Exploring the Genetic Potential of Thinopyrum Species in Wheat and Durum Wheat Improvement

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(A) T14-44 karyotype identified by genomic in situ hybridization (GISH) and fluorescence in situ hybridization (FISH). The genomic DNA of Th. intermedium (green) and the wheat centromeric retrotransposon 6C6 (red) were used as a probe. The genomic DNA of Chinese Spring (CS) was used as a blocker. Chromosomes were stained with 4, 6-diamidino-2-phenylindole (DAPI; blue). Scale bar = 10 μm. (B) Stripe rust evaluation on T14-44 and T14-42. Stripe rust tests were performed using three rust races CYR32, CYR33, and CYR34, respectively. Common wheat cultivar Mingxian 169 was used as the susceptible control.
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Homology modeling and molecular simulation of TtLEA2-1 protein. (A) Homology modeling of TtLEA2-1 protein using MODELLER9.22. (B) Ramachandran plot analysis. A, B, and L regions: most favored residues; a, b, l, and p regions: additional allowed residues; ∼a, ∼b, ∼l, and ∼p regions: generously allowed residues. (C) Molecular dynamics simulation. Backbone of root mean squared deviation (RMSD) plotted versus time in picoseconds (ps).
Original Research
04 April 2022

Late embryogenesis-abundant (LEA) proteins are critical in helping plants cope with salt stress. “Y1805” is a salt-tolerant Tritipyrum. We identified a “Y1805”-specific LEA gene that was expressed highly and sensitively under salt stress using transcriptome analysis. The novel group 2 LEA gene (TtLEA2-1) was cloned from “Y1805.” TtLEA2-1 contained a 453 bp open reading frame encoding an 151-amino-acid protein that showed maximum sequence identity (77.00%) with Thinopyrum elongatum by phylogenetic analysis. It was mainly found to be expressed highly in the roots by qRT-PCR analysis and was located in the whole cell. Forty-eight candidate proteins believed to interact with TtLEA2-1 were confirmed by yeast two-hybrid analysis. These interacting proteins were mainly enriched in “environmental information processing,” “glycan biosynthesis and metabolism,” and “carbohydrate metabolism.” Protein-protein interaction analysis indicated that the translation-related 40S ribosomal protein SA was the central node. An efficient wheat transformation system has been established. A coleoptile length of 2 cm, an Agrobacteria cell density of 0.55–0.60 OD600, and 15 KPa vacuum pressure were ideal for common wheat transformation, with an efficiency of up to 43.15%. Overexpression of TaLEA2-1 in wheat “1718” led to greater height, stronger roots, and higher catalase activity than in wild type seedlings. TaLEA2-1 conferred enhanced salt tolerance in transgenic wheat and may be a valuable gene for genetic modification in crops.

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