Cytogenetic, chromosome count optimization and automation of Neolamarckia cadamba (Rubiaceae) root tips derived from in vitro mutagenesis

Ho, Wei-Seng; Eng, Wee-Hiang; Ling, Kwong-Hung

doi:10.15835/nsb13310995

Cited by 2 publications

(2 citation statements)

References 31 publications

(55 reference statements)

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“…The staining process was carried out at room temperature. Feulgen stain was applied on the root tips for 30 min in dark condition before squashing with a metal bar in a drop of aceto-orcein 1% stain followed by thumb squash ( Eng, Ho & Ling, 2020 ).…”

Section: Methodsmentioning

confidence: 99%

In vitro induction and identification of polyploid Neolamarckia cadamba plants by colchicine treatment

Eng

Ling³

2021

PeerJ

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Polyploidization has played a crucial role in plant breeding and crop improvement. However, studies on the polyploidization of tropical tree species are still very scarce in this region. This paper described the in vitro induction and identification of polyploid plants of Neolamarckia cadamba by colchicine treatment. N. cadamba belongs to the Rubiaceae family is a natural tetraploid plant with 44 chromosomes (2n = 4x = 44). Nodal segments were treated with colchicine (0.1%, 0.3% and 0.5%) for 24 h and 48 h before transferring to shoot regeneration medium. Flow cytometry (FCM) and chromosome count were employed to determine the ploidy level and chromosome number of the regenerants, respectively. Of 180 colchicine-treated nodal segments, 39, 14 and 22 were tetraploids, mixoploids and octoploids, respectively. The highest percentage of polyploidization (20% octoploids; 6.7% mixoploids) was observed after treated with 0.3% colchicine for 48 h. The DNA content of tetraploid (4C) and octoploid (8C) was 2.59 ± 0.09 pg and 5.35 ± 0.24 pg, respectively. Mixoploid plants are made up of mixed tetraploid and octoploid cells. Chromosome count confirmed that tetraploid cell has 44 chromosomes and colchicine-induced octoploid cell has 88 chromosomes. Both octoploids and mixoploids grew slower than tetraploids under in vitro conditions. Morphological characterizations showed that mixoploid and octoploid leaves had thicker leaf blades, thicker midrib, bigger stomata size, lower stomata density, higher SPAD value and smaller pith layer than tetraploids. This indicates that polyploidization has changed and resulted in traits that are predicted to increase photosynthetic capacity of N. cadamba. These novel polyploid plants could be valuable resources for advanced N. cadamba breeding programs to produce improved clones for planted forest development.

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Section: Methodsmentioning

confidence: 99%

In vitro induction and identification of polyploid Neolamarckia cadamba plants by colchicine treatment

Eng

Ling³

2021

PeerJ

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“…From the morphological characters, N. cadamba is classified into subfamily Cinchonoideae, tribe Naucleeae 1 . Based on the cytogenetic study 35 , 36 , N. cadamba has 44 chromosomes (2n) and belongs to subfamily Cinchonoideae, tribe Naucleeae and Subtribes Neolamarckinae 37 . In this study, we determined the complete mtDNA sequence of N. cadamba and detailed its characteristics.…”

Section: Introductionmentioning

confidence: 99%

A complete sequence of mitochondrial genome of Neolamarckia cadamba and its use for systematic analysis

Wang

Chen

et al. 2021

Sci Rep

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Neolamarckia cadamba is an important tropical and subtropical tree for timber industry in southern China and is also a medicinal plant because of the secondary product cadambine. N. cadamba belongs to Rubiaceae family and its taxonomic relationships with other species are not fully evaluated based on genome sequences. Here, we report the complete sequences of mitochondrial genome of N. cadamba, which is 414,980 bp in length and successfully assembled in two genome circles (109,836 bp and 305,144 bp). The mtDNA harbors 83 genes in total, including 40 protein-coding genes (PCGs), 31 transfer RNA genes, 6 ribosomal RNA genes, and 6 other genes. The base composition of the whole genome is estimated as 27.26% for base A, 22.63% for C, 22.53% for G, and 27.56% for T, with the A + T content of 54.82% (54.45% in the small circle and 54.79% in the large circle). Repetitive sequences account for ~ 0.14% of the whole genome. A maximum likelihood (ML) tree based on DNA sequences of 24 PCGs supports that N. cadamba belongs to order Gentianales. A ML tree based on rps3 gene of 60 species in family Rubiaceae shows that N. cadamba is more related to Cephalanthus accidentalis and Hymenodictyon parvifolium and belongs to the Cinchonoideae subfamily. The result indicates that N. cadamba is genetically distant from the species and genera of Rubiaceae in systematic position. As the first sequence of mitochondrial genome of N. cadamba, it will provide a useful resource to investigate genetic variation and develop molecular markers for genetic breeding in the future.

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Cytogenetic, chromosome count optimization and automation of Neolamarckia cadamba (Rubiaceae) root tips derived from in vitro mutagenesis

Cited by 2 publications

References 31 publications

In vitro induction and identification of polyploid Neolamarckia cadamba plants by colchicine treatment

In vitro induction and identification of polyploid Neolamarckia cadamba plants by colchicine treatment

A complete sequence of mitochondrial genome of Neolamarckia cadamba and its use for systematic analysis

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