scholarly journals Oryzalin-induced Tetraploidy in Cryptomeria japonica (Cupressaceae)

HortScience ◽  
2010 ◽  
Vol 45 (2) ◽  
pp. 316-319 ◽  
Author(s):  
Ryan N. Contreras ◽  
John M. Ruter ◽  
Brian M. Schwartz
Keyword(s):  
Flow Cytometry ◽  
Environmental Conditions ◽  
Cryptomeria Japonica ◽  
Japanese Cedar ◽  
Phenotypic Marker ◽  
Ploidy Levels ◽  
Random Subset ◽  
Laboratory Conditions

Japanese-cedar [Cryptomeria japonica (L.f.) D. Don] represents an alternative to leyland cypress [×Cuprocyparis leylandii (A.B. Jacks. & Dallim.) Farjon] as an evergreen screen or specimen plant for landscapes. It performs well under a range of soil and environmental conditions but has been underused attributable, in part, to unsightly winter browning caused by photoinhibition. In previous studies, chance seedlings that did not exhibit winter browning were identified as tetraploids. The current study was conducted to induce polyploidy in japanese-cedar. Approximately 600 seedlings were sprayed with 150 μM oryzalin + 0.1% SilEnergy™ for 30 consecutive days under laboratory conditions. Two hundred thirty-seven seedlings with thickened and twisted leaves were selected, transplanted, and grown in a glasshouse for 120 days. Seedling ploidy levels were analyzed using flow cytometry 180 days after treatment (DAT), identifying 197 (83.1%) tetraploids, 22 (9.3%) cytochimeras, and 18 (7.6%) diploids. Morphology of induced tetraploids was similar to that previously described and provided a phenotypic marker during selection that was over 92% accurate. A random subset of 20 tetraploid individuals was analyzed 270 DAT and were found to contain only tetraploid cells in the leaves analyzed, confirming stability over this period. This study demonstrated the use of oryzalin for inducing tetraploids in japanese-cedar, which we predict will be effective in other gymnosperms.

scholarly journals Differences in Winter Browning among Japanese-cedar Cultivars Are Not Due to Variation in Ploidy Levels

HortScience ◽  
2011 ◽  
Vol 46 (11) ◽  
pp. 1465-1467
Author(s):  
Ryan N. Contreras ◽  
Ron Determann ◽  
Mara Friddle
Keyword(s):  
Flow Cytometry ◽  
Enzyme Activity ◽  
Quantitative Analysis ◽  
Growth Form ◽  
Japanese Cedar ◽  
Antioxidant Enzyme Activity ◽  
Flow Cytometry Analysis ◽  
Chromosome Counts ◽  
Ploidy Levels ◽  
Number Of Authors

There is a great deal of variation among japanese-cedar cultivars with regard to growth form, foliar characteristics, and winter browning. Differences in winter browning have been observed and documented by a number of authors. Previous research has established that there are differences in winter foliage color between cultivars included in the current study; however, no quantitative analysis under standardized conditions was conducted. Because of a previous report that tetraploid forms of japanese-cedar remain green during winter as a result of increased antioxidant enzyme activity, we hypothesized that cultivars that exhibit reduced winter browning were polyploids. We screened 56 accessions of japanese-cedar using flow cytometry analysis of 4′,6-diamidino-2-phenylindole (DAPI)-stained nuclei and performed chromosome counts on three cultivars. All accessions were diploid (2n = 2x = 22), although there were significant differences in genome sizes among the cultivars. Holoploid genome sizes ranged from 18.9 pg for var. sinensis JCRA to 22.3 pg for ‘Viridis’ with a mean of 20.1 pg. Chromosome counts for cultivars Ogon, Oye Keme, and Viridis supported the flow cytometry results. Although the underlying cause of the variability in morphology and winter browning among cultivars is unclear, our results show that differences in ploidy level are not responsible, because all tested genotypes were diploid. Chemical name: 4′,6-diamidino-2-phenylindole (DAPI).

scholarly journals Flow Cytometry-Based Determination of Ploidy from Dried Leaf Specimens in Genomically Complex Collections of the Tropical Forage Grass Urochloa s. l.

Genes ◽  
2021 ◽  
Vol 12 (7) ◽  
pp. 957
Author(s):  
Paulina Tomaszewska ◽  
Till K. Pellny ◽  
Luis M. Hernández ◽  
Rowan A. C. Mitchell ◽  
Valheria Castiblanco ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Germplasm Collection ◽  
Forage Grass ◽  
Breeding Programs ◽  
Robust Identification ◽  
Ploidy Levels ◽  
Sustainable Improvement ◽  
Tropical Forage ◽  
Relative Differences

Urochloa (including Brachiaria, Megathyrus and some Panicum) tropical grasses are native to Africa and are now, after selection and breeding, planted worldwide, particularly in South America, as important forages with huge potential for further sustainable improvement and conservation of grasslands. We aimed to develop an optimized approach to determine ploidy of germplasm collection of this tropical forage grass group using dried leaf material, including approaches to collect, dry and preserve plant samples for flow cytometry analysis. Our methods enable robust identification of ploidy levels (coefficient of variation of G0/G1 peaks, CV, typically <5%). Ploidy of some 348 forage grass accessions (ploidy range from 2x to 9x), from international genetic resource collections, showing variation in basic chromosome numbers and reproduction modes (apomixis and sexual), were determined using our defined standard protocol. Two major Urochloa agamic complexes are used in the current breeding programs at CIAT and EMBRAPA: the ’brizantha’ and ’humidicola’ agamic complexes are variable, with multiple ploidy levels. Some U. brizantha accessions have odd level of ploidy (5x), and the relative differences in fluorescence values of the peak positions between adjacent cytotypes is reduced, thus more precise examination of this species is required. Ploidy measurement of U. humidicola revealed aneuploidy.

scholarly journals CRISPR/Cas9-mediated targeted mutagenesis in Japanese cedar (Cryptomeria japonica D. Don)

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yoshihiko Nanasato ◽  
Masafumi Mikami ◽  
Norihiro Futamura ◽  
Masaki Endo ◽  
Mitsuru Nishiguchi ◽  
...  
Keyword(s):  
Genome Editing ◽  
Cryptomeria Japonica ◽  
Fluorescent Protein ◽  
Japanese Cedar ◽  
Targeted Mutagenesis ◽  
Embryogenic Tissue ◽  
Breeding Period ◽  
Single Mutation ◽  
Knock Out ◽  
Guide Rna

AbstractCryptomeria japonica (Japanese cedar or sugi) is one of the most important coniferous tree species in Japan and breeding programs for this species have been launched since 1950s. Genome editing technology can be used to shorten the breeding period. In this study, we performed targeted mutagenesis using the CRISPR/Cas9 system in C. japonica. First, the CRISPR/Cas9 system was tested using green fluorescent protein (GFP)-expressing transgenic embryogenic tissue lines. Knock-out efficiency of GFP ranged from 3.1 to 41.4% depending on U6 promoters and target sequences. The GFP knock-out region was mottled in many lines, indicating genome editing in individual cells. However, in 101 of 102 mutated individuals (> 99%) from 6 GFP knock-out lines, embryos had a single mutation pattern. Next, we knocked out the endogenous C. japonica magnesium chelatase subunit I (CjChlI) gene using two guide RNA targets. Green, pale green, and albino phenotypes were obtained in the gene-edited cell lines. Sequence analysis revealed random deletions, insertions, and replacements in the target region. Thus, targeted mutagenesis using the CRISPR/Cas9 system can be used to modify the C. japonica genome.

Digestion in the tsetse fly Glossina morsitans Westw.: the effect of feeding field-caught flies on guineapigs in the laboratory

1967 ◽  
Vol 57 (3) ◽  
pp. 447-450 ◽  
Author(s):  
P. A. Langley
Keyword(s):  
Natural Environment ◽  
Environmental Conditions ◽  
Tsetse Fly ◽  
Rapid Loss ◽  
Glossina Morsitans ◽  
The Third ◽  
Laboratory Conditions ◽  
Effect Of Feeding ◽  
Blood Meals

It has been shown that adults of Glossina morsitans Westw. that have fed from a bait ox in their natural environment digest their blood meals more rapidly than others that have emerged and been fed in the laboratory, even when both are maintained under identical environmental conditions after feeding.In further experiments with G. morsitans in Rhodesia, flies caught in the field and fed in the laboratory were found to lose their ability to digest their meals rapidly. Measurements, made throughout three hunger cycles, of the rate of digestion, as reflected in the rate of excretion, of blood meals by field-caught flies fed on guineapigs in the laboratory showed that this was not significantly different from that of the normal, flied-fed flies during the first two hunger cycles but that during the third it fell to a level comparable to that of flise that emerged and were fed in the laboratory.It is concluded that whatever may be the events that condition the field flies to digest their meals rapidly in the natural environment, these are repeated with the ingestion of each meal, and that laboratory conditions cause a rapid loss of this greater digestive capability.

Analysis of ploidy level of Artemisia annua L. based on flow cytometry and confocal laser scanning microscopy

2021 ◽  
Vol 50 (1) ◽  
pp. 29-35
Author(s):  
Shu-Gen Wei ◽  
Ling-Yun Wan ◽  
Ying Wei ◽  
Li-Li He ◽  
Jin-E Fu ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Confocal Laser Scanning Microscopy ◽  
Laser Scanning ◽  
Artemisia Annua ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Scanning Microscopy ◽  
Confocal Laser ◽  
Ploidy Levels ◽  
Artemisia Annua L

Eighty nine Artemisia samples treated with different concentrations of colchicine were used as breeding samples, with diploid Artemisia as the control. The ploidy levels of samples were determined by flow cytometry and confocal laser scanning microscopy (CLSM). An analysis of the flow cytometry results identified three suspected tetraploid plants and seven suspected triploid plants. The results of this study may be useful for breeding new Artemisia lines.

scholarly journals Ploidy Levels of Hardy Actinidia Accessions in the U.S. Determined by Flow Cytometry

HortScience ◽  
1997 ◽  
Vol 32 (3) ◽  
pp. 439D-439 ◽  
Author(s):  
Mary Ann Start ◽  
James Luby ◽  
Robert Guthrie ◽  
Debby Filler
Keyword(s):  
Flow Cytometry ◽  
Ploidy Level ◽  
Interspecific Variation ◽  
Root Tip ◽  
Interspecific Crosses ◽  
Ploidy Levels ◽  
Haploid Chromosome Number ◽  
Tip Cells ◽  
Root Tip Cells ◽  
The U.S

The hardy Actinidia species represent a source of genetic diversity for improving A. deliciosa (kiwifruit) as well as for creating new economically important cultivars through intra- and interspecific crosses. Attempts at breeding in Actinidia have been complicated by the existence of intraspecific as well as interspecific variation in ploidy. The haploid chromosome number in Actinidia is 29 and diploid (2n=2x=58), tetraploid (2n=4x=116), and hexaploid (2n=6x=174) levels have been identified. Because of the problems encountered when crossing parents differing in ploidy level, it is desirable to know the ploidy levels of plants to be used in breeding. We determined the ploidy levels of 61 Actinidia accessions currently available in the U.S., including primarily accessions of relatively winter-hardy species. The 61 accessions, representing eight species and three interspecific hybrids, were screened for ploidy using flow cytometry. Mitotic root tip cells from one plant from each putative ploidy level were examined microscopically to confirm the ploidy level derived from flow cytometry. There were 17 diploids, 40 tetraploids, and 4 hexaploids. Intraspecific variation was not found among accessions of the species arguta, callosa, deliciosa, kolomikta, melanandra, polygama, or purpurea. All kolomikta and polygama accessions were diploid. All arguta, callosa, melanandra, and purpurea accessions were tetraploid. Actinidia deliciosa was hexaploid. One chinensis accession was tetraploid. Two accessions (NGPR 0021.14 and 0021.3), acquired as chinensis, were hexaploid and may, in fact, be A. deliciosa based on their morphology. `Issai' (arguta × polygama) was hexaploid and `Ken's Red' and `Red Princess' (both melanandra × arguta) were tetraploid.

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