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Acta Agronomica Sinica ›› 2022, Vol. 48 ›› Issue (2): 511-517.doi: 10.3724/SP.J.1006.2022.11025

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Molecular cytogenetic identification of wheat-Thinopyrum intermedium 2A/6St substitution strain 014-459

TAO Jun1,2(), LAN Xiu-Jin1,*()   

  1. 1Triticeae Research Institute, Sichuan Agricultural University, Chengdu 611130, Sichuan, China
    2Mianyang Academy of Agricultural Sciences, Mianyang 621023, Sichuan, China
  • Received:2021-03-09 Accepted:2021-06-16 Online:2022-02-12 Published:2021-06-29
  • Contact: LAN Xiu-Jin E-mail:tj4bmy@163.com;lanxiujin@163.com

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Abstract:

Thinopyrum intermedium is a valuable resource for wheat genetic improvement, and numerous wheat-Th. intermedium chromosome addition and substitution lines and partial amphiploids have been developed. Zhong 4 is a wheat-Th. intermedium partial amphidiploid, which can easily hybridize with wheat and is extensively utilized for wheat improvement. The strain 014-459 was a progeny of partial amphidiploid Zhong 4 descendant of wheat and Th. intermedium with some special characteristics such as high crude protein and wet gluten content and partial F1 of common wheat cultivars and strain 014-459 were sterility while a few others were fertility despite reciprocal cross. Molecular cytogenetic identification of strain 014-459 were detected for speculated contained fragment of Th. intermedium chromosomes based on its special characters. Genomic composition of strain 014-459 was detected by FISH, GISH, and PLUG (polymerase chain reaction-based landmark unique gene) marker analysis. Combined FISH/GISH analysis confirmed that a pair of wheat 2A chromosomes in strain 014-459 of the hybrid progeny between wheat-Thinopyrum intermedium partial amphidiploid Zhong 4 and wheat was replaced by a pair of St chromosomes from Thinopyrum intermedium. PLUG marker analysis verified that this pair belonged to the sixth homologous group. The special characters of strain 014-459 might be attributable to the substitution of Th. intermedium St chromosome into wheat. The study of molecular cytogenetic identification of strain 014-459 was probably beneficial to quality improvement and utilization of 6St chromosome of Th. intermedium in wheat breeding.

Key words: GISH, FISH, substitution strain, Pseudoroegneria strigosa, cytogenetics

Fig. 1

Meiosis in wheat-Thinopyrum intermedium A-B: meiotic metaphase I; C-E: meiotic anaphase I; F-H: meiotic telophase I."

Fig. 2

In situ hybridization and pollen mother-cell meiosis in wheat-Thinopyrum intermedium A: FISH map of strain 014-459. Oligo-pSc119.2-2 (green), oligo-pTa535-2 (red); B: GISH map of strain 014-459. DNA of Th. intermedium used as the probe, and DNA of CS was used as the blocker; C: GISH map of strain 014-459. DNA of Pseudoroegneria strigosa was used as the probe and DNA of CS was used as the blocker; D: FISH map of strain 014-459. St2-80 was used as the probe and DNA of CS was used as the blocker; E: GISH map of pollen mother cells of strain 014-459 at the meiotic metaphase. DNA of Th. intermedium was used as the probe and DNA of CS was used as the blocker."

Fig. 3

In situ hybridization in wheat-Thinopyrum intermedium A: FISH map of strain 014-459. Oligo-pSc119.2-2 (green), oligo-pTa535-2 (red); B: GISH map of strain 014-459. DNA of Th. intermedium used as the probe, and DNA of CS was used as the blocker; C: GISH map of strain 014-459. DNA of rye was used as the probe and DNA of Chinese Spring (CS) wheat was used as the blocker."

Fig. 4

PLUG marker analysis in wheat-Thinopyrum intermedium A: TNAC1763 amplification product (Taq I digestion), with arrows indicating St-specific bands; B: TNAC1178 amplification product (Taq I digestion), with arrows indicating a CS-specific amplification band, which was absent from 014-459 and St (Pseudoroegneria strigosa)."

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