Estimation of the amount of pear pollen based on flowering stage detection using deep learning.

Pear pollination is performed by artificial pollination because the pollination rate through insect pollination is not stable. Pollen must be collected to secure sufficient pollen for artificial pollination. However, recently, collecting sufficient amounts of pollen in Japan has become difficult, resulting in increased imports from overseas. To solve this problem, improving the efficiency of pollen collection and strengthening the domestic supply and demand system is necessary. In this study, we proposed an Artificial Intelligence (AI)-based method to estimate the amount of pear pollen. The proposed method used a deep learning-based object detection algorithm, You Only Look Once (YOLO), to classify and detect flower shapes in five stages, from bud to flowering, and to estimate the pollen amount. In this study, the performance of the proposed method was discussed by analyzing the accuracy and error of classification for multiple flower varieties. Although this study only discussed the performance of estimating the amount of pollen collected, in the future, we aim to establish a technique for estimating the time of maximum pollen collection using the method proposed in this study.

Four mycelial strains of Entoloma clypeatum species complex form ectomycorrhiza-like roots with Pyrus betulifolia seedlings in vitro, and one develops fruiting bodies 2 months after inoculation.

Entoloma clypeatum species complex (ECSC) forms ectomycorrhiza-like roots (EMLR) with host plant species of Rosaceae or Ulmaceae. The EMLR colonized with ECSC are characterized by a thick fungal mantle, absence of a Hartig net structure, and collapse of the apical meristem caused by hyphal invasion. Some researchers have suggested parasitism of ECSC because of this unique mode of colonization; however, the nature of the interaction between ECSC and host plants has not been investigated in co-culture because of the difficulty of culturing this group of fungi. We established a procedure to synthesize EMLR of ECSC on pear seedlings using fungal cultures. Three conspecific strains of ECSC isolated from basidiospores and one strain isolated from EMLR were tested. Cultured mycelia were inoculated onto a modified Norkrans' C (MNC) or Hyponex-yeast-glucose (HYG) medium slant on the bottom of a polycarbonate jar and covered with autoclaved andosol or a vermiculite/sphagnum moss mixture (VSM); an axenically cultivated Pyrus betulifolia seedling was then planted in the jar. Five months after inoculation, the formation of EMLR with Hartig net-like hyphae was confirmed in all of the experimental plots. However, the rate of root colonization was significantly higher in experimental plots using andosol than in those using VSM. The growth of pear seedlings was similar irrespective of the level of root colonization, suggesting commensalism rather than parasitism of ECSC. One experimental plot using strain A3, an MNC slant, and andosol as a substrate produced ECSC fruiting bodies with mature basidia and basidiospores. The results suggested that our procedure enables the synthesis of EMLR of ECSC and cultivation of their fruiting bodies.

Gibberellin Promotes Fungal Entry and Colonization during Paris-Type Arbuscular Mycorrhizal Symbiosis in Eustoma grandiflorum.

Arbuscular mycorrhizas (AMs) are divided into two types according to morphology: Arum- and Paris-type AMs. Gibberellins (GAs) mainly inhibit the establishment of Arum-type AM symbiosis in most model plants, whereas the effects of GAs on Paris-type AM symbiosis are unclear. To provide insight into the mechanism underlying this type of symbiosis, the roles of GAs were investigated in Eustoma grandiflorum when used as the host plant for Paris-type AM establishment. Eustoma grandiflorum seedlings were inoculated with the model AM fungus, Rhizophagus irregularis, and the effects of GA and the GA biosynthesis inhibitor uniconazole-P on the symbiosis were quantitatively evaluated. Exogenous GA significantly increased hyphopodium formation at the epidermis, thus leading to the promotion of fungal colonization and arbuscule formation in the root cortex. By contrast, the suppression of GA biosynthesis and signaling attenuated fungal entry to E. grandiflorum roots. Moreover, the exudates from GA-treated roots strongly induced the hyphal branching of R. irregularis. Our results show that GA has an contrasting effect on Paris-type AM symbiosis in E. grandiflorum compared with Arum-type AM symbiosis. This finding could be explained by the differential regulation of the early colonization stage, where fungal hyphae make contact with and penetrate the epidermis.

Draft genome sequence of Taiwanese pear (Pyrus pyrifolia).

Pear (genus Pyrus) is one of the oldest temperate fruit crops, of which at least 22 primary species are recognized. This article documents the public availability of the partial draft genome sequence data of the Taiwanese pear 'Hengshanli' that is less dormant during the winter season. This dataset may be used to prepare molecular markers for the breeding of new cultivars that are subjected to chilling at low temperatures to overcome endodormancy. This data will also help analyze the process of evolution in the Pyrus species. We sequenced paired-end libraries using Illumina HiSeq. 2500 and generated approximately 210M reads. Data on the draft genome obtained in this study has been deposited to the DNA Data Bank of Japan (DDBJ). The read data were submitted to the DDBJ Read Archive (BioProject: PRJDB6877, BioSample: SAMD00117052).

Draft genome sequence of Japanese pear (Pyrus pyrifolia).

The Japanese pear (Pyrus pyrifolia), a member of the family Rosaceae, is one of the most important fruit trees in Japan. This article documents the public availability of the partial draft genome sequence data of the Japanese pear strain TH3, which is the S1 of 'Osa-Nijisseiki' and is homozygous for the S4sm gene. This dataset may be used to prepare molecular markers for breeding of new cultivars having a crisp texture and feel. This data will also help research on physiological disorders affecting Japanese pear fruit. We sequenced paired-end libraries using Illumina HiSeq. 2500 and generated approximately 212M reads. Data on the draft genome obtained in this study has been deposited to the DNA Data Bank of Japan (DDBJ). The read data were submitted to the DDBJ Read Archive (BioProject: PRJDB6878, BioSample: SAMD00117051).

Gene expression changes triggered by end-of-day far-red light treatment on early developmental stages of Eustoma grandiflorum (Raf.) Shinn.

To better understand the molecular mechanisms related to growth promotion in the early developmental stages of Eustoma grandiflorum (Raf.) Shinn. under end-of-day far-red light (EOD-FR) treatment, we analyzed the leaf transcriptome of treated (EOD) and untreated plants (Cont) by using RNA-seq technology. EOD-FR treatment for only about 2 weeks in regions with limited sunshine during winter resulted in significantly higher internode length between the 3rd and 4th nodes on the main stem in EOD than in Cont. Among the differentially expressed genes (DEGs) related to synthesis or transport of auxin, higher levels of YUCCA (CL6581) and PIN4 (CL6181) were noted after treatment in EOD than in Cont in the leaf. In addition, high expression levels of GA20ox (Unigene11862) related to gibberellin (GA) synthesis and transcription factor bHLH 135 (CL7761) were observed in the stem of EOD, 3 h after treatment. A vertical section of the stem showed that the pith length of cells at the 4th node was longer in EOD than in Cont. Collectively, these results suggested that EOD-FR treatment increased the expression of DEGs related to GA and auxin biosynthesis, bHLH transcription factor, and internodal cell elongation along the longitudinal axis of Eustoma plants.

Comparative Transcriptome Analysis of the Less-Dormant Taiwanese Pear and the Dormant Japanese Pear during Winter Season.

The flower bud transcriptome in the less dormant Taiwanese pear 'Hengshanli' and high-chilling requiring Japanese pear strain TH3 subjected to the same chilling exposure time were analyzed during winter using next-generation sequencing. In buds sampled on January 10th and on February 7th in 2014, 6,978 and 7,096 genes, respectively, were significantly differentially expressed in the TH3 and 'Hengshanli' libraries. A comparative GO analysis revealed that oxidation-reduction process (biological process) and ATP binding (molecular function), were overrepresented during the ecodormancy period (EP) when compared to the endodormancy deepest period (DP), indicating that ATP synthesis was activated during the transition between these dormancy stages. Among the 11 differently expressed genes (DEGs) annotated as probable dehydrins or LEA protein-related genes, 9 DEGs showed higher transcript levels in the DP than in the EP. In order to focus on transcription factors induced by low temperature or drought, 7 differently expressed genes (DEGs) annotated as probable ICE1 or DREB proteins were analyzed by real-time PCR. Expression levels of 3 genes were higher in TH3 than in 'Hengshanli' on all sampling days. Their expression increased during the endodormancy deepest period (DP) and then decreased before endodormancy breaking in TH3 buds. Taken together, these results suggest that these genes annotated as ICE1, DREB and ERF are involved in endodormancy maintenance and in the transition from endodormancy to ecodormancy.

Identification of the expressed protein and the impact of change in ascorbate peroxidase activity related to endodormancy breaking in Pyrus pyrifolia.

Endodormancy is an important feature of perennial deciduous fruit trees that survive in the extreme climates brought about by seasonal variation. To acquire a comprehensive knowledge of the biochemical processes occurring just before endodormancy breaking, the buds collected in the pre-breaking period (PP) phase were used as samples to identify the proteins related to the breaking of endodormancy in the Japanese pear (Pyrus pyrifolia Nakai). Using nano-ESI-LC-MS/MS analysis, 96 proteins were overlapped by analyses of three times and identified as expressed proteins at the PP stage. Among these proteins, dehydrin, several classes of heat shock proteins (HSP), auxin-binding protein, and auxin-induced protein were identified in the floral bud in the PP stage. The majority of these proteins were involved primarily in the oxidation-reduction process. We focused on catalase (CAT), peroxidase (POD), and ascorbate peroxidase (APX) as enzymes regulating the levels of hydrogen peroxide (H2O2) in the bud. From measurements taken during the deepest period (DP), PP, mid-breaking period (MP), and late-breaking period (LP) of endodormancy, CAT activity decreased gradually, while APX activity also decreased from DP to MP, but then increased rapidly during LP. Protein data for PP and the rapid increase in APX activity observed in LP provided knowledge of the biochemical processes that regulate the consecutive transition from endodormancy breaking to ecodormancy induction in the Japanese pear.

Myotonic dystrophy associated with variable circadian rhythms of serum cortisol and isolated thyrotropin deficiency.

We describe a case of myotonic dystrophy presenting with a disturbed circadian rhythm of the serum cortisol and an isolated thyrotropin deficiency. The diagnosis of myotonic dystrophy was based on clinical characteristics, positive electromyographic findings, and increased number of CTG repeats in the dystrophia myotonica protein kinase (DMPK) gene. The patient presented with a variable circadian rhythm of the serum cortisol, increased excretion of urinary free cortisol, and a high adrenocorticotropin hormone responses to corticotropin-releasing hormone. The basal serum thyrotropin concentration was low and did not increase after thyrotropin-releasing hormone stimulation. The protein encoded by the DMPK gene may act as a second messenger in signal transduction, like a protein kinase. The present patient had a diverse pattern of disturbances in the hypothalamus-pituitary-endocrine organ axis, probably mediated by differences in the action or expression of the gene products in each endocrine cell.

High glucose concentrations abolish the superoxide dismutase response of leukocytes to ascorbic acid or troglitazone in type 2 diabetes mellitus.

UNLABELLED: The hypoglycemic drug, troglitazone (TGZ) has antioxidant activity. Superoxide dismutase (SOD) removes superoxide produced by cells. We measured the response of SOD-like activity (deltaSOD) to ascorbic acid (AA) or TGZ using electron spin resonance at various glucose concentrations in polymorphonuclear leukocytes from 18 type 2 diabetic patients and 18 healthy controls. In control and diabetic subjects, ASOD in response to AA was dose-dependent (maximal effect at 100 ng/ml). Maximal response occurred 2 min after AA addition (50 ng/ml). In cells from diabetic patients, ASOD with 25 ng/ml AA was significantly less than for healthy controls. The deltaSOD with AA changed little at glucose concentration from 0 to 200 mg/dl. In patient and control cells, higher glucose concentrations (400 to 800 mg/dl) reduced ASOD with AA. Response patterns with TGZ resembled those with AA. deltaSOD with AA correlated positively with glycosylated hemoglobin A1c. CONCLUSIONS: The present data suggest that an amerioration of blood glucose on high levels in diabetic patients plays an important role in an antioxidant efficacy of TGZ and AA on leukocytes in patients.

Impaired fibrinolytic compensation for hypercoagulability in obese patients with type 2 diabetes: association with increased plasminogen activator inhibitor-1.

In patients with type 2 diabetes, fibrinolysis is considered impaired by increased plasma concentrations of plasminogen activator inhibitor (PAI)-1. However, several investigators found both coagulation and fibrinolysis to be activated in these patients. We further characterized the balance between coagulation and fibrinolysis in lean and obese patients with type 2 diabetes. We studied 112 type 2 diabetic patients (66 lean, 46 obese) and 69 age-matched healthy subjects (46 lean, 23 obese). We measured plasma concentrations of fibrinogen and prothrombin F1+2 (F1+2) as indicating coagulation activity and plasmin-antiplasmin complex (PAP) and D dimer as indicating fibrinolytic activity. Plasma PAI-1 concentrations also were determined. Plasma concentrations of F1+2, PAP, D dimer, and PAI-1 were higher in diabetic patients than in control subjects. Plasma fibrinogen and F1+2 were similar between lean and obese diabetic patients, but plasma PAP and D dimer were significantly lower in obese than lean diabetic patients (P <.0001, P =.0194, respectively). By multivariate analysis, plasma PAI-1 and body mass index (BMI) were independent factors in diabetic patients predicting PAP, while BMI and glycosylated hemoglobin (HbA(1c)) independently predicted D dimer. Plasma PAI-1 concentrations were significantly higher in obese than lean diabetic patients (P <.0001). In conclusions, both coagulation and fibrinolytic systems are enhanced in lean and obese type 2 diabetic patients compared with healthy subjects. Although the degree of activation of coagulation was similar between lean and obese diabetic patients, the fibrinolytic activity was lower in obese than lean patients. Fibrinolytic compensation for hypercoagulation is incomplete in obese patients with type 2 diabetes, partly because of elevated PAI-1 in the blood.