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Acta Agronomica Sinica ›› 2023, Vol. 49 ›› Issue (10): 2603-2612.doi: 10.3724/SP.J.1006.2023.21082

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

Creation and analysis of secondary translocation harbouring gene Pm21

ZHANG Lan-Yue1(), LUO Jiang-Tao2(), FAN Chao-Lan1, LI Ya-Zhou1, JIANG Bo1, CHEN Xue1, CHEN Xue-Jiao1, YUAN Zhong-Wei1, NING Shun-Zong1, ZHANG Lian-Quan3, LIU Deng-Cai3(), HAO Ming1()   

  1. 1Triticeae Research Institute, Sichuan Agricultural University, Wenjiang 611130, Sichuan, China
    2Crop Research Institute, Sichuan Academy of Agricultural Science, Jinjiang 610066, Sichuan, China
    3State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China, Sichuan Agricultural University, Wenjiang 611130, Sichuan, China
  • Received:2022-12-12 Accepted:2023-02-21 Online:2023-10-12 Published:2023-03-06
  • Contact: E-mail: haomingluo@foxmail.com; E-mail: dcliu7@sicau.edu.cn
  • About author:**Contributed equally to this study
  • Supported by:
    Sichuan Science and Technology Program(2022ZDZX0014);Sichuan Science and Technology Program(2022NSFSC1696);National Natural Science Foundation of China(31971884);National Natural Science Foundation of China(32172020)

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Abstract:

Wheat-Haynaldia villosa 6VS.6AL translocation harbouring the gene Pm21 has made a great contribution to powdery mildew resistance breeding in China. Based on the data of 55K SNP chip, 25 (15.4%) out of 162 Sichuan wheat varieties contained the translocation. In this study, recombination point and haplotype analysis on the 25 varieties showed that it was centric translocation. Combined with the pedigree information, 92R178 was the original donor of the 6VS.6AL translocation in these varieties. 6AS-6VS-6AS.6AL secondary recombinant containing Pm21 was generated by using primary recombinants 6VS-6AS.6AL and 6AS-6VS.6AL as the cross parents, which both formed by the induction of ph1b. The secondary recombinant had a much smaller 6VS chromatin than the primary recombinants. Based on the Chinese Spring reference genome, the crossover points of the secondary recombinant were located within 53.1-53.8 Mb and 90.7-92.2 Mb of chromosome 6A, with a 6VS fragment size about 36.9-39.1 Mb. Molecular cytological identification also detected the extensive recombinants among wheat endogenous homoeologs induced by ph1b, which was not only disadvantage for genetic stabilization of wheat-alien recombinants but also for wheat breeding. A proposed solution to reduce endogenous recombinants was to preserve the ph1b mutant line in a heterozygous condition and reduce the selfing times during the development of ph1b-mediated wheat-alien recombination. In breeding, it is necessary to eliminate endogenous recombinants as soon as possible.

Key words: wheat, Haynaldia villosa, powdery mildew, 6VS.6AL translocation, Pm21 gene, small fragment translocation line

Table 1

Twenty-five Sichuan wheat varieties (lines) carrying 6VS.6AL translocation"

品种(系)名
Accession name		 选育单位
Origin		 杂交组合
Pedigree		 审定年份
Released year		 细胞学*
Cytological*
绵阳27
Mianyang 27		 绵阳市农业科学院
Mianyang Academy of Agricultural Sciences		 81-5/81-24 1997 /
绵阳28
Mianyang 28		 绵阳市农业科学院
Mianyang Academy of Agricultural Sciences		 T79350-1-4/Mianyang 11 1997 /
川麦36
Chuanmai 36		 四川省农业科学院
Sichuan Academy of Agricultural Sciences		 Milan/SW5193 2002 ×
内麦8号
Neimai 8		 内江市农业科学院
Neijiang Academy of Agricultural Sciences		 Mianyang 26/92R178 2003 √
西科麦1号
Xikemai 1		 西南科技大学
SouthWest University of Science and Technology		 Mianyang 88-304/Mo-212 2003 ×
良麦2号
Liangmai 2		 四川农业大学
Sichuan Agricultural University		 Mianyang 26/// (10-A/88-1643//Chuanyu 12) 2004 ×
绵麦37
Mianmai 37		 绵阳市农业科学院
Mianyang Academy of Agricultural Sciences		 96EW37/Mianyang 90-100 2004 √
内麦9号
Neimai 9		 内江市农业科学院
Neijiang Academy of Agricultural Sciences		 Mianyang 26/92R178 2004 √
杏麦2号
Xingmai 2		 内江市农业科学院
Neijiang Academy of Agricultural Sciences		 Mianyang 26/92R178 2004 √
内麦11号
Neimai 11		 内江市农业科学院
Neijiang Academy of Agricultural Sciences		 Mianyang 26/92R178 2007 √
绵麦185
Mianmai 185		 绵阳市农业科学院
Mianyang Academy of Agricultural Sciences		 Mianyang 96-5/Liaochun 10 2008 √
内麦3416
Neimai 3416		 内江市农业科学院
Neijiang Academy of Agricultural Sciences		 R57/Ping 5 2010 /
绵麦228
Mianmai 228		 绵阳市农业科学院
Mianyang Academy of Agricultural Sciences		 1275-1/Nei 2938//99-1522 2011 ×
绵麦51
Mianmai 51		 绵阳市农业科学院
Mianyang Academy of Agricultural Sciences		 1275-1/99-1522 2012 √
西科麦7号
Xikemai 7		 西南科技大学
Southwest University of Science and Technology		 Chuanyu 11/Mo444 2012 √
Y11-1741 四川农业大学
Sichuan Agricultural University		 Zimai 1/Mianyang 2003-1 2012
绵麦1618
Mianmai 1618		 绵阳市农业科学院
Mianyang Academy of Agricultural Sciences		 1275-1//Nei 2938/99-1522 2013 √
西科麦8号
Xikemai 8		 西南科技大学
Southwest University of Science and Technology		 97-392/Yun 225747-5 2013 √
西科麦9号
Xikemai 9		 西南科技大学
Southwest University of Science and Technology		 Nei 4301/Mianyang 31 2014 √
宜麦9号
Yimai 9		 宜宾市农业科学院
Yibin Academy of Agricultural Sciences		 R59/Yi 97-24 2014 ×
川麦92
Chuanmai 92		 四川省农业科学院
Sichuan Academy of Agricultural Sciences		 Neimai 8/Jian 3//Chuanmai 42 2015 /
绵麦112
Mianmai 112		 绵阳市农业科学院
Mianyang Academy of Agricultural Sciences		 Mian 06-367/99-1522 2015 /
内麦366
Neimai 366		 内江市农业科学院
Neijiang Academy of Agricultural Sciences		 Balandal88/Nei 4344 2015 /
国豪麦3号
Guohaomai 3		 四川国豪种业
Sichuan Guohao Seeds Industry Co., Ltd.		 1227-185/99-1522//99-1572 2016 √
绵麦285
Mianmai 285		 绵阳市农业科学院
Mianyang Academy of Agricultural Sciences		 1275-1/99-1522 2016 √

Fig. 1

Genotypes of the chromosome 6A of 165 Sichuan wheats Each row represents a cultivar (line); each column represents a marker. The markers are arranged from left to right according their physical location on the Chinese spring reference genome of IWGSC v1.0. Markers are labeled in red with a genotype of A, in green with a genotype of C, in yellow with a genotype of G, in blue with a genotype of T, in grey with a genotype of heterozygote, and in pink with a genotype of missing. Hap1-3 represent haplotypes of centric region without recombination. Number in the bracket represents the cultivar (lines) number. Arrow points out the physical position on IWGSC v1.0."

Fig. 2

Flow diagram for creation and cytological identification of 6VS/6AS secondary translocation A: the crossing and selecting diagram to induce 6VS/6AS secondary translocation using their primary translocation; B: the confirmation the presence of Pm21 gene using its specific marker CINAU-NLR1 (from left to right: marker, Pm99915-1, CSph1b, HM782-15, HM780-7); C: genomic in situ hybridization (GISH) pattern of secondary translocation line Rec32; D: GSIH pattern of secondary translocation line Rec50; E: GISH pattern of 6VS.6AL translocation lines formed by homologous recombination between two 6VS/6AS primary translocation; F: the diagram of how primary translocation pair and exchange to generate secondary translocation. MI: metaphase I of meiosis; AI: anaphase I of meisosis. 6VS chromosome arm or fragment was filled in red, 6A chromosome or fragment in blue. Yellow line represents Pm21 gene. Yellow arrows represent C_6VS/6AS_AVA-Pm21-1 or 6VS.6AL. White arrows represent primary translocation without recombination."

Fig. 3

Graphical genotypes of Rec50 and HM887-9-51 Chromosomes on the left are from Rec50, right from HM887-9-51. Missing marker locations are labeled in green, others in red. The solid boxes indicate the deletion regions on nontarget wheat endogenous chromosomes. The dotted box indicates the deletion 6AS fragments on target primary and secondary recombined chromosome."

Fig. 4

Chromosome constitution of Rec50 a: genomic in situ hybridization on the root-tip chromosomes from A (pink), B (blue), and D (green) genomes. b: the fluorescence in situ hybridization using Oligo-pSc119.2 (green) and Oligo-pTa535 (pink) as the probes. White arrows indicate the nontarget wheat endogenous recombination chromosomes. Red arrows indicate the target primary and secondary 6VS/6AS recombined chromosomes."

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