Effect of herbicide stress on synchronization of carbon and nitrogen metabolism in lentil (Lens culinaris Medik.).

Weed invasion causes significant yield losses in lentil. Imazethapyr (IM), a broad-spectrum herbicide inhibits the biosynthesis of branched chain amino acids necessary for plant growth. Plant growth depends upon translocation of photo-assimilates and their partitioning regulated by carbon and nitrogen metabolism. This study aimed to investigate the impact of imazethapyr spray on carbon and nitrogen metabolism in tolerant (LL1397 and LL1612) and susceptible (FLIP2004-7L and PL07) lentil genotypes during vegetative and reproductive development. Significantly higher activities of invertases and sucrose synthase (cleavage) in leaves and in podwall and seeds during early phase of development in tolerant genotypes were observed as compared to susceptible genotypes under herbicide stress that might be responsible for providing hexoses required for their growth. Activities of sucrose synthesizing enzymes, sucrose phosphate synthase and sucrose synthase (synthesis) increased significantly in podwalls and seeds of LL1397 and LL1612 genotypes during later phase of development towards maturity while the activities decreased in FLIP2004-7L and PL07 genotypes under herbicide stress. Activities of nitrate and nitrite reductase, glutamine 2-oxoglutarate aminotransferase, glutamine synthetase and glutamate dehydrogenase were increased in leaves, podwalls and seeds of LL1397 and LL1612 under herbicide stress. A proper synchronization of carbon and nitrogen metabolism in tolerant lentil genotypes during vegetative and reproductive phase might be one of the mechanisms for their recovery from herbicide stress. This first ever comprehensive information will provide a basis for future studies on the molecular mechanism of source sink relationship in lentil under herbicide stress and will be utilized in breeding programmes.

Impact of post-emergent imazethapyr on morpho-physiological and biochemical responses in lentil (Lens culinaris Medik.).

Yield reduction in lentil crop due to weed infestation is a key hindrance to its growth due to poor weed-crop competition. Imazethapyr (IM), a selective herbicide, target acetolactate synthase (ALS) which catalyzes the first reaction in biosynthesis of branched chain amino acids, required for plant growth and development. The objective of the present study was to investigate the impact of IM treatment on weeds, ALS enzyme activity, antioxidant capacity, osmolyte accumulation, growth and yield related parameters in lentil genotypes. Two IM tolerant (LL1397 and LL1612) and two susceptible (FLIP2004-7L and PL07) lentil genotypes were cultivated under weed free, weedy check and IM treatments. Weed control efficiency reached its peak at 21 days after spray (DAS). Imazethapyr treatment decreased chlorophyll and carotenoid content up to 28 DAS with higher reduction in susceptible genotypes. FLIP2004-7L and PL07 had reduced plant height and lower number of pods under IM treatment which resulted in decreased seed yield. Higher ALS activity in LL1397 and LL1612 at 21 DAS, higher antioxidant capacity and glycine betaine content both at 21 and 28 DAS and lower decrease in relative leaf water content might be mediating herbicide tolerance in these genotypes that led to higher seed yield. The identified IM tolerance mechanism can be used to impart herbicide resistance in lentil. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01244-x.

Methylglyoxal detoxification pathway - Explored first time for imazethapyr tolerance in lentil (Lens culinaris L.).

Lentil is an important pulses crop but it's short stature and slow growth rate make it vulnerable to weed competition, limiting crop productivity. There is need to identify herbicide tolerant genotypes and their tolerance mechanism. The present investigation was conducted to understand the effect of imazethapyr (IM) treatment on accumulation of methylglyoxal (MG) and its detoxification mechanism in IM-tolerant (LL1397 and LL1612) susceptible (FLIP2004-7L and PL07) genotypes sown under control (weed free), weedy check (weeds were growing with crop) and sprayed with imazethapyr. The enzymes of glyoxalase pathway (glyoxalase I, II and III) and non glyoxalase pathway (methylglyoxal reductase), lactate dehydrogenase (LDH), glutathione content, gamma-glutamyl-cysteine synthetase (γ-GCS) were estimated in lentil genotypes at different days after spray. Higher activities of glyoxalase I, II and III and MGR along with the increased glutathione content (GSH) content in LL1397 and LL1612 under IM treatment as compared to FLIP2004-7L and PL07 might be responsible for lowering MG accumulation and increasing lactate content, which is end product of these pathways. Enhanced LDH activity in LL1397 and LL1612 might be responsible for energy production via TCA cycle that might be responsible for growth and recovery of tolerant genotypes after IM treatment. Higher γ-GCS activity in tolerant genotypes led to increased glutathione content required for glyoxalase pathway. However, decreased activities of glyoxalase enzymes and MGR in susceptible genotypes result in MG accumulation which limit plant growth. This is the first ever study elucidating the role of MG detoxification pathway conferring IM tolerance in lentil.

Impact of phosphorous-deficit conditions on morpho-physiological traits and phosphorous metabolism in chickpea genotypes.

Chickpea, an important food legume, is primarily grown on marginal soils with low soil fertility. Although chickpea can fix N, soil phosphorus (P) deficiency in crop growing areas is a major limiting factor for chickpea production. This study was undertaken to evaluate twenty-five chickpea cultivars for morpho-physiological traits and yield under low and normal phosphorous conditions. Based on morpho-physiological traits such as length and area of roots and shoots, root length density, root and shoot biomass, chlorophyll content, number of nodules and root tips, tolerance indices and yield, these cultivars were characterised into susceptible (ICC67, ICC1915, ICC2593, ICC5337, ICC5879, ICC8950, ICC13441, ICC1483, ICC15606 and ICC15888), tolerant (ICC10755, IG72070, ICCV97105, ICCV2, ICCV92809, ICCV92337 and ICCV95423) and the remaining cultivars were moderately tolerant to phosphorous-deficit conditions. Higher activities of enzymes of phosphorous metabolism such as acid phosphatase and phytase in roots and nodules of tolerant chickpea cultivars (ICCV97105, ICCV92337, ICCV95423) as compared to susceptible cultivars (ICC67, ICC15606, ICC15888) at different developmental stages might be attributing to their better performance for growth parameters and productivity traits under phosphorous-deficit conditions.

Hematological characteristics, cytogenetic features, and post-induction measurable residual disease in thymic stromal lymphopoietin receptor (TSLPR) overexpressed B-cell acute lymphoblastic leukemia in an Indian cohort.

The overexpression of cytokine receptor-like factor-2 (CRLF2) identified by anti-thymic stromal lymphopoietin receptor/TSLPR flow cytometry (FCM) has been reported as a screening tool for the identification of BCR-ABL1-like B-cell acute lymphoblastic leukemia/B-ALL with CRLF2 re-arrangement. TSLPR expression was studied prospectively in consecutive 478 B-ALLs (≤ 12 years (n = 244); 13-25 years (n = 129); > 25 years (n = 105)) and correlated with various hematological parameters and end-of-induction measurable residual disease (day 29; MRD ≥ 0.01% by 10-color FCM). TSLPR positivity in ≥ 10% leukemic cells was detected in 14.6% (n = 70) of B-ALLs. CRLF2 re-arrangement was detected in eight cases (11.4%) including P2RY8-CRLF2 (n = 6), and IgH-CRLF2 (n = 2) with a median TSLPR positivity of 48.8% and 99% leukemic cells, respectively. Recurrent gene fusions/RGF (BCR-ABL1 (17.1%); ETV6-RUNX1 (4.2%), TCF3-PBX1 (1.4%)), other BCR-ABL1-like chimeric gene fusions/CGFs (PDGFRB-rearrangement (2.9%), IgH-EPOR (1.4%)), CRLF2 extra-copies/hyperdiploidy (17.1%), and IgH translocation without a known partner (10%) were also detected in TSLPR-positive patients. CD20 positivity (52.9% vs 38.5%; p = 0.02) as well as iAMP21 (4.3% vs 0.5%; p = 0.004) was significantly more frequent in TSLPR-positive cases. TSLPR-positive patients did not show a significantly higher MRD, compared to TSLPR-negative cases (37% vs 33%). Increasing the threshold cut-off (from ≥ 10 to > 50% or > 74%) increased the specificity to 88% and 100% respectively in identifying CRLF2 translocation. TSLPR expression is not exclusive for CRLF2 translocations and can be seen with various other RGFs, necessitating their testing before its application in diagnostic algorithms. In patients with high TSLPR positivity (> 50%), the testing may be restricted to CRLF2 aberrancies, while patients with 10-50% TSLPR positivity need to be tested for both CRLF2- and non-CRLF2 BCR-ABL1-like CGFs.

NPM1 and FLT3-ITD/TKD Gene Mutations in Acute Myeloid Leukemia.

Background: A number of mutations have been reported to occur in patients with acute myeloid leukemia (AML), of which NPM1 and FLT3 genes mutations are the commonest and have important diagnostic and therapeutic implications. Material and Methods: Molecular testing for NPM1 and FLT3 genes was performed in 92 de-novo AML patients. The frequency and characteristics of NPM1 and FLT3 mutations were analyzed. Results: Nucleophosmin 1(NPM1) and fms-like tyrosine kinase 3 (FLT3) mutations were seen in 22.8% and 16.3% of patients, respectively. Amongst FLT3 mutations, FLT3-ITD mutation was seen in 8.7% cases, FLT3-TKD in 5.4%, and FLT3-ITD+TKD in 2.2% cases. Certain associations between the gene mutations and clinical characteristics were found, including in NPM1 mutated group- female preponderance, higher incidence in M4/M5 categories and decreased expression of CD34 and HLA-DR; and in FLT3-ITD mutated group- higher age of presentation, higher total leucocyte count and blast percentage. Conclusion- AML patients with NPM1 and FLT3 mutations have differences in clinical and hematological features, which might represent their different molecular mechanism in leukemogenesis. The frequency of NPM1 and FLT3 mutations in this study was comparable to reports from Asian countries but lower than that reported from western countries. However, as the number of patients in the study was less, a larger number of patients need to be studied to corroborate these findings.

Utility of cell population data (VCS parameters) as a rapid screening tool for Acute Myeloid Leukemia (AML) in resource-constrained laboratories.

INTRODUCTION: Despite advances in diagnostic techniques, many cases of acute myeloid leukemia (AML) remain underdiagnosed in remote centers unequipped with these technologies. We hypothesized that the automated cellular indices with scatter plots and flags may aid in rapid and cost-effective screening of AML. METHODS: Cell population data (CPD) parameters from 100 de novo AML samples were analyzed by Coulter LH 780 automated analyzer and were compared with 100 age-matched controls. Similar parameters were also compared with 100 and 50 reactive cases of neutrophilia and monocytosis, respectively. System-generated flags and scatter plot patterns were also analyzed. RESULTS: Results were compared between AML cases and normal controls; AML FAB M2, M3, M4 vs reactive neutrophilia; AML FAB M4, M5 vs reactive monocytosis. Significant parameters were selected from all comparison groups. Using appropriate statistical tools, we calculated the cutoff values of these parameters and were able to screen out AML cases with 94% sensitivity and 95% specificity. Three statistical equations were generated using two of the most significant parameters which improved the sensitivity to 98% and specificity to 99%. Five hypothetical scatter plot patterns were devised and were classified according to FAB categories of AML. Most common pattern was selected in AML which was seen in 56% of the cases. Output was analyzed combining these patterns and flags with CPD parameters. CONCLUSION: CPD either alone or in the form of statistical equations along with scatter plots and flags can provide rapid and economic tool in preliminary diagnosis of AML in cost-constrained settings.