Validation of spot urine in predicting 24-h sodium excretion at the individual level.

Background: Evidence for the effect of dietary sodium intake on the risk of cardiovascular disease has been controversial. One of the main explanations for the conflicting results lies in the great variability associated with measurement methods for sodium intake. Spot urine collection is a convenient method commonly used for sodium estimation, but its validity for predicting 24-h urinary sodium excretion at the individual level has not been well evaluated among the general population.Objective: The aim of this study was to evaluate the validity of the Kawasaki, the International Cooperative Study on Salt, Other Factors, and Blood Pressure (INTERSALT), and the Tanaka formulas in predicting 24-h urinary sodium excretion by using spot urine samples in Chinese adults.Design: We analyzed the relative and absolute differences and misclassification at the individual level from 3 commonly used methods for estimating sodium intake among 141 Chinese community residents.Results: The mean measured 24-h sodium excretion was 220.8 mmol/d. The median (95% CIs) differences between measured sodium and those estimated from the Kawasaki, INTERSALT, and Tanaka methods were 6.4 mmol/d (-17.5, 36.8 mmol/d), -67.3 mmol/d (-96.5, -46.9 mmol/d), and -42.9 mmol/d (-59.1, -24.8 mmol/d), respectively. The proportions of relative differences >40% with the Kawasaki, INTERSALT, and Tanaka methods were 31.2%, 41.1%, and 22.0%, respectively; and the absolute difference for the 3 methods was >51.3 mmol/d (3 g salt) in approximately half of the participants. The misclassification rate was 63.1% for the Kawasaki method, 78.7% for the INTERSALT method, and 66.0% for the Tanaka method at the individual level.Conclusion: The results from our study do not support the use of spot urine to estimate 24-h urinary sodium excretion at the individual level because of its poor performance with respect to misclassification. This trial was registered at www.chictr.org.cn as ChiCTR-IOR-16010278.

LATHYROIDES, encoding a WUSCHEL-related Homeobox1 transcription factor, controls organ lateral growth, and regulates tendril and dorsal petal identities in garden pea (Pisum sativum L.).

During organ development, many key regulators have been identified in plant genomes, which play a conserved role among plant species to control the organ identities and/or determine the organ size and shape. It is intriguing whether these key regulators can acquire diverse function and be integrated into different molecular pathways among different species, giving rise to the immense diversity of organ forms in nature. In this study, we have characterized and cloned LATHYROIDES (LATH), a classical locus in pea, whose mutation displays pleiotropic alteration of lateral growth of organs and predominant effects on tendril and dorsal petal development. LATH encodes a WUSCHEL-related homeobox1 (WOX1) transcription factor, which has a conserved function in determining organ lateral growth among different plant species. Furthermore, we showed that LATH regulated the expression level of TENDRIL-LESS (TL), a key factor in the control of tendril development in compound leaf, and LATH genetically interacted with LOBED STANDARD (LST), a floral dorsal factor, to affect the dorsal petal identity. Thus, LATH plays multiple roles during organ development in pea: it maintains a conserved function controlling organ lateral outgrowth, and modulates organ identities in compound leaf and zygomorphic flower development, respectively. Our data indicated that a key regulator can play important roles in different aspects of organ development and dedicate to the complexity of the molecular mechanism in the control of organ development so as to create distinct organ forms in different species.

Genetic analysis of ele mutants and comparative mapping of ele1 locus in the control of organ internal asymmetry in garden pea.

Previous study has shown that during zygomorphic development in garden pea (Pisum sativum L.), the organ internal (IN) asymmetry of lateral and ventral petals was regulated by a genetic locus, SYMMETRIC PETAL 1 (SYP1), while the dorsoventral (DV) asymmetry was determined by two CYC-like TCP genes or the PsCYC genes, KEELED WINGS (K) and LOBED STANDARD 1 (LST1). In this study, two novel loci, ELEPHANT EAR-LIKE LEAF 1 (ELE1) and ELE2 were characterized. These mutants exhibit a similar defect of IN asymmetry as syp1 in lateral and ventral petals, but also display pleiotropic effects of enlarged organ size. Genetic analysis showed that ELE1 and ELE2 were involved in same genetic pathway and the enlarged size of petals but not compound leaves in ele2 was suppressed by introducing k and lst1, indicating that the enlargement of dorsal petal in ele2 requires the activities of K and LST1. An experimental framework of comparative genomic mapping approach was set up to map and clone LjELE1 locus in Lotus japonicus. Cloning the ELE1 gene will shed light on the underlying molecular mechanism during zygomorphic development and further provide the molecular basis for genetic improvement on legume crops.

Wrinkled petals and stamens 1, is required for the morphogenesis of petals and stamens in Lotus japonicus.

Although much progress has been made in understanding how floral organ identity is determined during the floral development, less is known about how floral organ is elaborated in the late floral developmental stages. Here we describe a novel floral mutant, wrinkled petals and stamens1 (wps1), which shows defects in the development of petals and stamens. Genetic analysis indicates that wps1 mutant is corresponding to a single recessive locus at the long arm of chromosome 3. The early development of floral organs in wps1 mutant is similar to that in wild type, and the malfunction of the mutant commences in late developmental stages, displaying a defect on the appearance of petals and stamens. In the mature flower, petals and stamen filaments in the mutant are wrinkled or folded, and the cellular morphology under L1 layer of petals and stamen filaments is abnormal. It is found that the expression patterns of floral organ identity genes are not affected in wps1 mutants compared with that of wild type, consistent with the unaltered development of all floral organs. Furthermore, the identities of epidermal cells in different type of petals are maintained. The histological analysis shows that in wps1 flowers all petals are irregularly folded, and there are knotted structures in the petals, while the shape and arrangement of inner cells are malformed and unorganized. Based on these results, we propose that Wps1 acts downstream to the class B floral organ identity genes, and functions to modulate the cellular differentiation during the late flower developmental stages.

Different expression patterns of duplicated PHANTASTICA-like genes in Lotus japonicus suggest their divergent functions during compound leaf development.

Recent studies on leaf development demonstrate that the mechanism on the adaxial-abaxial polarity pattern formation could be well conserved among the far-related species, in which PHANTASTICA (PAHN)-like genes play important roles. In this study, we explored the conservation and diversity on functions of PHAN-like genes during the compound leaf development in Lotus japonicus, a papilionoid legume. Two PHAN-like genes in L. japonicus, LjPHANa and LjPHANb, were found to originate from a gene duplication event and displayed different expression patterns during compound leaf development. Two mutants, reduced leaflets1 (rel1) and reduced leaflets3 (rel3), which exhibited decreased adaxial identity of leaflets and reduced leaflet initiation, were identified and investigated. The expression patterns of both LjPHANs in rel mutants were altered and correlated with abnormalities of compound leaves. Our data suggest that LjPHANa and LjPHANb play important but divergent roles in regulating adaxial-abaxial polarity of compound leaves in L. japonicus.

Floral patterning in Lotus japonicus.

Floral patterning in Papilionoideae plants, such as pea (Pisum sativum) and Medicago truncatula, is unique in terms of floral organ number, arrangement, and initiation timing as compared to other well-studied eudicots. To investigate the molecular mechanisms involved in the floral patterning in legumes, we have analyzed two mutants, proliferating floral meristem and proliferating floral organ-2 (pfo-2), obtained by ethyl methanesulfonate mutagenesis of Lotus japonicus. These two mutants showed similar phenotypes, with indeterminate floral structures and altered floral organ identities. We have demonstrated that loss of function of LjLFY and LjUFO/Pfo is likely to be responsible for these mutant phenotypes, respectively. To dissect the regulatory network controlling the floral patterning, we cloned homologs of the ABC function genes, which control floral organ identity in Arabidopsis (Arabidopsis thaliana). We found that some of the B and C function genes were duplicated. RNA in situ hybridization showed that the C function genes were expressed transiently in the carpel, continuously in stamens, and showed complementarity with the A function genes in the heterogeneous whorl. In proliferating floral meristem and pfo-2 mutants, all B function genes were down-regulated and the expression patterns of the A and C function genes were drastically altered. We conclude that LjLFY and LjUFO/Pfo are required for the activation of B function genes and function together in the recruitment and determination of petals and stamens. Our findings suggest that gene duplication, change in expression pattern, gain or loss of functional domains, and alteration of key gene functions all contribute to the divergence of floral patterning in L. japonicus.

Cloning of LjCYC1 gene and nuclear localization of LjCYC1 protein in Lotus japonicus.

The LjCYC1 (Lotus japonicus Cycloidea-like 1) gene, a homolog of CYC (Cycloidea) belonging to the TCP [TB1(teosinte branched 1), CYC, PCFs (PCF1 and PCF2)] gene family and encoding a predicted transcription factor and being proposed controlling different aspects of plant development, was isolated from the papilionaceous plant Lotus japonicus by screening the genomic DNA library, in order to test the functional conservation and divarication of CYC-like genes in legume. Sequence analyses indicate that LjCYC1 gene contains two exons and one intron and encodes a 370-AA peptide LjCYC1. The putative protein, LjCYC1, contains a TCP domain and an R domain, being a member of the CYC/TB1 subfamily of TCP family, and has 39.0% identity with and 42.6% similarity to CYC. LjCYC1-cDNA was cloned through RT-PCR. Different regions of the LjCYC1-cDNA were fused with the report gene GUS and then the fused constructs were transiently expressed in the onion epidermal cells through particle bombardment. Results of GUS and DAPI staining showed that the chimeric proteins with TCP domain were localized within the nucleus, confirming that LjCYC1 may act as a transcription factor. But the TCP domain itself could not confer the nuclear localization because the chimeric proteins with TCP domain alone were dispersed all over the transformed cells.