10.5061/DRYAD.K98SF7M72
Fant, Jeremie
0000-0001-9276-1111
Chicago Botanic Garden
Walsh, Seana
0000-0002-4829-4488
National Tropical Botanical Garden
Foster, Jeremy
0000-0002-9426-4212
Northwestern University
Genetic and fitness measurements of Brighamia accessions for APPS
Dryad
dataset
2021
FOS: Biological sciences
microsatellite markers
parentage analysis
accession
Institute of Museum and Library Services
https://ror.org/030prv062
MG-30-16-0085-16
Institute of Museum and Library Services
https://ror.org/030prv062
MG-60-19-0064-19
National Geographic Society
https://ror.org/04bqh5m06
NGS-57037C-19
Botanic Garden*
2021-12-21T00:00:00Z
2021-12-21T00:00:00Z
en
41754 bytes
7
CC0 1.0 Universal (CC0 1.0) Public Domain Dedication
Premise of the study: Living collections maintained for generations are at
risk of diversity loss, inbreeding, and adaptation to cultivation. To
address these concerns the zoo community uses pedigrees to track
individuals and implement crosses that maximize founder contributions and
minimize inbreeding. Using a pedigree management approach in an
exceptional plant, we demonstrate how conducting such strategic crosses
can minimize genetic issues that have arisen under current practices.
Methods: We performed crosses between these collections and compared the
fitness of progeny, including plant performance and reproductive health.
We genotyped the progeny and paternal accessions to measure changes in
diversity and relatedness within and between accessions. Results: The mean
relatedness among individuals of an accession, suggests they are full
siblings. As a result there was high inbreeding and low diversity within
an accession, although less so among accessions. Progeny from the wider
crosses had increased genetic diversity, while selfed accessions were
smaller and less fertile. Discussion: Institutions which hold exceptional
species should consider how diversity is maintained within their
collections. Implementing a pedigree-based approach to managing
reproduction of ex situ plants will slow the inevitable loss of genetic
diversity and in turn, result in healthier collections.
Genomic DNA was extracted from parental accessions using the 2× CTAB
method (Doyle and Doyle, 1987) and the progeny of the aforementioned
crosses using DNeasy Plant Mini Kit (Cat 69104, Qiagen, Germantown, MD,
USA). For the parental accession plants, we extracted DNA from the
maternal accession (CBG), and for three of the four paternal accessions
(SDZ (Accession 2001-0273-022), NTBG (Accession 990833.3) and USBG1
(Accession 2014-0070)), however as the fourth accession (USBG2) died
before we could extract DNA, we used progeny derived (Accession 2019-000)
from a self of that plant to estimate paternal contribution. All samples
were genotyped using markers and protocols described in Fant et al. (2019)
which had already been tested for Hardy-weinberg and presence of null
alleles. For this study we used a total of 11 primers; eight primers (B05,
B08, B43, B44, B46, B47, B51, B57) were designed for B. insignis (Fant et
al., 2019) and three primers (L23, L33, L34) were designed for Lobelia
villosa (Rock) H. St. John & Hosaka. The eight B. insignis primers
were visualized using pre-labeled forward primers with either WellRed
Black (D2), Green (D3) or Blue (D4) fluorescent dye (Sigma-Proligo, St.
Louis, Missouri, USA), while for Lobelia villosa markers, the forward
primer was modified at the 5’ end (5’-CACGACGTTGTAAAACGAC-3’) so they
could be labelled separately (Schuelke, 2000). All products were analyzed
and scored using a CEQ 8000 Genetic Analysis System V9.0 (Beckman Coulter,
Brea, California, USA). Given that B. insignis is a paleotetraploid
(Lammers, 1988), four of the primer pairs (B44, B47, B51 and L23) produced
more than two bands. As the peaks were separated from other alleles by
large range (20-30bp), and segregated independently (Fant et al., 2019),
they were scored as independent loci.
Ready to be used in genalex.