Efficiency enhancement and technique refinement in chickpea (Cicer arietinum L.) hybridization using morphological and molecular markers

Abstract

Background and Aim. Chickpea (Cicer arietinum L.) is one of the most important legume crops cultivated in arid regions worldwide; however, its hybridization is complicated due to the small size of its flowers and high sensitivity of reproductive organs to mechanical damage. The aim of the study was to enhance the efficiency of chickpea hybridization under the conditions of southeastern Kazakhstan through an integrated approach combining morphological and molecular markers.
Materials and Methods. Thirty-three chickpea genotypes of the “Desi” and “Kabuli” ecotypes were studied in 2024–2025. Five STMS markers (TA37, TA110, TA27, TA96, and TA194) were used to assess the genetic polymorphism of the parental genotypes. The efficiency of four hybridization methods was evaluated based on pod set, while the hybrid origin of the progeny was confirmed using molecular
markers.
Results. STMS markers TA96 and TA194 were the most informative for differentiating the parental genotypes, detecting eight and nine alleles, respectively. The pod-set rate was 4.7% in crosses performed without emasculation and 2.0% in crosses performed with emasculation. The difference between the two methods was statistically significant (χ² = 4.027; p = 0.045). Among the individual experimental treatments, the highest pod-set rate (4.9%) was observed when crossing without emasculation was
combined with treatment with a mixture of the phytohormones GA3, NAA, and kinetin.
Conclusion. Chickpea hybridization without emasculation resulted in a higher pod-set percentage, although the rate remained low in practical terms, indicating the need for further methodological optimization. For breeding purposes, the non-emasculation method should be combined with mandatory molecular verification of hybrids because the proportion of true hybrids obtained using
this method is lower than that obtained with emasculation.