Diagnosis and management of congenital type D esophageal atresia.

This study was performed to describe the current clinical practice and outcomes of type D esophageal atresia. We retrospectively analyzed 10 patients who were diagnosed with type D esophageal atresia and underwent esophageal atresia and tracheoesophageal fistula repair in the Capital Institute of Pediatrics and Beijing Children's Hospital from January 2017 to May 2022. Ten patients include three newborns and seven non-newborns. Seven (70%) cases were misdiagnosed as type C esophageal atresia before the first operation. Three neonatal children underwent thoracoscopic distal tracheoesophageal fistula ligation and esophageal anastomosis: the proximal tracheoesophageal fistula was simultaneously repaired with thoracoscopy in one of these children, and the proximal tracheoesophageal fistula was not detected under thoracoscopy in the other two children. Among the seven non-neonatal children, one underwent repair of the proximal tracheoesophageal fistula through the chest and the other six underwent repair through the neck. Nine patients were cured, and one died of complications of severe congenital heart disease. Type D esophageal atresia lacks specific clinical manifestations. Misdiagnosis as type C esophageal atresia is the main cause of an unplanned reoperation. Patients without severe malformations have a good prognosis.

Wheat AGAMOUS LIKE 6 transcription factors function in stamen development by regulating the expression of Ta APETALA3.

Previous studies have revealed the functions of rice and maize AGAMOUS LIKE 6 (AGL6) genes OsMADS6 and ZAG3, respectively, in floral development; however, the functions of three wheat (Triticum aestivum) AGL6 genes are still unclear. Here, we report the main functions of wheat AGL6 homoeologous genes in stamen development. In RNAi plants, stamens showed abnormality in number and morphology, and a tendency to transform into carpels. Consistently, the expression of the B-class gene TaAPETALA3 (AP3) and the auxin-responsive gene TaMGH3 was downregulated, whereas the wheat ortholog of the rice carpel identity gene DROOPING LEAF was ectopically expressed in RNAi stamens. TaAGL6 proteins bind to the promoter of TaAP3 directly. Yeast one-hybrid and transient expression assays further showed that TaAGL6 positively regulates the expression of TaAP3 in vivo. Wheat AGL6 transcription factors interact with TaAP3, TaAGAMOUS and TaMADS13. Our findings indicate that TaAGL6 transcription factors play an essential role in stamen development through transcriptional regulation of TaAP3 and other related genes. We propose a model to illustrate the function and probable mechanism of this regulation. This study extends our understanding of AGL6 genes.

Correction: Arabidopsis WRKY6 Transcription Factor Acts as a Positive Regulator of Abscisic Acid Signaling during Seed Germination and Early Seedling Development.

[This corrects the article DOI: 10.1371/journal.pgen.1005833.].

Genome-Wide Analysis of the RAV Gene Family in Wheat and Functional Identification of TaRAV1 in Salt Stress.

RAV transcription factors (TFs) are unique to higher plants and contain both B3 and APETALA2 (AP2) DNA binding domains. Although sets of RAV genes have been identified from several species, little is known about this family in wheat. In this study, 26 RAV genes were identified in the wheat genome. These wheat RAV TFs were phylogenetically clustered into three classes based on their amino acid sequences. A TaRAV gene located on chromosome 1D was cloned and named TaRAV1. TaRAV1 was expressed in roots, stems, leaves, and inflorescences, and its expression was up-regulated by heat while down-regulated by salt, ABA, and GA. Subcellular localization analysis revealed that the TaRAV1 protein was localized in the nucleus. The TaRAV1 protein showed DNA binding activity in the EMSA assay and transcriptional activation activity in yeast cells. Overexpressing TaRAV1 enhanced the salt tolerance of Arabidopsis and upregulated the expression of SOS genes and other stress response genes. Collectively, our data suggest that TaRAV1 functions as a transcription factor and is involved in the salt stress response by regulating gene expression in the SOS pathway.

MYB77 regulates high-affinity potassium uptake by promoting expression of HAK5.

In Arabidopsis, the high-affinity K+ transporter HAK5 is the major pathway for root K+ uptake when below 100 µM; HAK5 responds to Low-K+ (LK) stress by strongly and rapidly increasing its expression during K+ -deficiency. Therefore, positive regulators of HAK5 expression have the potential to improve K+ uptake under LK. Here, we show that mutants of the transcription factor MYB77 share a LK-induced leaf chlorosis phenotype, lower K+ content, and lower Rb+ uptake of the hak5 mutant, but not the shorter root growth, and that overexpression of MYB77 enhanced K+ uptake and improved tolerance to LK stress. Furthermore, we demonstrated that MYB77 positively regulates the expression of HAK5, by binding to the HAK5 promoter and enhances high-affinity K+ uptake of roots. As such, our results reveal a novel pathway for enhancing HAK5 expression under LK stress, and provides a candidate for increasing the tolerance of plants to LK.

Comparison of Trihelix transcription factors between wheat and Brachypodium distachyon at genome-wide.

BACKGROUND: Plant Trihelix transcription factors, specifically bind to GT elements and play important roles in plant physiology and development. Wheat is a main cereal crop. Brachypodium distachyon is a close relative of wheat and has been described as a new model species for studying of grass functional genomics. Presently, little is known about wheat and B. distachyon Trihelix genes. RESULTS: In 51 species, 2387 Trihelix genes were identified, including 80 wheat Trihelix genes and 27 B. distachyon Trihelix genes. Consistent with the results of previous studies, these genes were classified into five subfamilies: GT-1, GT-2, SIP1, GTγ, and SH4. Members of the same subfamily shared similar gene structures and common motifs. Most TaGT and BdGT genes contained many kinds of cis-elements, such as development-, stress-, and phytohormone-related cis-acting elements. Additionally, 21 randomly selected TaGT genes were mainly expressed in the roots and flowers, while the expression of 19 selected BdGT genes was constitutive. These results indicate that the roles of Trihelix genes in wheat and B. distachyon might have diversified during the evolutionary process. The expression of the most selected TaGT and BdGT genes was down-regulated when exposed to low temperatures, NaCl, ABA, and PEG, implying that TaGT and BdGT genes negatively respond to abiotic stress. On the contrary, the expression of some genes was up-regulated under heat stress. CONCLUSIONS: Trihelix genes exist extensively in plants and have many functions. During the evolutionary process, this gene family expanded and their functions diversified. As a result, the expression pattern and functions of members of the same family might be different. This study lays a foundation for further functional analyses of TaGT and BdGT genes.

Arabidopsis WRKY6 Transcription Factor Acts as a Positive Regulator of Abscisic Acid Signaling during Seed Germination and Early Seedling Development.

The phytohormone abscisic acid (ABA) plays important roles during seed germination and early seedling development. Here, we characterized the function of the Arabidopsis WRKY6 transcription factor in ABA signaling. The transcript of WRKY6 was repressed during seed germination and early seedling development, and induced by exogenous ABA. The wrky6-1 and wrky6-2 mutants were ABA insensitive, whereas WRKY6-overexpressing lines showed ABA-hypersensitive phenotypes during seed germination and early seedling development. The expression of RAV1 was suppressed in the WRKY6-overexpressing lines and elevated in the wrky6 mutants, and the expression of ABI3, ABI4, and ABI5, which was directly down-regulated by RAV1, was enhanced in the WRKY6-overexpressing lines and repressed in the wrky6 mutants. Electrophoretic mobility shift and chromatin immunoprecipitation assays showed that WRKY6 could bind to the RAV1 promoter in vitro and in vivo. Overexpression of RAV1 in WRKY6-overexpressing lines abolished their ABA-hypersensitive phenotypes, and the rav1 wrky6-2 double mutant showed an ABA-hypersensitive phenotype, similar to rav1 mutant. Together, the results demonstrated that the Arabidopsis WRKY6 transcription factor played important roles in ABA signaling by directly down-regulating RAV1 expression.

BRS1 Function in Facilitating Lateral Root Emergence in Arabidopsis.

The BRS1 (BRI1 Suppressor 1) gene encodes a serine carboxypeptidase that plays a critical role in the brassinosteroid signaling pathway. However, its specific biological function remains unclear. In this study, the developmental role of BRS1 was investigated in Arabidopsis thaliana. We found that overexpressing BRS1 resulted in significantly more lateral roots in different Arabidopsis ecotypes (WS2 and Col-0) and in brassinosteroid mutants (bri1-5 and det2-28). Further research showed that BRS1 facilitates the process whereby lateral root primordia break through the endodermis, cortex, and epidermis. Consistent with this, BRS1 was found to be highly expressed in the root endodermis and accumulated in the extracellular space around the dome of the lateral root primordia. Taken together, these results highlight the role of BRS1 in the process of lateral root emergence and provide new insight into the role of serine carboxypeptidases in plant root development.

Arabidopsis RAV1 transcription factor, phosphorylated by SnRK2 kinases, regulates the expression of ABI3, ABI4, and ABI5 during seed germination and early seedling development.

The phytohormone abscisic acid (ABA) modulates a number of processes during plant growth and development. In this study, the molecular mechanism of Arabidopsis RAV (Related to ABI3/VP1) transcription factor RAV1 involving ABA signaling was investigated. RAV1-underexpressing lines were more sensitive to ABA than wild-type plants during seed germination and early seedling development, whereas RAV1-overexpressing lines showed strong ABA-insensitive phenotypes. Overexpression of RAV1 repressed ABI3, ABI4, and ABI5 expression, and RAV1 bound to the ABI3, ABI4, and ABI5 promoters in vitro and in vivo, indicating that RAV1 directly down-regulates the expression of ABI3, ABI4, and ABI5. The interruption of ABI5 function in RAV1-U abi5 plants abolished the ABA-hypersensitive phenotype of RAV1-U plants, demonstrating that ABI5 is epistatic to RAV1. RAV1 interacted with SNF1-RELATED PROTEIN KINASE SnRK2.2, SnRK2.3 and SnRK2.6 in the nucleus. In vitro kinase assays showed that SnRK2.2, SnRK2.3 and SnRK2.6 phosphorylated RAV1. Transient expression assays revealed that SnRK2.2, SnRK2.3 and SnRK2.6 reduced the RAV1-dependent repression of ABI5, and the ABA-insensitive phenotype of the RAV1-overexpressing line was impaired by overexpression of SnRK2.3 in the RAV1 OE3 plants. Together, these results demonstrated that the Arabidopsis RAV1 transcription factor plays an important role in ABA signaling by modulating the expression of ABI3, ABI4, and ABI5, and that its activity is negatively affected by SnRK2s.

A method for separating tonoplast from wheat.

Vacuoles account for 90% of plant cell volume and play important roles in maintaining osmotic pressure, storing metabolites and lysosomes, compartmentalizing harmful ions, and storing and reusing minerals. These functions closely relay on the ion channels and transporters located on the tonoplast. The separation of intact vacuoles from plant cells is the key technology utilized in the study of tonoplast-located ion channels and transporters. However, the current vacuole separation methods are available for Arabidopsis and some other dicotyledons but are lacking for monocot crops. In this study, we established a new method for the vacuole separation from wheat mesophyll cells and investigated the transmembrane proton flux of tonoplasts with non-invasive micro-test technology (NMT). Moreover, our study provides a technology for the study of vacuole functions in monocot crops.

Multiple ALMT subunits combine to form functional anion channels: A case study for rice ALMT7.

The Aluminum Activated Malate Transporter (ALMT) family members are anion channels that play important roles in organic acid transport, stress resistance, growth, development, fertilization and GABA responses. The rice malate permeable OsALMT7 influences panicle development and grain yield. A truncated OsALMT7 mutant, panicle apical abortion1 (paab1) lacking at least 2 transmembrane helices, mediates reduced malate efflux resulting in yield reducing. Here, we further investigated the contribution of OsALMT7 transmembrane helices to channel activity, using heterologous expression in Xenopus laevis oocytes. We further found that OsALMT7 formed as a homomer by co-expressing OsALMT7 and paab1 proteins in oocytes and detecting the physical interaction between two OsALMT7, and between OsALMT7 and paab1 mutant protein. Further study proved that not just OsALMT7, mutants of TaALMT1 inhibit wild-type TaALMT1 channel, indicating that ALMTs might perform channel function as homomers. Our discovery brings a light for ion channel structure and homomultimer regulation understanding for ALMT anion channels and potential for crop grain yield and stress response improvement in the context of the essential role of ALMTs in these plant processes.

Genome-wide identification of expansin genes in Brachypodium distachyon and functional characterization of BdEXPA27.

Plant expansin belongs to a group of cell wall proteins and functions in plant growth and development. However, limited data are available on the contributions of expansins in Brachypodium distachyon. In the present study, a total of 38 expansins were identified in B. distachyon genome. Phylogenetic analysis divided the expansins into four groups, namely EXPA, EXPB, EXLA, and EXLB. Chromosomal distribution showed that they were unevenly distributed on 4 chromosomes. A total of six tandem duplication pairs and four segmental duplication pairs were detected, which contributed to the expansion of the B. distachyon expansin gene family. Expansins in the same group shared similar gene structure and motif composition. Three types of cis-elements, development-related, hormone-related, and abiotic stresses-related elements were found in the B. distachyon expansin gene promoters. Expression profiles indicated that most of B. distachyon expansin genes participate in plant development and abiotic stress responses. Overexpression of BdEXPA27 increased seed width and length, root length, root hair number and length in Arabidopsis and showed higher germination rate in transgenic lines. This study establishes a foundation for further investigation of B. distachyon expansin genes and provides novel insights into their biological functions.