Differential gene expression analysis in germinating and dormant teliospores of Tilletia indica using RNA seq approach

Karnal bunt of wheat is an important quarantine disease that interrupts India’s wheat trade in the international market. The whole transcriptome of germinating and dormant teliospores of Tilletia indica was performed using the RNA Seq approach to identify germination-related genes. Approximately 63 million reads were generated using the RNA sequencing by the Illumina NextSeq500 platform. The high-quality reads were deposited in NCBI SRA database (accession: PRJNA522347). The unigenes from the pooled teliospores were 16,575 having unigenes length of 28,998,753 bases. The high-quality reads of germinating teliospores mapped on to 21,505 predicted CDSs. 9,680 CDSs were common between dormant and germinating teliospores of T. indica. 11,825 CDSs were found to be in germinating teliospores while only 91 were unique in dormant spores of pathogen. The pathway analysis showed the highest number of pathways was found in germinating spores than dormant spores. The highest numbers of CDSs were found to be associated with translation (431 in number), transport and catabolism (340), signal transduction (326), and carbohydrate metabolism (283). The differential expression analysis (DESeq) of germinating and dormant teliospores showed that 686 CDS were up-regulated and 114 CDS were down-regulated in the germinating teliospores. Significant germination-related genes in the spores were validated using qPCR analysis. Ten genes viz. Ti3931, Ti6828, Ti7098, Ti7462, Ti7522, Ti 9289, Ti 8670, Ti 7959, Ti 7809,and Ti10095 were highly up-regulated in germinated teliospores which may have role in germination of spores.Further, these differentially expressed genes provide insights into the molecular events. This first study of transcriptome will be helpful to devise better management strategies to manage Karnal bunt disease.

Utilization of fungal biocontrol agents against rice sheath blight disease provides insight into their role in plant defense responses

Biotic and abiotic factors have an effect on rice production all around the world. Diseases are regarded as major restrictions among the biotic stressors, and rice sheath blight (Rhizoctonia solani Kühn) is one of the most calamitous diseases that significantly damage the crop. Lately, biocontrol of fungal plant pathogens has appeared as an appealing approach. The present investigation was undertaken to evaluate different biocontrol agents like Talaromyces flavus, Chaetomium globosum, Pseudomonas fluorescens and Aspergillus niger against sheath blight disease. Prior to sowing, seeds were bioprimed with each isolate and sown in the nursery. After 21 days, seedlings were transplanted in-vivo and were inoculated with a virulent isolate of Rhizoctonia solani at maximum tillering stage. Observations on biochemical parameters and gene expression studies were carried out at 24, 48, 72, and 96 hpi. Enzymatic activity viz., chitinase, ?-1,3-glucanase, catalase, and PAL was observed maximum in Chaetomium globosum. PR-genes viz., IPT, BrD, HmPr, AMP, AldD, NIC and LisH showed up-regulation at 96 hpi. Chaetomium globosum had the highest yield, maximum number of tillers with least RLH% as compared to other treatments. However, results indicated biocontrol agents are helpful and they induce multitude of defence responses against R. solani in rice.

Vitamin D and COVID-19

The ongoing COVID -19 pandemic is caused by severe acute respiratory syndrome corona virus -2 (SARS-CoV-2). Since its emergence in Wuhan in Hubei province of China in December 2019, the virus has spread to every continent except Antartica. Currently, there is no registered treatment or vaccine for the disease. In the current scenario of the deadly virus spreading across continents and the absence of a specific treatment of novel corona virus, there is an urgent need to search for alternative strategies to prevent and control the rapid replication of virus. Vitamin D supplementation may reduce the incidence, severity and risk of death from pneumonia (consequent to the cytokine storm) in the current COVID pandemic. Through its effect on innate and adaptive immunity, vitamin D can reduce the risk of viral respiratory tract infections. 1, 25(OH) vitamin D directly stimulates the production of anti-microbial peptides like defensin and Cathelicidin that can reduce the rate of viral replication. In addition, it can also reduce the concentration of pro-inflammatory cytokines that are responsible for causing cytokine storm and resultant fatal pneumonia. In order to reduce the risk of infection especially in developing country like India, it is recommended that people at risk of COVDI19 may be considered for vitamin D supplementation.

Understanding the role of uncharacterized and hypothetical pathogenicity genes of Colletotrichum orbiculare to oxidative stress and phenolics of cucumber plant in anthracnose development

Uncharacterized and hypothetical pathogenicity genes of Colletotrichum orbiculare inciting anthracnose of cucumber are poorly annotated but could be potential antifungal targets in anthracnose control. Hence, to associate them with specific functions in fungal–host interactions for exploitation in anthracnose control, their responses to reactive oxygen species produced due to oxidative burst and constitutive or induced phenolics, the earliest known defence mechanisms of host plants were assessed. Among commercially available related phenolic compounds, at the lowest concentration of 1 mM, ferulic and chlorogenic acids were less toxic to the pathogen. Among the phenolics, a less significant expression of superoxide dismutase coupled with a strong expression of catalase in ferulic acid greater than that in menadione and hydrogen peroxide, respectively indicated its oxidative effect through generation of radicles similar to that of hydrogen peroxide. However, lack of expression of beta-ketoadipate pathway genes in response to the phenolic acids indicated the role of other phenol metabolizing genes of the pathogen. In planta expression followed by constitutive expression of uncharacterized and hypothetical pathogenicity genes of C. orbiculare in a minimal medium indicated that 6 of the genes are not redundant and may function under stress conditions. Among the genes, significant expression of ENH87556 in menadione (1.3-fold) together with a weak expression in hydrogen peroxide in relation to untreated control indicated its role in oxidative stress generated due to the superoxide radical of menadione. Additionally, a strong expression (3.8-fold) of the gene in ferulic acid greater than that of either control, minimal medium or minimal medium with leaf extract indicated its role in phenol metabolism. Thus, the gene could be a potential molecular target for anthracnose control upon validation by functional analysis.

Reversible thyrotoxic cardiomyopathy: a case report

Hyperthyroidism significantly affects the hemodynamics of the cardiovascular system. It is associated with development of atrial fibrillation, high output cardiac failure, pulmonary hypertension and dilated cardiomyopathy (DCM). In this paper we report a case of thyrotoxicosis induced cardiomyopathy. She was a 54 year old woman who presented with cardiomegaly(dilatation of all four chambers of the heart) secondary to Graves’ disease. She was treated with anti-thyroid drugs, β Blockers and diuretic and her euthyroid status was restored in 6 weeks. A repeat echocardiogram done at this point of time showed normal cardiac function with normalization of ejection fraction. So this was a case of reversible thyrotoxic cardiomyopathy

Vitamin B12 deficiency in patients with type 2 diabetes mellitus

Background: Vitamin B12 deficiency may occur consequent to long-term treatment with metformin but very few studies in India have assessed this risk. Metformin is considered the drug of first choice not only for the management of type 2 diabetes but also for prediabetes and polycystic ovary disease. The study was carried out to assess the serum levels of vitamin B12 in Indian patients with type 2 diabetes mellitus who were treated with metformin for a minimum duration of five years.Methods: Serum vitamin B12 level was estimated in 39 patients with type 2 diabetes mellitus, who were on metformin for more than five years. vitamin B12 deficiency was defined as serum concentration less than 200 pg/dl. Apart from vitamin B12, fasting blood sugar, post prandial sugar and glycosylated hemoglobin were also estimated.Results: 39 subjects in age group of 40-65 years, suffering from type 2 diabetes mellitus, for more than five years were recruited for the study. The mean serum level of vitamin B12 was 153.27±46.01. Vitamin B12 deficiency was found in 35.89% subjects.Conclusions: Patients suffering from long standing type 2 diabetes mellitus and taking large doses of metformin should be screened for Vitamin B 12 deficiency.

Thyrotoxic periodic paralysis: a short clinical review.

Thyrotoxic Periodic Paralysis (TPP) is a potentially lethal manifestation of hyperthyroidism which is characterized by hypokalemia and muscular weakness. It mainly affects Asian men in the age group of 20 to 40 years. Immediate supplementation with oral or intravenous potassium will help to not only abort the acute attack of paralysis but will also prevent serious and life threatening cardiac arrhythmias. Non selective beta blockers like propranolol can also be used to ameliorate and prevent subsequent paralytic attack. Acetazolamide has no role in the treatment of TPP.

Managing paediatric Graves’ disease.

Graves’ disease is the most common cause of hyperthyroidism in children. Anti-thyroid drug treatment with carbimazole or its active metabolite methimazole is offered as first line initial treatment but it induces remission in only 30%of children. Propylthiouracil is not recommended in children because of its association with severe hepatic toxicity. For those who relapse after ATD, radioactive iodine can be offered as definitive therapy except in cases with severe Graves’ ophthalmopathy or patients with large goitre who are the candidates for surgery. Total (or near total) thyroidectomy is the surgical procedure of choice for treating paediatric patients with Graves’ disease as it reduces the risk of recurrent hyperthyroidism which was seen in patients undergoing subtotal or partial thyroidectomy.

Cloning and expression of a small heat and salt tolerant protein (Hsp22) from Chaetomium globosum.

The present study reports molecular characterization of small heat shock protein gene in Indian isolates of Chaetomium globosum, C. perlucidum, C. reflexum, C. cochlioides and C. cupreum. Six isolates of C. globosum and other species showed a band of 630bp using specific primers. Amplified cDNA product of C. globosum (Cg 1) cloned and sequenced showed 603bp open reading frame encoding 200 amino-acids. The protein sequence had a molecular mass of 22 kDa and was therefore, named Hsp22. BlastX analysis revealed that the gene codes for a protein homologous to previously characterized Hsp22.4 gene from C. globosum (AAR36902.1, XP 001229241.1) and shared 95% identity in amino acid sequence. It also showed varying degree of similarities with small Hsp protein from Neurospora spp. (60%), Myceliophthora sp. (59%), Glomerella sp. (50%), Hypocrea sp. (52%), and Fusarium spp. (51%). This gene was further cloned into pET28a (+) and transformed E. coli BL21 cells were induced by IPTG, and the expressed protein of 30 kDa was analyzed by SDS-PAGE. The IPTG induced transformants displayed significantly greater resistance to NaCl and Na2CO3 stresses.

Molecular characterization of Chaetomium species using URP-PCR

Chaetomium spp. are common colonizers of soil and cellulose-containing substrates. Seventeen isolates of Chaetomium spp., which included 15 isolates of C. globosum and one each of C. reflexum and C. perlucidum, were genetically characterized with universal rice primers (URP - primers derived from DNA repeat sequences in the rice genome) using polymerase chain reaction (URP-PCR). Out of the 12 URP's used in the study, nine primers were effective in producing polymorphic fingerprint patterns from DNA of Chaetomium spp. Analysis of the entire fingerprint profile using the unweighted pair-group method with arithmetic averages (UPGMA) clearly differentiated C. globosum isolates from C. perlucidum and C. reflexum. One of the primers, URP-2R, produced a uniform DNA band of 1.9 kb in all the isolates of C. globosum but not in C. perlucidum and C. reflexum, which can be used as molecular marker to differentiate C. globosum from other species. Our results indicate that URP's are sensitive and give reproducible results for assaying the genetic variability in Chaetomium spp.

A study of growth parameters and prevalence of overweight and obesity in school children from delhi.

OBJECTIVE: (1) To assess the height, weight and body mass index (BMI) of school children from Delhi and generate percentile charts as appropriate for age, gender and socio-economic status. (2) To determine the prevalence of overweight and obesity in school children from low and upper socioeconomic status (LSES and USES respectively). DESIGN: Cross sectional evaluation of anthropometric parameters in Delhi school children (5-18 years) from different geographical zones. SETTING: Government schools (non-fee paying) and Private Schools (fee paying) in Delhi. SUBJECTS: 21485 children, 8840 (3566 boys, 5274 girls) from government schools and 12645 (6197 boys, 6448 girls) from private schools. Methods: Subjects underwent assessment of height and weight and calculation of BMI. Children were classified as normal, overweight and obese as per IOTF guidelines. Height, weight and BMI percentile charts specific for the socioeconomic status were generated using the LMS method. Prevalence of overweight and obesity was assessed and compared between the two socio-economic groups. RESULTS: A significant difference was noted in height, weight and BMI between LSES and USES. The prevalence of overweight and obesity in USES children was 16.75 % and 5.59 % in boys and 19.01 % and 5.03 % in girls respectively. CONCLUSIONS: There is a significant disparity in anthropometric parameters between children from USES and LSES, with a high prevalence of overweight and obesity in USES children.