Plant nuclear gene knockout reveals a role in plastid division for the homolog of the bacterial cell division protein FtsZ, an ancestral tubulin

Little is known about the division of eukaryotic cell organelles and up to now neither in animals nor in plants has a gene product been shown to mediate this process. A cDNA encoding a homolog of the bacterial cell division protein FtsZ, an ancestral tubulin, was isolated from the eukaryote Physcomitrella patens and used to disrupt efficiently the genomic locus in this terrestrial seedless plant. Seven out of 51 transgenics obtained were knockout plants generated by homologous recombination; they were specifically impeded in plastid division with no detectable effect on mitochondrial division or plant morphology. Implications on the theory of endosymbiosis and on the use of reverse genetics in plants are discussed.

Plastid Ontogeny during Petal Development in Arabidopsis1

Imaging of chlorophyll autofluorescence by confocal microscopy in intact whole petals of Arabidopsis thaliana has been used to analyze chloroplast development and redifferentiation during petal development. Young petals dissected from unopened buds contained green chloroplasts throughout their structure, but as the upper part of the petal lamina developed and expanded, plastids lost their chlorophyll and redifferentiated into leukoplasts, resulting in a white petal blade. Normal green chloroplasts remained in the stalk of the mature petal. In epidermal cells the chloroplasts were normal and green, in stark contrast with leaf epidermal cell plastids. In addition, the majority of these chloroplasts had dumbbell shapes, typical of dividing chloroplasts, and we suggest that the rapid expansion of petal epidermal cells may be a trigger for the initiation of chloroplast division. In petals of the Arabidopsis plastid division mutant arc6, the conversion of chloroplasts into leukoplasts was unaffected in spite of the greatly enlarged size and reduced number of arc6 chloroplasts in cells in the petal base, resulting in few enlarged leukoplasts in cells from the white lamina of arc6 petals.

Proteomic Analysis of Chloroplast-to-Chromoplast Transition in Tomato Reveals Metabolic Shifts Coupled with Disrupted Thylakoid Biogenesis Machinery and Elevated Energy-Production Components

A comparative proteomic approach was performed to identify differentially expressed proteins in plastids at three stages of tomato (Solanum lycopersicum) fruit ripening (mature-green, breaker, red). Stringent curation and processing of the data from three independent replicates identified 1,932 proteins among which 1,529 were quantified by spectral counting. The quantification procedures have been subsequently validated by immunoblot analysis of six proteins representative of distinct metabolic or regulatory pathways. Among the main features of the chloroplast-to-chromoplast transition revealed by the study, chromoplastogenesis appears to be associated with major metabolic shifts: (1) strong decrease in abundance of proteins of light reactions (photosynthesis, Calvin cycle, photorespiration) and carbohydrate metabolism (starch synthesis/degradation), mostly between breaker and red stages and (2) increase in terpenoid biosynthesis (including carotenoids) and stress-response proteins (ascorbate-glutathione cycle, abiotic stress, redox, heat shock). These metabolic shifts are preceded by the accumulation of plastid-encoded acetyl Coenzyme A carboxylase D proteins accounting for the generation of a storage matrix that will accumulate carotenoids. Of particular note is the high abundance of proteins involved in providing energy and in metabolites import. Structural differentiation of the chromoplast is characterized by a sharp and continuous decrease of thylakoid proteins whereas envelope and stroma proteins remain remarkably stable. This is coincident with the disruption of the machinery for thylakoids and photosystem biogenesis (vesicular trafficking, provision of material for thylakoid biosynthesis, photosystems assembly) and the loss of the plastid division machinery. Altogether, the data provide new insights on the chromoplast differentiation process while enriching our knowledge of the plant plastid proteome.

Natural variation in expression of genes associated with carotenoid biosynthesis and accumulation in cassava (Manihot esculenta Crantz) storage root.

ABSTRACT: BACKGROUND: Cassava (Manihot esculenta Crantz) storage root provides a staple food source for millions of people worldwide. Increasing the carotenoid content in storage root of cassava could provide improved nutritional and health benefits. Because carotenoid accumulation has been associated with storage root color, this study characterized carotenoid profiles, and abundance of key transcripts associated with carotenoid biosynthesis, from 23 landraces of cassava storage root ranging in color from white-to-yellow-to-pink. This study provides important information to plant breeding programs aimed at improving cassava storage root nutritional quality. RESULTS: Among the 23 landraces, five carotenoid types were detected in storage root with white color, while carotenoid types ranged from 1 to 21 in storage root with pink and yellow color. The majority of storage root in these landraces ranged in color from pale-to-intense yellow. In this color group, total ß-carotene, containing all-E-, 9-Z-, and 13-Z-ß-carotene isomers, was the major carotenoid type detected, varying from 26.13 to 76.72 %. Although no ?-carotene was observed, variable amounts of a ?-ring derived xanthophyll, lutein, was detected; with greater accumulation of ?-ring xanthophylls than of ß-ring xanthophyll. Lycopene was detected in a landrace (Cas51) with pink color storage root, but it was not detected in storage root with yellow color. Based on microarray and qRT-PCR analyses, abundance of transcripts coding for enzymes involved in carotenoid biosynthesis were consistent with carotenoid composition determined by contrasting HPLC-Diode Array profiles from storage root of landraces IAC12, Cas64, and Cas51. Abundance of transcripts encoding for proteins regulating plastid division were also consistent with the observed differences in total ß-carotene accumulation. CONCLUSIONS: Among the 23 cassava landraces with varying storage root color and diverse carotenoid types and profiles, landrace Cas51 (pink color storage root) had low LYCb transcript abundance, whereas landrace Cas64 (intense yellow storage root) had decreased HYb transcript abundance. These results may explain the increased amounts of lycopene and total ß-carotene observed in landraces Cas51 and Cas64, respectively. Overall, total carotenoid content in cassava storage root of color class representatives were associated with spatial patterns of secondary growth, color, and abundance of transcripts linked to plastid division. Finally, a partial carotenoid biosynthesis pathway is proposed.

Phylogenetic relationships in Solanaceae and related species based on cpDNA sequence from plastid trnE-trnT region

Intergenic spacers of chloroplast DNA (cpDNA) are very useful in phylogenetic and population genetic studies of plant species, to study their potential integration in phylogenetic analysis. The non-coding trnE-trnT intergenic spacer of cpDNA was analyzed to assess the nucleotide sequence polymorphism of 16 Solanaceae species and to estimate its ability to contribute to the resolution of phylogenetic studies of this group. Multiple alignments of DNA sequences of trnE-trnT intergenic spacer made the identification of nucleotide variability in this region possible and the phylogeny was estimated by maximum parsimony and rooted with Convolvulaceae Ipomoea batalas, the most closely related family. Besides, this intergenic spacer was tested for the phylogenetic ability to differentiate taxonomic levels. For this purpose, species from four other families were analyzed and compared with Solanaceae species. Results confirmed polymorphism in the trnE-trnT region at different taxonomic levels.

Wolbachia replication and host cell division in Aedes albopictus

Wolbachia pipientis is an obligate intracellular endosymbiont of a range of arthropod species. The microbe is best known for its manipulations of host reproduction that include inducing cytoplasmic incompatibility, parthenogenesis, feminization, and male-killing. Like other vertically transmitted intracellular symbionts, Wolbachiarsquos replication rate must not outpace that of its host cells if it is to remain benign. The mosquito Aedes albopictus is naturally infected both singly and doubly with different strains of Wolbachia pipientis. During diapause in mosquito eggs, no host cell division is believed to occur. Further development is triggered only by subsequent exposure of the egg to water. This study uses diapause in Wolbachia-infected Aedes albopictus eggs to determine whether symbiont replication slows or stops when host cell division ceases or whether it continues at a low but constant rate. We have shown that Wolbachia densities in eggs are greatest during embryonation and then decline throughout diapause, suggesting that Wolbachia replication is dependent on host cell replication.

The Rural Clinical Division: a student journey to rural medicine

Background: The University of Queensland has through an Australian Government initiative, established a Rural Clinical Division (RCD) at four regional sites in the southern and central Queensland. Over the fi rst four years of the existence of the RCD, an integrated package of innovative medical education has been developed. Method: The integrated aspects of the RCD program include: The Rural Medical Rotation: Every medical student undertakes an eight week rural rotation in Year 3. Year 3 and 4 MBBS - 100 students are currently spending one to two years in the rural school and demand is increasing. Interprofessional Education - Medical and Allied Health students attend lectures, seminars and workshops together and often share the same rural clinical placement. Rural health projects - allow students to undertake a project of benefi t to the rural community. Information Technology (IT) - the Clinical Discussion Board (CDB) and Personal Digital Assistants (PDA) demonstrate the importance of IT to medical students in the 21st century. Changing the Model of Medical Education - The Leichhardt Community Attachment Placement (LCAP), is a pilot study that resulted in the addition of three interns to the rural workforce. All aspects of the RCD are evaluated with surveys using both qualitative and quantitative free response questions, completed by all students regularly throughout the academic year. Results: Measures of impact include: Student satisfaction and quality of teaching surveys – 86-91% of students improved their clinical skills and understanding across all rotations. Academic results and progress – RCD students out-perform their urban colleagues. Intent to work in rural areas – 90% of students reported a greater interest in rural medicine. Intern numbers – rural / regional intern placements are increasing. Conclusions: The RCD proves to be a site for innovations all designed to help reach our primary goal of fostering increased recruitment of a rural medical workforce.

Post-familial families and the domestic division of labour

This paper takes as its starting point recent claims by Beck-Gernsheim (2002) that we are living in an era of post-familial families. Beck-Gernsheim (2002) argues that our lives are no longer structured as they once were by tradition, class, religion and kin. Instead the family has become a transitional phase as individuals strive for fulfillment of personal goals and personal life projects. The demographic evidence to support these claims is clearly evident in relation to changing patterns of family formation and dissolution, as well as the movement of married women into paid employment. But what is less evident is a decline in traditional patterns of gender stratification within families. This paper uses recent national data from Australia to examine the relationship between post-familial status, as indicated by marital status and employment, and time spent on housework. The results show that gender is still a clear predictor of time spent on housework, but that within gender there is evidence that gender inequality may be declining in non-traditional households.

Tooth-wear patterns in adolescents with normal occlusion and Class II Division 2 malocclusion

Introduction: In this study, we investigated tooth-wear patterns in adolescents with either normal occlusion or Class II Division 2 malocclusion. Methods: The sample consisted of dental casts from 165 subjects that were divided into 2 groups: 115 normal occlusion subjects (mean age, 14.3 years) and 50 complete Class II Division 2 subjects (mean age, 13.9 years). Dental wear was assessed by using a modified version of the tooth wear index. The 2 groups were compared with the Mann-Whitney test for the frequency and severity of wear on each surface of each group of teeth. The level of statistical significance was set at 5%. Results: The normal occlusion group statistically had greater tooth wear on the incisal surfaces of the maxillary lateral incisors and the incisal surfaces of the maxillary canines than did the Class II Division 2 malocclusion group. The malocclusion group showed statistically greater tooth wear on the labial surfaces of the mandibular lateral incisors, the occlusal surfaces of the maxillary premolars and first molars, the occlusal surfaces of the mandibular premolars, the palatal surfaces of the maxillary second premolars, and the buccal surfaces of the mandibular premolars and first molars than did the normal occlusion group. Conclusions: Subjects with normal occlusion and those with complete Class II Division 2 malocclusions have different tooth-wear patterns. Tooth wear on the malocclusion subjects should not be considered pathologic but, rather, the consequence of different interocclusal arrangements. (Am J Orthod Dentofacial Orthop 2010;137:730.e1-730.e5)

Tooth-wear patterns in subjects with Class II Division 1 malocclusion and normal occlusion

Introduction: The aim of this study was to investigate the prevalence of tooth wear in adolescents with Class II malocclusion, compared with those with normal occlusion. Methods: The sample consisted of dental casts obtained from 310 subjects, divided into 3 groups: group 1, 110 subjects with normal occlusion (mean age, 13.51 years); group 2, 100 complete Class II Division 1 patients (mean age, 13.44 years); and group 3, 100 half-cusp Class II Division 1 patients (mean age, 13.17 years). Dental wear was assessed by using a modified version of the tooth-wear index. The 3 groups were compared by means of the Kruskal-Wallis and Dunn tests, considering the frequency and the severity of wear on each surface of each group of teeth. The level of statistical significance was set at 5%. Results: The normal occlusion group had statistically greater tooth wear on the palatal surfaces of the maxillary central incisors and the incisal surfaces of the maxillary canines than the corresponding surfaces in both Class II malocclusion groups. The complete and half-cusp Class II Division 1 malocclusion groups had statistically greater tooth wear on the occlusal surfaces of the maxillary second premolar and first molar, the occlusal surfaces of the mandibular premolars, and the buccal surfaces of the mandibular posterior teeth compared with the normal occlusion group. The half-cusp Class II Division 1 malocclusion group had significantly greater tooth wear on the incisal surfaces of the mandibular incisors compared with the complete Class II Division 1 malocclusion group. Conclusions: Subjects with normal occlusion and complete or half-cusp Class II Division 1 malocclusions have different tooth-wear patterns. Tooth wear on the malocclusion subjects should not be considered pathologic but rather consequent to the different interocclusal tooth arrangement. (Am J Orthod Dentofacial Orthop 2010; 137: 14. e1-14.e7)

Stability and relapse of maxillary anterior crowding treatment in Class I and Class II Division 1 malocclusions

Introduction: The maxillary anterior teeth are the most important to facial esthetics because they are the first to show on a smile. Therefore, stability of the maxillary anterior teeth alignment is an important issue. The objective of this study was to compare the stability of maxillary anterior tooth alignment in Class I and Class II Division 1 malocclusions. Methods: The sample comprised dental casts of 70 patients with Class I and Class II Division 1 malocclusions and a minimum of 3 mm of maxillary anterior crowding measured by an irregularity index. The patients were treated with extractions and evaluated at pretreatment and posttreatment and at least 5 years after treatment. The sample was divided into 3 groups: group 1, Class I malocclusion treated with 4 first premolar extractions comprising 30 subjects, with an initial age of 13.16 years and 8.59 mm of initial maxillary irregularity; group 2, Class II malocclusion treated with 4 first premolar extractions comprising 20 subjects, with an initial age of 12.95 years and 11.10 mm of maxillary irregularity; and group 3, Class II malocclusion treated with 2 first maxillary premolar extractions comprising 20 subjects, with an initial age of 13.09 years and 9.68 mm of maxillary irregularity. Results: The decrease in the maxillary irregularity index was significantly greater in group 2 than in group 1 during treatment. The stability of maxillary anterior alignment was 88.12% over the long term; 77% of the linear displacement of the anatomic contact points tended to return to their original positions. Conclusions: Stability of maxillary anterior alignment between the 3 groups was similar. The stability of maxillary anterior alignment was high over the long term, but a high percentage of teeth tended to return to their original positions. (Am J Orthod Dentofacial Orthop 2011; 139: 768-74)

Occlusal outcomes and efficiency of 1-and 2-phase protocols in the treatment of Class II Division 1 malocclusion

Introduction: The purpose of this study was to compare the occlusal outcomes and the efficiency of 1-phase and 2-phase treatment protocols in Class II Division 1 malocclusions. Treatment efficiency was defined as a change in the occlusal characteristics in a shorter treatment time. Methods: Class II Division 1 subjects ( n = 139) were divided into 2 groups according to the treatment protocol for Class II correction. Group 1 comprised 78 patients treated with a 1-phase treatment protocol at initial and final mean ages of 12.51 and 14.68 years. Group 2 comprised 61 patients treated with a 2-phase treatment protocol at initial and final mean ages of 11.21 and 14.70 years. Lateral cephalometric radiographs were taken at the pretreatment stage to evaluate morphological differences in the groups. The initial and final study models of the patients were evaluated by using the peer assessment rating index. Chi-square tests were used to test for differences between the 2 groups for categorical variables. Variables regarding occlusal results were compared by using independent t tests. A linear regression analysis was completed, with total treatment time as the dependent variable, to identify clinical factors that predict treatment length for patients with Class II malocclusions. Results: Similar occlusal outcomes were obtained between the 1-phase and the 2-phase treatment protocols, but the duration of treatment was significantly shorter in the 1-phase treatment protocol group. Conclusions: Treatment of Class II Division 1 malocclusions is more efficient with the 1-phase than the 2-phase treatment protocol.