qPHR3A.2, was newly identified for plant height ratio in one environment and multi-environment joint analysis. Among 52 genes around qPHR3A.2, four genes (TraesCS3A01G101900, TraesCS3A01G102200, TraesCS3A01G104100, and TraesCS3A01G105400) were found to be differentially expressed in low-nitrogen-tolerant wheat genotypes, while TaCLH2 (TraesCS3A01G101900) was putatively involved in porphyrin metabolism in KEGG enrichment analyses. CONCLUSIONS: This study identified valuable candidate gene for low-N-tolerant wheat breeding and provides new insights into the genetic basis of low N tolerance in wheat.

Identification and validation of a novel locus, Qpm-3BL, for adult plant resistance to powdery mildew in wheat using multilocus GWAS.

BACKGROUND: Powdery mildew (PM), one of the major diseases in wheat, severely damages yield and quality, and the most economical and effective way to address this issue is to breed disease-resistant cultivars. Accordingly, 371 landraces and 266 released cultivars in Henan Province were genotyped by a 660 K microarray and phenotyped for adult plant resistance (APR) to PM from 2017 to 2020, and these datasets were used to conduct multilocus genome-wide association studies (GWASs). RESULTS: Thirty-six varieties showed stable APR in all the environments, and eleven quantitative trait nucleotides (QTNs) were found by multiple methods across multiple environments and best linear unbiased prediction (BLUP) values to be significantly associated with APR. Among these stable QTNs, four were previously reported, three were newly discovered in this study, and the others need to be further investigated. The major and newly discovered QTN, Qpm-3BL, was located at chr03BL_AX-109,052,670, while another newly discovered QTN, Qpm-1BL, was located between chr01BL_AX-108,771,002 and chr01BL_AX-110,117,322. Five and eight landraces were identified to be resistant based on Qpm-1BL (haplotype TC) and Qpm-3BL (allele T), respectively. To validate Qpm-3BL, a new kompetitive allele-specific PCR (KASP) marker was developed to scan 155 F2 individuals, and the average resistance score supported the value of Qpm-3BL in marker-assisted breeding. Near Qpm-3BL, PmBMYD was identified by KEGG, gene expression and comparative genomics analyses to be a candidate. Its resistance mechanism may involve gene tandem repeats. CONCLUSIONS: This study reveals a previously unknown gene for PM resistance that is available for marker-assisted breeding.

Mapping quantitative trait loci and developing their KASP markers for pre-harvest sprouting resistance of Henan wheat varieties in China.

Introduction: Pre-harvest Sprouting (PHS) seriously affects wheat quality and yield. However, to date there have been limited reports. It is of great urgency to breed resistance varieties via quantitative trait nucleotides (QTNs) or genes for PHS resistance in white-grained wheat. Methods: 629 Chinese wheat varieties, including 373 local wheat varieties from 70 years ago and 256 improved wheat varieties were phenotyped for spike sprouting (SS) in two environments and genotyped by wheat 660K microarray. These phenotypes were used to associate with 314,548 SNP markers for identifying QTNs for PHS resistance using several multi-locus genome-wide association study (GWAS) methods. Their candidate genes were verified by RNA-seq, and the validated candidate genes were further exploited in wheat breeding. Results: As a result, variation coefficients of 50% and 47% for PHS in 629 wheat varieties, respectively, in 2020-2021 and 2021-2022 indicated large phenotypic variation, in particular, 38 white grain varieties appeared at least medium resistance, such as Baipimai, Fengchan 3, and Jimai 20. In GWAS, 22 significant QTNs, with the sizes of 0.06% ~ 38.11%, for PHS resistance were stably identified by multiple multi-locus methods in two environments, e.g., AX-95124645 (chr3D:571.35Mb), with the sizes of 36.390% and 45.850% in 2020-2021 and 2021-2022, respectively, was detected by several multi-locus methods in two environments. As compared with previous studies, the AX-95124645 was used to develop Kompetitive Allele-Specific PCR marker QSS.TAF9-3D (chr3D:569.17Mb~573.55Mb) for the first time, especially, it is available in white-grain wheat varieties. Around this locus, nine genes were significantly differentially expressed, and two of them (TraesCS3D01G466100 and TraesCS3D01G468500) were found by GO annotation to be related to PHS resistance and determined as candidate genes. Discussion: The QTN and two new candidate genes related to PHS resistance were identified in this study. The QTN can be used to effectively identify the PHS resistance materials, especially, all the white-grained varieties with QSS.TAF9-3D-TT haplotype are resistant to spike sprouting. Thus, this study provides candidate genes, materials, and methodological basis for breeding wheat PHS resistance in the future.

Transgenic expression of rice OsPHR2 increases phosphorus uptake and yield in wheat.

Oryza sativa PHOSPHATE RESPONSE2 (OsPHR2) can promote the uptake and use of phosphorus (P) in rice. We introduced OsPHR2 into the winter wheat (Triticum aestivum L.) variety "Zhengmai0856." OsPHR2 was integrated into the wheat genome with two copy numbers and could be correctly transcribed and expressed. OsPHR2 was mainly expressed in the leaves at the seedling stage. From the jointing to filling stage, OsPHR2 was mainly expressed in the roots, followed by the leaves, with a low expression level in detected the tassels and stems. The transgenic lines exhibited higher P accumulation at each growth stage and increased P uptake intensity in comparison to the wild type under low P and high P conditions. Analysis of the root characteristics showed that the transgenic expression of OsPHR2 increased the maximum root length, total root length, root-to-shoot ratio, and root volume under the conditions of P deficiency or low P. A field experiment showed that the transgenic lines had a higher grain yield than the wild type under low P and high P conditions. The yield of the transgenic lines increased by 6.29% and 3.73% on average compared with the wild type under low P and high P conditions, respectively. Thus, the transgenic expression of OsPHR2 could increase P uptake and yield in wheat, but the effect was more prominent under low P conditions.

Joint expression of Zmpepc, Zmppdk, and Zmnadp-me is more efficient than expression of one or two of those genes in improving the photosynthesis of Arabidopsis.

This study assessed the effects of seven combinations of maize (Zea mays) genes phosphoenolpyruvate carboxylase (pepc), pyruvate phosphate dikinase (ppdk), and NADP-malic enzyme (nadp-me), on the photosynthesis of Arabidopsis. The photosynthetic rate, carboxylation efficiency, and shoot-dry-weight of Zmpepc (PC), Zmpepc + Zmppdk (PCK), Zmpepc + Zmnadp-me (PCM), and Zmpepc + Zmppdk + Zmnadp-me (PCKM) were significantly higher than those of the control wild-type (WT), with a trends to be PCKM > PCK > PC and PCM > WT. This indicated that Zmpepc was a prerequisite for improved photosynthetic performance; Zmppdk had a positive effect on Zmpepc, and the triple gene combination had the most significant synergistic effects. PCKM significantly enhanced activity of photosystem (PS)II (K, J phase) and PSI, light energy absorption (ABS/CSm) and conversion (TRo/ABS), and electron transfer (ETo/TRo). PCKM up-regulated 18 photosynthesis-related proteins, among which, 11 were involved in light reaction resulting in improved light-energy absorption and conversion efficiency, electron transfer, activity and stability of PSII and PSI, and the ATP and NADPH production. The remaining seven proteins were involved in dark reaction. The up-regulation of these proteins in PCKM improved the coordinated operation of light and dark reaction, increasing the photosynthesis and dry weight ultimately. These results also provide a promising strategy for the genetic improvement of the photosynthetic performance of C3 crops by inserting major C4 photosynthetic genes.

Response of transgenic Arabidopsis expressing maize C4 photosynthetic enzyme genes to high light.

This study assessed the responses of wild-type (WT) and transgenic Arabidopsis expressing seven combinations of maize (Zea mays) genes phosphoenolpyruvate carboxylase (pepc), pyruvate phosphate dikinase (ppdk), and NADP-malic enzyme (nadp-me) to high light. Our results showed that the net CO2 assimilation rate (Pn) and shoot dry weight of four of the transgenic Arabidopsis genotypes were significantly different from those of WT under high-light treatment, being in the order of Zmpepc+Zmppdk+Zmnadp-me (PC-K-M) > Zmpepc+Zmppdk (PC-K) > Zmpepc (PC), Zmpepc+Zmnadp-me (PC-M) > WT. The other genotypes did not differ from WT. This indicated that Zmpepc was essential for maintaining high photosynthetic performance under high light, Zmppdk had a positive synergistic effect on Zmpepc, and the combination of all three genes had the greatest synergistic effect. These four genotypes also maintained higher photosystem II (PSII) activity (K-phase, J-phase, RC/CSm), electron transfer capacity (J-phase), and photochemical efficiency (TRo/ABS), and accumulated less reactive oxygen species (O2·-, H2O2) and suffered less damage to the membrane system (MDA) than WT under high light. Collectively, PC, PC-K, PC-M, and PC-K-M used most of the absorbed energy for CO2 assimilation through a significantly higher Pn, which reduced the generation of excess electrons in the photosynthetic apparatus, thereby reducing damage to the membrane system and PSII. This ultimately resulted in improved high-light tolerance. Pn was the main reason for the significant difference in the high-light tolerance of the four genotypes. Joint expression of the three maize genes may be of great value in the genetic improvement of high-light tolerance in C3 crops.

A novel role of long non-coding RNAs in response to X-ray irradiation.

In the present study, the role of lncRNAs in response to radiation-induced DNA damage and oxidative stress were explored to improve our understanding of the biological pathways activated upon radiation-induced toxicity. The toxicity of X-ray radiation on human bronchial epithelial cell lines (HBE) was determined through a dose-dependent increase in ROS production and γ-H2AX formation and changes to lncRNA expression was observed and quantified using lncRNA-specific microarrays. 115 lncRNAs expression was increased in a dose-dependent manner following X-ray irradiation. Bioinformatic prediction algorithms determined that these lncRNAs significantly affect the p53 signaling pathway, and, more specifically, the BRCA 1 transcription factor and coding genes adjacent to BRCA 1. Our results highlight a previously uncharacterized role for lncRNAs to act via the p53-pathway in response to X-ray-induced DNA damage, and suggest lncRNAs may serve as novel indicators for radiation toxicity.