Drought stress is an important environmental factor limiting productivity of plants, especially fast growing species with high water consumption like poplar. Abscisic acid (ABA) is a phytohormone that positively regulates seed dormancy and drought resistance. The PYR1 (Pyrabactin Resistance 1)/ PYRL (PYR-Like)/ RCAR (Regulatory Component of ABA Receptor) (PYR/PYL/RCAR) ABA receptor family has been identified and widely characterized in Arabidopsis thaliana. However, their functions in poplars remain unknown. Here, we report that 2 of 14 PYR/PYL/RCAR orthologues in poplar (Populus trichocarpa) (PtPYRLs) function as a positive regulator of the ABA signal transduction pathway. The Arabidopsis transient expression and yeast two-hybrid assays showed the interaction among PtPYRL1 and PtPYRL5, a clade A protein phosphatase 2C, and a SnRK2, suggesting that a core signalling complex for ABA signaling pathway exists in poplars. Phenotypic analysis of PtPYRL1 and PtPYRL5 transgenic Arabidopsis showed that these two genes positively regulated the ABA responses during the seed germination. More importantly, the overexpression of PtPYRL1 and PtPYRL5 substantially improved ABA sensitivity and drought stress tolerance in transgenic plants. In summary, we comprehensively uncovered the properties of PtPYRL1 and PtPYRL5, which might be good target genes to genetically engineer drought-Resistant plants.

Significance Thrips are a group of piercing-sucking pest insects that do massive damage in agriculture and horticulture. The western flower thrip is a particularly notorious pest that has spread all over the world and is extremely difficult to control. In this work, we have shown that upon feeding, thrips take up substantial quantities of chloroplast RNA. When we expressed from the chloroplast genome long double-stranded RNAs that are targeted against essential thrip genes, the RNAs induced a potent RNA interference response and efficiently killed the insects. Our study demonstrates that an important group of nonchewing pest insects can be targeted by plant-mediated RNA interference and provides an efficient weapon to control thrips and other sucking plant pests.

Tripartite motif (TRIM) 31 has been implicated in diverse biological and pathological conditions. However, whether TRIM31 plays a role in ischemic stroke progression is not clarified. Here we demonstrated that TRIM31 was significantly downregulated in the ischemic brain and the deficiency of TRIM31 alleviated brain injury induced by middle cerebral artery occlusion by reducing reactive oxygen species production and maintaining mitochondrial homeostasis. Mechanistically, we found that TRIM31 is an E3 ubiquitin ligase for TP53-induced glycolysis and apoptosis regulator (TIGAR), which confers protection against brain ischemia by increasing the pentose phosphate pathway flux and preserving mitochondria function. TRIM31 interacted with TIGAR and promoted the polyubiquitination of TIGAR, consequently facilitated its degradation in a proteasome-dependent pathway. Furthermore, TIGAR knockdown effectively abolished the protective effect of TRIM31 deficiency after cerebral ischemia. In conclusion, we identified that TRIM31 was a novel E3 ubiquitin ligase for TIGAR, played a critical role in regulating its protein level, and subsequently involved in the ischemic brain injury, suggesting TRIM31 as a potential therapeutic target for ischemic stroke.

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Plastid transformation for the expression of recombinant proteins and entire metabolic pathways has become a promising tool for plant biotechnology. However, large-scale application of this technology has been hindered by some technical bottlenecks, including lack of routine transformation protocols for agronomically important crop plants like rice or maize. Currently, there are no standard or commercial plastid transformation vectors available for the scientific community. Construction of a plastid transformation vector usually requires tedious and time-consuming cloning steps. In this study, we describe the adoption of an in vivo Escherichia coli cloning (iVEC) technology to quickly assemble a plastid transformation vector. The method enables simple and seamless build-up of a complete plastid transformation vector from five DNA fragments in a single step. The vector assembled for demonstration purposes contains an enhanced green fluorescent protein (GFP) expression cassette, in which the gfp transgene is driven by the tobacco plastid ribosomal RNA operon promoter fused to the 5′ untranslated region (UTR) from gene10 of bacteriophage T7 and the transcript-stabilizing 3′UTR from the E. coli ribosomal RNA operon rrnB. Successful transformation of the tobacco plastid genome was verified by Southern blot analysis and seed assays. High-level expression of the GFP reporter in the transplastomic plants was visualized by confocal microscopy and Coomassie staining, and GFP accumulation was ~9% of the total soluble protein. The iVEC method represents a simple and efficient approach for construction of plastid transformation vector, and offers great potential for the assembly of increasingly complex vectors for synthetic biology applications in plastids.

BackgroundRNA interference (RNAi) has emerged as a promising technology for insect pest control. Because of the accumulation of high levels of long double‐stranded RNAs (dsRNAs) in plastids, it was previously shown that expression of dsRNAs from plastid genome led to higher mortality of some insect pests with chewing mouthparts than dsRNAs expression from nuclear genome. However, whether plastid‐expressed dsRNAs have effects on phloem sap‐sucking pests is unknown. In this study, we compared the RNAi effects of nuclear transgenic and transplastomic plants on the whitefly Bemisia tabaci, a serious sap‐sucking pest.ResultsNuclear transgenic and transplastomic tobacco plants were developed for the expression of dsRNA against BtACTB gene of Bemisia tabaci, respectively. Feeding nuclear transgenic plants to Bemisia tabaci resulted in reduced gene expression of BtACTB and survival rate, and impaired fecundity of Bemisia tabaci. We did not observe any effects of transplastomic plants on Bemisia tabaci fitness. Furthermore, we found that the inability of B. tabaci to obtain dsRNAs from plastids might restrict its RNAi responses.ConclusionOur study indicated that the expression of dsRNAs in nuclear transgenic plants was more effective than that in transplastomic plants for the control of Bemisia tabaci. The inaccessibility of Bemisia tabaci to plastids contributes to the inefficiency of plastid‐mediated RNAi. Our findings are of great significance to future optimization of transgenically delivered RNAi approaches for efficient controlling of sap‐sucking pests. © 2020 Society of Chemical Industry

‘Ganlv 1’ is a new cultivar of Actinidia eriantha selected from the wild natural population, which has the advantages of moderate taste, high yield, easy peeling and high ascorbic acid (AsA) content. In this study, ‘Ganlv 1’ was used to explore the changes in fruit quality, soluble sugar components, sucrose metabolism-related enzymes activities and sucrose metabolism-related enzyme genes’ expression during the fruit’s development. The results showed that, except for AsA, the changes in the fruit quality index and fruit growth and development during the development of ‘Ganlv 1’ basically exhibited the same trend. The fruit shape index was different in the different development stages of the fruit, and tended to be stable with fruit growth and development. The dynamic changes of the dry matter content indicated that the best time for fruit harvest was about 160 days after full bloom. The main sugar components in the fruit were fructose, glucose and sucrose, and sucrose and glucose were the main sugars in the soft-ripening stage. The trend of sucrose accumulation, the activities of the sucrose metabolism-related enzymes and the expression of the sucrose metabolism-related genes indicated that 130–145 days after full bloom (DAFB) might be the critical period of sucrose metabolism. The results are of great significance for clarifying the developmental characteristics and dynamic changes in the sugar components in A. eriantha fruits, and lay a foundation for further studying of the mechanism of sugar metabolism in A. eriantha.

Background The TIFY gene family is a group of plant-specific transcription factors involved in regulation of plant growth and development and a variety of stress responses. However, the TIFY family has not yet been well characterized in kiwifruit, a popular fruit with important nutritional and economic value. Results A total of 27 and 21 TIFY genes were identified in the genomes of Actinidia eriantha and A. chinensis, respectively. Phylogenetic analyses showed that kiwifruit TIFY genes could be classified into four major groups, JAZ, ZML, TIFY and PPD, and the JAZ group could be further clustered into six subgroups (JAZ I to JAZ VI). Members within the same group or subgroup have similar exon-intron structures and conserved motif compositions. The kiwifruit TIFY genes are unevenly distributed on the chromosomes, and the segmental duplication events played a vital role in the expansion of the TIFY genes in kiwifruit. Syntenic analyses of TIFY genes between kiwifruit and other five plant species (including Arabidopsis thaliana, Camellia sinensis, Oryza sativa, Solanum lycopersicum and Vitis vinifera) and between the two kiwifruit species provided valuable clues for understanding the potential evolution of the kiwifruit TIFY family. Molecular evolutionary analysis showed that the evolution of kiwifruit TIFY genes was primarily constrained by intense purifying selection. Promoter cis-element analysis showed that most kiwifruit TIFY genes possess multiple cis-elements related to stress-response, phytohormone signal transduction and plant growth and development. The expression pattern analyses indicated that TIFY genes might play a role in different kiwifruit tissues, including fruit at specific development stages. In addition, several TIFY genes with high expression levels during Psa (Pseudomonas syringae pv. actinidiae) infection were identified, suggesting a role in the process of Pas infection. Conclusions In this study, the kiwifruit TIFY genes were identified from two assembled kiwifruit genomes. In addition, their basic physiochemical properties, chromosomal localization, phylogeny, gene structures and conserved motifs, synteny analyses, promoter cis-elements and expression patters were systematically examined. The results laid a foundation for further understanding the function of TIFY genes in kiwifruit, and provided a new potential approach for the prevention and treatment of Psa infection.