Mating-type loci of Ustilago esculenta are essential for mating and development.

Ustilago esculenta is closely related to the smut fungus Ustilago maydis and, in an endophytic-like life in the plant Zizania latifolia, only infects host stems and causes swollen stems to form edible galls called Jiaobai in China. In order to study its different modes of invasion and sites of symptom development from other smut fungi at the molecular level, we first characterized the a and b mating-type loci of U. esculenta. The a loci contained three a mating-type alleles, encoding two pheromones and one pheromone receptor per allele. The pheromone/receptor system controlled the conjugation formation, the initial step of mating, in which each pheromone was specific for recognition by only one mating partner. In addition, there are at least three b alleles identified in U. esculenta, encoding two subunits of heterodimeric homeodomain transcription factors bE and bW, responsible for hyphal growth and invasiveness. Hyphal formation, elongation and invasion after mating of two compatible partners occurred, only when a heterodimer complex was formed by the bE and bW proteins derived from different alleles. We also demonstrated that even with only one paired pheromone-pheromone receptor, the active b locus heterodimer triggered hyphal growth and infection.

Transcriptome Comparison between Two Strains of Ustilago esculenta during the Mating.

Ustilago esculenta is a smut fungus that obligately infects Zizania latifolia and stimulates tissue swelling to form galls. Unlike T-type, MT-type U. esculenta can only proliferate within plant tissues and infect the offspring of their host. Production of telispores, haploid life, and plant cuticle penetration are not essential for it, which may lead to the degeneration in these processes. Transcriptome changes during the mating of T- and MT-type U. esculenta were studied. The functions of several secreted proteins were further confirmed by knock-out mutants. Our results showed that MT-type U. esculenta can receive environmental signals in mating and circumstance sensing as T-type does. However, MT-type U. esculenta takes a longer time for conjunction tube formation and cytoplasmic fusion. A large number of genes encoding secreted proteins are enriched in the purple co-expression module. They are significantly up-regulated in the late stage of mating in T-type U. esculenta, indicating their relationship with infecting. The knock-out of g6161 (xylanase) resulted in an attenuated symptom. The knock-out of g943 or g4344 (function unidentified) completely blocked the infection at an early stage. This study provides a comprehensive comparison between T- and MT-type during mating and identifies two candidate effectors for further study.

[Ectopic expression of the AmDREB1F gene from Ammopiptanthus mongolicus enhances stress tolerance of transgenic Arabidopsis].

Dehydration-responsive element binding proteins (DREBs) are an important class of transcription factors related to plant stress tolerance. Ammopiptanthus mongolicus is an evergreen broadleaf shrub endemic to desert areas of northwest China, and it has a very high tolerance to harsh environments. In order to reveal the functions and mechanisms of the AmDREB1F gene from this species in enduring abiotic stresses, we performed subcellular localization test, expression pattern analysis, and stress tolerance evaluation of transgenic Arabidopsis harboring this gene. The protein encoded by AmDREB1F was localized in the nucleus. In laboratory-cultured A. mongolicus seedlings, the expression of AmDREB1F was induced significantly by cold and drought but very slightly by salt and heat stresses, and undetectable upon ABA treatment. In leaves of naturally growing shrubs in the wild, the expression levels of the AmDREB1F gene were much higher during the late autumn, winter and early spring than in other seasons. Moreover, the expression was abundant in roots and immature pods rather than other organs of the shrubs. Constitutive expression of AmDREB1F in Arabidopsis induced the expression of several DREB-regulated stress-responsive genes and improved the tolerance of transgenic lines to drought, high salinity and low temperature as well as oxidative stress. The constitutive expression also caused growth retardation of the transgenics, which could be eliminated by the application of gibberellin 3. Stress-inducible expression of AmDREB1F also enhanced the tolerance of transgenic Arabidopsis to all of the four stresses mentioned above, without affecting its growth and development. These results suggest that AmDREB1F gene may play positive regulatory roles in response to abiotic stresses through the ABA-independent signaling pathways.

Functional Properties of the MAP Kinase UeKpp2 in Ustilago esculenta.

Ustilago esculenta undergoes an endophytic life cycle in Zizania latifolia. It induces the stem of its host to swell, forming the edible galls called jiaobai in China, which are the second most commonly cultivated aquatic vegetable in China. Z. latifolia raised for jiaobai can only reproduce asexually because the U. esculenta infection completely inhibits flowering. The infection and proliferation in the host plants during the formation of edible gall differ from those of conventional pathogens. Previous studies have shown a close relationship between mitogen-activated protein kinase (MAPK) and fungal pathogenesis. In this study, we explored the functional properties of the MAPK UeKpp2. Cross-species complementation assays were carried out, which indicated a functional complementation between the UeKpp2 of U. esculenta and the Kpp2 of Ustilago maydis. Next, UeKpp2 mutants of the UeT14 and the UeT55 sporidia background were generated; these showed an aberrant morphology of budding cells, and attenuated mating and filamentous growth in vitro, in the context of normal pathogenicity. Interestingly, we identified another protein kinase, UeUkc1, which acted downstream of UeKpp2 and may participate in the regulation of cell shape. We also found a defect of filamentous growth in UeKpp2 mutants that was not related to a defect of the induction of mating-type genes but was directly related to a defect in UeRbf1 induction. Overall, our results indicate an important role for UeKpp2 in U. esculenta that is slightly different from those reported for other smut fungi.

Rare spontaneous intrahepatic portosystemic shunt in hepatitis B-induced cirrhosis: A case report.

BACKGROUND: The portosystemic shunt is the pathway between the portal vein (PV) and systemic circulation. A spontaneous intrahepatic portosystemic shunt (SPISS) is a rare portosystemic shunt type. Here we report an extremely rare type of SPISS, a spontaneous intrahepatic PV-inferior vena cava shunt (SPIVCS). CASE SUMMARY: A 66-year-old woman was admitted to our hospital with the complaint of abdominal distention and a decreased appetite for 1 mo. The patient had a 20-year history of hepatitis B surface antigen positivity and a 5-year history of cirrhosis. She had been treated with Chinese herbal medicine for a long time. Liver function tests showed: alanine aminotransferase, 35 U/L; aspartate aminotransferase, 42 U/L; serum albumin (ALB) 32.2 g/L; and serum ascites ALB gradient, 25.2 g/L. Abdominal ultrasonography and enhanced computed tomography showed that the left branch of the PV was thin and occluded; the right branch of the PV was thick and showed a vermicular dilatation vein cluster in the upper pole of the right kidney that branched out and converged into the inferior vena cava from the bare area of the lower right posterior lobe of the liver. We diagnosed her with an extremely rare SPIVCS caused by portal hypertension and provided symptomatic treatment after admission. One week later, her symptoms disappeared and she was discharged. CONCLUSION: SPIVCS is a rare portosystemic shunt with a clear history of cirrhosis and portal hypertension. Clarifying the type PV shunt has important clinical significance.

AmDREB2C, from Ammopiptanthus mongolicus, enhances abiotic stress tolerance and regulates fatty acid composition in transgenic Arabidopsis.

Dehydration-responsive element-binding (DREB) transcription factors (TFs) play a vital role in plant response to abiotic stresses. However, little is known about DREB TFs in plants adapted to harsh environments and in the formation of polyunsaturated fatty acids (PUFAs), a major membrane component closely associated with plant stress tolerance. Here, we characterized AmDREB2C in Ammopiptanthus mongolicus (Maxim. ex kom.) Cheng F., a desert evergreen broadleaf shrub with a high tolerance to harsh environments. AmDREB2C encodes a canonical DREB2-type TF, and the protein was localized in the nucleus. AmDREB2C had the highest expression levels in leaves of naturally growing shrubs in the wild during the winter season of a year of sampling. The expression was also induced by cold, heat and drought stresses in laboratory-cultured seedlings. Moreover, AmDREB2C was most abundantly expressed in young leaves and immature seeds rather than other tissues of the shrubs. Constitutive expression of AmDREB2C in Arabidopsis enhanced freezing, heat and drought tolerances of the transgenic plants, likely through inducing the expression of important stress-responsive genes. The transgene also increased the level of linolenic acid (C18:3), a major PUFA in most plant species, in leaves and seeds of the transgenic plants. Correspondingly, the transcription of FAD3, FAD7 and FAD8, three genes encoding fatty acid desaturases (FADs) responsible for the production of C18:3, showed a differential up-regulation in these two organs. This study provides new insight into the underlying molecular mechanisms of A. mongolicus' ability to endure harsh environments and DREB TF regulation of fatty acid desaturation.