Crop and Pasture Science Crop and Pasture Science Society
Plant sciences, sustainable farming systems and food quality
RESEARCH ARTICLE

In vitro-assisted single-seed descent for breeding-cycle compression in subterranean clover (Trifolium subterraneum L.)

Maria Pazos-Navarro A C D , Marieclaire Castello A D , Richard G. Bennett A , Phillip Nichols A B and Janine Croser A
+ Author Affiliations
- Author Affiliations

A Centre for Plant Genetics and Breeding, UWA School of Agriculture and Environment, The University of Western Australia, 35 Stirling Highway, Crawley, WA 6009, Australia.

B Department of Agriculture and Food Western Australia, 3 Baron-Hay Court, South Perth, WA 6151, Australia.

C Corresponding author. Email: maria.pazosnavarro@uwa.edu.au

D These authors have joint first authorship.

Crop and Pasture Science 68(11) 958-966 https://doi.org/10.1071/CP17067
Submitted: 15 February 2017  Accepted: 5 May 2017   Published: 13 June 2017

Abstract

Subterranean clover (Trifolium subterraneum L.) is widely grown for its forage and ability to fix atmospheric nitrogen. Development of new varieties is constrained by the slow turnover time of generations, with only one generation per year possible under field conditions. We present an in vitro-assisted single-seed descent (IVASSD) technique, which enabled turnover of 2.7–6.1 generations per year across a diverse range of 27 T. subterraneum cultivars encompassing subspecies subterraneum, yanninicum and brachycalycinum. The IVASSD protocol accelerated the generation cycle in two ways: (i) time to floral initiation was minimised by growth under controlled temperature and extended photoperiod; and (ii) the seed-filling period was truncated and embryo and seed-coat dormancy avoided by the in vitro germination of immature seed on B5 medium plus L6KK overlay (0.525 mg gibberellic acid and 1.5 mg indole-butyric acid L–1). For the first time, an IVASSD system was validated on a full-scale breeding population with the production of 175 F7 recombinant inbred lines from an F4 population in less than one year. All F7 plants obtained were morphologically normal and fertile.

Additional keywords: annual pasture legumes, genetic resources, pasture breeding, rapid generation, RIL, seed dormancy.


References

Aitken Y (1955) Flower initiation in pasture legumes. II. Geographical implications of cold temperature requirements of varieties of Trifolium subterraneum L. Australian Journal of Agricultural Research 6, 245–257.
Flower initiation in pasture legumes. II. Geographical implications of cold temperature requirements of varieties of Trifolium subterraneum L.CrossRef |

Aitken Y (1974) ‘Flowering time, climate and genotype.’ (Melbourne University Press: Melbourne)

Asawaphan P, Mangkita W, Kachonpadungkitti Y, Matsuyama S, Satake T, Hisajima S (2005) Efficient flower induction from peanut (Arachis hypogaea L.) seedling in vitro. SABRAO. Journal of Breeding and Genetics 37, 131–140.

Berger JD, Buck R, Henzell JM, Turner NC (2005) Evolution in the genus Cicer—vernalisation response and low temperature pod set in chickpea (C. arietinum L.) and its annual wild relatives. Australian Journal of Agricultural Research 56, 1191–1200.
Evolution in the genus Cicer—vernalisation response and low temperature pod set in chickpea (C. arietinum L.) and its annual wild relatives.CrossRef |

Bermejo C, Gatti I, Cointry E (2016) In vitro embryo culture to shorten the breeding cycle in lentil (Lens culinaris Medik). Plant Cell, Tissue and Organ Culture 127, 585–590.
In vitro embryo culture to shorten the breeding cycle in lentil (Lens culinaris Medik).CrossRef | 1:CAS:528:DC%2BC28XhtlyjsLfK&md5=a0dd2366ee2940a57fb7e2554122d9d9CAS |

Bernier G, Périlleux C (2005) A physiological overview of the genetics of flowering time control. Plant Biotechnology Journal 3, 3–16.
A physiological overview of the genetics of flowering time control.CrossRef | 1:CAS:528:DC%2BD2MXhvFyqs7s%3D&md5=270c39e78140a09fe84de4d3835fa445CAS |

Castello M, Croser J, Lulsdorf M, Ramankutty P, Pradhan A, Nelson M, Real D (2015) Breaking primary dormancy in seeds of the perennial pasture legume tedera (Bituminaria bituminosa CH Stirt. vars albomarginata and crassiuscula). Grass and Forage Science 70, 365–373.
Breaking primary dormancy in seeds of the perennial pasture legume tedera (Bituminaria bituminosa CH Stirt. vars albomarginata and crassiuscula).CrossRef | 1:CAS:528:DC%2BC2MXmsFaktL4%3D&md5=cf973a4d4a74b671f121f95085989b73CAS |

Castello M, Stefanova K, Nichols P, Nutt B, Revell C, Croser J (2016) In vitro reproduction in the annual pasture legumes subterranean clover (Trifolium subterraneum L.) and French serradella (Ornithopus sativus Brot.). Grass and Forage Science 71, 79–89.
In vitro reproduction in the annual pasture legumes subterranean clover (Trifolium subterraneum L.) and French serradella (Ornithopus sativus Brot.).CrossRef | 1:CAS:528:DC%2BC28Xhtlygu7s%3D&md5=f023189c0f845781a7312d886ffae62cCAS |

Croser J, Pazos-Navarro M, Bennett RG, Tschirren S, Edwards K, Erskine W, Creasy R, Ribalta F (2016) Time to flower of temperate pulses in vivo and generation turnover in vivoin vitro of narrow-leaf lupin accelerated by low red to far-red ratio and high intensity in the far-red region. Plant Cell, Tissue and Organ Culture 127, 591–599.
Time to flower of temperate pulses in vivo and generation turnover in vivoin vitro of narrow-leaf lupin accelerated by low red to far-red ratio and high intensity in the far-red region.CrossRef | 1:CAS:528:DC%2BC28XhsFynt7rI&md5=61078355fc9699d865e0d126ac3cd048CAS |

Evans L (1959) Flower initiation in Trifolium subterraneum L. 1. Analysis of the partial processes involved. Crop & Pasture Science 10, 1–26.
Flower initiation in Trifolium subterraneum L. 1. Analysis of the partial processes involved.CrossRef |

Franklin G, Pius PK, Ignacimuthu S (2000) Factors affecting in vitro flowering and fruiting of green pea (Pisum sativum L.). Euphytica 115, 65–74.
Factors affecting in vitro flowering and fruiting of green pea (Pisum sativum L.).CrossRef | 1:CAS:528:DC%2BD3cXntFKqs7w%3D&md5=ebe88470337386c9f0c044943c1346b9CAS |

Gallardo K, Le Signor C, Vandekerckhove J, Thompson RD, Burstin J (2003) Proteomics of Medicago truncatula seed development establishes the time frame of diverse metabolic processes related to reserve accumulation. Plant Physiology 133, 664–682.
Proteomics of Medicago truncatula seed development establishes the time frame of diverse metabolic processes related to reserve accumulation.CrossRef | 1:CAS:528:DC%2BD3sXosVaqtL0%3D&md5=cc4c9010f4ee1e6971a83c9efae636b7CAS |

Gallardo K, Kurt C, Thompson Ochatt S (2006) In vitro culture of immature M. truncatula grains under conditions permitting embryo development comparable to that observed in vivo. Plant Science 170, 1052–1058.
In vitro culture of immature M. truncatula grains under conditions permitting embryo development comparable to that observed in vivo.CrossRef | 1:CAS:528:DC%2BD28XjvVWrsr0%3D&md5=fedad2cf5bb46a05494edb40f3f61c11CAS |

Gallardo K, Thompson R, Burstin J (2008) Reserve accumulation in legume seeds. Comptes Rendus Biologies 331, 755–762.
Reserve accumulation in legume seeds.CrossRef | 1:CAS:528:DC%2BD1cXht1Shtr7M&md5=9d280326b0a1646465208e6f5c5a0d44CAS |

Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Experimental Cell Research 50, 151–158.
Nutrient requirements of suspension cultures of soybean root cells.CrossRef | 1:CAS:528:DyaF1cXktVyqtLw%3D&md5=277335db89b7f0dc1043c39867df59b4CAS |

Ghamkhar K, Nichols PGH, Erskine W, Snowball R, Murillo M, Appels R, Ryan MH (2015) Hotspots and gaps in the world collection of subterranean clover (Trifolium subterraneum L.). Journal of Agricultural Science 153, 1069–1083.

Hirakawa H, Kaur P, Shirasawa K, Nichols P, Nagano S, Appels R, Erskine W, Isobe SN (2016) Draft genome sequence of subterranean clover, a reference for genus Trifolium. Scientific Reports 6, 30358
Draft genome sequence of subterranean clover, a reference for genus Trifolium.CrossRef | 1:CAS:528:DC%2BC28Xhtlyks7fL&md5=8924a454f3603bf92e674c5996b2f075CAS |

Kasten W, Paradise T, Kunert R, Straka P (1991) Progress in realization of interspecific hybrids in the genus Lupinus by means of an embryo rescue technique. Biologisches Zentralblatt 110, 301–309.

Kaur P, Bayer PE, Milec Z, Vrána J, Yuan Y, Appels R, Edwards D, Batley J, Nichols P, Erskine W, Doležel J (2017) An advance reference genome of Trifolium subterraneum L. revels genes related to agronomic performance. Plant Biotechnology Journal
An advance reference genome of Trifolium subterraneum L. revels genes related to agronomic performance.CrossRef |

Kim DH, Doyle MR, Sung S, Amasino RM (2009) Vernalization: Winter and the timing of flowering in plants. Annual Review of Cell and Developmental Biology 25, 277–299.
Vernalization: Winter and the timing of flowering in plants.CrossRef | 1:CAS:528:DC%2BD1MXhsVKisbvN&md5=370ccb231505fd752bd1a1f3a86d7b10CAS |

Kucera B, Cohn MA, Leubner-Metzger G (2005) Plant hormone interactions during seed dormancy release and germination. Seed Science Research 15, 281–307.
Plant hormone interactions during seed dormancy release and germination.CrossRef | 1:CAS:528:DC%2BD28XhsFGrtr0%3D&md5=09fd3b4783cee22fbeefed99de349ea1CAS |

Kumar AS, Gamborg OL, Nabors MW (1988) Plant regeneration from cell suspension cultures of Vigna aconitifolia. Plant Cell Reports 7, 138–141.
Plant regeneration from cell suspension cultures of Vigna aconitifolia.CrossRef | 1:CAS:528:DyaL1cXitFymur0%3D&md5=db8f0437ae840cc4197084f1c8dcf9d2CAS |

Landers KF (1995) Vernalization responses in narrow-leafed lupin (Lupinus angustifolius) genotypes. Australian Journal of Agricultural Research 46, 1011–1025.
Vernalization responses in narrow-leafed lupin (Lupinus angustifolius) genotypes.CrossRef |

Michaels SD, Amansino RM (2000) Memories of winter: vernalization and the competence to flower. Plant, Cell & Environment 23, 1145–1153.
Memories of winter: vernalization and the competence to flower.CrossRef |

Mobini SH, Lulsdorf M, Warkentin TD, Vandenberg A (2015) Plant growth regulators improve in vitro flowering and rapid generation advancement in lentil and faba bean. In Vitro Cellular & Developmental Biology - Plant 51, 71–79.
Plant growth regulators improve in vitro flowering and rapid generation advancement in lentil and faba bean.CrossRef | 1:CAS:528:DC%2BC2cXhsFKqsb%2FL&md5=1d07658eabde539d92cafeac1a72e28aCAS |

Nelson MN, Berger JD, Erskine W (2010) Flowering time control in annual legumes: prospects in a changing global climate. Plant Science Reviews 5, 49–62.

Nichols PGH, Revell CK, Humphries AW, Howie JH, Hall EJ, Sandral GA, Ghamkhar K, Harris CA (2012) Temperate pasture legumes in Australia—their history, current use and future prospects. Crop & Pasture Science 63, 691–725.
Temperate pasture legumes in Australia—their history, current use and future prospects.CrossRef |

Nichols P, Foster K, Piano E, Pecetti L, Kaur P, Ghamkhar K, Collins W (2013) Genetic improvement of subterranean clover (Trifolium subterraneum L.). 1. Germplasm, traits and future prospects. Crop & Pasture Science 64, 312–346.
Genetic improvement of subterranean clover (Trifolium subterraneum L.). 1. Germplasm, traits and future prospects.CrossRef | 1:CAS:528:DC%2BC3sXht12jt7nE&md5=6fbc60488e51a493330967888e195f1fCAS |

Nichols P, Jones RAC, Ridsdill-Smith TJ, Barbetti MJ (2014) Genetic improvement of subterranean clover (Trifolium subterraneum L.). 2. Breeding for disease and pest resistance. Crop & Pasture Science 65, 1207–1229.
Genetic improvement of subterranean clover (Trifolium subterraneum L.). 2. Breeding for disease and pest resistance.CrossRef | 1:CAS:528:DC%2BC2cXhvVOnsrvP&md5=01b16b14c4b7313c55dc99677da94dc2CAS |

Ochatt SJ, Sangwan RS (2010) In vitro flowering and seed set: Acceleration of generation cycles. In ‘Plant cell culture: essential methods’. (Eds MR Davey, P Anthony) pp. 97–110. (John Wiley & Sons: Chichester, UK)

Ochatt S, Sangwan R, Marget P, Ndong YA, Rancillac M, Perney P, Röbbelen G (2002) New approaches towards the shortening of generation cycles for faster breeding of protein legumes. Plant Breeding 121, 436–440.
New approaches towards the shortening of generation cycles for faster breeding of protein legumes.CrossRef |

Ochatt SJ, Benaddelmouna A, Marget P, Aubert G, Moussy F, Pontécaille C, Jacas L (2004) Overcoming hybridisation barriers between pea and some of its wild relatives. Euphytica 137, 353–359.
Overcoming hybridisation barriers between pea and some of its wild relatives.CrossRef | 1:CAS:528:DC%2BD2cXntlektbo%3D&md5=aff3f2e3281d5265ec9cac6fca3b4a62CAS |

Ribalta FM, Croser JS, Erskine W, Finnegan PM, Lulsdorf MM, Ochatt SJ (2014) Antigibberellin-induced reduction of internode length favors in vitro flowering and seed-set in different pea genotypes. Biologia Plantarum 58, 39–46.
Antigibberellin-induced reduction of internode length favors in vitro flowering and seed-set in different pea genotypes.CrossRef | 1:CAS:528:DC%2BC2cXht12ksbo%3D&md5=f8b2597e960c65acbfff6f7469c8662eCAS |

Ribalta FM, Pazos-Navarro M, Nelson K, Edwards K, Ross JJ, Bennett RG, Munday C, Erskine W, Ochatt SJ, Croser JS (2017) Precocious floral initiation and identification of exact timing of embryo physiological maturity facilitate germination of immature seeds to truncate the lifecycle of pea. Plant Growth Regulation 81, 345–353.
Precocious floral initiation and identification of exact timing of embryo physiological maturity facilitate germination of immature seeds to truncate the lifecycle of pea.CrossRef | 1:CAS:528:DC%2BC28XhsVSlsb7L&md5=49b67037e6aaf50f9c2260152ff26ab5CAS |

Sharma DR, Kaur R, Kumar K (1996) Embryo rescue in plants—a review. Euphytica 89, 325–337.

Surma M, Adamski T, Swiecicki W, Barzyk P, Kaczmarek Z, Kuczynska A, Krystkowiak K, Mikolajczak K, Ogrodowicz P (2013) Preliminary results of in vitro culture of pea and lupin embryos for the reduction of generation cycles in single seed descent technique. Acta Societatis Botanicorum Poloniae 82, 231–236.
Preliminary results of in vitro culture of pea and lupin embryos for the reduction of generation cycles in single seed descent technique.CrossRef |

Taylor G (2005) Hardseededness in Mediterranean annual pasture legumes in Australia: a review. Crop & Pasture Science 56, 645–661.
Hardseededness in Mediterranean annual pasture legumes in Australia: a review.CrossRef |

Tennant D (1965) The differential rate of seed development in Dwalganup and Geraldton varieties of subterranean clover. Australian Journal of Experimental Agriculture and Animal Husbandry 5, 46–48.
The differential rate of seed development in Dwalganup and Geraldton varieties of subterranean clover.CrossRef |



Rent Article (via Deepdyve) Export Citation