Cytogenetic analysis of Bienertia sinuspersici Akhani as the first step in genome sequencing.

BACKGROUND: C4 plants are efficient in suppressing photorespiration and enhancing carbon gain as compared to C3 plants. Bienertia sinuspersici Akhani is one of the few species in the family Amaranthaceae that can perform C4 photosynthesis within individual chlorenchyma cells, without the conventional Kranz anatomy in its leaf. This plant is salt-tolerant and is well-adapted to thrive in hot and humid climates. To date, there have been no reported cytogenetic analyses yet on this species. OBJECTIVE: This study aims to provide a cytogenetic analysis of B. sinuspersici as the first step in genome sequencing. METHODS: Fluorescence in situ hybridization (FISH) karyotype analysis was conducted using the metaphase chromosomes of B. sinuspersici probed with 5S and 45S rDNA and Arabidopsis-type telomeric repeats. RESULTS: Results of the cytogenetic analysis confirmed that B. sinuspersici has 2n = 2x = 18 consisting of nine pairs of metacentric chromosomes. Two loci of 45S rDNA were found on the distal regions of the short arm of chromosome 7. Nine loci of 5S rDNA were found in the pericentromeric regions of chromosomes 1, 3, 4, 6, and 8, which also colocalized with Arabidopsis-type telomeric repeats; while four loci in the interstitial regions of chromosome 5 and 8 can be observed. The single locus of 5S rDNA that was found in chromosome 8 appears to be hemizygous. CONCLUSION: The FISH karyotype analysis, based on the combination of rDNAs, telomeric tandem repeat markers and C0t DNA chromosome landmarks, allowed efficient chromosome identification and provided useful information in characterizing the genome of B. sinuspersici.

Comparative SEM and LM foliar epidermal and palyno-morphological studies of Amaranthaceae and its taxonomic implications.

Palynological features as well as comparative foliar epidermal using light and scanning electron microscope (SEM) of 17 species (10genera) of Amaranthaceae have been studied for its taxonomic significance. Different foliar and palynological micro-morphological characters were examined to explain their value in resolving the difficulty in identification. All species were amphistomatic but stomata on abaxial surface were more abundant. Taxonomically significant epidermal character including stomata type, trichomes (unicellular, multicellular, and capitate) and epidermal cells shapes (polygonal and irregular) were also observed. Pollens of this family are Polypantoporate, pores large, spheroidal, mesoporous region is sparsely to scabrate, densely psilate, and spinulose. All these characters can be active at species level for identification purpose. This study indicates that at different taxonomic levels, LM and SEM pollen and epidermal morphology is explanatory and significant to identify species and genera.

Developmental genetic mechanisms of C4 syndrome based on transcriptome analysis of C3 cotyledons and C4 assimilating shoots in Haloxylon ammodendron.

It is believed that transferring the C4 engine into C3 crops will greatly increase the yields of major C3 crops. Many efforts have been made since the 1960s, but relatively little success has been achieved because C4plant traits, referred to collectively as C4 syndrome, are very complex, and little is known about the genetic mechanisms involved. Unfortunately, there exists no ideal genetic model system to study C4 syndrome. It was previously reported that the Haloxylon species have different photosynthetic pathways in different photosynthetic organs, cotyledons and assimilating shoots. Here, we took advantage of the developmental switch from the C3 to the C4 pathway to study the genetic mechanisms behind this natural transition. We compared the transcriptomes of cotyledons and assimilating shoots using mRNA-Seq to gain insight into the molecular and cellular events associated with C4 syndrome. A total of 2959 differentially expressed genes [FDR ≤ 0.001 and abs (|log2(Fold change)| ≥ 1)] were identified, revealing that the transcriptomes of cotyledons and assimilating shoots are considerably different. We further identified a set of putative regulators of C4 syndrome. This study expands our understanding of the development of C4 syndrome and provides a new model system for future studies on the C3-to- C4 switch mechanism.

Protoplast isolation and transient gene expression in the single-cell C4 species, Bienertia sinuspersici.

Although transient gene expression using reporters such as green fluorescent protein is a versatile tool for examining gene functions and intracellular protein trafficking, the establishment of a highly efficient gene manipulation method remains a challenge in many plant species. A reliable transformation protocol has not yet been established for the three single-cell C(4) species, despite their potential of serving as model systems for their extraordinary C(4) photosynthetic metabolism. We report the first protocol optimized for isolating a large-scale and homogenous population of protoplasts from chlorenchyma cells of the single-cell C(4) species Bienertia sinuspersici. Cytochemical staining confirmed the preservation of the unusual subcellular compartmentation of organelles in chlorenchyma cells after cell wall digestion. Approximately 84% of isolated protoplasts expressed the reporter fluorescent protein following our optimized polyethylene glycol-mediated transfection procedures. Fluorescent fusion protein tagged with various intracellular sorting signals demonstrated potential use of the transient gene expression system in subcellular protein localization and organelle dynamics studies. Further applications of the current protoplast isolation and transfection techniques in understanding the novel single-cell C(4) photosynthetic mechanism are discussed.

Tolerance of combined submergence and salinity in the halophytic stem-succulent Tecticornia pergranulata.

BACKGROUND AND AIMS: Habitats occupied by many halophytes are not only saline, but are also prone to flooding. Few studies have evaluated submergence tolerance in halophytes. METHODS: Responses to submergence, at a range of salinity levels, were studied for the halophytic stem-succulent Tecticornia pergranulata subsp. pergranulata (syn. Halosarcia pergranulata subsp. pergranulata). Growth and total sugars in succulent stems were assessed as a function of time after submergence. Underwater net photosynthesis, dark respiration, total sugars, glycinebetaine, Na(+), Cl(-) and K(+), in succulent stems, were assessed in a NaCl dose-response experiment. KEY RESULTS: Submerged plants ceased to grow, and tissue sugars declined. Photosynthesis by succulent stems was reduced markedly when underwater, as compared with in air. Capacity for underwater net photosynthesis (P(N)) was not affected by 10-400 mM NaCl, but it was reduced by 30 % at 800 mM. Dark respiration, underwater, increased in succulent stems at 200-800 mM NaCl, as compared with those at 10 mM NaCl. On an ethanol-insoluble dry mass basis, K(+) concentration in succulent stems of submerged plants was equal to that in drained controls, across all NaCl treatments. Na(+) and Cl(-) concentrations, however, were elevated in stems of submerged plants, but so was glycinebetaine. Submerged stems increased in succulence, so solutes would have been 'diluted' on a tissue-water basis. CONCLUSIONS: Tecticornia pergranulata tolerates complete submergence, even in waters of high salinity. A 'quiescence response', i.e. no shoot growth, would conserve carbohydrates, but tissue sugars still declined with time. A low K(+) : Na(+) ratio, typical for tissues of succulent halophytes, was tolerated even during prolonged submergence, as evidenced by maintenance of underwater P(N) at up to 400 mM NaCl. Underwater P(N) provides O(2) and sugars, and thus should enhance survival of submerged plants.