The Genetic Diversity of Bletilla spp. Based on SLAF-seq and Oligo-FISH

Abstract

Bletilla spp.Rchb. F. is a traditional Chinese medicinal material. In this study, Bletilla striata (Thunb. ex A. Murray) Rchb F, Bletilla formosana (Hayata) Schltr, and Bletilla ochracea Schltr were collected to analyze the genetic diversity of 16 materials using specific site-amplified fragment sequencing (SLAF-seq) and fluorescence in situ hybridization (FISH). The results showed that the phylogenetic tree of the single-nucleotide polymorphism (SNP) data rendering system was correlated with the shape and geographical distribution of the material. The results of the population structural analysis showed that all the materials containing yellow labellum came from the same ancestor. The results of the principal component analysis were able to preliminarily judge the genetic distance and provided a reference for the selection of hybrid parents. The FISH analysis showed that the chromosomes of B. striata were 2n = 32 and the chromosomes of the B. striata (safflower) mutant were 2n = 34 and the chromosomes of B. ochracea and B. formosana were 2n = 34–36. The (AG3T3)3 non-terminal signal was different from the 5S rDNA signal. These results revealed that the 16 materials had rich genetic diversity, which can provide molecular and cytogenetic data for the study of the genus and its relatives and serve as a reference for the breeding of new genus varieties and improve breeding efficiency and cost.

Karyotype Description and Comparative Chromosomal Mapping of 5S rDNA in 21 species

“…Based on this, the karyotype formula and cytotype are obtained to compare whether the species is evolving (Alberts et al 2002). In this study, the chromosome numbers in (Miduno 1954, He et al 2022c, Huan et al 2022, Yang et al 2023; iv) J. regia and J. sigillata (2n = 34) (Luo and Chen 2020), Juglans (2n = 32) (Woodworth 1930, Hans 1970, Tulecke et al 1988, Mu and Xi 1988, Mu et al 1990 sigillata (all 2n = 34). These results contradict previous studies.…”

“…(2n = 28), P. cyrtonema (2n = 22) (Sun 1996, Chen and Zhou 2005); ii ) Zanthoxylum acanthopodium Candelle (2n = 64), Zanthoxylum dimorphophyllum Hemsley (2n = 36/68), Zanthoxylum scandens Blume (2n = 68), Zanthoxylum oxyphyllum Edgeworth (2n = 72), Zanthoxylum tomentellum J.D. Hance (2n = 72), Zanthoxylum simulans Hance (2n =∼132, Z. nitidum (2n = 68), Z. armatum (2n = 66/98/128/132/136), Z. bungeanum (2n = 132/136) (Zhang and Hartley 2008, Chen et al 2009, Yu et al 2010, Luo et al 2018d, Luo et al 2022b, He et al 2023, Hu et al 2023); iii ) Bletilla formosana (2n = 32/36), B. striata (2n = 32/34/36/48/51/64/76), B. ochracea (2n = 34/36) (Miduno 1954, He et al 2022c, Huan et al 2022, Yang et al 2023); iv ) J. regia and J. sigillata (2n = 34) (Luo and Chen 2020), Juglans (2n = 32) (Woodworth 1930, Hans 1970, Tulecke et al 1988, Mu and Xi 1988, Mu et al 1990). In this study, P. cyrtonema (2n = 18), Z. bungeanum (2n = 76/134/136/160), Z. armatum (2n = 96/100/102/104/132), Z. nitidum (2n = 66), B. formosana, B. striata , B. ochracea , J. regia, and J. sigillata (all 2n = 34).…”

“…On the contrary, it was different from the 5S rDNA signal pattern of F. pennsylvanica , R. pseudoacacia , S. japonicum , T. wallichiana var. mairei , Z. bungeanum ‘Hanyuanhuajiao’, two types of B. ochracea , and two varieties of Z. armatum (Luo and Liu 2019, He et al 2022c, Huan et al 2022, He et al 2023) from previous studies. The possible causes of the 5S rDNA signal pattern discrepancy are i ) lost satellite chromosome with signal and ii ) variation in different batches of materials in the same species (i.e., intraspecific variation).…”

“…The copyright holder for this this version posted February 9, 2024. ; https://doi.org/10.1101/2024.02.08.579556 doi: bioRxiv preprint 9 Z. armatum (Luo and Liu 2019, He et al 2022c, Huan et al 2022, He et al 2023) from previous studies. The possible causes of the 5S rDNA signal pattern discrepancy are i) lost satellite chromosome with signal and ii) variation in different batches of materials in the same species (i.e., intraspecific variation).…”

“…sinensis and three Hippophaë rhamnoides L. cultivars of Elaeagnaceae (Luo et al 2022a); in Fabaceae of six Hedysarum L. species (Yurkevich et al 2022); in Juglans regia L. and Juglans sigillata Dode (Luo and Chen) of Juglandaceae; in four Glechoma L. species of Lamiaceae (Jang et al 2016); in three Bletilla Rchb. f. species of Orchidaceae (Huan et al 2022); in three Zea L. species of Poaceae (Han et al 2003); in four Citrus L. species of Rutaceae (He et al 2020), Zanthoxylum bungeanum Maxim. and Z. armatum (Hu et al 2023); in four Populus L. species of Salicaceae (Xin et al 2019); in five Taxus L. species of Taxaceae (He et al 2022b).…”