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Australian Journal of Botany Australian Journal of Botany Society
Southern hemisphere botanical ecosystems
RESEARCH ARTICLE

Whole-chloroplast analysis as an approach for fine-tuning the preservation of a highly charismatic but critically endangered species, Wollemia nobilis (Araucariaceae)

Abigail Greenfield A B , Hannah McPherson B E , Tony Auld C , Sven Delaney A , Catherine A. Offord D , Marlien van der Merwe B , Jia-Yee S. Yap B and Maurizio Rossetto B
+ Author Affiliations
- Author Affiliations

A School of Biotechnology and Biomolecular Biosciences, University of New South Wales, Kensington, NSW 2033, Australia.

B National Herbarium of NSW, Royal Botanic Garden Sydney, Mrs Macquaries Road, Sydney, NSW 2000, Australia.

C NSW Office of Environment and Heritage, 43 Bridge Street, Hurstville, NSW 2220, Australia.

D The Australian PlantBank, Australian Botanic Garden, Narellan Road, Mount Annan, NSW, 2567, Australia.

E Corresponding author. Email: hannah.mcpherson@rbgsyd.nsw.gov.au

Australian Journal of Botany 64(8) 654-658 https://doi.org/10.1071/BT16105
Submitted: 20 May 2016  Accepted: 19 September 2016   Published: 31 October 2016

Abstract

The critically endangered Wollemia nobilis W.G. Jones, K.D. Hill & J.M. Allen is endemic to Wollemi National Park north of Sydney (Australia). All known wild individuals are restricted to four sites in a single canyon system. W. nobilis can reproduce sexually but at all sites individual clumps can be multi-stemmed from a common base. In the first genetic study of this species, no genetic variation was found across multiple genetic marker types representing hundreds of nuclear loci, indicating this species is characterised by very low genetic variation. In this study we searched for variation across the chloroplast using shotgun sequencing, bioinformatic extraction of chloroplast DNA and variant detection. Six chloroplast single nucleotide polymorphisms were detected, producing three chlorotypes. Chlorotype 1 is found in every individual surveyed at Sites 1 and 3, and in individuals from Sites 2 and 4. Chlorotype 2 – the most distinct chlorotype – was found in two individuals from Site 4. Chlorotype 3 consists of a single difference from Chlorotype 1 and may represent a somaclonal mutant. These findings will guide management and translocation of this critically endangered species. This study provides a practical template that is highly informative and easily applicable to other taxa in similar circumstances.

Additional keywords: Araucariaceae, chloroplast genome, conservation genetics, endangered species, rainforest tree.


References

Bullock S, Summerell BA, Gunn LV (2000) Pathogens of the Wollemi pine, Wollemia nobilis. Australasian Plant Pathology 29, 211–214.
Pathogens of the Wollemi pine, Wollemia nobilis.Crossref | GoogleScholarGoogle Scholar |

Burrows GE (1999) Wollemi pine (Wollemia nobilis, Araucariaceae) possesses the same unusual leaf axil anatomy as the other investigated members of the family. Australian Journal of Botany 47, 61–68.
Wollemi pine (Wollemia nobilis, Araucariaceae) possesses the same unusual leaf axil anatomy as the other investigated members of the family.Crossref | GoogleScholarGoogle Scholar |

Burrows GE, Meagher PF, Heady RD (2007) An anatomical assessment of branch abscission and branch-base hydraulic architecture in the endangered Wollemia nobilis. Annals of Botany 99, 609–623.
An anatomical assessment of branch abscission and branch-base hydraulic architecture in the endangered Wollemia nobilis.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD2s7nvFyhug%3D%3D&md5=441c55351b8b741a866a5ce06febe29aCAS | 17272303PubMed |

Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin 19, 1–15.

Durzan D (2012) Female parthenogenetic apomixis and androsporogenetic parthenogenesis in embryonal cells of: interpolation of progenesis and asexual heterospory in an artificial sporangium. Sexual Plant Reproduction 25, 227–246.
Female parthenogenetic apomixis and androsporogenetic parthenogenesis in embryonal cells of: interpolation of progenesis and asexual heterospory in an artificial sporangium.Crossref | GoogleScholarGoogle Scholar | 22669467PubMed |

Ebert D, Peakall R (2009) A new set of universal de novo sequencing primers for extensive coverage of noncoding chloroplast DNA: new opportunities for phylogenetic studies and cpSSR discovery. Molecular Ecology Resources 9, 777–1075.
A new set of universal de novo sequencing primers for extensive coverage of noncoding chloroplast DNA: new opportunities for phylogenetic studies and cpSSR discovery.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXls1Ggur0%3D&md5=6e9c3390f1aa0d300755c651561c4bffCAS | 21564742PubMed |

Gilmore S, Hill KD (1997) Relationships of the Wollemi pine (Wollemia nobilis) and a molecular phylogeny of the Araucariaceae. Telopea 7, 275–291.
Relationships of the Wollemi pine (Wollemia nobilis) and a molecular phylogeny of the Araucariaceae.Crossref | GoogleScholarGoogle Scholar |

Heady RD, Banks JG, Evans PD (2002) Wood anatomy of Wollemi pine. The International Association of Wood Anatomists 23, 339–357.

Hill KD (1997) Architecture of the Wollemi pine (Wollemia nobilis, Araucariaceae), a unique combination of model and reiteration. Australian Journal of Botany 45, 817–826.
Architecture of the Wollemi pine (Wollemia nobilis, Araucariaceae), a unique combination of model and reiteration.Crossref | GoogleScholarGoogle Scholar |

Li X, Yang Y, Henry R, Rossetto M, Wang Y, Chen S (2015) Plant DNA barcoding: from gene to genome. Biological Reviews of the Cambridge Philosophical Society 90, 157–166.
Plant DNA barcoding: from gene to genome.Crossref | GoogleScholarGoogle Scholar | 24666563PubMed |

Macphail M, Carpenter RJ, Iglesias A, Wilf P (2013) First evidence for Wollemi pine-type pollen (Dilwynites: Araucariaceae) in South America. PlosOne 8, e69281
First evidence for Wollemi pine-type pollen (Dilwynites: Araucariaceae) in South America.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3sXht1aju7rO&md5=dd47fa430a72f043f5f51a7e068e8b6cCAS |

McGee PA, Bullock S, Summerell BA (1999) Structure of mycorrhizae of the Wollemi pine (Wollemia nobilis) and related Araucariaceae. Australian Journal of Botany 47, 85–95.
Structure of mycorrhizae of the Wollemi pine (Wollemia nobilis) and related Araucariaceae.Crossref | GoogleScholarGoogle Scholar |

McLoughlin S, Vajda V (2005) Ancient Wollemi pines resurgent ten years after its discovery, a vanishingly rare tree from the Cretaceous Period is a scientific darling and may soon become a commercial success too. American Scientist 93, 540–547.
Ancient Wollemi pines resurgent ten years after its discovery, a vanishingly rare tree from the Cretaceous Period is a scientific darling and may soon become a commercial success too.Crossref | GoogleScholarGoogle Scholar |

McPherson H, van der Merwe M, Delaney SK, Edwards MA, Henry RJ, McIntosh E, Rymer PD, Milner ML, Siow J, Rossetto M (2013) Capturing chloroplast variation for molecular ecology studies: a simple next generation sequence approach applied to a rainforest tree. Molecular Ecology 13, 8
Capturing chloroplast variation for molecular ecology studies: a simple next generation sequence approach applied to a rainforest tree.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3sXmslSjsbw%3D&md5=69e9375bae9141125f1ea751f10e2ceaCAS |

Mogie M, Mogie M (1992) ‘The evolution of asexual reproduction in plants.’ (Chapman & Hall: London)

NSW Department of Environment and Conservation (2006) Wollemi Pine (Wollemia nobilis) Rocovery Plan. NSW Department of Environment and Conservation, Hurstville NSW.

Offord CA, Porter CL, Meagher PF, Errington G (1999) Sexual reproduction and early plant growth of the Wollemi pine (Wollemia nobilis), a rare and threatened Australian conifer. Annals of Botany 84, 1–9.
Sexual reproduction and early plant growth of the Wollemi pine (Wollemia nobilis), a rare and threatened Australian conifer.Crossref | GoogleScholarGoogle Scholar |

Peakall R, Ebert D, Scott LJ, Meagher PF, Offord CA (2003) Comparative genetic study confirms exceptionally low genetic variation in the ancient and endangered relictual conifer, Wollemia nobilis (Araucariaceae). Molecular Ecology 12, 2331–2343.
Comparative genetic study confirms exceptionally low genetic variation in the ancient and endangered relictual conifer, Wollemia nobilis (Araucariaceae).Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXnvVSgt7w%3D&md5=bcd3ef7dfc4045edc519badc01a239deCAS | 12919472PubMed |

Rossetto M, McPherson H, Siow J, Kooyman R, van der Merwe M, Wilson PD (2015) Where did all the trees come from? A novel multispecies approach reveals the impacts of biogeographical history and functional diversity on rain forest assembly. Journal of Biogeography 42, 2172–2186.
Where did all the trees come from? A novel multispecies approach reveals the impacts of biogeographical history and functional diversity on rain forest assembly.Crossref | GoogleScholarGoogle Scholar |

Schlögl PS, de Souza AP, Nodari RO (2007) PCR-RFLP analysis of non-coding regions of cpDNA in Araucaria angustifolia (Bert.) O. Kuntze. Genetics and Molecular Biology 30, 423–427.
PCR-RFLP analysis of non-coding regions of cpDNA in Araucaria angustifolia (Bert.) O. Kuntze.Crossref | GoogleScholarGoogle Scholar |

Slippers B, Summerell B, Crous PW, Coutinho TA, Wingfield BD, Wingfield MJ (2005) Preliminary studies on Botryosphaeria species from Southern Hemisphere conifers in Australasia and South Africa. Australasian Plant Pathology 34, 213–220.
Preliminary studies on Botryosphaeria species from Southern Hemisphere conifers in Australasia and South Africa.Crossref | GoogleScholarGoogle Scholar |

Stewart CN (1997) Rapid DNA extraction from plants. In ‘Fingerprinting methods based on arbitrarily primed PCR’. (Eds MR Micheli, R Bova) pp. 25–28. (Springer Verlag: Heidelberg)

Straub SCK, Parks M, Weitemier K, Fishbein M, Cronn R, Liston A (2012) Navigating the tip of the genomic iceberg: next-generation sequencing for plant systematics. American Journal of Botany 99, 349–364.
Navigating the tip of the genomic iceberg: next-generation sequencing for plant systematics.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XksValtbo%3D&md5=4d16a090239b010ed88444724f56d1aaCAS |

Wagner DB (1992) Nuclear, chloroplast, and mitochondrial DNA polymorphisms as biochemical markers in population genetic analyses of forest trees. In ‘Population genetics of forest trees. Proceedings of the international symposium on population genetics of forest trees’. (Eds WT Adams, SH Strauss, DL Copes, AR Griffin) pp. 373–390. (Springer: Berlin)

Yap J-YS, Rohner T, Greenfield A, van der Merwe M, McPherson H, Glenn W, Kornfeld G, Marendy E, Pan AYH, Wilton A, Wilkins MR, Rossetto M, Delaney SK (2015) Complete chloroplast genome of the Wollemi pine (Wollemia nobilis): structure and evolution. PLoS One 10, e0128126
Complete chloroplast genome of the Wollemi pine (Wollemia nobilis): structure and evolution.Crossref | GoogleScholarGoogle Scholar |