Crops ›› 2023, Vol. 39 ›› Issue (5): 24-29.doi: 10.16035/j.issn.1001-7283.2023.05.004

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Gene Differential Expression Analysis of TMS5 in the Fertility Conversion of Wheat BNS Sterile Line

Liu Shuhan1,2(), Chen Lei1, Zhang Jianchao2, Hu Gan2, Sun Junyan1, Liu Dongtao3(), Wang Junwei2()   

  1. 1School of Agriculture, Xinyang Agriculture and Forestry University, Xingyang 464000, Henan, China
    2College of Agronomy, Northwest A & F University/National Yangling Agricultural Biotechnology & Breeding Center/Yangling Branch of State Wheat Improvement Center/Wheat Breeding Engineering Research Center, Ministry of Education/Key Laboratory of Crop Heterosis of Shaanxi Province, Yangling 712100, Shaanxi, China
    3Xuzhou Institute of Agricultural Science, Xuhuai District, Jiangsu Province, Xuzhou 221131, Jiangsu, China
  • Received:2022-03-22 Revised:2022-07-22 Online:2023-10-15 Published:2023-10-16

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Abstract:

The TMS5 is a thermosensitive sterility gene of rice, which encodes a RNase Z protein. We analyzed the expression differences of wheat TMS5 gene in wheat BNS sterile line and fertile line at different developmental stages of anthers, aiming to provide a theoretical basis for revealing the mechanism of wheat BNS thermosensitive sterility. Using the sterile and fertile anthers of wheat BNS as test materials, a 408bp sequence of wheat TMS5 was cloned, which was 80% identical to the sequence of the TMS5 in rice. Compared the TMS5 expression in stages of wheat anther development including tetrad, early mononuclear, late mononuclear, dinuclear and trinuclear, the results showed that the TMS5 was mainly expressed in the tetrad stage, and the TMS5 expression level in anthers of fertile lines was much higher than that of sterile lines. The wheat TMS5 gene was closely related to the pollen abortion of wheat BNS thermosensitive sterile line, and the tetrad stage was the key period of TMS5’s regulation.

Key words: Wheat, Photo-thermo-sensitive male sterility, BNS, TMS5 gene, qRT-PCR

Table 1

Sequence of the primers"

引物Primer 序列（5′-3′）Sequence (5′-3′)
TMS5-1F CCCCTCCCCAGCCATAAAG
TMS5-1R AAGGCCCTGACCTTGAGATC
TMS5-2F CCATAAAGCGTAAGGCGAAGC
TMS5-2R GCGTGCGAGATGAAGAGGAA
18S-F CGTCCCTGCCCTTTGTACAC
18S-R AACACTTCACCGGACCATTCA

Fig.1

Alignment result of TMS5 sequence TMS5-CDs indicates the sequence of this gene in rice"

Fig.2

Semi-quantitative PCR analysis of TMS5 gene STe: the tetrad stage in sterile line; SUe: early mononuclear in sterile line; SUl: late mononuclear in sterile line; SBi: dinuclear in sterile line; STr: trinuclear in sterile line; FTe: the tetrad stage in fertile line; FUe: early mononuclear in fertile line; FUl: late mononuclear in fertile line; FBi: dinuclear in fertile line; FTr: trinuclear in fertile line"

Fig.3

Melting curves of 18S and TMS5"

Fig.4

Relative expressions of TMS5 gene during the various periods of anther development The different letters indicate significant difference at 0.05 level"

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