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Acta Agronomica Sinica ›› 2020, Vol. 46 ›› Issue (11): 1743-1749.doi: 10.3724/SP.J.1006.2020.01016

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Analysis of the stripe rust resistance in a wheat line CB037 with high regeneration and transformation efficiency

ZHENG Yan-Yan(), HUANG De-Hua, LI Ji-Long, ZHANG Hui-Fei, BAO Yin-Guang, NI Fei, WU Jia-Jie*()   

  1. State Key Laboratory of Crop Biology / College of Agronomy, Shandong Agricultural University, Tai'an 271018, Shandong, China
    State Key Laboratory of Crop Biology / College of Agronomy, Shandong Agricultural University, Tai’an 271018, Shandong, China
  • Received:2020-02-24 Accepted:2020-06-02 Online:2020-11-12 Published:2020-06-15
  • Contact: Jia-Jie WU E-mail:zyy18854889722@163.com;jiajiewu@sdau.edu.cn
  • Supported by:
    This study was supported by the National Major Project for Developing New GM Crops(2016ZX08009003001);the Youth Innovation and Scientific Program of Higher Education of Shandong(2019KJF026)

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Abstract:

Genetic transformation is used for efficient gene cloning, gene editing and gene engineering, etc. Obtaining recipient lines amenable to transformation and with pure genetic background is critical for high efficiency transformations. For recent years, the wheat inbreed line CB037 had been widely used as a recipient for transgenes and obtain its high transformation potential. Despite having stable agronomic traits, the CB037 is genetically heterogeneous for resistance to wheat stripe rust (Puccinia striiformis f. sp. tritici, Pst). In this study, the Pst-resistant line CB037-PstR and Pst-susceptible line CB037-PstS were isolated, and their F2 population was created. Genetic analysis showed that the CB037-PstR carried a single dominant resistance gene. The identified resistance gene was mapped on the short arm of chromosome 1B using BSR-seq and molecular marker analysis. GISH results further revealed that CB037-PstR is a 1BL/1RS translocation line and likely carried Yr9. This study segregated genetic heterogeneity in CB037 for stripe rust resistance and isolated its Pst-susceptible and resistant lines. These results will facilitate trait-oriented use of CB037 in genetic engineering of wheat.

Key words: wheat transformation, 1BL/1RS translocation line, stripe rust resistance, BSR-Seq, Yr9

Fig. 1

Characteristic stripe rust infection types (ITs) of two different lines segregated from CB037 1, 2: seedlings leaf; 3: adult plant leaf."

Fig. 2

Chromosomal distribution of polymorphisms between stripe rust resistant line CB037-PstR and susceptible line CB037-PstS and the susceptible bulk in F2 individuals Vertical lines and green dots represent wheat chromosomes and centromeres, respectively. Numbers and letters on top identify each chromosome. Numbers below each chromosome mark the amount of the SNPs and InDels mapped on each chromosome. Short horizontal lines indicate positions of SNPs or InDels."

Table 1

Molecular markers developed in this study"

分子标记
Molecular marker		 染色体及位置
Chromosome & position (Mb)		 正向引物
Forward primer
(5′-3′)		 反向引物
Reverse primer
(5′-3′)		 退火温度
Annealing temperature (℃)		 内切酶
Restriction enzyme		 多态性
Polymorphism
(bp)
CBSR-53/54 a 1B, 69.9 GAACGGCTCGGATCTGATC TGTACAACAACAAGAAGGAGGAG 55 Dde I CBS: 79+83
CBR: 163
CBSR-1/2 b 1B, 98.7 CCGGAGTTGAAGATAAGTAGTTG CTTCCCTGTATTCGAACAGACTCA 56 N/A CBS: 242
CBR: 0
CBSR-35/36 a 1B, 115.3 ACACCGAGGTGCGTCTAAT TATCCCATGCGGTAAATGGAC 55 Msp I CBS: 150+57
CBR: 207
InDel-3/4 b 1B, 376.6 TCCATGGCAGTACCGCTTGCTT CCGGAGCAGCAATGAAACGTC 56 N/A CBS: 240
CBR: 207
CBSR-25/26 b 4A, 723.7 GCCGCGTTTTGCATGTTTTCCG TGGGTTGAATTTTGGGTTGTTTGAA 56 N/A CBS: 408
CBR: 378

Fig. 3

Detection of molecular markers (iag95, AF1/4, H20) of the stripe rust resistance gene Yr9 Letters S and R indicate phenotypes of susceptible and resistant of F2 individuals; CK is the blank control."

Fig. 4

GISH and FISH results of two different lines isolated from CB037 A, B: CB037-PstS; C, D: CB037-PstR; A & C: GISH patterns; B & D: FISH patterns. Bar = 10 μm."

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