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Large scale propagation and in vitro weaning for the restoration of Viola palustris to support assisted colonisation of a threatened butterfly


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Figure 1

Flow chart explaining stages from seedling production (highlighted in green)to transplantation to the field (highlighted in red). Substrates/conditions in which the plants are kept at the various stages are marked (agar, liquid medium, plug system subtrate and field conditions) along with the duration of each stage in days, weeks (wks) or months. Average multiplication rates are shown for agar, liquid and repeated bioreactor cycle stages.
Flow chart explaining stages from seedling production (highlighted in green)to transplantation to the field (highlighted in red). Substrates/conditions in which the plants are kept at the various stages are marked (agar, liquid medium, plug system subtrate and field conditions) along with the duration of each stage in days, weeks (wks) or months. Average multiplication rates are shown for agar, liquid and repeated bioreactor cycle stages.

Figure 2

Multiplication rate for Viola palustris cultured in semi-solid agar and liquid immersion system. Agar multiplication alone for 16 weeks; multiplication using RITA® for 10 days followed by 16 weeks on agar; 3 – multiplication using a second RITA® cycle for 10 days followed by 16 weeks on agar. Bars represent means of at least 28 explants +/- standard error. Bars with the same letter are not significantly different (P>0.05).
Multiplication rate for Viola palustris cultured in semi-solid agar and liquid immersion system. Agar multiplication alone for 16 weeks; multiplication using RITA® for 10 days followed by 16 weeks on agar; 3 – multiplication using a second RITA® cycle for 10 days followed by 16 weeks on agar. Bars represent means of at least 28 explants +/- standard error. Bars with the same letter are not significantly different (P>0.05).

Figure 3

Improved rooting of in vitro Viola palustris propagules using photoautotrophic plug substrate and sucrose-free liquid nutrient medium. a – agar culture; b – roots of agar-grown propagule; c – photoautotrophic plug propagule; d – photoautotrophic propagule with substrate removed to show robust root growth. Scale bars 1 cm.
Improved rooting of in vitro Viola palustris propagules using photoautotrophic plug substrate and sucrose-free liquid nutrient medium. a – agar culture; b – roots of agar-grown propagule; c – photoautotrophic plug propagule; d – photoautotrophic propagule with substrate removed to show robust root growth. Scale bars 1 cm.

Figure 4

Establishment rating measure for Viola palustris five months after potting up from photoautotrophic plug trays. A = 2, b = 3, c = 4, d = 5. No photograph is available for rating 1 because no pots achieved an establishment rating of 1, i.e. very poor or no establishment.
Establishment rating measure for Viola palustris five months after potting up from photoautotrophic plug trays. A = 2, b = 3, c = 4, d = 5. No photograph is available for rating 1 because no pots achieved an establishment rating of 1, i.e. very poor or no establishment.

Figure 5

Growth and establishment of ex vitro Viola palustris propagules in field sites after 16 months (2014 plantings) and 4 months (2015 plantings). Bars represent the average from a minimum of five colonies +/- standard error. b = significantly (P<0.05) lower ground cover than all other sites.
Growth and establishment of ex vitro Viola palustris propagules in field sites after 16 months (2014 plantings) and 4 months (2015 plantings). Bars represent the average from a minimum of five colonies +/- standard error. b = significantly (P<0.05) lower ground cover than all other sites.

Figure 6

Establishment of modules planted out in the field. a – 16 months after planting; b – 4 months after planting. Scale bar 5 cm.
Establishment of modules planted out in the field. a – 16 months after planting; b – 4 months after planting. Scale bar 5 cm.
eISSN:
2564-615X
Language:
English
Publication timeframe:
4 times per year
Journal Subjects:
Life Sciences, Genetics, Biotechnology, Bioinformatics, other