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Assessment of phenotypic diversity and determination of cytoplasmic male sterility types in some commercial forage maize hybrids cultivated in Iran using morphological–agronomic traits and PCR markers

Document Type : Research Paper

Authors

1 Department of Agronomy and Plant Breeding Sciences, Faculty of Agricultural Technology (College of Aburaihan), University of Tehran, Tehran, Iran

2 Department of Agronomy Sciences and Plant Breeding, Faculty of Agricultural Technology of Aburaihan, University of Tehran, Pakdasht, Iran

3 Department of Agronomy and Plant Breeding, Islamic Azad University, Khorasgan Branch, Isfahan, Iran

4 Department of Agronomy Sciences and Plant Breeding, Faculty of Agricultural Technology of Aburaihan, University of Tehran, Pakdasht, Iran.

10.22059/jci.2026.409816.2972

Abstract

Objective: Commercial forage maize hybrids widely cultivated in Iran may serve not only as important materials for forage production but also as potential germplasm resources for identifying cytoplasmic male sterility (CMS) sources and developing breeding populations for future hybrid breeding programs. This study aimed to evaluate the phenotypic diversity and cytoplasmic status of seven commercial forage maize hybrids cultivated in Iran and to determine their CMS type using CMS-specific PCR markers. In addition, the agreement between molecular identification and phenotypic observations, including tassel characteristics, pollen production, and self-pollination response, was investigated.
Methods: The experiment was conducted in a randomized complete block design (RCBD) with three replications. Agronomic, morphological, phenological, and yield-related traits were recorded, including fresh forage yield, dry matter yield, plant height, stem diameter, leaf length, leaf width, number of leaves, days to tasseling, days to silking, anthesis–silking interval (ASI), days to milk-stage maturity, ear length, ear diameter, number of kernel rows per ear, and number of kernels per row. Analysis of variance, mean comparison, correlation analysis, path analysis, cluster analysis, factor analysis, principal component analysis (PCA), and estimation of genetic parameters were performed. CMS-specific PCR markers were used to identify cytoplasmic types, and molecular results were compared with field observations and self-pollination tests.
Results: Significant differences (P≤0.01) were observed among the hybrids for most measured traits, indicating substantial phenotypic diversity within the evaluated germplasm. Mean comparison identified hybrid P5 (Moghan 704) as the superior genotype for several agronomic and yield-related traits. Correlation and path analyses revealed that dry matter yield exerted the greatest direct effect on forage yield, whereas plant height and the number of leaves above the ear influenced forage production mainly through indirect effects. Cluster analysis classified the hybrids into three phenotypic groups, with hybrid P7 forming a distinct cluster because of its unique morphological and phenological characteristics. The Kaiser–Meyer–Olkin (KMO) value of 0.63 confirmed the adequacy of the data for factor analysis, and four factors explained 85.88% of the total phenotypic variation. Principal component analysis further summarized trait variability within the first two principal components. Broad-sense heritability estimates were high for most traits, suggesting that a large proportion of the observed variation was genetically controlled. Molecular analysis revealed that hybrids P2, P4, P5, P6, and P7 possessed CMS-C cytoplasm, whereas P1 and P3 carried normal (N) cytoplasm. These molecular findings were consistent with tassel phenotype, pollen production, and self-pollination responses observed in both F₁ and F₂ generations.
Conclusion: The evaluated commercial forage maize hybrids exhibited considerable variation in agronomic, morphological, phenological, and cytoplasmic traits. The identification of CMS-C cytoplasm in several hybrids provides valuable preliminary information for the selection of potential CMS sources in maize breeding programs. Nevertheless, the development of male-sterile lines, maintainer lines, and new hybrid parents requires further studies involving segregating populations, inbred line development, CMS stability assessment, and combining ability evaluation across subsequent generations.

Keywords

References
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Journal of Crops Improvement
Volume 28, Issue 2
June 2026
Pages 169-197
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