Co-elevation of atmospheric [CO2] and temperature alters photosynthetic capacity and instantaneous water use efficiency in rice cultivars in a cold-temperate region.

Crop photosynthetic capacity in response to climate change likely constrains crop productivity and adaptability to changing environments, which requests the investigation on the dynamics of photosynthetic parameters over growth season among varieties, especially in cold-temperate regions. Three Japonica rice cultivars i.e., Shoubaimao (SH), Hejiang 19 (HJ); Longjing 31, (LJ). were planted under the control, e[CO2] (700 µmol mol-1), warming (2°C above the air temperature) and the co-elevation of [CO2] and temperature in open-top chambers (OTC). The objective of this study is to examine the rice photosynthetic parameters, water use efficiency (WUE) and yield formation in responses to the co-elevation of [CO2] and temperature which is the main predicted features of future climate. e[CO2] significantly increased An of SH, HJ and LJ by 37%, 39% and 23% in comparison to 34%, 34% and 27% under elevated [CO2] plus warming, respectively. However, An had a weak response to warming for three cultivars. [CO2] and temperature co-elevation significantly decreased the stomatal conductance, resulting in a significant increase of the WUE. e[CO2] significantly increased Vc, max , Jmax and Jmax /Vc, max . e[CO2] significantly increased grain yield and grain number of all cultivars. The positive effect of co-elevation of [CO2] and temperature on grain yield was less than e[CO2]. Warming is likely to partially offset the increased photosynthetic rate caused by e[CO2]. The [CO2] and temperature co-elevation may be favorable to rice crop with increasing the photosynthetic ability of rice crop and improving water use efficiency. The present study provided evidence that the rice genotypic difference in photosynthetic potential under [CO2] and temperature co-elevation. Therefore, it is crucial to explore a broader range of phenotypes and cultivars to be applied to climate change response research, advancing the knowledge that climate change impacts rice crop under the cold-temperate climate region.

Genome-Wide Identification of Wild Soybean Mitochondrial Calcium Uniporter Family Genes and Their Responses to Cold and Carbonate Alkaline Stresses.

The mitochondrial calcium uniporter (MCU), as an important component of the Ca2+ channel uniporter complex, plays a regulatory role in intracellular Ca2+ signal transduction. However, only a few studies to date have investigated plant MCU genes. In this study, we identified the MCU family genes in wild soybean and investigated their expression under cold and carbonate alkaline stresses. Eleven Glycine soja MCU genes (GsMCUs) were identified and clustered into two subgroups (subgroups I and II), and subgroup II could be further divided into two branches (MCU5 and MCU6). A total of 21 pairs of GsMCUs were characterized as duplicated genes, and displayed a similar exon-intron architecture. All GsMCU proteins contained one conserved MCU domain, within which two transmembrane domains were found. An analysis of the conserved motifs further supported that the GsMCUs showed high conservation in protein sequence and structure. Moreover, we found that all GsMCUs were expressed ubiquitously in different tissues and organs, and GsMCUs from the same subgroup displayed varied tissue expression profiles. In addition, based on RNA-seq and qRT-PCR assays, six and nine GsMCUs were differentially expressed under cold and carbonate alkaline stress, respectively. Promoter analysis also uncovered the existence of two canonical cold-related cis-acting elements, LTR and DRE/CRT, as well as stress-related phytohormone-responsive elements. Our results provide valuable information about the MCU family in soybean responses to cold and carbonate alkaline stress, which will be helpful in further characterizing their biological roles in response to abiotic stress.

Wild soybean SNARE proteins BET1s mediate the subcellular localization of the cytoplasmic receptor-like kinases CRCK1s to modulate salt stress responses.

Plants have evolved numerous receptor-like kinases (RLKs) that modulate environmental stress responses. However, little is known regarding soybean (Glycine max) RLKs. We have previously identified that Glycine soja Ca2+ /CAM-binding RLK (GsCBRLK) is involved in salt tolerance. Here, we report that soluble NSF attachment protein receptor proteins BET1s mediate subcellular localization of calmodulin-binding receptor-like cytoplasmic kinases CRCK1s to modulate salt stress responses. Direct interaction between GsCBRLK and GsBET11a was initially identified via yeast two-hybrid and bimolecular fluorescence complementation assays. Further analysis demonstrated conserved interaction between BET1s and CRCK1s. GsCBRLK interacted with all BET1 proteins in wild soybean (Glycine soja) and Arabidopsis, and GsBET11a strongly associated with GsCRCK1a-1d, but slightly with AtCRCK1. In addition, GsBET11a interacted with GsCBRLK via its C-terminal transmembrane domain (TMD), where the entire TMD, not the sequence, was critical for the interaction. Moreover, the N-terminal variable domain (VD) of GsCBRLK was responsible for interacting with GsBET11a, and the intensity of interaction between GsCBRLK/AtCRCK1 and GsBET11a was dependent on VD. Furthermore, GsBET11a was able to mediate the GsCBRLK subcellular localization via direct interaction with VD. Additionally, knockout of AtBET11 or AtBET12 individually did not alter GsCBRLK localization, while GsBET11a expression caused partial internalization of GsCBRLK from the plasma membrane (PM). We further suggest the necessity of GsCBRLK VD for its PM localization via N-terminal truncation assays. Finally, GsBET11a was shown to confer enhanced salt stress tolerance when overexpressed in Arabidopsis and soybean. These results revealed the conserved and direct interaction between BET1s and CRCK1s, and suggested their involvement in salt stress responses.

Plant grafting relieves asymmetry of jasmonic acid response induced by wounding between scion and rootstock in tomato hypocotyl.

Plant grafting is a sequential wound healing process. However, whether wounding induces a different jasmonic acid (JA) response within half a day (12 h) after grafting or non-grafting remains unclear. Using the tomato hypocotyl grafting method, we show that grafting alleviates the asymmetrical accumulation of JA and jasmonic acid isoleucine conjugate (JA-Ile) in scion and rootstock caused by wounding, and from 2 h after tomato micrografting, grafting obviously restored the level of JA-Ile in the scion and rootstock. Meanwhile, five JA-related genes, SlLOX11, SlAOS, SlCOI1, SlLAPA and SlJA2L, are detected and show significant changes in transcriptional expression patterns within 12 h of grafting, from asymmetrical to symmetrical, when the expression of 30 JA- and defense-related genes were analyzed. The results indicated that grafting alleviates the asymmetrical JA and defense response between scion and rootstock of the tomato hypocotyl within 12 h as induced by wounding. Moreover, we demonstrate that in the very early hours after grafting, JA-related genes may be involved in a molecular mechanism that changes asymmetrical expression as induced by wounding between scion and rootstock, thereby promoting wound healing and grafting success.

The antimicrobial activity of protein elicitor AMEP412 against Streptomyces scabiei.

In this paper, we report the antimicrobial activity of AMEP412 (a protein elicitor from Bacillus subtilis) against Streptomyces scabiei, which is the potato common scab pathogen. The purified protein samples showed an obvious inhibition zone on an S. scabiei agar plate, and the minimum inhibition concentration detected was 50 µg mL-1. The fluorescence localization assay revealed that AMEP412 could bind to aerial mycelia and spores. The stability test showed that AMEP412 was stable at 60 °C for 30 min and in pH values from 5.0 to 10.0. Its antimicrobial activity was not sensitive to metal cations. However, its activity declined by 23% when treated with Proteinase K, and was completely abrogated with Tween 80 treatment. Three antimicrobial peptides (GS21, GY20 and GY23) were identified from AMEP412, which further verified its antimicrobial activity. This research reveals the antimicrobial function of AMEP412, which not only enriches the function of the protein elicitor, but also provides a candidate for the biocontrol of potato common scab.

Warming and elevated CO2 alter the transcriptomic response of maize (Zea mays L.) at the silking stage.

Exploring the transcriptome of crops in response to warming and elevated CO2 (eCO2) is important to gaining insights of botanical adaption and feedback to climate change. This study deployed Illumina sequencing technology to characterize transcriptomic profile of maize plants at the silking stage, which were grown under warming (2 °C higher than ambient temperature) and eCO2 (550 ppm) conditions. The treatment of ambient temperature and ambient CO2 concentration was considered as control (CK). Warming, eCO2 and warming plus eCO2 resulted in 2732, 1966 and 271 genes expressing differently (DEGs) compared to the CK, respectively. Among the DEGs, 48, 47 and 36 gene ontology (GO) terms were enriched in response to warming, eCO2 and warming plus eCO2 compared to the CK, respectively. The majority of genes were assigned to the biological process category and the cellular component category. Elevated CO2 significantly inhibited gene expressions in terms of photosynthesis and carbohydrate biosynthesis pathways. Warming not only negatively affected expressions of these genes, but also secondary pathways of nitrogen (N) metabolism, including key enzymes of GST30, GST7, GST26, GST15, GLUL and glnA. These results indicated the negative biochemical regulation and physiological functions in maize in response to warming and eCO2, highlighting the necessity to improve the genetic adaptability of plant to future climate change.

Isolation and identification of a novel protein elicitor from a Bacillus subtilis strain BU412.

Here, we report a novel protein elicitor from Bacillus subtilis BU412 which could cause hypersensitive response (HR) and systemic acquired resistance (SAR) in tobacco. The purification was executed by ion-exchange and size exclusion chromatography. The target band on SDS-PAGE was analyzed by mass spectrometry, and the peptide mass fingerprinting matched an uncharacterized protein (WP_017418614.1), which was then named AMEP412. AMEP412 could cause a clearly defined HR necrosis in tobacco leaves, which was less affected by thermal treatment. The sub-cellular localization assay revealed that AMEP412 localized on the cell surface. This protein could also trigger early defense events such as the generation of reactive oxygen species (H2O2 and O2-) and the induction of defense enzymes, including superoxide dismutase (SOD), peroxidase (POD), polyphenol oxidase (PPO) and phenylalanine ammonia-lyase (PAL). Moreover, AMEP412 could stimulate plant systemic resistance against Pseudomonas syringae pv. tomato DC3000.

A late embryogenesis abundant protein GsPM30 interacts with a receptor like cytoplasmic kinase GsCBRLK and regulates environmental stress responses.

A Glycine soja receptor like cytoplasmic kinase GsCBRLK was previously characterized as a positive regulator of salt tolerance. However, how GsCBRLK regulates stress responses remains obscure. Here, we report the interaction between GsCBRLK and a group 3 late embryogenesis abundant protein GsPM30, and suggest its role in stress responses. GsPM30 was found to physically associate with GsCBRLK through yeast two hybrid assays, which was verified by bimolecular fluorescence complementation analysis. Deletion analyses showed that the N-terminal variable domain of GsCBRLK was sufficient for GsPM30 interaction. Besides GsPM30, GsCBRLK could associate with several group 3 LEAs, of which the N-terminus sequences show high identity with GsPM30. Lower binding affinity or even no interaction was observed between GsCBRLK and other group 3 LEAs, which are less closely related to GsPM30. Furthermore, we observed that GsPM30 could localize surrounding the internal circumference of plant cells, as well as in cytoplasm and nucleus. In addition, GUS staining and quantitative real-time PCR results suggested the ubiquitous expression in different tissues and induced expression by NaCl and mannitol treatments for GsPM30. Consistently, GsPM30 overexpression in Arabidopsis caused increased tolerance to high salinity and dehydration/water deficit at both the young and adult seedling stages. Our results demonstrated the interaction between GsCBRLK and LEAs, and revealed the positive role of GsPM30 in stress responses.

The Glycine soja NAC transcription factor GsNAC019 mediates the regulation of plant alkaline tolerance and ABA sensitivity.

KEY MESSAGE: Overexpression of Gshdz4 or GsNAC019 enhanced alkaline tolerance in transgenic Arabidopsis. We proved that Gshdz4 up-regulated both GsNAC019 and GsRD29B but GsNAC019 may repress the GsRD29B expression under alkaline stress. Wild soybean (Glycine soja) has a high tolerance to environmental challenges. It is a model species for dissecting the molecular mechanisms of salt-alkaline stresses. Although many NAC transcription factors play important roles in response to multiple abiotic stresses, such as salt, osmotic and cold, their mode of action in alkaline stress resistance is largely unknown. In our study, we identified a G. soja NAC gene, GsNAC019, which is a homolog of the Arabidopsis AtNAC019 gene. GsNAC019 was highly up-regulated by 50 mM NaHCO3 treatment in the roots of wild soybean. Further investigation showed that a well-characterized transcription factor, Gshdz4 protein, bound the cis-acting element sequences (CAATA/TA), which are located in the promoter of the AtNAC019/GsNAC019 genes. Overexpression of Gshdz4 positively regulated AtNAC019 expression in transgenic Arabidopsis, implying that AtNAC019/GsNAC019 may be the target genes of Gshdz4. GsNAC019 was demonstrated to be a nuclear-localized protein in onion epidermal cells and possessed transactivation activity in yeast cells. Moreover, overexpression of GsNAC019 in Arabidopsis resulted in enhanced tolerance to alkaline stress at the seedling and mature stages, but reduced ABA sensitivity. The closest Arabidopsis homolog mutant plants of Gshdz4, GsNAC019 and GsRD29B containing athb40, atnac019 and atrd29b were sensitive to alkaline stress. Overexpression or the closest Arabidopsis homolog mutant plants of the GsNAC019 gene in Arabidopsis positively or negatively regulated the expression of stress-related genes, such as AHA2, RD29A/B and KIN1. Moreover, this mutation could phenotypically promoted or compromised plant growth under alkaline stress, implying that GsNAC019 may contribute to alkaline stress tolerance via the ABA signal transduction pathway and regulate expression of the downstream stress-related genes.

A novel AP2/ERF family transcription factor from Glycine soja, GsERF71, is a DNA binding protein that positively regulates alkaline stress tolerance in Arabidopsis.

KEY MESSAGE: Here we first found that GsERF71, an ERF factor from wild soybean could increase plant alkaline stress tolerance by up-regulating H+-ATPase and by modifing the accumulation of Auxin. Alkaline soils are widely distributed all over the world and greatly limit plant growth and development. In our previous transcriptome analyses, we have identified several ERF (ethylene-responsive factor) genes that responded strongly to bicarbonate stress in the roots of wild soybean G07256 (Glycine soja). In this study, we cloned and functionally characterized one of the genes, GsERF71. When expressed in epidermal cells of onion, GsERF71 localized to the nucleus. It can activate the reporters in yeast cells, and the C-terminus of 170 amino acids is essential for its transactivation activity. Yeast one-hybrid and EMSA assays indicated that GsERF71 specifically binds to the cis-acting elements of the GCC-box, suggesting that GsERF71 may participate in the regulation of transcription of the relevant biotic and abiotic stress-related genes. Furthermore, transgenic Arabidopsis plants overexpressing GsERF71 showed significantly higher tolerance to bicarbonate stress generated by NaHCO3 or KHCO3 than the wild type (WT) plants, i.e., the transgenic plants had greener leaves, longer roots, higher total chlorophyll contents and lower MDA contents. qRT-PCR and rhizosphere acidification assays indicated that the expression level and activity of H+-ATPase (AHA2) were enhanced in the transgenic plants under alkaline stress. Further analysis indicated that the expression of auxin biosynthetic genes and IAA contents were altered to a lower extent in the roots of transgenic plants than WT plants under alkaline stress in a short-term. Together, our data suggest that GsERF71 enhances the tolerance to alkaline stress by up-regulating the expression levels of H+-ATPase and by modifying auxin accumulation in transgenic plants.

Role of the Arabidopsis PIN6 auxin transporter in auxin homeostasis and auxin-mediated development.

Plant-specific PIN-formed (PIN) efflux transporters for the plant hormone auxin are required for tissue-specific directional auxin transport and cellular auxin homeostasis. The Arabidopsis PIN protein family has been shown to play important roles in developmental processes such as embryogenesis, organogenesis, vascular tissue differentiation, root meristem patterning and tropic growth. Here we analyzed roles of the less characterised Arabidopsis PIN6 auxin transporter. PIN6 is auxin-inducible and is expressed during multiple auxin-regulated developmental processes. Loss of pin6 function interfered with primary root growth and lateral root development. Misexpression of PIN6 affected auxin transport and interfered with auxin homeostasis in other growth processes such as shoot apical dominance, lateral root primordia development, adventitious root formation, root hair outgrowth and root waving. These changes in auxin-regulated growth correlated with a reduction in total auxin transport as well as with an altered activity of DR5-GUS auxin response reporter. Overall, the data indicate that PIN6 regulates auxin homeostasis during plant development.

[Isolation, expression analysis of a chilling induced cDNA from rice root with differential display: an evidence role for caffeine-sensitive calcium signal].

Chilling-sensitive rice varieties acquire chilling tolerance when their roots are exposed to water stress for short time. Caffeine-sensitive calcium signal was involved in this procedure. By using total RNA differential display, a chilling induced cDNA(ICT: induction of chilling treatment) was isolated from roots of chilling-sensitive rice variety. It was determined that it is a novel cDNA by homology searching. The transcript level of ict mRNA is up-regulated under chilling stress, it is decreased to low level when the samples were transferred to standard culture conditions. Pre-treated with mannitol for two hours is beneficial to inducing ICT level of expression. This chilling induction was inhibited by caffeine, suggesting that it may play a putative role in signal transduction of caffeine-sensitive calcium.