Enhancing plant adaptation to challenging climates through breeding techniques requires studying plant systems with diverse genetic architectures. Comprehensive understanding of the genetic architecture of root traits is crucial for analyzing overall plant development and incorporating these insights into crop breeding programs on challenging climate adaptation. To dissect genetic architecture of root traits in soybean, we applied genome-wide association study (GWAS) in soybean germplasm population. Phenotyping of six root-related traits was performed at two plant growth stages, V1 (two-weeks growth stage) and V2 (three-weeks growth stage), under hydroponic culture and GWAS was performed to identify key SNPs and genes associated with root traits. Total 58 single nucleotide polymorphisms (SNPs) associated with six root-related traits were detected for two growth stages using three GWAS models, Mixed Linear Model (MLM), the Fixed and Random Model Circulating Probability Unification (FarmCPU) and 3 V multi-locus random-SNP-effect Mixed Linear Model (3VmrMLM). A total of 35 SNPs were detected for six root traits at V1 stage, while 23 SNPs were detected for the same traits at V2 stage. Quantitative trait locus (QTL) qRoot10.1 represented by three significant SNPs, was identified for primary root length (PRL) at V1 and V2 stage, and for root tips (RT) at V2 stage. Further, QTL qRoot10.1 was validated for PRL and total root length (TRL) in a separate set of soybean population. Candidate gene analysis in genomic regions of 58 SNPs identified 63 candidate genes, with annotations associated with various pathways of root development. Differential gene expression analysis of the candidate gene Glyma.10g273000 at qRoot10.1 revealed a significant difference in expression between long-rooting and short-rooting genotypes. In this study, we offer new insights into the root architecture of soybean, identifying key SNPs and genes that could be instrumental in future breeding programs aimed at developing highly efficient root systems in soybean.

Clusterbean (Cyamopsis tetragonoloba L.), an important legume crop of arid and semi-arid regions, plays a vital role in sustaining the livelihoods of farmers in Rajasthan due to its drought tolerance, soil-enriching properties, and industrial applications. This study was undertaken to assess the yield potential and economic viability of an improved clusterbean variety, RGC-1033, compared to local farmer varieties under rainfed conditions in the Jhunjhunu district through Front Line Demonstrations (FLDs) during Kharif 2023 and 2024. The results revealed that RGC-1033 consistently outperformed the control varieties, recording an average yield of 17.68 quintal/ha compared to 15.01 quintal/ha under farmer practices, with yield increases of 17.57% and 18.12% in 2023 and 2024, respectively. The extension and technology gaps indicated the need for improved dissemination and adoption of recommended practices. Economically, the improved variety demonstrated higher profitability with a gross return of ₹98,450/ha and ₹78,350/ha, and a net return of ₹62,825/ha and ₹42,650/ha in 2023 and 2024, respectively, compared to lower returns from local varieties. The benefit-cost (B:C) ratio also improved under demonstration plots (2.76 and 2.19) over farmer practices (2.43 and 1.91). These findings underscore the significance of adopting improved clusterbean varieties and scientific practices to bridge the yield and technology gaps, thereby enhancing farmer income under rainfed agro-ecologies.

The present study was conducted during rabi season (October to April) of four consecutive years from 2018–19 to 2021–22 at 175 selected farmer’s field in Dhani Gangajal, Kalana Tal, Bas Mamraj, Bhamra, Nethwa, Sahawa and Dingli villages of Churu District, Rajasthan, India. A total of 175 Cluster Front Line Demonstrations (CFLD) on chickpea was conducted. The farming situation was both irrigated and rainfed while, soil was sandy to sandy loam low in nitrogen, medium in phosphorus and medium to high in potash. Gap assessment was done and on the basis of gap assessment, improved recommended technologies were demonstrated. On four-year average basis, higher grain yield was recorded under demonstration field (1714 kg ha-1) as compared to farmers practices (1379 kg ha-1) which was 24.30% higher than farmer’s practices (Local check). The average extension gap, technology gap and technology index were 336 kg ha-1, 849 kg ha-1 and 20.12%, respectively. The higher average total returns was also recorded in demonstration plot which was ` 86721 ha-1 as compared to farmer’s practices (`Rs. 86721 ha-1). There was no much difference in cost of cultivation for demonstration plot (` 24980 ha-1) and farmers practices (`Rs. 22980 ha-1). An additional investment of `Rs. 2000 ha-1 consists with scientific monitoring of demonstration and non–monetary factors resulted in additional return of ` Rs. 16,939 ha-1. Due to higher additional return farmers got `Rs. 14939 ha-1 as effective gain. On four-year average basis, incremental benefit:cost ratio was recorded 8.45.

Soybean is one of the largest sources of protein and oil in the world and is also considered a “super crop” due to several industrial advantages. However, enhanced acreage and adoption of monoculture practices rendered the crop vulnerable to several diseases. Phytophthora root and stem rot (PRSR) caused by Phytophthora sojae is one of the most prevalent diseases adversely affecting soybean production globally. Deployment of genetic resistance is the most sustainable approach for avoiding yield losses due to this disease. PRSR resistance is complex in nature and difficult to address by conventional breeding alone. Genetic mapping through a cost-effective sequencing platform facilitates identification of candidate genes and associated molecular markers for genetic improvement against PRSR. Furthermore, with the help of novel genomic approaches, identification and functional characterization of Rps (resistance to Phytophthora sojae) have also progressed in the recent past, and more than 30 Rps genes imparting complete resistance to different PRSR pathotypes have been reported. In addition, many genomic regions imparting partial resistance have also been identified. Furthermore, the adoption of emerging approaches like genome editing, genomic-assisted breeding, and genomic selection can assist in the functional characterization of novel genes and their rapid introgression for PRSR resistance. Hence, in the near future, soybean growers will likely witness an increase in production by adopting PRSR-resistant cultivars. This review highlights the progress made in deciphering the genetic architecture of PRSR resistance, genomic advances, and future perspectives for the deployment of PRSR resistance in soybean for the sustainable management of PRSR disease.

Secondary ion mass spectrometry (SIMS)-based imaging techniques have become effective tools for studies of elements and molecules in biological samples.

SummaryRecently, a report raised the possibility of shrapnel‐induced chronic beryllium disease from long‐term exposure to the surface of retained aluminum shrapnel fragments in the body. Since the shrapnel fragments contained trace beryllium, methodological developments were needed for beryllium quantification and to study its spatial distribution in relation to other matrix elements, such as aluminum and iron, in metallurgic samples. In this work, we developed methodology for quantification of trace beryllium in samples of shrapnel fragments and other metallurgic sample‐types with main matrix of aluminum (aluminum cans from soda, beer, carbonated water and aluminum foil). Sample preparation procedures were developed for dissolving beryllium for its quantification with the fluorescence detection method for homogenized measurements. The spatial distribution of trace beryllium on the sample surface and in 3D was imaged with a dynamic secondary ion mass spectrometry instrument, CAMECA IMS 3f secondary ion mass spectrometry ion microscope. The beryllium content of shrapnel (∼100 ppb) was the same as the trace quantities of beryllium found in aluminum cans. The beryllium content of aluminum foil (∼25 ppb) was significantly lower than cans. SIMS imaging analysis revealed beryllium to be distributed in the form of low micron‐sized particles and clusters distributed randomly in X–Y‐ and Z dimensions, and often in association with iron, in the main aluminum matrix of cans. These observations indicate a plausible formation of Be–Fe or Al–Be alloy in the matrix of cans. Further observations were made on fluids (carbonated water) for understanding if trace beryllium in cans leached out and contaminated the food product. A direct comparison of carbonated water in aluminum cans and plastic bottles revealed that beryllium was below the detection limits of the fluorescence detection method (∼0.01 ppb). These observations indicate that beryllium present in aluminum matrix was either present in an immobile form or its mobilization into the food product was prevented by a polymer coating on the inside of cans, a practice used in food industry to prevent contamination of food products. The lack of such coating in retained shrapnel fragments renders their surface a possible source of contamination for long‐term exposure of tissues and fluids and induction of disease, as characterized in a recent study. Methodological developments reported here can be extended to studies of beryllium in electronics devices and components.

Chronic beryllium disease (CBD) is an exposure-related granulomatous disease mimicking sarcoidosis. Beryllium exposure-associated disease occurs mainly via inhalation, but skin may also be a source of sensitization. A 65-year-old male with a history of war-related shrapnel wounds was initially diagnosed with pulmonary sarcoidosis. Twenty years later, the possibility of a metal-related etiology for the lung disease was raised. A beryllium lymphocyte proliferation test, elemental analysis of removed shrapnel, and genetic studies were consistent with a diagnosis of CBD. This case demonstrates that retained beryllium-containing foreign bodies can be linked to a pathophysiologic response in the lung consistent with CBD.

Boron delivery characteristics of cis and trans isomers of a boronated unnatural amino acid, 1-amino-3-boronocyclopentanecarboxylic acid (ABCPC) were tested in the B16 mouse model for human melanoma. Both ABCPC isomers delivered comparable boron to B16 melanoma tumor cells as l-p-boronophenylalanine (BPA). Secondary ion mass spectrometry (SIMS) analysis revealed the presence of boron throughout the tumor from these compounds, and a near homogeneous distribution between the nucleus and cytoplasm of B16 cells grown in vitro. These encouraging observations support further studies of these new boron carriers in BNCT.

There is a pressing need for new and more efficient boron delivery agents to tumor cells for use in boron neutron capture therapy (BNCT). A class of boronated unnatural cyclic amino acids has demonstrated a remarkable selectivity toward tumors in animal and cell culture models, far superior to currently used agents in clinical BNCT. One of these amino acids, 1-amino-3-boronocyclopentanecarboxylic acid (ABCPC), has shown a tumor to blood ratio of 8 and a tumor to normal brain ratio of nearly 21 in a melanoma bearing mouse model. This work represents further biological characterization of this compound for tumor targeting in an EMT6 murine mammary carcinoma mouse model and a T98G human glioblastoma cell line. Female BALB/c mice bearing EMT6 tumors were injected with the fructose complex form of racemic mixtures of cis and trans isomers of ABCPC in identical concentrations. Boron concentrations were measured in the tumor, blood, brain, skin, and liver tissues at 1, 3, and 5h post-injection. These observations revealed a remarkable difference in racemic mixtures of cis and trans isomers in tumor targeting by boron. This implies that further separation of the L and D forms of this compound may enhance tumor targeting to an even higher degree than that provided by the racemic mixtures. Since the uptake measurements were made in homogenized tumor and normal tissues, little is known about the subcellular location of the boron arising from the various isomeric forms of the amino acid. To study subcellular delivery of boron from ABCPC in T98G human glioblastoma cells, we employed secondary ion mass spectrometry (SIMS) based technique of ion microscopy, which is capable of quantitatively imaging isotopic (elemental) gradients in cells and tissues at 500nm spatial resolution. The T98G cells were exposed to the nutrient medium containing 100ppm boron equivalent of a mixture of both L and D isomers of ABCPC in the form of a fructose complex for 1h. Following this treatment, the cells were fast frozen, freeze-fractured, and freeze-dried for SIMS analysis. Within an hour of exposure, ABCPC provided partitioning of intracellular to extracellular boron of 3/1. SIMS imaging revealed that boron from ABCPC was distributed throughout the cell, including the nucleus. This level of boron delivery within an hour of exposure is superior to p-boronophenylalanine (BPA) and sodium borocaptate (BSH), which have been previously studied by SIMS in the same cell line. These encouraging observations provide compelling support for further isomeric separations of ABCPC into the D and L forms for enhanced tumor targeting and continued testing of these compounds as new boron carriers in BNCT.

The development of cryogenic sample preparations, subcellular image quantification schemes, and correlative confocal laser scanning microscopy and ion microscopy have made dynamic SIMS a versatile tool in biology and medicine. For example, ion microscopy can provide much needed, novel information on calcium influx and intracellular calcium stores at organelle resolution in normal and transformed cells in order to better understand the altered calcium signaling in malignant cells. 3-D SIMS imaging of cells revealed dynamic gradients of calcium in cells undergoing mitosis and cytokinesis. Studies of subcellular localization of anticancer drugs is another area of research where ion microscopy can provide novel observations in many types of cancers. Ion microscopy is already an essential tool in boron neutron capture therapy (BNCT) of brain cancer as it can be used to quantitatively image the subcellular location of boron in cells and tissues. This information is critically needed for testing the efficacy of boronated agents and for calculations of radiation dosimetry.

Seed yield data from 16 genotypes (G) of chickpea (Cicer arietinum L.), tested in 49 transcontinental locations (L) were analyzed using residual maximum likelihood (REML) procedure and pattern analysis methods of classification and ordination to gein an understanding of the structure of Genotype x Environment (GE) interactions. The genotypes included the two major types of chickpea-desi and kabuli drawn from two main areas for chickpea adaptation, the Indian subcontinent and the Mediterranean region. Using hierarchical classification, the 16x49 GL mean data matrix was reduced to 8x9 (G-group x L-group) matrix capturing 50% of GL interaction sums of squares (SS). The genotypes fell into three discernible groups according to their geographic proximity. The location grouping suggested the presence of two mega-environments. Sub grouping of locations showed that generally the tropical (<23°) locations grouped together and separate from sub-tropical locations. The performance plots of G-groups indicated that two low yielding genotypes, Giza and Harigantars, contril?uted to large variation from one L-group to another. In general the medium-duration G-group (K 850, L 550, ICCC 8 and ICCC 4) from India was consistently better in most L-groups. In contrast, the G-group comprising Rabat (Morocco) and ILC 482 (Turkey), generally performed well in higher latitudes. The principal component biplot results were essentially similar to those obtained from the performance plots. It appears that development of early to medium-duration varieties may increase ahd stabilize seed yields. Based on genotype grouping, separate breeding programs for subtropical, tropical and the Mediterranean regions appear appropriate. This study showed that breeding for kabuli type and relatively bold seed might not necessarily affect varietal productivity adversely. Implications of these findings for chickpea improvement are discussed.

Ion microscopy observations on animal and cell culture models of glioblastoma multiforme revealed that a 6 hr. long infusion of boronophenylalanine (BPA) resulted in a significant increase in boron content of infiltrating tumor cells in the normal brain (or T98G human glioblastoma cells in cultures) as compared to the 2 hr. infusion. Subcellular imaging observations on fluorinated-boronophenylalanine (F-BPA) indicated that both boron and fluorine were co-compartmentalized in T98G cells. The F-BPA was comparable to BPA in delivering boron to cells. A preliminary ion microscopy study revealed that Gd-157 from Gd-DTPA concentrated mainly in the cytoplasm of T98G cells. This multitude of different studies demonstrates the versatility and great potential of ion microscopy in BNCT.

Clinical protocols of boron neutron capture therapy (BNCT) using a 2-h infusion of p-boronophenylalanine (BPA) have been performed with the assumption that infiltrating tumor cells contain equivalent amounts of B-10 as the main tumor mass. Ion microscopy studies of rat brain tumor and cultured cell models suggest that this is not the case and that a 6h or longer infusion of BPA may be necessary. In other studies, ion microscopy was used to relate the microdistribution of B-10 from BPA and sodium borocaptate (BSH) in a rat oral mucosa model to the biological effectiveness of BNC-radiation. We have been able to confirm the cellular co-compartmentalization of fluorine and boron from fluorinated BPA, a drug being developed for positron emission tomography. Preliminary studies of a potential agent for GdNCT are currently underway in our laboratory.

Several N-3 substituted carboranyl Thd analogs were synthesized. These agents as well as some non-boronated nucleosides were evaluated in, phosphoryl transfer assays with recombinant human TK1 and TK2. For some carboranyl thymidine analogs, TK1 phosphorylation rates approached 38% that of thymidine. Their in vitro cytotoxicty appeared to correlate with the TK1 levels in the tested cells. In some cases increased uptake in tumor cell nuclei compared with the surrounding cytoplasm was detected in vitro.

Several lines of evidence suggest that the Golgi apparatus is involved in Ca2+ regulation in renal epithelial LLC-PK1 cells. Laser scanning confocal microscopy (LSCM) was employed to establish that a prominent perinuclear region is occupied mainly by the Golgi apparatus in this cell line. LSCM measurements in individual cells with the ionized Ca2+ indicator calcium green revealed that stimulation of LLC-PK1 cells with arginine vasopressin (AVP) resulted in the elevation of ionized Ca2+ levels. However, the vasopressin-induced rise in ionized Ca2+ was attenuated if the Golgi apparatus was disassembled by pretreating the cells with brefeldin A (BFA). Subcellular measurements of total Ca2+ with ion microscopy in cryogenically prepared cells indicated that 1) within 1 min of AVP treatment significant quantities of sequestered Ca2+ were released from the perinuclear Golgi region and 2) the BFA treatment reduced the total Ca2+ stored in the Golgi region. These observations indicate that the Golgi apparatus is sensitive to hormonal stimulation and may play important roles in intracellular Ca2+ regulation in LLC-PK1 cells.