Hybridity Assessment of Chickpea Genotypes to Identify True Hybrids Using SSR Markers

Abstract

Chickpea (Cicer arietinum L.) is one of the most important food legumes globally. However, its production is significantly affected by Ascochyta blight (AB), caused by Ascochyta rabiei, and Botrytis gray mold (BGM), caused by Botrytis cinerea. Breeding programs have been effective in developing disease-resistant cultivars, but closely related germplasm often lacks morphological markers required to confirm hybridity, a crucial step in breeding programs. This study aimed to identify SSR markers suitable for confirming true hybridity in desi chickpea. To improve desi chickpea cultivars resistant to AB and BGM through cross breeding, disease-susceptible genotypes (C214, PDG3, and JG14) were crossed with disease-resistant genotypes (GLWP61, GLWP147 and GLWP63). Four crosses-PDG3 × GLWP61, PDG3 × GLWP147, C214 × GLWP63 and JG14 × GLWP61-were performed to improve resistance further. In order to identify true F₁ hybrids, Simple Sequence Repeat (SSR) markers were employed. A total of 101 SSR markers were screened across the crosses. Among them, 10 markers in PDG3 × GLWP147, 4 markers in C214 × GLWP63, 10 markers in PDG3 × GLWP61 and 10 markers in JG14 × GLWP61 exhibited parental polymorphism, leading to the identification of 21 unique polymorphic SSR markers that were able to identify true F₁ hybrid. Notably, marker CaM2049 exhibited polymorphism across all the crosses. A total of 37 putative F₁ plants from the four crosses were examined and 33 plants (~89%) were confirmed as true hybrids. These results demonstrate that SSR markers are effective for molecular characterization and hybridity assessment in chickpea breeding programs.