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Acta Agronomica Sinica ›› 2021, Vol. 47 ›› Issue (7): 1402-1414.doi: 10.3724/SP.J.1006.2021.01072

• RESEARCH NOTES • Previous Articles    

Creation of disomic addition, substitution and translocation lines of durum wheat-Thinopyrum elongatum

DUAN Ya-Mei1,2, LUO Xian-Lei2, CHEN Shi-Qiang3, GAO Yong2, CHEN Jian-Min1,2,*(), DAI Yi1,*()   

  1. 1Institute of Agricultural Science and Technology Development / Joint International Research Laboratory of Agriculture & Agri-Product Safety of MOE, Yangzhou University, Yangzhou 225009, Jiangsu, China;
    2College of Bioscience and Biotechnology / Co-Innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou 225009, Jiangsu, China
    3Institute of Agricultural Sciences, Lixia River Region, Yangzhou 225009, Jiangsu, China
  • Received:2020-09-04 Accepted:2020-12-02 Online:2021-07-12 Published:2020-12-28
  • Contact: CHEN Jian-Min,DAI Yi E-mail:jmchen@yzu.edu.cn;daiyi@yzu.edu.cn
  • Supported by:
    This research was supported by the Natural Science Foundation of Jiangsu(BK20180908);This research was supported by the Natural Science Foundation of Jiangsu(BK20181213);the Natural Science Foundation of Yangzhou(YZ2018097)

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Abstract:

The establishment of chromosome addition, substitution and translocation lines through distant hybridization between wheat and Thinopyrum elongatum is an important approach to utilize useful genes of Th. elongatum in wheat improvement. Based on chromosome-specific molecular markers, chromosome counting, genome in situ hybridization (GISH), and non-denaturing fluorescence in situ hybridization (ND-FISH), several stable addition, substitution or translocation lines were identified from a cross between Triticum turgidum ssp. durum Langdon (AABB) and Trititrigia 8801 (AABBEE). Two 3E and 6E disomic addition lines Du-DA3E and Du-DA6E of durum wheat-Th. elongatum, a 1E (1B) disomic substitution line Du-DS1E (1B), and a 1AS-1EL disomic translocation line Du-T1AS·1EL were created in this study. The chromosome-segments of Th. elongatum could be inherited stably in these new lines, which not only increased the types of addition and substitution lines of durum wheat-Th. elongatum, but also provided special germplasm resources for the follow-up use of excellent genes of Th. elongatum in wheat improvement.

Key words: Triticum turgidum, Thinopyrum elongatum, addition line, substitution line, translocation line

Table 1

Plant materials used in this study"

材料名称
Material name		 染色体数
Chromosome number		 染色体组成
Chromosome composition		 缩写
Abbreviations
硬粒小麦(Langdon)
Triticum turgidum ssp. durum (Langdon)		 2n=28 AABB Ld
六倍体小偃麦8801
Hexaploid Trititrigia		 2n=28+14=42 AABBEE 8801
二倍体长穗偃麦草
Diploid Th. elongatum		 2n=14 EeEe 2X
四倍体长穗偃麦草
Tetraploid Th. elongatum		 2n=28 Ee1Ee1Ee2Ee2 4X
普通小麦中国春
Common wheat Chinese spring		 2n=42 AABBDD CS
中国春-长穗偃麦草1E-7E附加系
Chinese spring-Th. elongatum 1E-7E addition lines		 2n=44 AABBDD+II1E-7E DA1E~7E
中国春-长穂偃麦草1EL端体附加系
Chinese spring-Th. elongatum 1EL telosomic addition line		 2n=44 AABBDD+II1EL DA1EL
中国春-长穂偃麦草1ES端体附加系
Chinese spring-Th. elongatum 1ES telosomic addition line		 2n=44 AABBDD+II1ES DA1ES
硬粒小麦-长穗偃麦草1E(1B)代换系
Durum wheat-Th. elongatum 1E(1B) substitution line		 2n=26+2=28 AABB+II1E(1B) Du-DS1E(1B)
硬粒小麦-长穗偃麦草3E附加系
Durum wheat-Th. elongatum 3E addition line		 2n=28+2=30 AABB+II3E Du-DA3E
硬粒小麦-长穗偃麦草6E附加系
Durum wheat-Th. elongatum 6E addition line		 2n=28+2=30 AABB+II6E Du-DA6E
硬粒小麦-长穗偃麦草1AS-1EL易位系
Durum wheat-Th. elongatum 1AS-1EL translocation line		 2n=26+2T=28 AABB+ II1AS·1EL Du-T1AS·1EL

Table 2

Molecular markers and their sequences of Th. elongatum"

标记名称
Molecular marker		 引物序列Primer sequence (5'-3') 扩增染色体
Amplified chromosome		 参考文献
Reference
正向Forward 反向Reverse
GSM-1 AATCAGTCATAGTATCCAAAGTCTCCG TGTTTAGAATAGGATGGGTAGAGC 1E-7E This study
GSM-2 TCTTACAGATAGGTCCCTGGCAT TGGTAGTCACCTATCCAAGAAT 1E-7E This study
GSM-3 TGTTTCTTAGTTGTTTTGTT GCCTTGACCACCATAC 1E-7E Chen et al. [37]
CSM-1E-1 GATCTGTTTACGTCGATGCCG TGCCGAACTATTTGCTCCTTG 1E, 1EL This study
CSM-1E-2 TGAGAGGCGTGAGGACCTTGCTAAT GGTTTCCCAAGTCTTGTGATGC 1E, 1ES This study
CSM-1E-3 AGGTTCATAAGAATCCATC TACTGGATTTGTCACTTGCT 1E, 1EL This study
CSM-1E-4 ATGTCAGCCTATCCTACGGGTGT ATTCGTTGGGTCACTTGTCTCCT 1E, 1ES This study
CSM-1E-5 TCCCAACCACCTTCGCATTCATT GGCGAAAGACCATCAAAAGGAGTG 1E, 1EL This study
CSM-1E-6 CGTCACCAGAGAATAACAGGAG GTTCTATGTCAAGCAGTTCCG 1E, 1EL This study
CSM-1E-7 CCCGTGTAAGGAACAGAAAC TTAAGGTCGTCCCAACAGG 1E, 1EL This study
CSM-2E-1 TCAATGTCAAGGAGGAGGGTC TGGTCAAGAAGACAAGGGAAG 2E This study
CSM-3E-1 TATTAGTCGAGCACCTGTCG ATCTTCCTTGGCATCCTTC 3E This study
CSM-3E-2 CTGAGGGTTTAATTCGTAGCG TGATGGTTTCCCACTGTTCC 3E This study
CSM-3E-3 AGAGGATTTGAGGCTGTTCG AGATGTGCCGTGTAAAGGAGT 3E This study
CSM-3E-4 TGATGAGTCGAGCAAACAACC ATGTCCCTAAAGCCAAGTCC 3E This study
CSM-3E-5 TACTCCATAGAGGCTAGAAGAAC TGTCAAAGGCAAGCAAAGC 3E This study
CSM-4E-1 GAGAAGCCTCTGCCTTTAAG ACGGCGAGTCCTTGGTC 4E Luo et al. [38]
CSM-5E-1 ACCATTAGGTCACCGCAGTCT CGTTCTCGTGTCCCTACTCTTGT 5E This study
CSM-6E-1 GAGTGCTGGGATGAAGAAG GACGAAGACTAGGCACCAG 6E This study
CSM-6E-2 GCTAATGAGGGTTACAGAGT TAAAGAGCATCGGGGAC 6E This study
CSM-6E-3 AGCTGGCTCATCCCCATTTCT CGTGTTTACTGCCCATTTGTCC 6E This study
CSM-6E-4 TTTTTCGCACCACCCTTTC TTGTTCGAGAACGGGTTGC 6E Luo et al. [38]
CSM-6E-5 ACGAGGTTTGGCCGGTAT TACTCTTTGCTGATGATAGGCTC 6E Luo et al. [38]
CSM-7E-1 CTGCGGTTCTAAAATACTGAGG AAAGCTTGCCAAATGCC 7E This study

Fig. 1

Amplification of F2 (a) and F4 (b) generations of 8801 × Langdon by genome-specific molecular markers in Th. elongatum a1-a3: amplification of markers in F2 generation. M: DL2501 marker; 1-22: different plants of F2 generation; 23: Ld; 24: 4X; 25: 8801. a1: markers GSM-1; a2: marker GSM-2; a3: marker GSM-3. b1-b4: amplification of marker GSM-3 to F4 generation. M: DL2501 marker; 1-13: different plants of F4 generation; b1: line Du_No.1; b2: line Du_No.3; b3: line Du_No.6; b4: line Du_No.8; 14: Ld; 15: 4X; 16: 8801."

Table 3

Amplification of Th. elongatum by chromosome-specific molecular markers"

标记名称
Molecular marker		 扩增染色体
Amplified chromosome		 Du_No.1 Du_No.3 Du_No.6 Du_No.8
CSM-1E-1 1E, 1EL + - - +
CSM-2E-1 2E - - - -
CSM-3E-1 3E - + - -
CSM-4E-1 4E - - - -
CSM-5E-1 5E - - - -
CSM-6E-1 6E - - + -
CSM-7E-1 7E - - - -
CSM-1E-2 1E, 1ES + * * -
CSM-1E-3 1E, 1EL + * * +
CSM-1E-4 1E, 1ES + * * -
CSM-1E-5 1E, 1EL + * * +
CSM-1E-6 1E, 1EL + * * +
CSM-1E-7 1E, 1EL + * * +
CSM-3E-2 3E * + * *
CSM-3E-3 3E * + * *
CSM-3E-4 3E * + * *
CSM-3E-5 3E * + * *
CSM-6E-2 6E * * + *
CSM-6E-3 6E * * + *
CSM-6E-4 6E * * + *
CSM-6E-5 6E * * + *

Fig. 2

Amplification of F5 generation of 8801×Langdon by the chromosome-specific molecular markers in Th. elongatum M: DL2501 marker; a: line Du_No.1, marker CSM-1E-1; b: line Du_No.3, marker CSM-3E-1; c: line Du_No.6, marker CSM-6E-4; d: line Du_No.8, marker CSM-1E-1; 1: Ld; 2: CS; 3: 4X; 4: 8801; 5-14: different plants of F5 generation."

Fig. 3

Amplification of line Du_No.8 by the 1EL and 1ES chromosome arm-specific markers M: DL2501 marker; a: CSM-1E-3, amplified the long arm of 1E chromosome (1EL); b: CSM-1E-4, amplified the short arm of 1E chromosome (1ES); 1: Ld; 2: CS; 3: 4X; 4: 8801. 5-9: randomly selected individual plant."

Fig. 4

Mitotic metaphase chromosomes of root tips of four lines from F4 generation a: line Du_No.1, 2n=28; b: line Du_No.3, 2n=30; c: line Du_No.6, 2n=30; d: line Du_No.8, 2n=28."

Fig. 5

GISH and ND-FISH patterns of four lines a1: GISH of line Du_No.3; a2: ND-FISH on the same metaphase; a3: the E chromosomes in a2; a4: the 3E chromosome of 8801. b1: GISH of line Du_No.6; b2: ND-FISH on the same metaphase; b3: the E chromosomes in b2; b4: the 6E chromosome of 8801. c1: GISH of line Du_No.1; c2: ND-FISH on the same metaphase; c3: the E chromosomes in c2; c4: the 1E chromosome of 8801; c5: the 1B chromosome of 8801. d1: GISH of line Du_No.8; d2: ND-FISH on the same metaphase; d3: the E chromosomes in d2; d4: the 1E chromosome of 8801; c5: the 1A chromosome of 8801. The green fluorescence signals show the chromosomes of Th. elongatum in GISH. The arrows refer to chromosomes of Th. elongatum in ND-FISH. Bar: 10 μm."

Table 4

Agronomic traits of durum wheat-Th. elongatum addition, substitution lines and their parents"

材料
Materials		 调查株数
Number of plants		 株高
Plant height
(cm)		 穗长
Spike length
(cm)		 小穗粒数
Kernel number per spikelet		 每穗小穗数
Spikelet number per spike		 每穗粒数
Kernel number per spike		 千粒重
1000-kernel weight (g)		 旗叶面积
Flag leaf area (cm2)
Langdon 22 169.8±10.4 d 9.7±1.0 ab 2.3±0.2 bc 24.7±1.9 bc 56.1±7.3 c 37.2±1.4 d 30.6±4.7 a
8801 18 178.6±13.3 d 18.4±1.6 e 2.1±0.3 ab 20.9±2.7 a 43.6±4.5 ab 18.0±3.1 a 47.4±8.8 bc
Du-DA3E 10 95.1±15.1 a 9.3±0.5 a 2.6±0.4 c 21.0±2.5 a 53.9±7.7 bc 25.2±4.9 b 42.3±13.9 b
Du-DA6E 10 117.1±10.3 b 12.8±0.9 d 3.5±0.3 d 26.4±2.5 c 93.1±10.0 d 30.7±3.1 c 57.2±8.0 c
Du-DS1E (1B) 10 109.1±11.8 b 12.0±2.0 cd 1.8±0.6 a 23.7±2.1 b 42.2±16.9 a 20.1±3.3 a 48.1±9.6 bc
Du-T1AS·1EL 12 134.8±7.7 c 10.7±0.8 bc 2.5±0.4 bc 23.6±1.4 b 58.9±9.9 c 22.5±2.6 ab 54.1±4.2 c

Fig. 6

Phenotypes of seeds and spikes of durum-Th. elongatum addition lines and substitution line, and their parents A, a: Langdon; B, b: 8801; C, c: addition line Du-DA3E; D, d: addition line Du-DA6E; E, e: substitution line Du-DS1E(1B); F, f: translocation line Du-T1AS·1EL. Bar: 10 mm."

Fig. S1

Selection of chromosomal-specific molecular markers based on the amplification results in CS-Th. elongatum addition lines M: DL2501 marker; 1-7: CS-Th. elongatum 1E-7E addition lines; 8: Th. elongatum (2x); 9: Ld; 10: CS; 11: 8801. Specific markers: CSM-1E-1, CSM-2E-1, CSM-3E-5, CSM-4E-1, CSM-5E-1, CSM-6E-2, CSM-7E-1, GSM-2."

Fig. S2

PCR amplification of the 1E chromosome-specific molecular markers in CS-Th. elongatum 1E telodisomic addition lines M: DL2501 marker; a: CSM-1E-3, amplified the long arm of the 1E chromosome (1EL); b: CSM-1E-3, amplified the short arm of the 1E chromosome (1ES). 1: CS-Th. elongatum 1E addition line; 2: CS-Th. elongatum 1EL telodisomic addition line (DA1EL); 3: CS-Th. elongatum 1ES telodisomic addition line (DA1ES); 4: Th. elongatum (2x); 5: Ld."

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