Abstract
This paper describes the micropropagation of Douglas-fir via somatic embryogenesis using media and methods available in the public domain. Douglas-fir is a conifer species with high and increasing interest for forest industries in both New Zealand and France. Delivery of the best trees to the forest from breeding programmes is currently constrained by an inability to effectively and reliably multiply selections through vegetative propagation. Somatic embryogenesis coupled with cryopreservation is essential biotechnology tool in conifers for scaling up variety design and production. The aim of this work was therefore to develop protocols for the initiation and proliferation of Douglas-fir embryogenic cell lines based on the modifications of previously published techniques and media available in the public domain, especially successful methods developed for P. radiata at Scion. Three years of initiation experiments have resulted in a simple and efficient protocol. Disinfection of whole cones (instead of seeds) was sufficient to prevent contamination. Immature zygotic embryos were excised from megagametophytes and placed onto a modified Litvay medium (Plant Cell Rep 4(6):325–328, 1985***). At optimal development stages of zygotic embryos, the average initiation percentage of embryogenic tissue was 69.3% when our most effective protocol was used. Proliferation of initiated cell lines on a Glitz formulation was challenging, with a high percentage of cell lines composed of a mixture of both embryonal masses and callus. 2,4-D at a lower concentration reduced the number of such mixed lines. Repeated liquid suspension and subculture of the embryogenic parts of the tissue was an efficient means to increase the number of embryogenic cell lines with sustained proliferation. Maltose added to the proliferation medium in place of sucrose improved fresh mass gain and consistently increased early somatic embryo patterning and growth. A sample of proliferating cell lines was successfully cryopreserved, thawed and somatic embryos were matured and germinated from these lines. The method may be of practical interest and provide new opportunities to realize increased genetic gain in this species through the clonal-assisted deployment of the best genetic material in both New Zealand and France.
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Claims 10
Claim · model-written summary
A simple and efficient protocol for initiating Douglas-fir embryogenic tissue was developed from modified previously published techniques.
- Taxon
- Douglas-fir
Pseudotsuga menziesiit0 - Outcome
- simple and efficient protocole0
- Comparator
- previously published techniquese1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Simple and efficient protocols for the initiation and proliferation of embryogenic tissue of Douglas-fir”
e0“Three years of initiation experiments have resulted in a simple and efficient protocol”
e1“based on the modifications of previously published techniques”
Claim · model-written summary
Disinfection of whole cones was sufficient to prevent contamination, instead of disinfecting seeds.
- Taxon
- Douglas-firPseudotsuga menziesiit0
- Treatment
- Disinfection of whole conese2
- Outcome
- prevent contaminatione2
- Comparator
- seedse2
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Simple and efficient protocols for the initiation and proliferation of embryogenic tissue of Douglas-fir”
e2“Disinfection of whole cones (instead of seeds) was sufficient to prevent contamination”
Claim · model-written summary
At optimal zygotic embryo development stages, the average initiation percentage of embryogenic tissue was 69.3% with the most effective protocol.
- Taxon
- Douglas-firPseudotsuga menziesiit0
- Organ / stage
- zygotic embryose3
- Treatment
- our most effective protocole3
- Outcome
- average initiation percentage of embryogenic tissuee3
- Value
- 69.3%e3
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Simple and efficient protocols for the initiation and proliferation of embryogenic tissue of Douglas-fir”
e3“At optimal development stages of zygotic embryos, the average initiation percentage of embryogenic tissue was 69.3% when our most effective protocol was used”
Claim · model-written summary
Proliferating Douglas-fir cell lines were challenging to maintain, with a high percentage composed of a mixture of embryonal masses and callus.
- Taxon
- Douglas-firPseudotsuga menziesiit0
- Outcome
- cell lines composed of a mixturee4
- Qualifier
- of both embryonal masses and calluse4
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Simple and efficient protocols for the initiation and proliferation of embryogenic tissue of Douglas-fir”
e4“Proliferation of initiated cell lines on a Glitz formulation was challenging, with a high percentage of cell lines composed of a mixture of both embryonal masses and callus”
Claim · model-written summary
2,4-D at a lower concentration reduced the number of mixed cell lines.
- Taxon
- Douglas-firPseudotsuga menziesiit0
- Treatment
- 2,4-D at a lower concentratione5
- Outcome
- the number of such mixed linese5
- Direction
- reducede5
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Simple and efficient protocols for the initiation and proliferation of embryogenic tissue of Douglas-fir”
e5“2,4-D at a lower concentration reduced the number of such mixed lines”
Claim · model-written summary
Repeated liquid suspension and subculture increased the number of embryogenic cell lines with sustained proliferation.
- Taxon
- Douglas-firPseudotsuga menziesiit0
- Treatment
- Repeated liquid suspension and subculturee6
- Outcome
- the number of embryogenic cell linese6
- Direction
- increasee6
- Qualifier
- with sustained proliferatione6
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Simple and efficient protocols for the initiation and proliferation of embryogenic tissue of Douglas-fir”
e6“Repeated liquid suspension and subculture of the embryogenic parts of the tissue was an efficient means to increase the number of embryogenic cell lines with sustained proliferation”
Claim · model-written summary
Maltose in place of sucrose in the proliferation medium improved fresh mass gain.
- Taxon
- Douglas-firPseudotsuga menziesiit0
- Treatment
- Maltose added to the proliferation mediume7
- Outcome
- fresh mass gaine7
- Direction
- improvede7
- Comparator
- sucrosee7
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Simple and efficient protocols for the initiation and proliferation of embryogenic tissue of Douglas-fir”
e7“Maltose added to the proliferation medium in place of sucrose improved fresh mass gain and consistently increased early somatic embryo patterning and growth”
Claim · model-written summary
Maltose in place of sucrose in the proliferation medium consistently increased early somatic embryo patterning and growth.
- Taxon
- Douglas-firPseudotsuga menziesiit0
- Treatment
- Maltose added to the proliferation mediume7
- Outcome
- early somatic embryo patterning and growthe7
- Direction
- increasede7
- Comparator
- sucrosee7
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Simple and efficient protocols for the initiation and proliferation of embryogenic tissue of Douglas-fir”
e7“Maltose added to the proliferation medium in place of sucrose improved fresh mass gain and consistently increased early somatic embryo patterning and growth”
Claim · model-written summary
Proliferating cell lines were successfully cryopreserved and thawed.
- Taxon
- Douglas-firPseudotsuga menziesiit0
- Outcome
- cryopreserved, thawede7
- Qualifier
- successfullye7
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Simple and efficient protocols for the initiation and proliferation of embryogenic tissue of Douglas-fir”
e7“A sample of proliferating cell lines was successfully cryopreserved, thawed and somatic embryos were matured and germinated from these lines”
Claim · model-written summary
Somatic embryos from cryopreserved cell lines were matured and germinated.
- Taxon
- Douglas-firPseudotsuga menziesiit0
- Organ / stage
- somatic embryose7
- Outcome
- were matured and germinatede7
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Simple and efficient protocols for the initiation and proliferation of embryogenic tissue of Douglas-fir”
e7“A sample of proliferating cell lines was successfully cryopreserved, thawed and somatic embryos were matured and germinated from these lines”
The same questions in other papers
in vitro · growth · Pseudotsuga menziesii (Douglas-fir)Results agree
1 paper in the same direction as this paper
- positive
Claims are grouped by organism, treatment and what was measured or targeted. A paper counts as positive when any of its claims in the group is positive.