Near-isogenic Lines Research Articles

Identification of qTGW2, a Minor-Effect QTL Controlling Grain Weight in Rice

Grain weight and grain shape are key traits affecting grain yield and quality in rice. In this research, a quantitative trait locus (QTL), qTGW2, that controls 1000-grain weight (TGW), grain length (GL), and grain width (GW) in rice, was fine-mapped within an 84.7 kb region on chromosome 2 using three sets of near isogenic lines (NILs) originated from the indica rice cross, Teqing (TQ)/IRBB52. In the NIL populations, the TGW, GL, and GW of the IRBB52 homozygous lines increased by 0.22 g, 0.020 mm, and 0.009 mm compared with the TQ homozygous lines. Four annotated genes showed nucleotide polymorphisms between the two parental lines in the qTGW2 region. Only one annotated gene, LOC_Os02g57660, exhibited significant expression differences between NILTQ and NILIRBB52 in the young panicles performed by RNA sequencing and the quantitative real-time polymerase chain reaction. These results indicated that LOC_Os02g57660, which encodes phosphatidylinositol-4-phosphate 5-kinase (PIP5K), was the candidate gene of qTGW2. Then, one insertion-deletion (InDel) was found in the LOC_Os02g57660 coding region. The haplotype analysis was performed based on the phenotypic data of 4720 rice accessions from RiceVarMap V2.0. Two haplotypes, Hap1 (TQ-type) and Hap2 (IRBB52-type), were classified according to one InDel. Significant differences in grain weight traits were identified between Hap1 and Hap2. Hap2 has greater GL and RLW but lower GW, thus exhibiting potential to simultaneously improve grain yield and quality.

Dough properties and baking quality of near isogenic lines with single HMW-GS null at Glu-A1, Glu-B1 and Glu-D1 loci in soft wheat

High molecular weight glutenin subunits (HMW-GSs) encoded by the Glu-1 loci are key factors in determining wheat processing quality. However, the contributions of single HMW-GS to dough viscoelastic properties and end-use quality have not been well understood, especially in soft wheat. In this study, the kernel samples of five Near Isogenic Lines (NILs) of a soft wheat each with individual HMW-GS null were investigated. Significant lower SDS-sedimentation volume and lactic acid solvent retention capacity (SRC) were observed in the NILs, compared to the wild type (WT). Except for the NIL with Glu-D1y12 null, the other NILs were associated with significantly lower dough mixing tolerance compared to the WT. Also, the deletion of individual HMW-GS was associated with reduction in dough elasticity and strength, compared to the WT, with the exception of the Glu-A1x1 null. Moreover, Glu-B1x7 was the most important HMW-GS in determining dough mixing tolerance and elasticity. Compared to the WT, the deletions of Glu-D1x2, Glu-B1x7 and Glu-B1y8 significantly increased alveograph L, and decreased alveograph P and P/L ratios. All NILs with single HMW-GS null showed better sugar snap cookie baking quality than the WT. New germplasm and tools are identified for improving dough viscoelasticity and baking quality in soft wheat.

Identification and segregation of two closely linked major QTLs for kernel row number in advanced maize-teosinte populations.

Two closely linked novel loci, qKRN2-1 and qKRN2-2, associated with kernel row number were fine-mapped on chromosome 2, and a key candidate gene for qKRN2-1 was identified through expression analysis. Kernel row number (KRN) is a crucial factor influencing maize yield and serves as a significant target for maize breeding. The use of wild progenitor species can aid in identifying the essential traits for domestication and breeding. In this study, teosinte (MT1) served as the donor parent, the inbred maize line of Mo17 was used as the recurrent parent, we identified a major quantitative trait locus (QTL) for KRN, designated qKRN2, into two closely linked loci, qKRN2-1 and qKRN2-2. Here, fine mapping was performed to investigate two QTLs, qKRN2-1 and qKRN2-2, within a genomic range of 272kb and 775kb, respectively. This was achieved using a progeny test strategy in an advanced backcross population, with the two QTLs explaining 33.49% and 35.30% of the phenotypic variance. Molecular marker-assisted selection resulted in the development of two nearly isogenic lines (NILs), qKRN2-1 and qKRN2-2, which differed only in the segment containing the QTL. Notably, the maize (Mo17) alleles increased the KRN relative to teosinte by approximately 1.4 and 1.2 rows for qKRN2-1 and qKRN2-2, respectively. Zm00001d002989 encodes a cytokinin oxidase/dehydrogenase and its expression in the immature ears exhibited significant differences among the qKRN2-1 NILs. In situ hybridization localized Zm00001d002989 to the primordia of the inflorescence meristem and spikelet pair meristems, is predicted to be the causal gene of qKRN2-1. The findings of this study deepen our understanding of the genetic basis of KRN and hold significant potential for improving maize grain yields.

Fine mapping and candidate gene mining of QSc/Sl.cib-7H for spike compactness and length and its pleiotropic effects on yield-related traits in barley (Hordeum vulgare L.).

A major locus for spike compactness and length was mapped on chromosome 7H and its pleiotropic effects, candidate genes and transcriptional regulatory network were analyzed. Spike compactness (SC) and length (SL) are important traits of barley (Hordeum vulgare L.) due to their close association with grain yield. In this study, a major SC and SL locus QSc/Sl.cib-7H was primarily identified on chromosome 7H by bulked segregant analysis, and further fine mapped to a recombination cold spot expanding 244.36-388.09Mb by developing a secondary population using residual heterozygous lines. This region is much more accurate than previously reported spike compactness loci on chromosome 7H.The strong effects of QSc/Sl.cib-7H on SL and SC were validated in two pair of near isogenic lines (NILs) and diverse genetic backgrounds. QSc/Sl.cib-7H exhibited pleiotropic effects on plant height (PH), thousand grain weight and grain length, and did not significantly influence the spikelet number of main spike (SMS) and grain width. Transcriptome analysis based on NILs showed that regulation of SC and SL might be related to the plant circadian rhythm pathway. The candidate genes were mined by analyzing variants and expression patterns of genes in the target region employing multiple genome and transcriptome data. This study takes a further step towards cloning of QSc/Sl.cib-7H, and the data obtained and the developed molecular markers will facilitate its utilization in barley breeding.

Characterization of genetic resistance to cucumber mosaic virus (CMV) in spinach (Spinacia oleracea L.)

ObjectiveCucumber mosaic virus (CMV) is a significant pathogen causing quality loss in spinach. Although host genetic resistance is the primary method of managing CMV infection in this crop, CMV resistance genes are not widely utilized in spinach breeding programs as the genetic and molecular mechanisms underlying resistance are not yet fully understood. CMV infections were therefore studied in different lines of spinach plants, and their progeny, to develop a model of the genetic basis of CMV resistance.ResultsVisual observations and RT-PCR assays revealed that three monoecious lines (03-258, 03-263, and 03-336) were susceptible to CMV, while three traditional resistant cultivars and a near-isogenic line (NIL-M) exhibited resistance. A dioecious line (03–009) consisted of susceptible and resistant plants. Notably, resistant plants did not exhibit the lesions typical of the hypersensitive response. Genetic analysis of progeny from the cross NIL-M × 03-336 indicated that a single dominant allele (designated SRCm1, standing for Spinach Resistance to CMV 1) controlled CMV resistance; analysis of sib-cross progeny populations derived from line 03–009 supported this conclusion. These results offer a valuable model for CMV resistance in spinach and will enhance future breeding programs.

Transcriptome Analysis of Rice Near-Isogenic Lines Inoculated with Two Strains of Xanthomonas oryzae pv. oryzae, AH28 and PXO99A.

Rice bacterial blight (BB), caused by Xanthomonas oryzae pv. oryzae (Xoo), is a major threat to rice production and food security. Exploring new resistance genes and developing varieties with broad-spectrum and high resistance has been a key focus in rice disease resistance research. In a preliminary study, rice cultivar Fan3, exhibiting high resistance to PXO99A and susceptibility to AH28, was developed by enhancing the resistance of Yuehesimiao (YHSM) to BB. This study performed a transcriptome analysis on the leaves of Fan3 and YHSM following inoculation with Xoo strains AH28 and PXO99A. The analysis revealed significant differential expression of 14,084 genes. Among the transcription factor (TF) families identified, bHLH, WRKY, and ERF were prominent, with notable differences in the expression of OsWRKY62, OsWRKY76, and OsbHLH6 across samples. Over 100 genes were directly linked to disease resistance, including nearly 30 NBS-LRR family genes. Additionally, 11 SWEET family protein genes, over 750 protein kinase genes, 63 peroxidase genes, and eight phenylalanine aminolysase genes were detected. Gene ontology (GO) analysis showed significant enrichment in pathways related to defense response to bacteria and oxidative stress response. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis indicated that differentially expressed genes (DEGs) were enriched in phenylpropanoid biosynthesis and diterpenoid biosynthesis pathways. Gene expression results from qRT-PCR were consistent with those from RNA-Seq, underscoring the reliability of the findings. Candidate genes identified in this study that may be resistant to BB, such as NBS-LRR family genes LOC_Os11g11960 and LOC_Os11g12350, SWEET family genes LOC_Os01g50460 and LOC_Os01g12130, and protein kinase-expressing genes LOC_Os01g66860 and LOC_Os02g57700, will provide a theoretical basis for further experiments. These results suggest that the immune response of rice to the two strains may be more concentrated in the early stage, and there are more up-regulated genes in the immune response of the high-resistant to PXO99A and medium-resistant to AH28, respectively, compared with the highly susceptible rice. This study offers a foundation for further research on resistance genes and the molecular mechanisms in Fan3 and YHSM.

Fine mapping of QGPC.caas-7AL for grain protein content in bread wheat.

A major stable QTL, QGPC.caas-7AL, for grain protein content of wheat, was narrowed down to a 1.82-Mb inter on chromosome 7AL, and four candidate genes were predicated. Wheat grain protein content (GPC) is important for end-use quality. Identification of genetic loci for GPC is helpful to create new varieties with good processing quality and nutrients. Zhongmai 578 (ZM578) and Jimai 22 (JM22) are two elite wheat varieties with different contents of GPC. In the present study, 262 recombinant inbred lines (RILs) derived from a cross between ZM578 and JM22 were used to map the GPC with high-density wheat Illumina iSelect 50K single-nucleotide polymorphism (SNP) array. Seven quantitative trait loci (QTLs) were identified for GPC on chromosomes 3AS, 3AL, 3BS, 4AL, 5BS, 5DL and 7AL by inclusive composite interval mapping, designated as QGPC.caas-3AS, QGPC.caas-3AL, QGPC.caas-3BS, QGPC.caas-4AL, QGPC.caas-5BS, QGPC.caas-5DL and QGPC.caas-7AL, respectively. Among these, alleles for increasing GPC at QGPC.caas-3AS, QGPC.caas-3BS, QGPC.caas-4AL and QGPC.caas-7AL loci were contributed by ZM578, whereas those at the other three loci were from JM22. The stable QTL QGPC.caas-7AL was fine mapped to a 1.82-Mb physical interval using secondary populations from six heterozygous recombinant plants obtained by selfing a residual RIL. Four genes were predicted as candidates of QGPC.caas-7AL based on sequence polymorphism and expression patterns. The near-isogenic lines (NILs) with the favorable allele at the QGPC.caas-7AL locus increased Farinograph stability time, Extensograph extension area, extensibility and maximum resistance by 19.6%, 6.3%, 6.0% and 20.3%, respectively. Kompetitive allele-specific PCR (KASP) marker for QGPC.caas-7AL was developed and validated in a diverse panel of 166 Chinese wheat cultivars. These results provide further insight into the genetic basis of GPC, and the fine-mapped QGPC.caas-7AL will be an attractive target for map-based cloning and marker-assisted selection in wheat breeding programs.

Identification of powdery mildew resistance quantitative trait loci in melon and development of resistant near-isogenic lines through marker-assisted backcrossing

BackgroundMelon (Cucumis melo L.), an important cucurbit crop, faces production limitations due to powdery mildew (PM). Developing resistant varieties offers a sustainable, genetics-based alternative to chemical treatments. Therefore, identifying PM resistance quantitative trait loci (QTL) and creating trait-associated markers are essential for efficient melon PM resistance improvement through marker-assisted backcrossing (MABC).ResultsThree F2 populations, A6, B2, and C4, were generated for QTL mapping of PM resistance. Major QTL were identified on chromosome 2 in A6, chromosome 5 in B2, and chromosomes 5 and 12 in C4. A series of TaqMan® assays targeting regions on chromosomes 2, 5, and 12 were developed and validated for foreground and recombinant selection, complemented by the double digest restriction-site associated DNA genotyping system to evaluate the recurrent parent genome recovery. Three MABC programs using resistant donor parents from A6 and C4 crossed with elite susceptible recurrent parents with green and orange fruit flesh were implemented. After two to three cycles of MABC, individual QTL was successfully introgressed into elite genetic backgrounds, giving six PM resistance lines in each green- and orange-fleshed background. PM inoculation on the twelve near-isogenic lines confirmed their resistance to PM.ConclusionsWe have identified major PM resistance QTL for melon on chromosomes 2, 5, and 12 and have introgressed individual QTL to elite genetic backgrounds using MABC in three and a half years. This study demonstrates the power of combining high-throughput genotyping with breeding efforts and showcases the efficiency of molecular breeding.

Comparative RNA-seq analysis reveals transposable element-mediated transcriptional reprogramming under phosphorus-starvation stress in rice (Oryza sativa L.)

Phosphorus (P) deficiency hinders crop productivity of 50 % of the rice grown in Asia, Africa, and South America. About 90 % of the phosphate in fertilizers applied to the crops gets fixed in the soil, reducing its availability to plants. This necessitates increased use of phosphatic fertilizers leading to higher cost of cultivation and environmental pollution. Although molecular mechanisms of P-deficiency tolerance in rice are being deciphered, the role of transposable elements (TEs) in transcriptional reprogramming under P-starvation/deficiency stress has not yet been reported. To investigate the role of Pup1 QTL in controlling TE-mediated reprogramming of gene expression, a pair of contrasting rice [Pusa-44 and its Near-Isogenic Line (NIL)-23] genotypes were grown hydroponically under control and stressed (0 ppm Pi) conditions. Comparative RNA-seq analysis of root and shoot tissues from 45-day-old plants of the rice genotypes revealed TE-mediated transcriptional reprogramming affecting biological processes and cellular components. Significantly up-regulated expression of several TEs under P-starvation stress, controlled by Pup1 QTL, particularly in shoots of NIL-23 indicates their crucial role in P homeostasis. Moreover, comparative physio-biochemical analyses confirmed the stress tolerance of NIL-23. Several biological processes including DNA replication/repair, metabolism, signaling, and phosphorylation were modulated through differential (mainly up-regulated) expression of TEs (controlled by Pup1 QTL) in shoots of NIL-23 under P-starvation. To the best of our knowledge, this is a pioneer study on the role of TEs in reprogramming biological processes/molecular functions/cellular components involved in P-use efficiency in rice under stress. The findings advance our understanding of the functions of Pup1 to improve the P-use efficiency/productivity of rice in P-deficient soils.

RETRACTION: Genotypic and phenotypic characterization of a large, diverse population of maize near-isogenic lines.

L. Morales, A. C. Repka, K. L. Swarts, W. C. Stafstrom, Y. He, S. M. Sermons, Q. Yang, L. O. Lopez-Zuniga, E.Rucker, W. E. Thomason, R. J. Nelson, P. J. Balint-Kurti. "Genotypic and phenotypic characterization of a large, diverse population of maize near-isogenic lines," The Plant Journal 103, no. 3 (2020): 1246-1255. https://doi.org/10.1111/tpj.14787. The above article, published online on 29 April 2020, in Wiley Online Library (wileyonlinelibrary.com), has been retracted by agreement between the authors; the journal Editor-in-Chief, Katherine Denby; Society for Experimental Biology (SEB); and John Wiley & Sons Ltd. The retraction has been agreed following a report submitted by the authors indicating that the imputation of genotypes outlined in the article caused errors in the reported positions of introgressions in the lines. The authors also declared that some of the pedigree information is inaccurate. As a result, all authors agree that the paper must be retracted. The authors have stated their intention to publish a corrected report with improved analyses of introgression positions.

Identification and molecular marker development for peel color gene in melon (Cucumis melo L.)

Peel color is an important appearance quality of melons that greatly affects consumer preferences. In this study, a near-isogenic line NIL-G (dark green peel) was generated from B8 (grey-green peel) and B15 (white peel). The F2 population constructed by crossing NIL-G and B15 was used to study the inheritance pattern of peel color, and bulked-segregant analysis sequencing (BSA-seq) was employed to identify the interval in which the target gene was located. Genetic analysis results showed that the dark green peel trait at maturity is controlled by a dominant gene. BSA-seq and molecular markers were used to localize the candidate gene in a 263.7 kb interval of chromosome 4, which contained the CmAPRR2 gene with known functions. Moreover, allelic sequence analysis revealed four SNP variations of the CmAPRR2 gene in B15, of which SNP.G614331A was located at the junction of the 6th exon and 6th intron. The G-to-A mutation caused alternative splicing of the transcript of CmAPRR2 in B15, generating two transcripts (CmAPRR2-A and CmAPRR2-B) with premature termination codons. Furthermore, the Kompetitive Allele Specific PCR (KASP) marker, APRR2-G/A, was developed based on this SNP and shown to co-segregate with the peel color phenotype in the F2 population. Compared to white-peel B15, the expression level of CmAPRR2 in dark green peel NIL-G was higher at each growth stage. Therefore, CmAPRR2 may be the key gene controlling the fruit color of melons. Overall, this study identified a novel allelic variant of CmAPRR2 that leads to white peel formation in mature melons. The study also provides a theoretical basis for further research on the gene regulatory mechanism of melon peel colors and may promote the future use of molecular marker-assisted selection to modify melon peel colors.

Structural, molecular, and physicochemical properties of starch in high-amylose durum wheat lines

There is growing interest in the development of high-amylose cereals such as wheat due to their functional properties and nutritional value in foods. The increase in amylose content is associated with significant changes in the physicochemical, molecular, and structural characteristics of wheat starch which could affect its processing quality. The present study aimed to analyze the physicochemical, molecular, and structural characteristics of high-amylose starch from three durum wheat near isogenic lines (NILs), each obtained using three different recurrent parents. Morphology, granule size distribution, pasting and thermal properties, X-ray diffraction, and structural features were determined. The NILs showed the highest amylose content (65.3 and 69.8%), granules with an elongated shape, size between the wild-type and the mutant, restricted swelling in the pasting profile, the highest average and final gelatinization temperature, and B-polymorphism. The starches from the NILs showed the highest molar mass of amylopectin (AP) (7.0–7.6 × 107 Da) and the lowest amylose (AM) (1.1–1.8 × 106 Da), an issue associated with the biosynthesis of both components. The study on the debranched sample by high-performance size-exclusion chromatography showed that the high-amylose starch had the highest content of long B2 and B3 chains of AP (23.9–26.6%), which was corroborated with the high-performance anion-exchange chromatography analysis where the chains with degree of polymerization >37 had the highest content (18.3–19.1%). The PC1 is highly associated with the MwAM and the AM fraction in the debranched starch analyzed by HPSEC. Characterizing high-amylose durum wheat starch provides information to determine the structure-function relationship and design strategies to produce crosses with structural characteristics of AM and AP for specific applications.

Bell pepper endornavirus alters green peach aphid (Hemiptera: Aphididae) host choice and population dynamics.

Bell pepper endornavirus (BPEV) Alphaendornavirus capsici (Endornaviridae) is an RNA virus that infects many pepper (Capsicum annuum) horticultural types and is seed transmitted. BPEV does not cause apparent symptoms and is found at every plant developmental stage. During the domestication of bell pepper, plant breeders, unaware of the existence of endornaviruses in the germplasm, selected endornavirus-infected genotypes. This could be an indication that the presence of endornaviruses in this crop is beneficial. Among the possible beneficial effects that endornaviruses may provide to their host could include tolerance or resistance to biotic and abiotic agents and, therefore, may have evolved a symbiotic relationship with their hosts. With this in mind, we set out to determine host preference, host suitability, and population dynamics of green peach aphid Myzus persicae (Sulzer) on BPEV-infected and virus-free bell pepper near-isogenic lines. During choice bioassay experiments, we observed that a higher proportion of M. persicae adults settled on BPEV noninfected leaves as compared to BPEV-infected leaves. Life table analysis revealed that M. persicae performed less well on BPEV-infected leaf tissues, with reductions in longevity, progeny, and intrinsic rate of increase. These results indicate BPEV is beneficial to its host, protecting against an important generalist pest.

CmSN Regulates Fruit Skin Netting Formation in Melon

Melon (Cucumis melo) includes more than ten botanical groups, many of which feature netting ornamentation on the surface of mature fruit. Ripe melons display a netted skin that signifies their ripeness and readiness for consumption. Previously, we identified SKIN NETTING (CmSN), which encodes an EamA-like transporter family protein, as the candidate gene controlling fruit skin netting formation in melon, while its biological functions remain unclear. In this study, we demonstrated that the expression of the CmSN gene was considerably lower in netted melons compared to smooth-skinned melons, indicating a negative correlation between CmSN expression and netting formation. Subsequently, we employed transient overexpression and virus-induced gene silencing (VIGS) experiments to explore the role of CmSN gene during fruit development. Overexpression of the CmSN gene inhibited netting development, whereas silencing it promoted netting formation. Using heterologous transformation in tomato, we further confirmed the effect of the CmSN gene on rind texture and toughness, as these tomatoes exhibited rougher and tougher skins. Analysis with near-isogenic lines (NILs) revealed that CmSN gene-bearing fruits (NIL_CmSN) possessed significantly harder rinds than the control smooth-skinned variety HB42, underscoring the role of CmSN in enhancing rind protection. Together, our research offers essential insights into the netting formation and genetic improvement of melon fruits.

The Tetratricopeptide repeat protein TaTPR-B1 regulates spike compactness in bread wheat.

Spike compactness (SC) is strongly associated with wheat (Triticum aestivum L.) grain yield. In this study, we conducted a quantitative trait locus (QTL) analysis using a doubled haploid (DH) population derived from a cross between two common wheat varieties with contrasting spike morphology, revealing 16 stable QTLs associated with SC. The effect of a major QTL, QSc.cau-6B.1, was validated in 231 F7 recombinant inbred lines (RILs) derived from the same cross as the DH population. Using two residual heterozygous lines (RHLs), we delimited QSc.cau-6B.1 to an approximately 0.5-Mbp physical interval containing four high-confidence genes. The tetratricopeptide repeat-TraesCS6B03G1214400 (TaTPR-B1) was the priority candidate gene according to sequence and expression variations between near-isogenic lines. Accordingly, TaTPR-B1 knockout in the common wheat variety 'CB037' significantly increased SC compared to the wild type (WT). Conversely, TaTPR-B1 overexpression in the common wheat variety 'Fielder' significantly decreased SC compared to the WT. Moreover, we developed a PCR-based marker targeting the 32-bp insertion/deletion (InDel) between the two TaTPR-B1 alleles, which could be practical and valuable in modern wheat breeding programs for diagnostic purposes. Collectively, these findings provide insight into the genetic basis of SC in common wheat and present a valuable target with a breeder-friendly diagnostic marker for gene pyramid breeding.

Disruption of Energy Metabolism and Reactive Oxygen Species Homeostasis in Honglian Type-Cytoplasmic Male Sterility (HL-CMS) Rice Pollen

Honglian type-cytoplasmic male sterility (HL-CMS) is caused by the inter-communication between the nucleus and mitochondria. However, the mechanisms by which sterility genes regulate metabolic alterations and changes in mitochondrial morphology in the pollen of HL-CMS remains unclear. In this study, we compared the morphological differences between the pollen of the male sterile line YA and the near-isogenic line NIL-Rf6 using hematoxylin and eosin staining as well as 4ʹ,6-diamidino- 2-phenylindole (DAPI) staining. HL-CMS is characterized by gametophytic sterility, where the aborted pollen grains are empty and the tapetal layer remains intact. Transmission electron microscopy was employed to observe mitochondrial morphological changes at the microspore stage, revealing that significant mitochondrial alternations, characterized by the formation of 'large spherical mitochondria', occurred at the binucleate stage in the YA line. Additionally, metabolomics analysis revealed decreased levels of metabolites associated with the carbohydrate and flavonoid pathways. Notably, the decrease in flavonoids was found to contribute to an elevation in reactive oxygen species (ROS) levels. Therefore, we propose a model in which rice fertility is modulated by the levels of pollen carbohydrates and flavonoid metabolites, with impaired mitochondrial energy production and reduced flavonoid biosynthesis being the main causes of ROS accumulation and pollen abortion in rice.

Phosphorus uptake 1 (Pup1) QTL performs major regulatory functions under phosphorus starvation/deficiency stress in rice (Oryza sativa L.)

Phosphorus (P) is an essential macronutrient required for respiration, photosynthesis/carbohydrate metabolism, redox homeostasis, signaling, and synthesis/function of nucleic acids, cellular membranes, enzymes, etc. To cope with P-deficiency, plants reprogram gene expression for necessary alterations in metabolic/signaling pathways. To attain P homeostasis, plants readjust metabolism through transcriptional, post-transcriptional, and/or post-translational machinery involving biochemical, physiological, genomic, epigenomic, proteomic, and metabolomic functions. However, the underlying molecular mechanisms/pathways/genes for P-deficiency tolerance in crop plants remain elusive. To decipher the mechanisms/pathways adopted by rice under P-starvation/deficiency stress, a pair of contrasting rice [Pusa-44 (high-yielding, P-deficiency sensitive) and its near-isogenic line (NIL)-23, P-deficiency tolerant) for Pup1 QTL] genotype was used for comparative analyses. Omics analyses of shoot and root tissues from 45-day-old plants grown hydroponically in P-sufficient (16 ppm Pi), P-deficient (4 ppm Pi) or P-starved (0 ppm Pi) medium revealed important roles of P transporters, transcription factors (TFs), Transposable elements (TEs), auxin-responsive proteins, cell wall modulation, fatty acid metabolism, and chromatin architecture/epigenetic modifications in making NIL-23 tolerant to stress. The proteins involved in photosynthesis, sucrose-/starch-/energy-metabolism, transcription factors, and phytohormone signaling were observed to be differentially expressed in NIL-23. Since only a few coding genes are located on the QTL, modulations in morpho-physio-biochemical and molecular parameters under P-starvation/deficiency stress were attributed to the regulatory functions of Pup1 through TFs, TEs, epigenetic/chromatin architectural changes, etc. introgressed in Pusa-44 genetic background. Thus, the study provides new insights into molecular/regulatory functions of Pup1 under P-starvation/deficiency stress in rice, which might be useful to improve P-use efficiency in rice for better productivity in P-deficient soils.

Management and rhizosphere microbial associations modulate genetic-driven nitrogen fate

The interplay between plant genotype and nutrient management affects rhizodeposition, which in turn modulates the rhizosphere-microbiome and microbe-mediated functions. Substituting mineral nitrogen (N) with an N-fixing inoculant reduces reliance on N fertilizer while supplying N to crops. We evaluated the effectiveness of integrating maize near-isogenic lines (NIL 1 and NIL 2) with the biological nitrification inhibition (BNI) trait into management practices aimed at optimizing N provisioning. Management strategies included mineral N inputs (0 and 67 kg ha⁻¹) with and without an N-fixing inoculant. Our approach synthesized insights from amplicon sequencing data and evaluated nitrification rates, rhizosphere N content, maize N uptake, and N use efficiency (NUE). Genotypes and management structured prokaryotic communities, while the developmental stages of genotypes further refined both fungal and prokaryotic communities. The N-fixing inoculant increased N availability, triggering the BNI capacity without increasing the nitrification rate. This was reflected in lower NO₃⁻ and higher NH₄⁺ levels in BNI-NIL leachate compared to B73, suggesting improved N retention. NIL2, characterized by distinct fungal biomarkers, exhibited higher N content (72.3 kg ha⁻¹) and superior NUE compared to NIL1 (65.0 kg ha⁻¹). NIL2’s enhanced N uptake was associated with a robust microbial network, featuring Archangium (prokaryote) and Trichoderma (eukaryote) as keystone taxa. Notably, Archangium was linked to rhizosphere N dynamics Synergizing BNI with diazotroph inoculants reduces N fertilizer reliance and increases maize N supply for sustainable agroecosystems.

Identification of molecular markers linked to leaf rust resistance gene Lr13 in wheat

Aim. Identification of codominant molecular markers closely linked to the Lr13 leaf rust resistance gene. Methods. PCR with detection in a polyacrylamide gel. Hybridological analysis. Phytopathology evaluation. Statistical analysis. Results. PCR analysis of varieties carrying the Lr13 gene and near-isogenic lines of the Thatcher variety with genes for resistance to leaf rust Lr13 (TcLr13), Lr16 (TcLr16), Lr23 (TcLr23) and the F2 population TcLr13 x TcLr34 by the microsatellite loci Xbarc13, Xbarc55, Xgwm148, Xwmc261, Xgwm271, Xwmc272, Xwmc344, Xbarc361, Xgwm410, Xwmc474, Xwmc477 and Xgwm630 was performed. To test the two microsatellite loci Xgwm148 and Xwmc344 as candidate markers for the Lr13 gene, we used the F2 population, which was obtained from crossing two near-isogenic lines of the Thatcher variety with leaf rust resistance genes Lr13 (TcLr13) and Lr34 (TcLr34). Conclusions. When comparing the genotyping data of F2 hybrids and the phenotypic manifestation of resistance to leaf rust, it was determined that the microsatellite locus Xwmc344 is closely linked to the Lr13 gene.

Do anthocyanins affect acrylamide formation in wholemeal bakery products? A study on near-isogenic wheat lines

Near-isogenic wheat lines (NILs) derived from the variety Novosibirskaya 67, differing in grain color, were used to analyze the content of acrylamide in bakery products, as well as its precursors (glucose, fructose, asparagine) and anthocyanins, in key steps of bread preparation. The total anthocyanin content of the flour averaged 2.49 and 30.8 mg/kg for the purple and blue NILs, respectively. However, purple NILs showed higher anthocyanin stability during processing compared to blue NILs, with the total content changing to 113 and 37% of the initial value in the flour. The acrylamide content of the product ranged from 83.3 to 132 μg/kg and was moderately positively correlated with the dough's asparagine (r = 0.529) and fructose (r = 0.441) content. The non-significant negative correlation found between acrylamide and anthocyanins (r = −0.268) indicates that anthocyanins do not significantly affect the formation of this process contaminant and therefore, their presence in wheat grain does not pose a health risk to consumers.