Unraveling the genetics of heat tolerance in chickpea landraces (Cicer arietinum L.) using genome-wide association studies.

Chickpea, being an important grain legume crop, is often confronted with the adverse effects of high temperatures at the reproductive stage of crop growth, drastically affecting yield and overall productivity. The current study deals with an extensive evaluation of chickpea genotypes, focusing on the traits associated with yield and their response to heat stress. Notably, we observed significant variations for these traits under both normal and high-temperature conditions, forming a robust basis for genetic research and breeding initiatives. Furthermore, the study revealed that yield-related traits exhibited high heritability, suggesting their potential suitability for marker-assisted selection. We carried out single-nucleotide polymorphism (SNP) genotyping using the genotyping-by-sequencing (GBS) method for a genome-wide association study (GWAS). Overall, 27 marker-trait associations (MTAs) linked to yield-related traits, among which we identified five common MTAs displaying pleiotropic effects after applying a stringent Bonferroni-corrected p-value threshold of <0.05 [-log10(p) > 4.95] using the BLINK (Bayesian-information and linkage-disequilibrium iteratively nested keyway) model. Through an in-depth in silico analysis of these markers against the CDC Frontier v1 reference genome, we discovered that the majority of the SNPs were located at or in proximity to gene-coding regions. We further explored candidate genes situated near these MTAs, shedding light on the molecular mechanisms governing heat stress tolerance and yield enhancement in chickpeas such as indole-3-acetic acid-amido synthetase GH3.1 with GH3 auxin-responsive promoter and pentatricopeptide repeat-containing protein, etc. The harvest index (HI) trait was associated with marker Ca3:37444451 encoding aspartic proteinase ortholog sequence of Oryza sativa subsp. japonica and Medicago truncatula, which is known for contributing to heat stress tolerance. These identified MTAs and associated candidate genes may serve as valuable assets for breeding programs dedicated to tailoring chickpea varieties resilient to heat stress and climate change.

A Multi-Model Based Stability Analysis Employing Multi-Environmental Trials (METs) Data for Discerning Heat Tolerance in Chickpea (Cicer arietinum L.) Landraces.

Identifying a congenially targeted production environment and understanding the effects of genotype by environmental interactions on the adaption of chickpea genotypes is essential for achieving an optimal yield stability. Different models like additive main effect and multiplicative interactions (AMMI 1, AMM2), weighted average absolute scores of BLUPs (WAASB), and genotype plus genotype-environment (GGE) interactions were used to understand their suitability in the precise estimation of variance and their interaction. Our experiment used genotypes that represent the West Asia-North Africa (WANA) region. This trial involved two different sowing dates, two distinct seasons, and three different locations, resulting in a total of 12 environments. Genotype IG 5871(G1) showed a lower heat susceptibility index (HSI) across environments under study. The first four interactions principal component axis (IPCA) explain 93.2% of variations with significant genotype-environment interactions. Considering the AMMI stability value (ASV), the genotypes IG5862(G7), IG5861(G6), ILC239(G40), IG6002(G26), and ILC1932(G39), showing ASV scores of 1.66, 1.80, 2.20, 2.60, and 2.84, respectively, were ranked as the most stable and are comparable to the weighted average absolute scores of BLUPs (WAASB) ranking of genotypes. The which-won-where pattern of genotype plus genotype-environment (GGE) interactions suggested that the target environment consists of one mega environment. IG5866(G10), IG5865(G9), IG5884(G14), and IG5862(G7) displayed higher stability, as they were nearer to the origin. The genotypes that exhibited a superior performance in the tested environments can serve as ideal parental lines for heat-stress tolerance breeding programs. The weighted average absolute scores of BLUPs (WAASB) serve as an ideal tool to discern the variations and identify the stable genotype among all methods.

Genetic mapping of quantitative trait loci associated with drought tolerance in chickpea (Cicer arietinum L.).

Elucidation of the genetic basis of drought tolerance is vital for genomics-assisted breeding of drought tolerant crop varieties. Here, we used genotyping-by-sequencing (GBS) to identify single nucleotide polymorphisms (SNPs) in recombinant inbred lines (RILs) derived from a cross between a drought tolerant chickpea variety, Pusa 362 and a drought sensitive variety, SBD 377. The GBS identified a total of 35,502 SNPs and subsequent filtering of these resulted in 3237 high-quality SNPs included in the eight linkage groups. Fifty-one percent of these SNPs were located in the genic regions distributed throughout the genome. The high density linkage map has total map length of 1069 cm with an average marker interval of 0.33 cm. The linkage map was used to identify 9 robust and consistent QTLs for four drought related traits viz. membrane stability index, relative water content, seed weight and yield under drought, with percent variance explained within the range of 6.29%-90.68% and LOD scores of 2.64 to 6.38, which were located on five of the eight linkage groups. A genomic region on LG 7 harbors quantitative trait loci (QTLs) explaining > 90% phenotypic variance for membrane stability index, and > 10% PVE for yield. This study also provides the first report of major QTLs for physiological traits such as membrane stability index and relative water content for drought stress in chickpea. A total of 369 putative candidate genes were identified in the 6.6 Mb genomic region spanning these QTLs. In-silico expression profiling based on the available transcriptome data revealed that 326 of these genes were differentially expressed under drought stress. KEGG analysis resulted in reduction of candidate genes from 369 to 99, revealing enrichment in various signaling pathways. Haplotype analysis confirmed 5 QTLs among the initially identified 9 QTLs. Two QTLs, qRWC1.1 and qYLD7.1, were chosen based on high SNP density. Candidate gene-based analysis revealed distinct haplotypes in qYLD7.1 associated with significant phenotypic differences, potentially linked to pathways for secondary metabolite biosynthesis. These identified candidate genes bolster defenses through flavonoids and phenylalanine-derived compounds, aiding UV protection, pathogen resistance, and plant structure.The study provides novel genomic regions and candidate genes which can be utilized in genomics-assisted breeding of superior drought tolerant chickpea cultivars.

Reverse Takotsubo Cardiomyopathy Following COVISHIELD Vaccination: A Rare Case of an Anaphylactic Reaction.

This case report describes a 47-year-old female healthcare worker who developed reverse takotsubo cardiomyopathy (TTC) following the administration of the COVISHIELD COVID-19 vaccine. Within minutes of receiving the vaccine, she experienced acute shortness of breath, nausea, and restlessness, along with a significant drop in blood pressure. She was diagnosed with an anaphylactic reaction and promptly treated with adrenaline and fluids. In the intensive cardiac care unit, she exhibited hypotension, had vision loss, and developed pulmonary edema. Further evaluation revealed abnormal ECG findings, elevated troponin levels, and reduced left ventricular ejection fraction (LVEF). Coronary angiography ruled out obstructive coronary artery disease. The patient gradually improved over several days and was discharged with a recovered left ventricular function. This case highlights the occurrence of TTC triggered by anaphylaxis to the COVID-19 vaccine and emphasizes the need for preparedness to manage such emergencies in vaccination centers.

The interaction between drought stress and nodule formation under multiple environments in chickpea.

Environmental stresses, particularly drought, limit symbiotic nitrogen fixation in legumes, resulting in decreased yielding capacity. Drought is one of the most important constraints limiting yield potential in crops and it is the major abiotic stress that can cause more than 70% yield loss in chickpea. In this study, a total of two hundred four chickpea (Cicer arietinum L.) genotypes were selected to study the interaction between drought stress and nodule formation. This interaction was assessed by using morphological, yield and yield components. The field experiments were laid out in two locations (Terbol and Kfardan stations, Bekaa valley, Lebanon) using Alpha lattice design with two replications and two watering treatments (irrigation and rainfed) during 2016 and 2017 seasons. Parameters that were measured include days to 50% flowering (DFL), day to maturity (DM), plant height (PLH), nodule biomass (NB), nodule fresh weight (NFW), nodule dry weight (NDW), grain yield (GY), Biological yield (BY), 100 seed weight (100SW) and drought tolerance stress (DTS). The results indicated a significant variation between genotypes, environments and other morphological, yield and yield components traits. Drought stress reduced significantly the yield and the nodule's characteristics, biological and grain yield. The genotypes with the highest levels of drought tolerance, such as IG70399, IG8256, IG71832, IG70270, and IG70272, showed a minimal decrease in yield and nodule biomass. Nodule observations significantly and positively correlated with GY (0.36-0.38) under drought stress treatment. The correlation values for nodule characteristics with DFL and DM were higher under drought stress compared to irrigated conditions. This is a comparative study between drought stress and nodule formation traits associated with morphological, yield and yield components traits.

A comprehensive analysis of Trehalose-6-phosphate synthase (TPS) gene for salinity tolerance in chickpea (Cicer arietinum L.).

Soil salinity affects various crop cultivation but legumes are the most sensitive to salinity. Osmotic stress is the first stage of salinity stress caused by excess salts in the soil on plants which adversely affects the growth instantly. The Trehalose-6-phosphate synthase (TPS) genes play a key role in the regulation of abiotic stresses resistance from the high expression of different isoform. Selected genotypes were evaluated to estimate for salt tolerance as well as genetic variability at morphological and molecular level. Allelic variations were identified in some of the selected genotypes for the TPS gene. A comprehensive analysis of the TPS gene from selected genotypes was conducted. Presence of significant genetic variability among the genotypes was found for salinity tolerance. This is the first report of allelic variation of TPS gene from chickpea and results indicates that the SNPs present in these conserved regions may contribute largely to functional distinction. The nucleotide sequence analysis suggests that the TPS gene sequences were found to be conserved among the genotypes. Some selected genotypes were evaluated to estimate for salt tolerance as well as for comparative analysis of physiological, molecular and allelic variability for salt responsive gene Trehalose-6-Phosphate Synthase through sequence similarity. Allelic variations were identified in some selected genotypes for the TPS gene. It is found that Pusa362, Pusa1103, and IG5856 are the most salt-tolerant lines and the results indicates that the identified genotypes can be used as a reliable donor for the chickpea improvement programs for salinity tolerance.

Identification of promising chickpea interspecific derivatives for agro-morphological and major biotic traits.

The wild Cicer species is well-known for having climate-resilient and productivity-enhancing traits of interest. Therefore, wide hybridization could be used as a realistic strategy for introgressing prospective traits from wild species into the cultivated gene pool. The present study was, thus, undertaken to evaluate F7 chickpea interspecific derivatives derived from Cicer reticulatum Ladiz. and C. echinospermum P. H. Davis wild annual Cicer species. As a result, a set of six interspecific crosses were advanced using the single seed descent (SSD) method of breeding. The F7 generation of these crosses was assessed in two diverse agro-ecological regions of India. The data revealed a wide range of variation with respect to seed yield and its important component traits, which resulted in the identification of the most promising derivatives carrying desirable characters as indicated by range, mean, and coefficient of variation. Further, fruitful heterosis was also estimated as promising selection criteria for identifying superior lines for earliness and high seed yield, including resistance against prevailing stresses (ascochyta blight, botrytis gray mold, dry root rot, and fusarium wilt). The superior derivatives carrying putative characters could be recommended for further breeding and selection of genetic materials for developing suitable genotypes.

A case of "very" very late stent thrombosis: More than 12 years after DES.

A rare but possibly catastrophic consequence of drug-eluting stents (DES) is very late stent thrombosis. We report a case of 74-year-old male who sustained a ST elevation myocardial infarction (STEMI) 12 years after initial Paclitaxel eluting stent implantation (PES). This is the longest time between stent placement and the development of an acute coronary event due to very late stent thrombosis that we are aware of (VLST). The implications for prognosis and therapy are significant because they highlight the uncertainty around the recommended duration of antiplatelet medication in patients with DES. Clinicians face challenges in treating those patients particularly when competing medical conditions demand the discontinuation of antiplatelet therapy. VLST is concerning since the underlying pathophysiology is unknown, and the best preventive treatments and duration of antiplatelet medication after stent implantation are unknown.

Identification and morphological characterization of promising kabuli chickpea genotypes for short-season environment in central India.

Chickpea (Cicer arietinum L.) is an important food legume crop grown in arid and semi-arid regions of the world. In India, kabuli chickpea is grown in central India in ~0.5 million ha, predominantly under short winter (<110 days). Efforts are underway to select promising genotypes at the Food Legume Research Platform (FLRP), Amlaha, located in intensive kabuli chickpea growing area of India. Sixty-four kabuli chickpea lines were evaluated for agronomic traits during 2017-2018 and 2018-2019 crop seasons at FLRP following simple 8 X 8 lattice design with two replications. The analysis of variance over two years revealed significant variation exists for days to flowering, plant height, maturity period, biomass, seed size and seed yield. It was observed that with similar maturity time (106 days), FLIP09-432C produced 2273 kg/ha, which out-yielded the popular variety in central India, JGK-3 by 15%. The breeding lines, FLIP09- 436C, FLIP09-171C, FLIP09-373C and FLIP09-247C were also found promising for earliness (104-110 days), and high yielding with the good yield ability (1003-2273 kg/ha). These promising genotypes for a short duration with good yield have been selected and can be used for various chickpea breeding programmes to develop high yielding varieties in central India.

Evaluation of disease resistant and high yielding faba bean germplasm in India.

Faba bean (Vicia faba L.) is one of the earliest domesticated food legumes after chickpea and pea in the world. It is been produced in many countries including China, Ethiopia, Egypt, northern Europe, the Mediterranean region, central Asia, East Asia, Latin America and as a minor crop in India. The crop is affected by many diseases and alternaria leaf blight (Alternaria spp.) is one of the serious threat to faba bean production. Twenty-five lines of faba bean were selected from three international nurseries and were evaluated at ICARDA-FLRP-Amlaha during 2016-2017 and 2017-2018, to identify resistant lines against alternaria blight disease. A wide range of variation to disease reaction was observed among faba bean genotypes. One faba bean line (S2011-134) found tolerant, six genotypes (S2011-116, FLIP15-139, FLIP15-156, FLIP15-159, FLIP15-164-S2 and FLIP15-169) were found moderately tolerant and 16 genotypes were found susceptible to alternaria blight. The faba bean genotypes showed resistance to the disease scoring (0-9) with high yield as compared to the checks, Giza and Gwalior local. The identified sources of resistance can be utilized in faba bean breeding programmes for the development of disease tolerant cultivars with high yield.

Identification of Allelic Variation in Drought Responsive Dehydrin Gene Based on Sequence Similarity in Chickpea (Cicer arietinum L.).

Chickpea (Cicer arietinum L.) is an economically important food legume grown in arid and semi-arid regions of the world. Chickpea is cultivated mainly in the rainfed, residual moisture, and restricted irrigation condition. The crop is always prone to drought stress which is resulting in flower drop, unfilled pods, and is a major yield reducer in many parts of the world. The present study elucidates the association between candidate gene and morpho-physiological traits for the screening of drought tolerance in chickpea. Abiotic stress-responsive gene Dehydrin (DHN) was identified in some of the chickpea genotypes based on the sequence similarity approach to play a major role in drought tolerance. Analysis of variance revealed a significant effect of drought on relative water content, membrane stability index, plant height, and yield traits. The genotypes Pusa1103, Pusa362, and ICC4958 were found most promising genotypes for drought tolerance as they maintained the higher value of osmotic regulations and yield characters. The results were further supported by a sequence similarity approach for the dehydrin gene when analyzed for the presence of single nucleotide polymorphisms (SNPs) and indels. Homozygous indels and single nucleotide polymorphisms were found after the sequencing in some of the selected genotypes.

Potassium: A key modulator for cell homeostasis.

Potassium (K) is the most vital and abundant macro element for the overall growth of plants and its deficiency or, excess concentration results in many diseases in plants. It is involved in regulation of many crucial roles in plant development. Depending on soil-root interactions, complex soil dynamics often results in unpredictable availability of the elements. Based on the importance index, K is considered to be the second only to nitrogen for the overall growth of plants. More than 60 enzymes within the plant system depend on K for its activation, in which K act as a key regulator. K helps plants to resist several abiotic and biotic stresses in the environment. We have reviewed the research progress about K's role in plants covering various important considerations of K highlighting the effects of microbes on soil K+; K and its contribution to adsorbed dose in plants; the importance of K+ deficiency; physiological functions of K+ transporters and channels; and interference of abiotic stressor in the regulatory role of K. This review further highlights the scope of future research regarding K.

Spontaneous coronary artery dissection involving all major arteries LAD, LCX and RCA -A case report.

Spontaneous coronary artery dissection (SCAD) is a rare cause of acute coronary syndrome and sudden cardiac death. Amongst various etiologies thought to be responsible for this condition, atherosclerosis seems to be the most common. There are various case reports on (SCAD) of single coronary artery but the presence of SCAD of all three arteries is not known. We describe a case of a 45-year-old gentleman without any conventional coronary risk factors, who was referred to us at Tata Main hospital, Jamshedpur, for cardiac evaluation prior to his urethral surgery. Although asymptomatic cardiac-wise, his ECG incidentally revealed healed inferior wall myocardial infarction. Echocardiogram confirmed hypokinetic inferior wall with left ventricular ejection fraction of 50%. His coronary angiography showed spontaneous dissection of LAD, LCX and RCA which was managed conservatively.

Development and validation of a high throughput LC–MS/MS method for simultaneous quantitation of pioglitazone and telmisartan in rat plasma and its application to a pharmacokinetic study / 药物分析学报

Management of cardiovascular risk factors in diabetes demands special attention due to their co-existence. Pioglitazone (PIO) and telmisartan (TLM) combination can be beneficial in effective control of cardiovascular complication in diabetes.In this research,we developed and validated a high throughput LC–MS/MS method for simultaneous quantitation of PIO and TLM in rat plasma. This developed method is more sensitive and can quantitate the analytes in relatively shorter period of time compared to the previously reported methods for their individual quantification. Moreover, till date, there is no bioanalytical method available to simultaneously quantitate PIO and TLM in a single run. The method was validated according to the USFDA guidelines for bioanalytical method validation.A linear response of the analytes was observed over the range of 0.005–10μg/mL with satisfactory precision and accuracy. Accuracy at four quality control levels was within 94.27%–106.10%. The intra-and inter-day precision ranged from 2.32% to 10.14% and 5.02% to 8.12%,respectively.The method was reproducible and sensitive enough to quantitate PIO and TLM in rat plasma samples of a preclinical pharmacokinetic study.Due to the potential of PIO-TLM combination to be therapeutically explored,this method is expected to have significant usefulness in future.

High-density linkage map construction and mapping of seed trait QTLs in chickpea (Cicer arietinum L.) using Genotyping-by-Sequencing (GBS).

This study reports the use of Genotyping-by-Sequencing (GBS) for large-scale SNP discovery and simultaneous genotyping of recombinant inbred lines (RILs) of an intra-specific mapping population of chickpea contrasting for seed traits. A total of 119,672 raw SNPs were discovered, which after stringent filtering revealed 3,977 high quality SNPs of which 39.5% were present in genic regions. Comparative analysis using physically mapped marker loci revealed a higher degree of synteny with Medicago in comparison to soybean. The SNP genotyping data was utilized to construct one of the most saturated intra-specific genetic linkage maps of chickpea having 3,363 mapped positions including 3,228 SNPs on 8 linkage groups spanning 1006.98 cM at an average inter marker distance of 0.33 cM. The map was utilized to identify 20 quantitative trait loci (QTLs) associated with seed traits accounting for phenotypic variations ranging from 9.97% to 29.71%. Analysis of the genomic sequence corresponding to five robust QTLs led to the identification of 684 putative candidate genes whose expression profiling revealed that 101 genes exhibited seed specific expression. The integrated approach utilizing the identified QTLs along with the available genome and transcriptome could serve as a platform for candidate gene identification for molecular breeding of chickpea.

Development of gene-based markers for use in construction of the chickpea (Cicer arietinum L.) genetic linkage map and identification of QTLs associated with seed weight and plant height.

Seed weight and plant height are important agronomic traits and contribute to seed yield. The objective of this study was to identify QTLs underlying these traits using an intra-specific mapping population of chickpea. A F11 population of 177 recombinant inbred lines derived from a cross between SBD377 (100-seed weight--48 g and plant height--53 cm) and BGD112 (100-seed weight--15 g and plant height--65 cm) was used. A total of 367 novel EST-derived functional markers were developed which included 187 EST-SSRs, 130 potential intron polymorphisms (PIPs) and 50 expressed sequence tag polymorphisms (ESTPs). Along with these, 590 previously published markers including 385 EST-based markers and 205 genomic SSRs were utilized. Of the 957 markers tested for analysis of parental polymorphism between the two parents of the mapping population, 135 (14.64%) were found to be polymorphic. Of these, 131 polymorphic markers could be mapped to the 8 linkage groups. The linkage map had a total length of 1140.54 cM with an average marker density of 8.7 cM. The map was further used for QTL identification using composite interval mapping method (CIM). Two QTLs each for seed weight, qSW-1 and qSW-2 (explaining 11.54 and 19.24% of phenotypic variance, respectively) and plant height, qPH-1 and qPH-2 (explaining 13.98 and 12.17% of phenotypic variance, respectively) were detected. The novel set of genic markers, the intra-specific linkage map and the QTLs identified in the present study will serve as valuable genomic resources in improving the chickpea seed yield using marker-assisted selection (MAS) strategies.

Microfilaria in breast aspiration--an uncommon finding.

Filariasis is a major public health problem in many tropics and subtropics of Africa, Western Pacific and parts of the Americas, affecting over 83 countries. It is also a public health problem in India. A large majority of the cases found in India are attributed to infection by Wuchereria bancrofti which mainly affects the lymph nodes and the lymphatic channels. Breast is an unusual site for filariasis. In the present case report the patient presented with a gradually increasing breast lump, clinically diagnosed as fibroadenoma.

Forensic diagnosis and classification of snakebite (neurotoxic and haemotoxic) from renal changes at autopsy: morphological and histological determinants using a semi-quantitative method.

Mortality due to poisonous snakebite is an emerging public health concern. Of late, snakebite has drawn global attention. Snakebite is a well-known cause of renal pathology especially in the haemotoxic group. Deaths due to renal failure are very common in envenomation due to poisonous snakebite. In this investigation our aim was to devise autopsy-proven diagnostic criteria of snakebite from the renal changes to confirm and classify the type of snakebite in forensic casework. That would be of practical help to the forensic pathologist with access to limited resources. An arbitrary scoring system from histology was adopted for the present semi-quantitative analysis. The scoring system (with scores from 0 to 5) developed by the present study was effective, valid and was able to distinguish between snakebite and control. The score obtained at histology in cases of snakebite was significantly different from the control group (Mann-Whitney U = 71.5; P = 0.011). Using stepwise linear discriminant function (DF) analysis, the score (Mukhopadhyay's score [MS]) was seen to be the only predictor of the type of snakebite. This model (DF = 1.05 x MS-1.78) could correctly classify 93.3% of the cases into neurotoxic and haemotoxic variety of offending snake. This is an effective, cheap and easy semi-quantitative method of distinguishing neurotoxic and haematoxic snakes from the renal changes at autopsy. It would be useful for practical application in centres where sophisticated biochemical or immunological analyses are not available due to limited resources.