Marker-assisted Selection Research Articles

Advancements in breeding for high oil content in maize (Zea mays. L)

Maize is one of the world’s most important food crops, supplying>5% deity energy. Due to its wider adaptability, high yield potential, maize is widely utilized as food and animals world worldwide. It enriched with caloric starch and the high energy in its oil. The kernel oil content in commercial high-oil maize hybrids averages ∼8%, far lower than that in developed high-oil maize lines (as high as 20%). The maize seed/kernel consisted of endosperm (82%) and the germ (embryo and scutellum) (12%). The germ region makes up 80–84% of kernel oil, with the aleurone and endosperm making up 12% and 5%, respectively. High-oil maize may serve as a valuable forage maize germplasm for breeding. Maize oil is also considered as high-quality oil for human health due to the high proportion of polyun-saturated fatty acids. Selection for high oil increases the proportion of germ and further content of germ oil. The problems that over-shadow the successful production of high-oil corn are low grain yield potential, physiological cost of oil synthesis, low seed vigor, low kernel weight, shorter seed longevity, and poor germination of high-oil corn lines. It is necessary to make efforts to generate nested populations or use marker-assisted recurrent selection (MARS) to accumulate the advantageous quantitative trait loci (QTLs) in the pa-rental lines. The oil content of maize inbreeds and hybrids could be further improved with the use of transgenes, genomics, and metabolic engineering technologies in new insight for the development of high oil maize breeding and strategies.

PSIV-5 Prioritizing candidate genes for seminal traits and residual feed intake in cattle: Integrating QTLome Analysis and RNAseq Datasets

Abstract The QTLome represents the collection of QTLs (quantitative trait loci) documented for each trait within a specific species, detailing the position, effect, and mode of inheritance of each locus. Despite the abundance of known QTLs for productive traits in cattle, only a few genes have been identified as selection markers. This study aimed to prioritize genes within validated QTLs associated with crucial traits in cattle by integrating this information with RNAseq data. Initially, bovine QTLs (n = 161,730) were sourced from the AnimalQTLdb database (version 44), and both graphical and quantitative analyses were conducted using R software. Only annotations within the UMD 3.1 genome were considered, with overlapping QTLs from the same scientific report grouped together. Following quality control measures, the number of QTLs was reduced to 37,146. Validated QTLs were determined by identifying overlapping annotations from various studies or those within a distance of < 1 Mb for the same phenotypic trait, resulting in a total of 3,856 validated QTLs. For gene prioritization, validated QTLs were selected based on semen traits and residual feed intake (RFI). Transcriptomes corresponding to these traits were sourced from NCBI (PRJNA516089 and PRJEB7696). Sequence filtering was performed using the fastp program, and transcript counts were obtained with SALMON software. Sequences were aligned to the Ensembl UMD 3.1 reference genome (version 94), and differential expression analysis was conducted using the R package DESeq2, considering only the group factor (high/low) for each trait (fertility/RFI). Integration of QTLs and transcriptomes utilized the methodology of the cageminer R package, which prioritizes candidate genes within validated QTLs, identifies those grouped in co-expression modules enriched by reference genes, and correlates with the study phenotype (biserial r > 0.2). Reference genes were obtained from Ignatieva et al. (2021 PMID 34290736) and Chen et al. (2021; PMID 33664767). Among the 18 validated QTLs for semen traits, four candidate genes (TSPAN6, QSER1, ADRB2, AKAP4) were identified. Except for ADRB2, these genes exhibited significant differential expression (P < 0.05) in contrasting groups of Holstein cattle. They are associated with semen traits and perform functions such as regulation of spermatogenesis, development, activation, and cell growth. Regarding RFI, 127 validated QTLs yielded 205 prioritized genes, of which 123 showed significant differential expression (P < 0.05) in divergent groups of the Nelore breed. These genes are involved in the immune response and the polymerization and organization of actin filaments, processes that have been reported as biologically relevant to RFI. The implemented strategy was efficient for gene prioritization through the integration of QTLs and RNAseq.

Genetic analysis of QTLs for lysine content in four maize DH populations

BackgroundLow levels of the essential amino acid lysine in maize endosperm is considered to be a major problem regarding the nutritional quality of food and feed. Increasing the lysine content of maize is important to improve the quality of food and feed nutrition. Although the genetic basis of quality protein maize (QPM) has been studied, the further exploration of the quantitative trait loci (QTL) underlying lysine content variation still needs more attention.ResultsEight maize inbred lines with increased lysine content were used to construct four double haploid (DH) populations for identification of QTLs related to lysine content. The lysine content in the four DH populations exhibited continuous and normal distribution. A total of 12 QTLs were identified in a range of 4.42–12.66% in term of individual phenotypic variation explained (PVE) which suggested the quantitative control of lysine content in maize. Five main genes involved in maize lysine biosynthesis pathways in the QTL regions were identified in this study.ConclusionsThe information presented will allow the exploration of candidate genes regulating lysine biosynthesis pathways and be useful for marker-assisted selection and gene pyramiding in high-lysine maize breeding programs.

Identification and analysis of low light responsive yield enhancing QTLs in rice

Rice is one of the major food crops grown globally. However, during the wet season, rice suffers significant yield loss due to reduced light intensity caused by overcast clouds when the light intensity is only around 450–500 µmol/m2/s, compared to 1400–1800 µmol/m2/s in summer. This reduction in light intensity leads to a decrease in seed yield, mainly by limiting tiller or panicle numbers. Yield and its attributing parameters were recorded in one hundred thirty RILs for four consecutive wet seasons in ambient light (AL) and low light (LL, 35% light-cut using white shade net). QTL analysis was performed using Inclusive Composite Interval Mapping (ICIM) with all the phenotypic data and 927 polymorphic SNPs identified by the 7 K Infinium chip. The study identified a large QTL influencing panicle numbers and yield exclusively in lowlight on chromosome 1 (qPNLL1.1, qGYLL1.1) in four consecutive seasons with LOD > 10 and PVE > 30%. The favourable alleles are from the tolerant parent, Swarnaprabha. Another grain yield improving QTL was identified on chromosome 6 (qGYLL6.1), with LOD > 3 in three consecutive seasons. In a diverse rice panel of one hundred seventeen genotypes with five different models, association analysis identified the associated marker for panicle numbers and grain yield in LL, which is also the left marker of the newly identified QTLs for the traits under LL condition. A shade-responsive gene, monoculm 2 (MOC2, LOC_Os01g64660) inside the QTL on chromosome 1, upregulated in the tolerant parent and its QTL-carrying RILs, whereas repressed in the susceptible one. Therefore, due to its significant additive effect and validation across various genotypes, the yield-improving QTL on chromosome 1 can be directly utilised in marker-assisted selection (MAS) for developing shade-tolerant rice. This can also help reduce the yield gap between wet and dry-season rice.

Physio-biochemical and molecular characterization of wheat cultivars (Triticum aestivum L.) under post-anthesis heat stress

BackgroundHeat stress is one of the abiotic stresses that make wheat crops vulnerable, which significantly impacts crop production around the world. An increase in temperature during the reproductive phase (anthesis) beyond the optimal range of 15–20 °C leads to decreased crop production, poor quality of the grain, and altered physiological and biochemical processes. To study the association between high temperature and physio-biochemical traits under normal and late sown, a set of fifteen genotypes was utilized.ResultsRelative water content under high temperatures had an overall decrease of 8.7%. However, grain protein and malondialdehyde content were higher in the stressed conditions than in the control, with increases of 20.2% and 38.9%, respectively. Marker Xgwm67, located on chromosome 5B, was found to be significantly associated with malondialdehyde content (R2 = 21%) and Xgwm570, located on 6A, was closely linked to relative water content as well as grain protein content (R2 = 16%) revealed by regression analysis. The correlation matrix displays a positive association between the control and stressed condition by R2 = 0.92, 0.82, and 0.53 in malondialdehyde, relative water content, and grain protein, respectively. However, there was a negative correlation between water content–malondialdehyde and malondialdehyde–grain protein, though there was only a 4% correlation between grain protein content (control) and relative water content (stressed). Based on the tolerance matrix, WH730 and RAJ4079 were heat tolerant, and DBW173 and HD3086 were sensitive.ConclusionsThese findings indicate that to identify tolerant genotypes, physiological and biochemical traits can be utilized as an alternate criterion, and these closely associated markers can be applied for improved late-planted wheat production through MAS. The breeding scheme and genome editing by recognizing novel genes through physio-biochemical parameters, marker-assisted selection, and prospective screening of tolerant genotypes are proclaimed by the study.

Multi-trait modeling and machine learning discover new markers associated with stem traits in alfalfa

Alfalfa biomass can be fractionated into leaf and stem components. Leaves comprise a protein-rich and highly digestible portion of biomass for ruminant animals, while stems constitute a high fiber and less digestible fraction, representing 50 to 70% of the biomass. However, little attention has focused on stem-related traits, which are a key aspect in improving the nutritional value and intake potential of alfalfa. This study aimed to identify molecular markers associated with four morphological traits in a panel of five populations of alfalfa generated over two cycles of divergent selection based on 16-h and 96-h in vitro neutral detergent fiber digestibility in stems. Phenotypic traits of stem color, presence of stem pith cells, winter standability, and winter injury were modeled using univariate and multivariate spatial mixed linear models (MLM), and the predicted values were used as response variables in genome-wide association studies (GWAS). The alfalfa panel was genotyped using a 3K DArTag SNP markers for the evaluation of the genetic structure and GWAS. Principal component and population structure analyses revealed differentiations between populations selected for high- and low-digestibility. Thirteen molecular markers were significantly associated with stem traits using either univariate or multivariate MLM. Additionally, support vector machine (SVM) and random forest (RF) algorithms were implemented to determine marker importance scores for stem traits and validate the GWAS results. The top-ranked markers from SVM and RF aligned with GWAS findings for solid stem pith, winter standability, and winter injury. Additionally, SVM identified additional markers with high variable importance for solid stem pith and winter injury. Most molecular markers were located in coding regions. These markers can facilitate marker-assisted selection to expedite breeding programs to increase winter hardiness or stem palatability.

Genome-Wide Association Studies and QTL Mapping Reveal a New Locus Associated with Resistance to Bacterial Pustule Caused by Xanthomonas citri pv. glycines in Soybean.

Bacterial pustule (BP), caused by Xanthomonas citri pv. glycines, is an important disease that, under favorable conditions, can drastically affect soybean production. We performed a genome-wide association study (GWAS) with a panel containing Brazilian and American cultivars, which were screened qualitatively and quantitatively against two Brazilian X. citri isolates (IBS 333 and IBS 327). The panel was genotyped using a genotyping by sequencing (GBS) approach, and we identified two main new regions in soybeans associated with X. citri resistance on chromosomes 6 (IBS 333) and 18 (IBS 327), different from the traditional rxp gene located on chromosome 17. The region on chromosome 6 was also detected by QTL mapping using a biparental cross between Williams 82 (R) and PI 416937 (S), showing that Williams 82 has another recessive resistance gene besides rxp, which was also detected in nine BP-resistant ancestors of the Brazilian cultivars (including CNS, S-100), based on haplotype analysis. Furthermore, we identified additional SNPs in strong LD (0.8) with peak SNPs by exploring variation available in WGS (whole genome sequencing) data among 31 soybean accessions. In these regions in strong LD, two candidate resistance genes were identified (Glyma.06g311000 and Glyma.18g025100) for chromosomes 6 and 18, respectively. Therefore, our results allowed the identification of new chromosomal regions in soybeans associated with BP disease, which could be useful for marker-assisted selection and will enable a reduction in time and cost for the development of resistant cultivars.

Biotechnological frontiers in harnessing allelopathy for sustainable crop production.

Allelopathy, the phenomenon in which plants release biochemical compounds that influence the growth and development of neighbouring plants, presents promising opportunities for revolutionizing agriculture towards sustainability. This abstract explores the role of biotechnological advancements in unlocking the potential of allelopathy for sustainable crop production and its applications in agriculture, ecology, and natural resource management. By combining molecular, genetic, biochemical, and bioinformatic tools, researchers can unravel the complexities of allelopathic interactions and their potential for sustainable crop production and environmental stewardship. The development of novel management methods for weed control is getting a lot of attention with the introduction of new genetic technologies such as Gene drive, Transgene technologies, Gene silencing, Marker-assisted selection (MAS), and Clustered regularly interspaced short palindromic repeats (CRISPR-Cas9). By strengthening competitive characteristics these tools hold great promise for boosting crops' ability to compete with weeds. Considering recent literature, this review highlights the genetic, transcriptomics, and metabolomics approaches to allelopathy. Employing allelopathic properties in agriculture offer sustainable benefits like natural weed management, pest management, and reduced chemical pollution, but challenges include environmental factors, toxicity, regulatory hurdles, and limited resources. Effective integration requires continued research, regulatory support, and farmer education​. Also, we aimed to identify the biotechnological domains requiring more investigation and to provide the basis for future advances through this assessment.

Multi-Population Analysis for Leaf and Neck Blast Reveals Novel Source of Neck Blast Resistance in Rice.

Rice blast is one of the most devastating biotic stresses that limits rice productivity. The North Eastern Hill (NEH) region of India is considered to be one of the primary centres of diversity for both rice and pathotypes of Magnaporthe grisea. Therefore, the present study was carried out to elucidate the genetic basis of leaf and neck blast resistance under Meghalaya conditions. A set of 80 diverse genotypes (natural population) and 2 F2 populations involving resistant parent, a wildtype landrace, LR 5 (Lal Jangali) and susceptible genotypes Sambha Mahsuri SUB 1 (SMS) and LR 26 (Chakhao Poireiton) were used for association analysis of reported major gene-linked markers with leaf and neck blast resistance to identify major effective genes under local conditions. Genotyping using twenty-five gene-specific markers across diverse genotypes and F2 progenies revealed genes Pi5 and Pi54 to be associated with leaf blast resistance in all three populations. Genes Pib and qPbm showed an association with neck blast resistance in both natural and LR 5 × SMS populations. Additionally, a set of 184 genome-wide polymorphic markers (SSRs and SNPs), when applied to F2-resistant and F2-susceptible DNA bulks derived from LR 5 × LR 26, suggested that Pi20(t) on chromosome 12 is one of the major genes imparting disease resistance. Markers snpOS318, RM1337 and RM7102 and RM247 and snpOS316 were associated with leaf blast and neck blast resistance, respectively. The genotypes, markers and genes will help in marker-assisted selection and development of varieties with durable resistance.

Phase 1b/2 study of the liposomal formulation of eribulin (E7389-LF) in combination with nivolumab: Results from the phase 2 esophageal cancer cohort

BackgroundEsophageal cancer is one of the most common types of cancer in Japan. Herein, we report the efficacy and safety of E7389-LF plus the immune checkpoint inhibitor, nivolumab, from the esophageal cancer cohort of the phase 2 part of Study 120.MethodsEligible patients received E7389-LF 2.1 mg/m2 plus nivolumab 360 mg intravenously Q3W. The primary objective was to evaluate the objective response rate (ORR); other objectives included safety, progression-free survival (PFS), and overall survival (OS).ResultsOf the 35 Japanese patients enrolled, 7 (20.0%) had a partial response as their best overall response, and 14 (40.0%) had stable disease. The ORR was 20.0% (95% CI 8.4–36.9). The duration of response was 5.6 months (95% CI 1.7–not estimable [NE]). The median PFS was 2.81 months (95% CI 1.31–4.17). The median OS was not reached (95% CI 6.54 months–NE). The most common treatment-emergent adverse events were neutropenia (65.7%), pyrexia (60.0%), and leukopenia (57.1%). Select plasma endothelial cell markers levels increased from day 1 of cycle 1 and changes were pronounced between days 8–15 of each cycle.ConclusionsE7389-LF plus nivolumab showed antitumor activity in patients with unresectable and pretreated esophageal cancer and should be evaluated further in a broader population.Clinical Trial RegistrationNCT04078295.

Identification and validation of oxidative stress-related diagnostic markers for recurrent pregnancy loss: insights from machine learning and molecular analysis.

It has been recognized that oxidative stress (OS) is implicated in the etiology of recurrent pregnancy loss (RPL), yet the biomarkers reflecting oxidative stress in association with RPL remain scarce. The dataset GSE165004 was retrieved from the Gene Expression Omnibus (GEO) database. From the GeneCards database, a compendium of 789 genes related to oxidative stress-related genes (OSRGs) was compiled. By intersecting differentially expressed genes (DEGs) in normal and RPL samples with OSRGs, differentially expressed OSRGs (DE-OSRGs) were identified. In addition, four machine learning algorithms were employed for the selection of diagnostic markers for RPL. The Receiver Operating Characteristic (ROC) curves for these genes were generated and a predictive nomogram for the diagnostic markers was established. The functions and pathways associated with the diagnostic markers were elucidated, and the correlations between immune cells and diagnostic markers were examined. Potential therapeutics targeting the diagnostic markers were proposed based on data from the Comparative Toxicogenomics Database and ClinicalTrials.gov. The candidate biomarker genes from the four models were further validated in RPL tissue samples using RT-PCR and immunohistochemistry. A set of 20 DE-OSRGs was identified, with 4 genes (KRAS, C2orf69, CYP17A1, and UCP3) being recognized by machine learning algorithms as diagnostic markers exhibiting robust diagnostic capabilities. The nomogram constructed demonstrated favorable predictive accuracy. Pathways including ribosome, peroxisome, Parkinson's disease, oxidative phosphorylation, Huntington's disease, and Alzheimer's disease were co-enriched by KRAS, C2orf69, and CYP17A1. Cell chemotaxis terms were commonly enriched by all four diagnostic markers. Significant differences in the abundance of five cell types, namely eosinophils, monocytes, natural killer cells, regulatory T cells, and T follicular helper cells, were observed between normal and RPL samples. A total of 180 drugs were predicted to target the diagnostic markers, including C544151, D014635, and CYP17A1. In the validation cohort of RPL patients, the LASSO model demonstrated superiority over other models. The expression levels of KRAS, C2orf69, and CYP17A1 were significantly reduced in RPL, while UCP3 levels were elevated, indicating their suitability as molecular markers for RPL. Four oxidative stress-related diagnostic markers (KRAS, C2orf69, CYP17A1, and UCP3) have been proposed to diagnose and potentially treat RPL.

A user-friendly CRISPR/Cas9 system for mutagenesis of Neurosporacrassa

As a widely used eukaryotic model organism, Neurosporacrassa offers advantages in genetic studies due to its diverse biology and rapid growth. Traditional genetic manipulation methods, such as homologous recombination, require a considerable amount of time and effort. In this study, we present an easy-to-use CRIPSR/Cas9 system for N.crassa, in which the cas9 sequence is incorporated into the fungal genome and naked guide RNA is introduced via electroporation. Our approach eliminates the need for constructing multiple vectors, speeding up the mutagenesis process. Using cyclosporin-resistant-1 (csr-1) as a selectable marker gene, we achieved 100% editing efficiency under selection conditions. Furthermore, we successfully edited the non-selectable gene N-acylethanolamineamidohydrolase-2 (naa-2), demonstrating the versatility of the system. Combining gRNAs targeting csr-1 and naa-2 simultaneously increased the probability of finding mutants carrying the non-selectable mutation. The system is not only user-friendly but also effective, providing a rapid and efficient method for generating loss-of-function mutants in N.crassa compared to traditional methods.

InDel mutations within the bovine PER2 gene are significantly associated with reproductive traits.

Functioning as a key regulator of circadian rhythms, the PER2 gene exerts a substantial impact on the reproductive traits of animals. However, the effect of the PER2 gene on ovarian development remains unclear. In order to examine the relationship between bovine reproductive trait and the PER2 gene, a total of 901 ovarian samples were collected, categorized into different oestrus cycles (proestrus, oestrus, post-oestrus, anoestrous), and subjected to analysis for two potential insertion/deletions (InDels) in the PER2 gene. Through agarose gel electrophoresis and DNA sequencing, two polymorphic deletion mutations (P2-D5-bp, P3-D13-bp) were identified. Furthermore, a significant association between mature follicle diameter and P2-D5-bp was found (P < 0.05). Additionally, several significant correlations with ovarian length, width, height, and white body diameter were found for P3-D13-bp (P < 0.05). These findings suggested that the bovine PER2 gene plays an important role in above-mentioned reproductive traits, offering new avenues for improving cow fertility through marker-assisted selection (MAS).

QTL mapping for branch- and leaf-related traits with a high-density SNP genetic map in litchi (Litchi chinensis Sonn.)

Litchi (Litchi chinensis Sonn.), an important fruit tree in tropical and subtropical regions, possesses substantial economic value. The branch- and leaf-related traits of litchi have a significant impact on litchi yield and quality. However, due to limitations such as the density of the genetic linkage map, there have been few studies on mapping QTLs of branch- and leaf-related traits. In this study, a high-density genetic map was constructed by next-generation sequencing (NGS) using an F1 population of 264 progenies, derived from the cross between the cultivars ‘Sanyuehong’ and ‘Ziniangxi’. A total of 2 574 high-quality BINs (binomial intervals) were obtained, and a genetic linkage map was constructed with a total length of 1 753.3 cM and an average marker distance of 0.68 cM. With the genetic map and the phenotyping of single leaf length (SLL), single leaf width (SLW), leaf shape index (LSI), weight of specific leaf (WSL), petiole length (PL) and compound leaf length (CLL) measured in three seasons, 11, 9, 9, 10, 9 and 12 QTLs were detected for SLL, SLW, WSL, LSI, PL and CLL traits, respectively. Among these QTLs, five QTLs were consistently detected in two seasons and 12 pleiotropic QTLs were identified for at least two traits. These findings will provide new insights for the gene cloning for branch- and leaf-related traits as well as marker-assisted selection (MAS).

Genetic Basis and Exploration of Major Expressed QTL qLA2-3 Underlying Leaf Angle in Maize

Leaf angle (LA) is closely related to plant architecture, photosynthesis and density tolerance in maize. In the current study, we used a recombinant inbred line population constructed by two maize-inbred lines to detect quantitative trait loci (QTLs) controlling LA. Based on the average LA in three environments, 13 QTLs were detected, with the logarithm of odds ranging from 2.7 to 7.21, and the phenotypic variation explained by a single QTL ranged from 3.93% to 12.64%. A stable QTL, qLA2-3, on chromosome 2 was detected and was considered to be the major QTL controlling the LA. On the basis of verifying the genetic effect of qLA2-3, a fine map was used to narrow the candidate interval, and finally, the target segment was located at a physical distance of approximately 338.46 kb (B73 RefGen_v4 version), containing 16 genes. Re-sequencing and transcriptome results revealed that five candidate genes may be involved in the regulation of LA. The results enrich the information for molecular marker-assisted selection of maize LA and provide genetic resources for the breeding of dense planting varieties.

Exploration of the pneumocandin biosynthetic gene cluster based on efficient CRISPR/Cas9 gene editing strategy in Glarea lozoyensis

Pneumocandin B0 (PB0) is a lipopeptide produced by the fungus Glarea lozoyensis. The existing challenges with the low-yield and the extended-fermentation cycle emphasize necessity for strain improvement. In this study, we optimized conditions to obtain high-quality protoplasts and screened effective selection markers, leading to the construction of three CRISPR/Cas9 gene editing systems. Utilizing a constitutive Cas9 expression recipient strain, combined with dual sgRNAs targeting, we achieved highly efficient editing of target genes. We successfully knocked out 10 genes within the pneumocandin putative biosynthetic gene cluster and analyzed their roles in PB0 production. Our findings reveal that 4 of 10 genes are directly involved in PB0 production. Specially, the deletion of gltrt or gl10050 resulted in reduced PB0 production, while the absence of glhyp or glhtyC led to the complete loss of PB0 biosynthesis. Notably, the deletion of glhyp caused the silencing of nearly all cluster genes, whereas overexpression of glhyp led to a 2.38-fold increase in PB0 production. Therefore, this study provides the first comprehensive exploration of the functions of 10 genes within the pneumocandin putative biosynthetic gene cluster. Our findings provide valuable technical strategies for constructing bioengineering strains with purposefully enhanced PB0 production.

Fruquintinib in metastatic colorectal cancer: a multicenter real-world analysis on efficacy, safety, and predictive and prognostic factors.

Randomized trials have shown a survival benefit for fruquintinib over placebo in patients with metastatic colorectal cancer (mCRC) that progressed after standard therapies, but real-world prognostic analyses have been seldom reported. We evaluated survival, safety outcomes, and predictive and prognostic factors in patients treated with fruquintinib in a real-life setting. We conducted a multi-center study by collecting relevant data on patients with advanced colorectal cancer (CRC) who received fruquintinib, focusing on progression-free survival (PFS), overall survival (OS), and L3 skeletal muscle index (SMI), including safety follow-up. From January 2020 to January 2022, a total of 140 patients were selected and included in this study. The cut-off date was 30 July 2022. The median follow-up time was 18.3 months (range, 6-29.3 months) and the median age of included cases was 63 years (range, 32-81 years). The median PFS and OS for the 140 patients was 6.3 and 12.6 months, respectively. The median PFS and OS for the 76 patients who were included in SMI analysis was 6.0 and 12.0 months, respectively. Multivariate analysis suggested brain metastasis {hazard ratio (HR) [95% confidence interval (CI)]: 2.779 (1.162-6.646), P=0.02}, decrease in SMI of >5% [HR (95% CI): 9.732 (2.201-43.028), P=0.003], and baseline carcinoembryonic antigen (CEA) level [HR (95% CI): 4.061 (1.391-11.858), P=0.01] as independent predictors of OS. The most common treatment-related adverse events (TRAEs) were hypertension (24, 17.1%), fatigue (21, 15%), and hand-foot syndrome (20, 14.3%); 9 (13.6%) and 15 (10.7%) patients had dose reduction and treatment discontinuation due to TRAEs respectively. The real-world efficacy and safety of fruquintinib in advanced CRC patients are numerically superior to that in the previous phase III studies. SMI, brain metastasis and CEA could serve as potential markers for patient selection.

Virulence Adaptation by Rice Planthoppers and Leafhoppers to Resistance Genes and Loci: A Review

In recent decades, research on developing and deploying resistant rice has accelerated due to the availability of modern molecular tools and, in particular, advances in marker-assisted selection. However, progress in understanding virulence adaptation has been relatively slow. This review tracks patterns in virulence adaptation to resistance genes (particularly Bph1, bph2, Bph3, and bph4) and examines the nature of virulence based on selection experiments, responses by virulent populations to differential rice varieties (i.e., varieties with different resistance genes), and breeding experiments that interpret the genetic mechanisms underlying adaptation. The review proposes that varietal resistance is best regarded as a combination of minor and major resistance traits against which planthoppers develop partial or complete virulence through heritable improvements that are reversable or through evolutionary adaptation, respectively. Agronomic practices, deployment patterns, and herbivore population pressures determine the rates of adaptation, and there is growing evidence that pesticide detoxification mechanisms can accelerate virulence adaptation. Research to delay adaptation has mainly focused on gene pyramiding (i.e., including ≥ two major genes in a variety) and multilines (i.e., including ≥ two resistant varieties in a field or landscape); however, these strategies have not been adequately tested and, if not managed properly, could inadvertently accelerate adaptation compared to sequential deployment. Several research gaps remain and considerable improvements in research methods are required to better understand and manage virulence adaptation.

Applications of Marker-Assisted Gene Pyramiding in Rice Crop Improvement

In order to give wide-ranging and long-lasting disease resistance that reduces rice output, breeding techniques are used such as the importance of selection assisted by markers and gene pyramiding. Bacterial leaf blight (BLB), which is carried on by the pathogen Xanthomonas oryzae pv. oryzae, is one such disease that significantly reduces rice yield. Numerous molecular marker techniques have already been developed because molecular markers are crucial for both marker-assisted selection and gene pyramiding. Currently, DNA-based markers, sometimes referred to as molecular markers, are the most frequently employed ones. Based on the methods used to find them, the molecular markers are divided into two main types. These markers are based on hybridization and polymerase chain reaction. The morphological (traditionally based) and biochemical (enzyme-based) markers are two more types of markers that are available. The best method for preventing rice BLB disease is host plant/varietal resistance. Gene pyramiding with the help of markers offers the potential to speed up breeding operations and ensure that the host plant’s acquired resistance will last. This research explores the use, economic significance, constraints, and potential of marker-assisted selection and gene pyramiding for rice BLB disease resistance.

Identifying the physiological traits associated with DNA marker using genome wide association in wheat under heat stress

Heat stress poses a significant environmental challenge that profoundly impacts wheat productivity. It disrupts vital physiological processes such as photosynthesis, by impeding the functionality of the photosynthetic apparatus and compromising plasma membrane stability, thereby detrimentally affecting grain development in wheat. The scarcity of identified marker trait associations pertinent to thermotolerance presents a formidable obstacle in the development of marker-assisted selection strategies against heat stress. To address this, wheat accessions were systematically exposed to both normal and heat stress conditions and phenotypic data were collected on physiological traits including proline content, canopy temperature depression, cell membrane injury, photosynthetic rate, transpiration rate (at vegetative and reproductive stage and ‘stay-green’. Principal component analysis elucidated the most significant contributors being proline content, transpiration rate, and canopy temperature depression, which exhibited a synergistic relationship with grain yield. Remarkably, cluster analysis delineated the wheat accessions into four discrete groups based on physiological attributes. Moreover, to explore the relationship between physiological traits and DNA markers, 158 wheat accessions were genotyped with 186 SSRs. Allelic frequency and polymorphic information content value were found to be highest on genome A (4.94 and 0.688), chromosome 1A (5.00 and 0.712), and marker Xgwm44 (13.0 and 0.916). Population structure, principal coordinate analysis and cluster analysis also partitioned the wheat accessions into four subpopulations based on genotypic data, highlighting their genetic homogeneity. Population diversity and presence of linkage disequilibrium established the suitability of population for association mapping. Additionally, linkage disequilibrium decay was most pronounced within a 15–20 cM region on chromosome 1A. Association mapping revealed highly significant marker trait associations at Bonferroni correction P < 0.00027. Markers Xwmc418 (located on chromosome 3D) and Xgwm233 (chromosome 7A) demonstrated associations with transpiration rate, while marker Xgwm494 (chromosome 3A) exhibited an association with photosynthetic rates at both vegetative and reproductive stages under heat stress conditions. Additionally, markers Xwmc201 (chromosome 6A) and Xcfa2129 (chromosome 1A) displayed robust associations with canopy temperature depression, while markers Xbarc163 (chromosome 4B) and Xbarc49 (chromosome 5A) were strongly associated with cell membrane injury at both stages. Notably, marker Xbarc49 (chromosome 5A) exhibited a significant association with the 'stay-green' trait under heat stress conditions. These results offers the potential utility in marker-assisted selection, gene pyramiding and genomic selection models to predict performance of wheat accession under heat stress conditions.