Creation of Early Maturity Mutants by Editing GmphyA2 Using CRISPR/Cas9 Technology
Author:
Affiliation:

1.School of Agronomy,Anhui Agricultural University,Hefei 230036;2.Institute of Crop Sciences,Chinese Academy of Agricultural Sciences/State Key Laboratory of Crop Gene Resources and Breeding,Beijing 100081

Fund Project:

Foundation project: The Major Science and Technology Project of the Chinese Academy of Agricultural Sciences(CAAS-ZDRW202301)

  • Article
  • | |
  • Metrics
  • |
  • Reference [47]
  • | | | |
  • Comments
    Abstract:

    The photoperiod sensitivity of soybean limits the popularization of excellent soybean varieties for planting. In this study, we created new soybean varieties with early maturity and stable yield by reducing the sensitivity of soybean to photoperiod and expanding the planting range of good varieties. Three soybean GmphyA2 pure mutants were obtained using CRISPR/Cas9 technology. The created GmphyA2 mutants encodes a truncated protein without PHY structural domain due to a base deletion resulting in a code-shift mutation. Comparisons were made to characterize the fertility, plant size, and yield of the GmphyA2 mutant and the WT ‘Jack’. The results showed that because of significantly increased expression of GmFT2a and GmFT5a during the pre-flowering period, the GmphyA2 mutant flowered 5-6 d earlier and matured 6-11 d earlier compared with the WT. Although the fertility period was shortened, grains weight per plant was not significantly different from that of the WT due to a significant increase in both the effective number of branches and the number of grains per plant in the GmphyA2 mutant. These results indicate that the editing of GmphyA2 using CRISPR/Cas9 technology has created a new soybean germplasm with unchanged yield per plant with shortened fertility period, which provides a new genetic resource for breeding for early maturity and high yield.

    Reference
    [1] Zhang L, Liu W, Tsegaw M, Xu X, Qi Y, Sapey E, Liu L, Wu T, Sun S, Han T. Principles and practices of the photo-thermal adaptability improvement in soybean. Journal of Integrative Agriculture, 2020, 19(2): 295-310
    [2] Han Y, Zhao X, Liu D, Li Y, Lightfoot D A, Yang Z, Zhao L, Zhou G, Wang Z, Huang L, Zhang Z, Qiu L, Zheng H, Li W. Domestication footprints anchor genomic regions of agronomic importance in soybeans. New Phytologist, 2016, 209(2): 871-884
    [3] Lin X, Liu B, Weller J, Abe J, Kong F. Molecular mechanisms for the photoperiodic regulation of flowering in soybean. Journal of Integrative Plant Biology, 2021, 63(6): 981-994
    [4] Cober E R, Morrison M J. Regulation of seed yield and agronomic characters by photoperiod sensitivity and growth habit genes in soybean. Theoretical and Applied Genetics, 2010, 120(5): 1005-1012
    [5] Garner W W, Allard H A. Effect of the relative length of day and night and other factors of the environment on growth and reproduction in plants. Monthly Weather Review, 1920, 48(7): 415-415
    [6] Dong L, Li S, Wang L, Su T, Zhang C, Bi Y, Lai Y, Kong L, Wang F, Pei X, Li H, Hou Z, Du H, Du H, Li T, Cheng Q, Fang C, Kong F, Liu B. The genetic basis of high-latitude adaptation in wild soybean. Current Biology, 2023, 33(2): 252-262
    [7] Carpentieri-Pípolo V, Almeida L A, Kiihl R A. Inheritance of a long juvenile period under short-day conditions in soybean. Genetics and Molecular Biology, 2002,25 (4):463-469
    [8] 王源才. 超早熟、高蛋白大豆新品种“东农36”. 种子, 1983 (3): 84Wang Y C. New ultra-early-maturing, high-protein soybean variety ‘Dongnong 36’. Seed, 1983 (3): 84
    [9] Dong L, Cheng Q, Fang C, Kong L, Yang H, Hou Z, Li Y, Nan H, Zhang Y, Chen Q, Zhang C, Kou K, Su T, Wang L, Li S, Li H, Lin X, Tang Y, Zhao X, Lu S, Liu B, Kong F. Parallel selection of distinct Tof5 alleles drove the adaptation of cultivated and wild soybean to high latitudes. Molecular Plant, 2022, 15(2): 308-321
    [10] Xia Z, Watanabe S, Yamada T, Tsubokura Y, Nakashima H, Zhai H, Anai T, Sato S, Yamazaki T, Lü S, Wu H, Tabata S, Harada K. Positional cloning and characterization reveal the molecular basis for soybean maturity locus E1 that regulates photoperiodic flowering. Proceedings of the National Academy of Sciences of the United States of America, 2012, 109(32): E2155-E2164
    [11] Watanabe S, Xia Z, Hideshima R, Tsubokura Y, Sato S, Yamanaka N, Takahashi R, Anai T, Tabata S, Kitamura K, Harada K. A map-based cloning strategy employing a residual heterozygous line reveals that the GIGANTEA gene is involved in soybean maturity and flowering. Genetics, 2011, 188(2): 395-407
    [12] Watanabe S, Hideshima R, Xia Z, Tsubokura Y, Sato S, Nakamoto Y, Yamanaka N, Takahashi R, Ishimoto M, Anai T, Tabata S, Harada K. Map-based cloning of the gene associated with the soybean maturity locus E3. Genetics, 2009, 182(4): 1251-1262
    [13] Liu B, Kanazawa A, Matsumura H, Takahashi R, Harada K, Abe J. Genetic redundancy in soybean photoresponses associated with duplication of the phytochrome a gene. Genetics, 2008, 180(2): 995-1007
    [14] Zhao C, Takeshima R, Zhu J, Xu M, Sato M, Watanabe S, Kanazawa A, Liu B, Kong F, Yamada T, Abe J. A recessive allele for delayed flowering at the soybean maturity locus E9 is a leaky allele of FT2a, a FLOWERING LOCUS T ortholog. BMC Plant Biology, 2016, 16(1): 20
    [15] Samanfar B, Molnar S J, Charette M, Schoenrock A, Dehne F, Golshani A, Belzile F, Cober E R. Mapping and identification of a potential candidate gene for a novel maturity locus, E10, in soybean. Theoretical and Applied Genetics, 2017, 130(2): 377-390
    [16] Lu S, Dong L, Fang C, Liu S, Kong L, Cheng Q, Chen L, Su T, Nan H, Zhang D, Zhang L, Wang Z, Yang Y, Yu D, Liu X, Yang Q, Lin X, Tang Y, Zhao X, Yang X, Tian C, Xie Q, Li X, Yuan X, Tian Z, Liu B, Weller J L, Kong F. Stepwise selection on homeologous PRR genes controlling flowering and maturity during soybean domestication. Nature Genetics, 2020, 52(4): 428-436
    [17] Cao D, Li Y, Lu S, Wang J, Nan H, Li X, Shi D, Fang C, Zhai H, Yuan X, Anai T, Xia Z, Liu B, Kong F. GmCOL1a and GmCOL1b function as flowering repressors in soybean under long-day conditions. Plant and Cell Physiology, 2015, 56(12): 2409-2422
    [18] Xia Z J, Zhai H, Liu B H, Kong F J, Yuan X H, Wu H Y, Cober E R, Harada K. Molecular identification of genes controlling flowering time, maturity, and photoperiod response in soybean. Plant Systematics and Evolution, 2012, 298(7): 1217-1227
    [19] Casal J J, Candia A N, Sellaro R. Light perception and signalling by phytochrome A. Journal of Experimental Botany, 2014, 65(11): 2835-2845
    [20] Zhang Y, Lin X, Ma C, Zhao J, Shang X, Wang Z, Xu B, Gao N, Deng X W, Wang J. Structural insights into plant phytochrome A as a highly sensitized photoreceptor. Cell Research, 2023, 33(10): 806-809
    [21] Sheerin D J, Hiltbrunner A. Molecular mechanisms and ecological function of far-red light signalling. Plant, Cell & Environment, 2017, 40(11): 2509-2529
    [22] Casal J, Sanchez R, Yanovsky M. The function of phytochrome A. Plant, Cell & Environment, 1997, 20: 813-819
    [23] Cober E R, Tanner J W, Voldeng H D. Genetic control of photoperiod response in early-maturing, near-isogenic soybean lines. Crop Science, 1996, 36(3): 601-605
    [24] Kong F, Liu B, Xia Z, Sato S, Kim B M, Watanabe S, Yamada T, Tabata S, Kanazawa A, Harada K, Abe J. Two coordinately regulated homologs of FLOWERING LOCUS T are involved in the control of photoperiodic flowering in soybean. Plant Physiology, 2010, 154(3): 1220-1231
    [25] Singh N, Wu S, Tiwari V, Sehgal S, Raupp J, Wilson D, Abbasov M, Gill B, Poland J. Genomic analysis confirms population structure and identifies inter-lineage hybrids in Aegilops tauschii. Frontiers in Plant Science, 2019, 10:9
    [26] Martínez-Lüscher J, Brillante L, Kurtural S K. Flavonol profile is a reliable indicator to assess canopy architecture and the exposure of red wine grapes to solar radiation. Frontiers in Plant Science, 2019, 10:10
    [27] Zhang X, Zhai H, Wang Y, Tian X, Zhang Y, Wu H, Lü S, Yang G, Li Y, Wang L, Hu B, Bu Q, Xia Z. Functional conservation and diversification of the soybean maturity gene E1 and its homologs in legumes. Scientific Reports, 2016, 6(1): 29548
    [28] Thakare D, Kumudini S, Dinkins R D. The alleles at the E1 locus impact the expression pattern of two soybean FT-like genes shown to induce flowering in Arabidopsis. Planta, 2011, 234(5): 933-943
    [29] Takano M, Kanegae H, Shinomura T, Miyao A, Hirochika H, Furuya M. Isolation and characterization of rice phytochrome a mutants. The Plant Cell, 2001, 13(3): 521-534
    [30] Takano M, Inagaki N, Xie X, Yuzurihara N, Hihara F, Ishizuka T, Yano M, Nishimura M, Miyao A, Hirochika H, Shinomura T. Distinct and cooperative functions of phytochromes A, B, and C in the control of deetiolation and flowering in rice. The Plant Cell, 2005, 17(12): 3311-3325
    [31] Lin C. Photoreceptors and regulation of Flowering Time1. Plant Physiology, 2000, 123(1): 39-50
    [32] Weller J L, Murfet I C, Reid J B. Pea mutants with reduced sensitivity to far-red light define an important role for phytochrome A in day-length detection. Plant Physiology, 1997, 114(4): 1225-1236
    [33] Lin X, Dong L, Tang Y, Li H, Cheng Q, Li H, Zhang T, Ma L, Xiang H, Chen L, Nan H, Fang C, Lu S, Li J, Liu B, Kong F. Novel and multifaceted regulations of photoperiodic flowering by phytochrome A in soybean. Proceedings of the National Academy of Sciences of the United States of America, 2022, 119(41): e2208708119
    [34] Cober E R, Tanner J W, Voldeng H D. Soybean photoperiod-sensitivity loci respond differentially to light quality. Crop Science, 1996, 36(3): 606-610
    [35] Kanazawa A, Liu B, Kong F, Arase S, Abe J. Adaptive evolution involving gene duplication and insertion of a novel Ty1/copia-Like retrotransposon in soybean. Journal of Molecular Evolution, 2009, 69(2): 164-175
    [36] Tsubokura Y, Matsumura H, Xu M, Liu B, Nakashima H, Anai T, Kong F, Yuan X, Kanamori H, Katayose Y, Takahashi R, Harada K, Abe J. Genetic variation in soybean at the maturity locus E4 is involved in adaptation to long days at high latitudes. Agronomy, 2013, 3(1):117-134
    [37] Wan Z, Liu Y, Guo D, Fan R, Liu Y, Xu K, Zhu J, Quan L, Lu W, Bai X, Zhai H. CRISPR/Cas9-mediated targeted mutation of the E1 decreases photoperiod sensitivity, alters stem growth habits, and decreases branch number in soybean. Frontiers in Plant Science, 2022, 13:1066820
    [38] Zhao F, Lyu X, Ji R, Liu J, Zhao T, Li H, Liu B, Pei Y. CRISPR/Cas9-engineered mutation to identify the roles of phytochromes in regulating photomorphogenesis and flowering time in soybean. The Crop Journal, 2022, 10(6): 1654-1664
    [39] Xing H L, Dong L, Wang Z P, Zhang H Y, Han C Y, Liu B, Wang X C, Chen Q J. A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biology, 2014, 14(1): 327
    [40] Wang Y, Li Z, Chen X, Gu Y, Zhang L, Qiu L. An efficient soybean transformation protocol for use with elite lines. Plant Cell, Tissue and Organ Culture, 2022, 151(3): 457-466
    [41] Wang X,Chang X, Jing Y, Zhao J, Fang Q, Sun M, Zhang Y, Li W, Li Y. Identification and functional prediction of soybean CircRNAs involved in low-temperature responses. Journal of Plant Physiology, 2020, 250: 153188
    [42] Guo S, Zhang Z, Guo E, Fu Z, Gong J, Yang X. Historical and projected impacts of climate change and technology on soybean yield in China. Agricultural Systems, 2022, 203: 103522
    [43] Tan Q, Liu Y, Dai L, Pan T. Shortened key growth periods of soybean observed in China under climate change. Scientific Reports, 2021, 11(1): 8197
    [44] Bueno A F, Sutil W P, Jahnke S M, Carvalho G A, Cingolani M F, Colmenarez Y C, Corniani N. Biological control as part of the soybean integrated pest management (IPM): Potential and challenges. Agronomy, 2023,13(10): 2532
    [45] Kofsky J, Zhang H, Song B-H. Novel resistance strategies to soybean cyst nematode (SCN) in wild soybean. Scientific Reports, 2021, 11(1): 7967
    [46] Qin C, Li H, Zhang S, Lin X, Jia Z, Zhao F, Wei X, Jiao Y, Li Z, Niu Z, Zhou Y, Li X, Li H, Zhao T, Liu J, Li H, Lu Y, Kong F, Liu B. GmEID1 modulates light signaling through the evening complex to control flowering time and yield in soybean. Proceedings of the National Academy of Sciences of the United States of America, 2023, 120(15): e2212468120
    [47] Xu M, Xu Z, Liu B, Kong F, Tsubokura Y, Watanabe S, Xia Z, Harada K, Kanazawa A, Yamada T, Abe J. Genetic variation in four maturity genes affects photoperiod insensitivity and PHYA-regulated post-flowering responses of soybean. BMC Plant Biology, 2013, 13: 91
    Related
    Cited by
    Comments
    Comments
    分享到微博
    Submit
Get Citation
Share
Article Metrics
  • Abstract:0
  • PDF: 1
  • HTML: 0
  • Cited by: 0
History
  • Received:June 04,2024
  • Online: March 07,2025
Article QR Code
You are the 556203th visitor 京ICP备09069690号-23
® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.
Firefox, Chrome, IE10, IE11 are recommended. Other browsers are not recommended.