Genetic Analysis and Molecular Marker Development for a Wheat—Thinopyrum ponticum Substitution Line WTS135 with Leaf Rust Resistance

doi:10.11983/CBB25014

Abstract

Abstract:

INTRODUCTION: The genetic diversity of common wheat (Triticum aestivum) has decreased sharply due to the artificial domestication and modern breeding operations, making it more vulnerable to the threats from pests and diseases. Leaf rust, caused by the fungal pathogen Puccinia triticina Eriks. (Pt), is a devastating disease in wheat. Over 80 leaf rust resistance (Lr) genes have been formally identified, with nearly half originating from wheat wild relatives. However, the rapid evolution of Pt physiological races has rendered many Lr genes ineffective against prevalent Pt races. Consequently, identifying novel sources of resistance in wild relatives remains an urgent priority for sustainable wheat breeding.

RATIONALE: As one of the most widely used relatives in the genetic improvement of wheat, decaploid Thinopyrum ponticum (Podp.) Barkworth and D. R. Dewey shows excellent resistance to multiple diseases including leaf rust. By wild hybridization and chromosome engineering, we created a wheat-Th. ponticum germplasm WTS135. We evaluated its disease resistance with Pt race THTT, developed Th. ponticum specific markers by specific-locus amplified fragment sequencing technology and assessed its agronomic traits by phenotypic investigation. Sequential genomic in situ hybridization (GISH)-fluorescence in situ hybridization analysis (FISH) and liquid chip have been used to discover its chromosome composition.

RESULTS: WTS135 is immune to the Pt race THTT. Pedigree analysis showed that this resistance originated from the exogenous chromosome of Th. ponticum. GISH-FISH analysis revealed that the wheat chromosomes 7D were replaced by the Th. ponticum-derived chromosomes. Liquid chip showed that the alien chromosomes belonged to the homoeologous group 7, and the density and abundance of the signals in the peri-centromeric region along them were significantly lower, which was consistent with the GISH results. Therefore, it is indicated that WTS135 is a 7St (7D) disomic substitution line. After detected by the molecular markers related to known Lr genes on wheat 7D chromosome, it is presumed that WTS135 could probably carry a novel resistance gene that is not identical to genes Lr19 and Lr29. Ten primers specific to Th. ponticum were developed to rapidly trace the exogenous chromatin in WTS135. Phenotypic investigation showed that the yield of WTS135 was not significantly different from that of the recurrent parent Jimai 22, suggesting that this line can be useful for improving disease resistance in wheat.

CONCLUSION: Introducing resistance genes from wild relatives into wheat through wide hybridization can broaden the genetic base of wheat and provide new sources for breeding disease-resistant varieties. We developed a wheat-Th. ponticum 7St (7D) substitution line, which possibly has a novel alien resistance gene and could be used in wheat disease resistance breeding.

Chromosome composition and leaf rust resistance evaluation of WTS135 (A) GISH analysis using Th. ponticum gDNA as a probe and CS gDNAs as a block; (B) mc-FISH analysis using combined probes (bars=20 μm). (C) The liquid chip analysis of WTS135. (D) WTS135 (1), Jimai 22 (2), Xiaoyan 81 (3), Zhongnong 28 (4), Th. Ponticum (5). The white arrows indicate exogenous chromosomes, purple boxes represent chromosome additions or deletions.

Key words: wheat, Thinopyrum ponticum, leaf rust, wide hybridization, substitution line, in situ hybridization, molecular markers

Gaiya Jia, Na Zhang, Hongwei Li, Bin Li, Zhensheng Li, Zhaosheng Kong, Qi Zheng. Genetic Analysis and Molecular Marker Development for a Wheat—Thinopyrum ponticum Substitution Line WTS135 with Leaf Rust Resistance [J]. Chinese Bulletin of Botany, 2025, 60(5): 1-0.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB25014

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