Search Results (102)

(1) Background: Backcross (BC) breeding is a key technology of crop improvement, yet its efficiency largely depends on the precise assessment of the genetic background recovery. Conventional molecular marker-assisted techniques suffer from inadequate genomic coverage or an inability to resolve true chromosomal structure. (2) Methods: To address major issues in maize BC breeding, we devised a G2H block-scanning strategy. This approach converts high-density point markers into haplotype blocks, enabling precise evaluation of the genetic background in backcross progenies. A key innovation is the CFDI, which quantifies the distribution of unrecovered fragments, allowing for visual tracking of chromosomal recombination and identification of ideal individuals with both a high genetic background recovery rate and few small fragments retention. (3) Results: We validated the accuracy and effectiveness of the G2H strategy across multiple backcross generations. Through enabling a precise “point-to-line-to-area” panoramic assessment of genetic background, G2H provides a powerful tool for developing ideal breeding materials with pure genetic background and minimized linkage drag. (4) Conclusions: Notably, this strategy significantly shortens the breeding cycle by 2–3 generations compared to conventional background assessment methods, thereby accelerating precision molecular design breeding in crops. Full article

Doubled haploid (DH) technology accelerates maize breeding by generating completely homozygous lines within two generations, but its efficiency depends on reliable haploid identification markers. The Purple plant 1 (Pl-1) root pigmentation marker has emerged as a promising alternative to R1-Navajo (R1-nj), which suffers from frequent suppression in tropical germplasm. This study evaluated Pl-1 expression and seedling vigor in 298 diverse Thai maize genotypes across four market classes under controlled growth chamber conditions (24 ± 1 °C, 85–90% humidity, and standardized lighting), followed by testcross analysis with 89 representative genotypes crossed with BHI306 to distinguish between allelic absence and epistatic suppression mechanisms. Complete absence of Pl-1 expression was observed in 99.3% of Thai genotypes, contrasting with consistent expression in the temperate-derived control (BHI306). Testcross F₁ progeny from 89 Thai × BHI306 crosses exhibited intermediate expression levels (1.57–2.05) across all market classes, confirming allelic absence rather than suppressor-mediated inhibition. Substantial genetic diversity was detected in seedling vigor traits independent of Pl-1 status, with root length varying 43-fold and fresh weight 20-fold, showing highly significant genotypic effects. The uniform lack of Pl-1 expression across Thai germplasm eliminates background interference, highlighting its utility as a complementary marker when introgressed from inducer lines. These findings establish the genetic foundation for implementing optimized DH breeding strategies in tropical maize through marker-assisted backcrossing approaches. Full article

Xiangwanxian 13 (XWX13), an elite fragrant indica rice, is highly susceptible to rice blast and accumulates cadmium (Cd) in grain above the food safety limit in Cd-contaminated paddies, severely constraining its commercial use. Despite these shortcomings, the variety is widely grown for its high yield and superior grain quality. To overcome these limitations, we conducted marker-assisted backcrossing (MABC) complemented by genome-wide background selection. Four major genes, namely Pi1, Pi2, OsHMA3, and OsNramp5, were precisely introduced into XWX13. Two preferable BC₃MF₅ improved lines iXWX13-1 (stacking Pi1 + Pi2 + OsHMA3) and iXWX13-2 (stacking Pi1 + Pi2 + OsNramp5) were obtained with genomic background recovery rates of 94.44% and 94.63%, evaluated by using the RICE 1K SNP array, respectively. Seedling resistance spectrum assays demonstrated more than a 97% blast resistance rate against 39 Magnaporthe oryzae isolates, and both lines showed enhanced leaf and panicle neck blast resistance in natural nurseries. Multi-site field trials revealed grain Cd concentrations of 0.009–0.077 mg kg⁻¹ in iXWX13-2, 90.98–98.87% lower than those in XWX13. Importantly, yield, major agronomic traits, and grain quality remained indistinguishable from the original variety. This study provides the first demonstration that MABC coupled with SNP array background selection can simultaneously enhance blast resistance and reduce grain Cd in XWX13 without yield or quality penalties, offering a robust strategy for pyramiding multiple desirable genes into elite cultivars. Full article

The fungal pathogen Puccinia striiformis f. sp. tritici, which causes yellow rust disease, poses a significant economic threat to wheat production not only in Uzbekistan but also globally, leading to substantial reductions in grain yield. This study aimed to develop yellow rust-resistance wheat lines by introgressing Yr10 and Yr15 genes into high-yielding cultivar Grom using the marker-assisted backcrossing (MABC) method. Grom was crossed with donor genotypes Yr10/6*Avocet S and Yr15/6*Avocet S, resulting in the development of F₁ generations. In the following years, the F₁ hybrids were advanced to the BC₂F₁ and BC₂F₂ generations using the MABC approach. Foreground and background selection using microsatellite markers (Xpsp3000 and Barc008) were employed to identify homozygous Yr10- and Yr15-containing genotypes. The resulting BC₂F₂ lines, designated as Grom-Yr10 and Grom-Yr15, retained key agronomic traits of the recurrent parent cv. Grom, such as spike length (13.0–11.9 cm) and spike weight (3.23–2.92 g). Under artificial infection conditions, the selected lines showed complete resistance to yellow rust (infection type 0). The most promising BC₂F₂ plants were subsequently advanced to homozygous BC₂F₃ lines harboring the introgressed resistance genes through marker-assisted selection. This study demonstrates the effectiveness of integrating molecular marker-assisted selection with conventional breeding methods to enhance disease resistance while preserving high-yielding traits. The newly developed lines offer valuable material for future wheat improvement and contribute to sustainable agriculture and food security. Full article

Bacterial blight (BB) caused by Xanthomonas oryzae pv. oryzae (Xoo) is a major threat to global rice productivity. Although hybrid rice breeding has significantly enhanced yields, persistent genetic vulnerabilities within restorer lines continue to compromise BB resistance. This study addresses this challenge by implementing functional marker-assisted selection (FMAS) to pyramid two broad-spectrum resistance (R) genes, Xa21 and Xa23, into the elite, yet BB-susceptible, restorer line K608R. To enable precise Xa23 genotyping, we developed a novel three-primer functional marker (FM) system (IB23/CB23/IR23). This system complements the established U1/I2 markers used for Xa21. This recombination-independent FMAS platform facilitates simultaneous, high-precision tracking of both homozygous and heterozygous alleles, thereby effectively circumventing the linkage drag limitations typical of conventional markers. Through six generations of marker-assisted backcrossing followed by intercrossing, we generated K608R2123 pyramided lines harboring both R genes in homozygous states, achieving a recurrent parent genome recovery rate of 96.93%, as determined by single nucleotide polymorphism (SNP) chip analysis. The pyramided lines exhibited enhanced resistance against six virulent Xoo pathogenic races while retaining parental yield performance across key agronomic traits. Our FMAS strategy overcomes the historical trade-off between broad-spectrum resistance and the preservation of elite phenotypes, with the developed lines exhibiting resistance coverage complementary to that of both introgressed R genes. This integrated approach provides breeders with a reliable molecular tool to accelerate the development of high-yielding, disease-resistant varieties, demonstrating significant potential for practical deployment in rice improvement programs. The K608R2123 germplasm represents a dual-purpose resource suitable for both commercial hybrid seed production and marker-assisted breeding programs, and it confers synergistic resistance against diverse Xoo races, thereby providing a pivotal breeding resource for sustainable BB control in epidemic regions. Full article

Morkhor 60-3 is an upland rice variety primarily cultivated in northeastern Thailand. This glutinous rice is valued for its adaptability and rich aroma but remains susceptible to significant diseases, particularly blast and bacterial blight. Using resistant varieties represents the most cost-effective approach to address this limitation. This study incorporated the QTLs/genetic markers qBl1, qBl2, and xa5 from Morkhor 60-1 through marker-assisted backcrossing. From the BC₁F₃ population, ten lines were selected based on their parentage and evaluated for blast resistance using a spray inoculation method with 12 isolates of Pyricularia oryzae, and for bacterial blight (BB) resistance using a leaf-clipping method with nine isolates of Xanthomonas oryzae pv. oryzae. Broad-spectrum resistance (BSR) was also assessed in the lines for both diseases. Subsequently, BC₁F₄ lines were evaluated for field performance, including agronomic traits and aroma. Results identified three superior lines, BC₁F₄ 22-7-140-4, BC₁F₄ 22-7-322-5, and BC₁F₄ 22-7-311-9, that demonstrated resistance to both BB and blast pathogens with average BSR values of 0.61 and 1.00, 0.66 and 1.00, and 0.55 and 0.87, respectively. These lines also exhibited enhanced performance in flowering date, plant height, panicle number per plant, grain number per plant, and grain weight. These findings demonstrate the effectiveness of marker-assisted selection (MAS) for gene pyramiding in rice improvement. Full article

Background/Objectives: To overcome the limitations imposed by bacterial blight on widely adopted indica–japonica hybrid rice, this study employed molecular design breeding strategies to develop a resistant germplasm. Methods: Through conventional backcross breeding combined with molecular-marker-assisted selection, the Xa23-carrying material XR39 was hybridized with the wide-compatibility restorer line R5315 harboring the S5n gene. Progeny selection integrated evaluations of agronomic traits, disease resistance identification, and test-crossing with sterile lines. Results: Five wide-compatibility restorer lines simultaneously incorporating the Xa23 and S5n genes were successfully developed, demonstrating outstanding bacterial blight resistance and restoration ability. The selected hybrid combinations, A3/RP1, A1/RP4, and A4/RP4, exhibited yield increases of 2.6–8.6% compared to the control. Conclusions: This study not only established a novel germplasm for developing bacterial blight-resistant indica–japonica hybrid rice varieties, but also established a model for gene design breeding for rice improvement. Full article

The Yangtze River Delta (YRD) is one of the most important japonica rice planting areas in China. Balancing the resistance, yield, and quality has always been a core issue in rice breeding due to the negative correlation among these three factors, while the broad-spectrum blast resistance gene Piz is closely linked with Hd1, the major gene regulating days to heading (DTH), and a precise combination of their beneficial alleles plays a key role in synchronously improving blast resistance and the regional adaptability of japonica rice in YRD. In this study, using the backcross progeny population derived from backbone parent ZD9471 and W1063, two alleles of Hd1 were identified. Then, through molecular marker-assisted selection combined with Green Super Rice 40K (GSR40K) chip-based screening, six introgression lines (ILs) with two different alleles combinations of Hd1 and Pigm were obtained. An evaluation of the blast resistance, yield, and quality traits showed that compared with the recipient parent, the panicle blast resistance of ILs was significantly enhanced; the grain number per panicle increased consistently with the delaying of the growth period, leading to higher yield in the ILs; the grain quality were synchronously improved. Two representative lines with similar genetic backgrounds but a significantly different regional adaptability, exhibiting a good blast resistance, high yield, and prominent quality were approved and demonstrated promising application prospects. Full article

Bacterial leaf streak (BLS) is one of the internationally significant quarantine diseases in rice. Effectively utilizing BLS resistance genes from wild rice (Oryza rufipogon Griff.) to breed new varieties offers a fundamental solution for BLS control. This study focused on the fine mapping of the BLS resistance gene bls2 and the development of closely linked molecular markers for breeding BLS-resistant lines. Using a Guangxi common wild rice accession DY19 (carrying bls2) as the donor parent and the highly BLS-susceptible indica rice variety 9311 as the recipient parent, BLS-resistant rice lines were developed through multiple generations of backcrossing and selfing, incorporating molecular marker-assisted selection (MAS), single nucleotide polymorphism(SNP) chip genotyping, pathogen inoculation assays, and agronomic trait evaluation. The results showed that bls2 was delimited to a 113 kb interval between the molecular markers ID2 and ID5 on chromosome 2, with both markers exhibiting over 98% accuracy in detecting bls2. Four stable new lines carrying the bls2 segment were obtained in the BC₅F₄ generation. These four lines showed highly significant differences in BLS resistance compared with 9311, demonstrating moderate resistance or higher with average lesion lengths ranging from 0.69 to 1.26 cm. Importantly, no significant differences were observed between these resistant lines and 9311 in key agronomic traits, including plant height, number of effective panicles, panicle length, seed setting rate, grain length, grain width, length-to-width ratio, and 1000-grain weight. Collectively, two molecular markers closely linked to bls2 were developed, which can be effectively applied in MAS, and four new lines with significantly enhanced resistance to BLS and excellent agronomic traits were obtained. These findings provide technical support and core germplasm resources for BLS resistance breeding. Full article

The direct-seeded rice (DSR) system is poised to become the dominant rice cultivation method due to its advantages, including reduced water usage, less labor requirements, decreased greenhouse gas emissions, and improved adaptation to climate change. However, weeds, particularly jungle rice (Echinochloa colona), significantly hinder DSR and cause substantial yield losses. This study aimed to develop rice cultivars competitive against jungle rice through selective breeding, focusing on early seed germination (ESG) and seedling vigor (ESV). We utilized 181 early-backcross selective introgression breeding lines (EB-SILs) developed using Green Super Rice (GSR) technology by backcrossing Weed Tolerant Rice1 (WTR1) with three donor parents, Haoannong, Cheng Hui 448, and Y134. Using the tunable genotyping-by-sequencing (tGBS^®, Data2Bio Technologies, Ames, IA, USA) method, we identified 3971 common single nucleotide polymorphisms (SNPs) that facilitated the mapping of 19 novel quantitative trait loci (QTLs) associated with weed competitiveness—eight linked to ESG traits and eleven to ESV traits. Notably, all QTLs were novel except qRPH1, linked to relative plant height at 14 and 21 days after sowing. Key QTLs were located on chromosomes 2, 3, 5, 6, 8, 9, 10, and 12. Candidate genes identified within these QTLs are implicated in the plant’s response to various abiotic and biotic stresses. Our findings enhance the understanding of the genetic basis for ESG and ESV traits critical for weed competitiveness, supporting marker-assisted and genomic selection approaches for breeding improved rice varieties. Furthermore, this research lays the groundwork for employing gene expression, cloning, and CRISPR editing strategies to combat jungle rice, with potential applications for other weed species and contributing to effective integrated weed management in the DSR system. Full article

Maize is highly sensitive to water deficit but has high transpiration and biomass production, leading to a substantial water demand. Genetic engineering can overcome reproductive isolation and utilize drought-tolerant genes from distant species. Ammopiptanthus nanus is a relic of the Tertiary glaciation that can adapt to harsh environmental conditions. In our previous study, five maize homozygous T₈ lines overexpressing the AnVP1 gene from Ammopiptanthus nanus were generated and showed the enhancement of drought tolerance. However, the recipient inbred line Zh-1 was poor in yield and agronomic performance. In the present study, the AnVP1 gene was backcrossed from donor parent L10 (one of the five T₈ lines) into recurrent parent Chang 7-2 (one of the elite parents of the commercial hybrid Zhengdan 958). In total, 103 InDel markers were developed and used for assisted background selection. After two generations of foreground selection through glufosinate spraying, the detection of CP4 EPSP MAb1 strips, and the PCR amplification of the AnVP1 gene, along with the similarity of agronomic traits to the recurrent parent, and background selection assisted by these InDel markers, the transgenic AnVP1 gene became homozygous in the BC₂ lines. The average recovery rate of the genetic background of the recurrent parent reached 74.80% in the BC₁ population and 91.93% in the BC₂ population, respectively. The results of RT-PCR and RT-qPCR indicated the stable expression of the AnVP1 gene in the two ultimately selected BC₂F₃ lines, BC₂-36-12 and BC₂-5-15. The drought tolerance of these two BC₂F₃ lines were significantly improved compared to the recurrent parent Chang 7-2, as revealed by their wilting phenotype and survival rate of seedlings. This improvement was related to the enhancement of water-retention ability, as indicated by higher RWC and the reduction in damage, as shown by the decrease in REL, MDA, and H₂O₂ under drought stress. The result of field evaluation in two arid and semi-arid environments indicated that the drought tolerance of Chang 7-2 was significantly improved. This study suggests that the improved Chang 7-2 can be crossed with Zheng 58 to develop the transgenic commercial hybrid Zhengdan 958. Full article

Rice blast is one of the most important diseases of rice, causing significant economic losses to agricultural production. A new gene, Pigm-1, which is allelic to Pigm, was cloned from Shuangkang 77009 using map based cloning. However, it is unclear whether there is a difference in the resistance spectrum between Pigm and Pigm-1. In this study, using 195 rice blast isolates collected from different areas of the Fujian Province, the Pigm-1 and Pigm single gene lines were inoculated to test their resistance. There was only one blast fungus JL-37 that showed a differential response in Pigm and Pigm-1 single gene lines, while the remaining 194 showed no difference. To further explore the application range of Pigm-1, the resistant rice R20-4 containing Pigm-1 was used as the donor, and a sensitive sticky rice S19-118 was used as the receptor. The hybrid F₁ was first backcrossed with S19-118 using a molecular marker-assisted selection breeding method, and a strain containing the Pigm-1 gene was selected to continue to backcross with S19-118 until BC₃F₁. A new blast resistance rice material, Xiannuo 23, containing Pigm-1 was developed and confirmed by laboratory and field tests. This material can be broadly used for the future breeding of rice blast resistant cultivars to reduce the loss of rice production. Full article

With its controlled environment, protected cultivation is advantageous and effective for breeding programs. This distinct setting also guarantees that fresh vegetables meet high quality standards. The controlled environment allows for precise monitoring and tuning of breeding efforts, a critical factor in continuously improving the quality of fresh vegetable production. Classical breeding strategies include hybridization, pedigree selection, backcrossing, recombination, and marker-assisted breeding. However, advanced techniques like phenomics and genome editing are revolutionizing the field. These methods accelerate phenotyping and aid in identifying traits and genetic variants linked to quality characteristics. Modern biotechnological tools, specifically genetic engineering and gene editing methods like CRISPR/Cas, have enhanced a wide array of traits in numerous vegetable species. These technological advancements have the potential to effectively address challenges associated with stress resistance, product quality, and shelf-life, thereby presenting promising prospects for the advancement of agriculture. The protracted process of developing new vegetable cultivars with reduced physiological issues through contemporary techniques is an enduring endeavor. Full article

Traditional maize possesses low concentrations of provitamin-A and vitamin-E, leading to various health concerns. Mutant alleles of crtRB1 and vte4 that enhance β-carotene (provitamin-A) and α-tocopherol (vitamin-E), respectively, in maize kernels have been explored in several biofortification programs. For genetic improvement of these target nutrients, uniplex-PCR assays are routinely used in marker-assisted selection. However, due to back-to-back breeding seasons, the time required for genotyping individually for each target gene in large backcross populations becomes a constraint for advancing the generations. Additionally, multiple PCR assays for various genes increase the required costs and resources. Here, we aimed to develop a multiplex-PCR assay to simultaneously identify different allelic forms of crtRB1 and vte4 genes and validate them in a backcross-based segregating population. The PCR assay was carried out using newly developed primers for crtRB1 and a gene-specific primer for vte4. The uniplex-PCR assay was standardized for selected primer pairs in the BC₁F₁ population segregating for crtRB1 and vte4 genes. Subsequently, a multiplex-PCR assay for crtRB1 and vte4 genes was developed and employed for genotyping in the BC₁F₁ population. The assay differentiated among four possible genotypic classes, namely crtRB1⁺crtRB1/vte4⁺vte4, crtRB1crtRB1/vte4⁺vte4, crtRB1⁺crtRB1/vte4⁺vte4⁺, and crtRB1crtRB1/vte4⁺vte4⁺. This newly developed multiplex-PCR assay saved 41.7% of the cost and 35.6% of the time compared to two individual uniplex-PCR assays. The developed assay could accelerate maize nutritional quality breeding programs through rapid and cost-effective genotyping for the target genes. This is the first report of a multiplex-PCR assay specific to crtRB1 and vte4 genes for its use in genomics-assisted breeding in maize. Full article

Maize is the second most important staple food crop in the world after wheat. For maize hybrid seed production, a prominent step is detasseling in the female parent, which is laborious, time consuming, and increases the hybrid seed cost by 15 to 20 percent. Hence, to overcome this problem, exploitation of male sterility in maize crops gains special significance. In this direction, the research was conducted to transfer cytoplasmic male sterility (CMS-C) from a CMS donor (VL192114) into the female parents (CAL1514 and ZL153493) of heat- and drought-resilient maize hybrids (RCRMH-2 and RCRMH-3) by a marker-assisted backcross scheme. The present research used Diversity Array Technology (DArTag) and Kompetitive Allele Specific PCR (KASP)-based single-nucleotide polymorphic markers for background selection in backcross populations. Genome recovery percentage ranged from 64.25 to 72.70, 78.94 to 87.69 and 82.28 to 90.77 percent in the BC₁F₁, BC₂F₁ and BC₃F₁ population, respectively, in the CAL1514 population, while it was 63.47 to 73.55, 78.16 to 88.76 and 83.96 to 91.81 percent in the BC₁F₁, BC₂F₁ and BC₃F₁ population, respectively, in the ZL153493 population. When the near-isogenic CMS lines of both populations are compared for agro-morphological traits with their recurrent parents, the agronomic qualities of recurrent parents, as well as the attributes of distinctness, uniformity and stability, are shown. Therefore, male sterility-transferred, female lines of RCRMH-2 and RCRMH-3 maize hybrids can be used directly to produce maize hybrid seed without the need of the detasseling process. Full article