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To explore the possible novel microRNA (miRNA) regulatory pathways in Zhengmai 1860, a newly cultivated drought-tolerant wheat ( Triticum aestivum L.) cultivar, miRNA transcriptome sequencing of the flag leaves of Zhengmai 1860, drought-sensitive variety Zhoumai 18, and drought-resistant variety Bainong 207 was performed during the grain filling stage. We also observed changes in the chloroplast ultrastructure, phytohormone levels, and antioxidant- and photosynthesis-related physiological indicators in three wheat varieties. The results showed that the flag leaves of the drought-tolerant variety Zhengmai 1860 had higher chlorophyll contents and net photosynthetic rates than those of Zhoumai 18 under drought stress during the grain filling stage; in addition, the chloroplast structure was more complete. However, there was no significant difference between Zhengmai 1860 and Bainong 207. MiRNA transcriptome analysis revealed that the differential expression of the miRNAs and mRNAs exhibited variable specificity. The KEGG pathway enrichment results indicated that most of the genes were enriched in the MAPK signaling pathway, plant hormone signal transduction, photosynthetic antennae protein, and amino acid and carbohydrate metabolism. In the drought-tolerant cultivar Zhengmai 1860, tae-miR408 was targeted to regulate the allene oxide synthase (AOS) gene, inhibit its expression, reduce the AOS content, and decrease the synthesis of jasmonic acid (JA) and abscisic acid (ABA). The results of this study suggest that Zhengmai 1860 could improve the photosynthetic performance of flag leaves by inhibiting the expression of genes involved in the JA pathway through miRNAs under drought conditions. Moreover, multiple miRNAs may target chlorophyll, antioxidant enzymes, phytohormone signal transduction, and other related pathways; thus, it is possible to provide a more theoretical basis for wheat molecular breeding.

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 CO 2 assimilation rate ( P n ) 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 (O 2 · - , H 2 O 2 ) 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 CO 2 assimilation through a significantly higher P n , 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. P n 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 C 3 crops.

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 C 3 crops by inserting major C 4 photosynthetic genes., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

Using particle bombardment transformation, we introduced maize pepc cDNA encoding phosphoenolpyruvate carboxylase (PEPC) and ppdk cDNA encoding pyruvate orthophosphate dikinase (PPDK) into the C3 crop wheat to generate transgenic wheat lines carrying cDNA of pepc (PC lines), ppdk (PK lines) or both (PKC lines). The integration, transcription, and expression of the foreign genes were confirmed by Southern blot, Real-time quantitative reverse transcription PCR (Q-RT-PCR), and Western blot analysis. Q-RT-PCR results indicated that the average relative expression levels of pepc and ppdk in the PKC lines reached 10 and 4.6, respectively, compared to their expressions in untransformed plants (set to 1). The enzyme activities of PEPC and PPDK in the PKC lines were 4.3- and 2.1-fold higher, respectively, than in the untransformed control. The maximum daily net photosynthetic rates of the PKC, PC, and PK lines were enhanced by 26.4, 13.3, and 4.5%, respectively, whereas the diurnal accumulations of photosynthesis were 21.3, 13.9, and 6.9%, respectively, higher than in the control. The Fv/Fm of the transgenic plants decreased less than in the control under high temperature and high light conditions (2 weeks after anthesis), suggesting that the transgenic wheat transports more absorbed light energy into a photochemical reaction. The exogenous maize C4-specific pepc gene was more effective than ppdk at improving the photosynthetic performance and yield characteristics of transgenic wheat, while the two genes showed a synergistic effect when they were transformed into the same genetic background, because the PKC lines exhibited improved photosynthetic and physiological traits.

With 7 figures and 2 tables Abstract Attempts have been made to transform C3 crops with C4 genes to improve the photosynthetic rate of C3 plants. In our study, a full-length cDNA for phosphoenolpyruvate carboxylase gene ( pepc) was isolated from Zea mays by PCR. The pepc gene was introduced into wheat plants ( Triticum aestivum) by particle bombardment transformation. The integration, transcription and expression of the pepc gene were confirmed by Southern blot, RT-PCR and Western blot analysis. Physiological studies showed that phosphoenolpyruvate carboxylase activity in the transgenic plants was 140% higher than that of the untransformed plants. The highest rate of photosynthesis in the transgenic plants was 31.95 μmol CO2/m2/s, which was 26% greater than the rate in untransformed plants. The light saturation point and carboxylation efficiency of pepc transgenic wheat were 20% and 22.57% higher, respectively, than those of untransformed wheat. As a result, the weight of seed per spike and thousand-grain weight were 0.23 g and 1.21 g higher than those of untransformed wheat. Our research will facilitate the further study of the function of the pepc gene in wheat plants. [ABSTRACT FROM AUTHOR]