Summary• The petals of the lower eudicot family Ranunculaceae are thought to have been derived many times independently from stamens. However, investigation of the genetic basis of their identity has suggested an alternative hypothesis: that they share a commonly inherited petal identity program. This theory is based on the fact that an ancient paralogous lineage of APETALA3 (AP3) in the Ranunculaceae appears to have a conserved, petal-specific expression pattern.• Here, we have used a combination of approaches, including RNAi, comparative gene expression and molecular evolutionary studies, to understand the function of this petal-specific AP3 lineage.• Functional analysis of the Aquilegia locus AqAP3-3 has demonstrated that the paralog is required for petal identity with little contribution to the identity of the other floral organs. Expanded expression studies and analyses of molecular evolutionary patterns provide further evidence that orthologs of AqAP3-3 are primarily expressed in petals and are under higher purifying selection across the family than the other AP3 paralogs.• Taken together, these findings suggest that the AqAP3-3 lineage underwent progressive subfunctionalization within the order Ranunculales, ultimately yielding a specific role in petal identity that has probably been conserved, in stark contrast with the multiple independent origins predicted by botanical theories.

BackgroundEnteric parasites are the most common cause of parasitic diseases and cause significant morbidity and mortality, particularly in developing countries like Nepal. The objective of this study was to estimate the prevalence and risk factors of intestinal parasitic infections among school going children of Lalitpur district of Nepal.MethodsA total of 1392 stool samples were collected from school children of two government, two private and two community schools of the same district. The stool samples were examined for evidence of parasitic infections by direct microscopy and confirmed by concentration methods (formal ether sedimentation technique or floatation technique by using Sheather’s sugar solution). Modified Ziehl-Neelsen (ZN) staining was performed for the detection of coccidian parasites.ResultsPrevalence of intestinal parasitosis was found to be 16.7%. The highest prevalence rate was seen with Giardia lamblia (7.4%) followed by Entamoeba histolytica (3.4%) and Cyclospora cayetanensis (1.6%). Children aged 11–15 years and the ones belonging to family of agriculture workers were most commonly affected. Hand washing practice and type of drinking water also showed significant difference.ConclusionsThe burden of parasitic infections among the school children, coupled with the poor sanitary conditions in the schools, should be regarded as an issue of public health priority and demands for effective school health programs involving periodic health education and screening.

SummaryPrevious studies of the lower eudicot model Aquilegia have revealed differential expression patterns of two APETALA3 (AP3) paralogs that appear to coincide with the development of a distinct fifth floral organ type, the staminodium. The AqAP3-1 locus quickly becomes limited to the staminodia while AqAP3-2 becomes stamen-specific.We used transient RNAi-based methods to silence each of these loci individually and in combination, followed by detailed studies of the resultant morphologies and the effects on gene expression patterns.Silencing of AqAP3-1 had a strong effect on the staminodia, causing transformation into carpeloid organs, while silencing of AqAP3-2 only affected the stamens, resulting in sterility, stunting or weak transformation towards carpel identity. Much more dramatic phenotypes were obtained in the doubly silenced flowers, where all stamens and staminodia were transformed into carpels. Quantitative reverse-transcription polymerase chain reaction analyses of B gene homolog expression in these flowers are consistent with complex patterns of regulatory feedback among the loci.These findings suggest that the presence of ancient AP3 paralogs in the Ranunculaceae has facilitated the recent evolution of a novel organ identity program in Aquilegia. Specifically, it appears that downregulation of AqAP3-2 in the innermost whorl of stamens was a critical step in the evolution of elaborated sterile organs in this position.

SUMMARYThe APETALA1/FRUITFULL (AP1/FUL) MADS box transcription factors are best known for the role of AP1 in Arabidopsis sepal and petal identity, the canonical A function of the ABC model of flower development. However, this gene lineage underwent multiple duplication events during angiosperm evolution, providing different taxa with unique gene complements. One such duplication correlates with the origin of the core eudicots, and produced the euAP1 and euFUL clades. Together, euAP1 and euFUL genes function in proper floral meristem identity and repression of axillary meristem growth. Independently, euAP1 genes function in floral meristem and sepal identity, whereas euFUL genes control phase transition, cauline leaf growth and fruit development. To investigate the impact of the core eudicot duplication on the functional diversification of this gene lineage, we studied the role of pre-duplication FUL-like genes in columbine (Aquilegia coerulea). Our results show that AqcFL1 genes are broadly expressed in vegetative and reproductive meristems, leaves and flowers. Virus-induced gene silencing of the loci results in plants with increased branching, shorter inflorescences with fewer flowers, and dramatic changes in leaf shape and complexity. However, aqcfl1 plants have normal flowers and fruits. Our results show that, in contrast to characterized AP1/FUL genes, the AqcFL1 loci are either genetically redundant or have been decoupled from the floral genetic program, and play a major role in leaf morphogenesis. We analyze the results in the context of the core eudicot duplication, and discuss the implications of our findings in terms of the genetic regulation of leaf morphogenesis in Aquilegia and other flowering plants.

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