Yaxiong Meng

Physiological and proteomic analyses of salt stress response in the halophyte Halogeton glomeratus

Very little is known about the adaptation mechanism of Chenopodiaceae Halogeton glomeratus, a succulent annual halophyte, under saline conditions. In this study, we investigated the morphological and physiological adaptation mechanisms of seedlings exposed to different concentrations of NaCl treatment for 21 d. Our results revealed that H. glomeratus has a robust ability to tolerate salt; its optimal growth occurs under approximately 100 mm NaCl conditions. Salt crystals were deposited in water-storage tissue under saline conditions. We speculate that osmotic adjustment may be the primary mechanism of salt tolerance in H. glomeratus, which transports toxic ions such as sodium into specific salt-storage cells and compartmentalizes them in large vacuoles to maintain the water content of tissues and the succulence of the leaves. To investigate the molecular response mechanisms to salt stress in H. glomeratus, we conducted a comparative proteomic analysis of seedling leaves that had been exposed to 200 mm NaCl for 24 h, 72 h and 7 d. Forty-nine protein spots, exhibiting significant changes in abundance after stress, were identified using matrix-assisted laser desorption ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF MS/MS) and similarity searches across EST database of H. glomeratus. These stress-responsive proteins were categorized into nine functional groups, such as photosynthesis, carbohydrate and energy metabolism, and stress and defence response.

Comparative Proteomic Analysis of Cultured Suspension Cells of the Halophyte Halogeton glomeratus by iTRAQ Provides Insights into Response Mechanisms to Salt Stress

Soil salinity severely threatens land use capability and crop yields worldwide. An analysis of the molecular mechanisms of salt tolerance in halophytes will contribute to the development of salt-tolerant crops. In this study, a combination of physiological characteristics and iTRAQ-based proteomic approaches was conducted to investigate the molecular mechanisms underlying the salt response of suspension cell cultures of halophytic Halogeton glomeratus. These cells showed halophytic growth responses comparable to those of the whole plant. In total, 97 up-regulated proteins and 192 down-regulated proteins were identified as common to both 200 and 400 mM NaCl concentration treatments. Such salinity responsive proteins were mainly involved in energy, carbohydrate metabolism, stress defense, protein metabolism, signal transduction, cell growth, and cytoskeleton metabolism. Effective regulatory protein expression related to energy, stress defense, and carbohydrate metabolism play important roles in the salt-tolerance of H. glomeratus suspension cell cultures. However, known proteins regulating Na+ efflux from the cytoplasm and its compartmentalization into the vacuole did not change significantly under salinity stress suggesting our existing knowledge concerning Na+ extrusion and compartmentalization in halophytes needs to be evaluated further. Such data are discussed in the context of our current understandings of the mechanisms involved in the salinity response of the halophyte, H. glomeratus.

Identification and selection of low-phosphate-tolerant germplasm in barley (Hordeum vulgare L.)

ABSTRACT Phosphorus (P) deficiency is one of the major constraints to crop yield worldwide, and genotypes or cultivars with high phosphate use efficiency (PUE) sustain growth when exposed to phosphate stress. Therefore, it is imperative to develop the genotypes or cultivars with high PUE. A pot experiment was conducted to evaluate the PUE among 150 barley (Hordeum vulgare L.) genotypes. Two high-tolerant and -sensitive accessions were selected. These two candidate materials were used to investigate the differences among the root morphology characteristics, antioxidant enzyme activity, inorganic phosphate (Pi) content and gene expression of HvPT5 under P-deficiency and P-sufficiency conditions. The values of these parameters were higher in the low-P-tolerant genotype than in the sensitive one. In pot experiment 1, all genotypes showed a significant difference in low-P tolerance, with variety GN121 achieving the highest tolerance, and GN42 being most sensitive. The results of this study may provide elite genetic germplasms for future work on isolation of P-related genes, and the improvement of PUE in barley. Abbreviations: PUE: phosphate use efficiency; CAT: catalase; POD: peroxidase; SOD: superoxide dismutase; DMSO: dimethyl sulphoxide; MDA: malondialdehyde; TOPSIS: technique for order preference by similarity to an ideal solution; MCDM/MADM: multi-criteria (or attribute) decision making

Transcriptome sequencing and comparative analysis of differentially-expressed isoforms in the roots of Halogeton glomeratus under salt stress

Although Halogeton glomeratus (H. glomeratus) has been confirmed to have a unique mechanism to regulate Na+ efflux from the cytoplasm and compartmentalize Na+ into leaf vacuoles, little is known about the salt tolerance mechanisms of roots under salinity stress. In the present study, transcripts were sequenced using the BGISEQ-500 sequencing platform (BGI, Wuhan, China). After quality control, approximately 24.08 million clean reads were obtained and the average mapping ratio to the reference gene was 70.00%. When comparing salt-treated samples with the control, a total of 550, 590, 1411 and 2063 DEIs were identified at 2, 6, 24 and 72h, respectively. Numerous differentially-expressed isoforms that play important roles in response and adaptation to salt condition are related to metabolic processes, cellular processes, single-organism processes, localization, biological regulation, responses to stimulus, binding, catalytic activity and transporter activity. Fifty-eight salt-induced isoforms were common to different stages of salt stress; most of these DEIs were related to signal transduction and transporters, which maybe the core isoforms regulating Na+ uptake and transport in the roots of H. glomeratus. The expression patterns of 18 DEIs that were detected by quantitative real-time polymerase chain reaction were consistent with their respective changes in transcript abundance as identified by RNA-Seq technology. The present study thoroughly explored potential isoforms involved in salt tolerance on H. glomeratus roots at five time points. Our results may serve as an important resource for the H. glomeratus research community, improving our understanding of salt tolerance in halophyte survival under high salinity stress.

Genetic Diversity and Association Analysis of Agronomic Characteristics with SSR Markers in Hulless Barley

The objectives of this study were to find molecular markers associated with yield-related traits and guide parental combination in molecular marker-assisted breeding and hybrid breeding of hulless barley (Hordeum vulgare L. var. nudum HK. f.). A natural hulless barley population composed of 108 parental varieties/lines was screened with 92 SSR markers, in which 48 markers were polymorphic. Population structure was analyzed based on the polymorphic SSR data and association between markers and five agronomic traits were performed in TASSEL GLM (general linear model) and MLM (mixed linear model) programs. A total of 156 alleles were detected in the 108 varieties/lines with 2–6 alleles per locus. The Shannon’s index of the population ranged from 0.6727 to 1.1368 and the genetic similarity between varieties ranged from 0.2250 to 1.0000, with the mean of 0.7585. Structure analysis revealed four genetic subpopulations for the entire materials tested. Based on GLM analysis, 12 SSR markers were found to be associated with plant height, spike length, grain number per spike and tiller number, with phenotypic contributions of 11.5–17.6%, 19.4–45.4%, 15.4–22.1% and 29.2%, respectively. Based on MLM analysis, 8 SSR markers were associated with plant height, awn length, and spikelet compactness, with the phenotypic contributions of 31.7–49.9%, 28.1–37.2%, and 22.7–32.7%, respectively. These associated markers were distributed on 6 chromosomes of the barley genome. 第 2期 孟亚雄等: 青稞遗传多样性及其农艺性状与 SSR标记的关联分析 181

Comparative transcriptome analysis of genes involved in Na+ transport in the leaves of halophyte Halogeton glomeratus

Compartmentalization of Na+ into vacuoles is considered to be the most critical aspect of salt tolerance in H. glomeratus, an annual, succulent halophyte. Previous analysis of transcriptome involved in the H. glomeratus salt stress response relied on next-generation sequencing technologies that limit the capture of accurately spliced, full-length isoforms. To gain deeper insights into its salt stress response, we used the H. glomeratus Iso-Seq transcriptome database as a reference, and subsequent next-generation sequencing was subjected to various NaCl concentrations of leaves from plants revealed 115 upregulated and 87 downregulated differentially expressed isoforms (core DEIs). The majority of the core DEIs were involved in carbohydrate metabolism and energy production and conversion. In contrast, levels of known isoforms encoding Na+ transporters did not change significantly under salt stress. However, 16 core DEIs of unknown function were predicted to possess transmembrane domains, suggesting that these candidate isoforms could be involved in Na+ transport in H. glomeratus. These results suggest a potential means for identification of novel Na+ transporters, in addition to providing a foundation for further investigation of Na+ transport networks in halophytes.

Molecular Mechanisms of Acclimatization to Phosphorus Starvation and Recovery underlying full-length transcriptome profiling in Barley (Hordeum vulgare L.)

A lack of phosphorus (P) in plants can severely constrain growth and development. Barley, one of the earliest domesticated crops, is extensively planted in poor soil around the world. To date, the molecular mechanisms of enduring low phosphorus, at the transcriptional level, in barley are still unclear. In the present study, two different barley genotypes (GN121 and GN42)—with contrasting phosphorus efficiency—were used to reveal adaptations to low phosphorus stress, at three time points, at the morphological, physiological, biochemical, and transcriptome level. GN121 growth was less affected by phosphorus starvation and recovery than that of GN42. The biomass and inorganic phosphorus concentration of GN121 and GN42 declined under the low phosphorus-induced stress and increased after recovery with normal phosphorus. However, the range of these parameters was higher in GN42 than in GN121. Subsequently, a more complete genome annotation was obtained by correcting with the data sequenced on Illumina HiSeq X 10 and PacBio RSII SMRT platform. A total of 6,182 and 5,270 differentially expressed genes (DEGs) were identified in GN121 and GN42, respectively. The majority of these DEGs were involved in phosphorus metabolism such as phospholipid degradation, hydrolysis of phosphoric enzymes, sucrose synthesis, phosphorylation/dephosphorylation and post-transcriptional regulation; expression of these genes was significantly different between GN121 and GN42. Specifically, six and seven DEGs were annotated as phosphorus transporters in roots and leaves, respectively. Furthermore, a putative model was constructed relying on key metabolic pathways related to phosphorus to illustrate the higher phosphorus efficiency of GN121 compared to GN42 under low phosphorus conditions. Results from this study provide a multi-transcriptome database and candidate genes for further study on phosphorus use efficiency (PUE).