Abstract
The recent discovery of aquaporins - proteins that mediate bidirectional passage of water over membranes - provides a molecular basis for the underlying processes in plant water relations. In plants, aquaporins are found in plasma membranes and in vacuolar membranes (tonoplasts) and are suggested to participate in dynamic operations such as cell elongation, osmotic regulation, and long-distance transport. Focusing on the role of aquaporins or homologous proteins during seed development, the gene expression patterns and the accumulation of their products were investigated in the conifer Norway spruce (Picea abies (L.) Karst), which belongs to a plant group not studied before in this respect. In plant propagation programs, also the germination of seeds or somatic embryos derived from tissue cultures, as well as the early plant establishment are critical steps as regards to water limiting conditions. Involvement of aquaporins or homologues also in these processes was therefor investigated. Using a broad-selective antiserum raised against a plant seed aquaporin, immunoblot analyses indicated the presence of at least two isoforms in seeds during development. One isoform was abundant in mature seeds and another was detected in extracts of young seedlings, particularly in the roots. The protein from mature seeds was purified and partially sequenced. That information was used in primer design for PCR-based methods and resulted in the identification of full-length genes in both seeds (mipfg) and roots (mipr). These two genes showed high homology to the tonoplast sub-class of aquaporins. In addition, in mature seeds, immunolocalization experiments gave a high labelling in the tonoplast of protein storage vesicles (PSVs). Contrary, in the early stages of seeds, before PSV formation, labelling was seen in the plasmamembrane of the jacket cells lining the archegonia in young female gametophytes. In situ hybridization experiments agreed with these results, as strong expression signals were present in the jacket cells as well as in the maturing seeds. In the latter the hybridization signal was found in the entire female gametophyte. A strong signal was also seen in the spongy tissue surrounding the female gametophyte of early ovules. The high signal corresponded well with results from Northern analyses. Similarly a strong hybridization signal was seen in root tips from plants of various ages, including lateral roots and adventitious roots formed on hypocotyl cuttings. The columella cells of the root cap stained intensely, as did guard cells of stomata in needles. A gene transcript identical to mipfg was also identified in somatic embryos of Norway spruce. The in situ hybridization staining patterns in plantlets derived from somatic embryos were similar to the staining signals seen in seedlings grown from seeds. The results indicate important roles of aquaporin function in Norway spruce, in regulating transmembrane water and solute transport and in response to stress reactions.
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Claims 17
Claim · model-written summary
Immunoblot analyses indicated at least two aquaporin isoforms in Norway spruce seeds during development.
- Taxon
- Norway spruce
Picea abiest0 - Organ / stage
- seeds during developmente1
- Outcome
- presence of at least two isoformse1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Aquaporin homologues in Norway spruce (Picea abies)”
e1“Using a broad-selective antiserum raised against a plant seed aquaporin, immunoblot analyses indicated the presence of at least two isoforms in seeds during development”
Claim · model-written summary
One aquaporin isoform was abundant in mature Norway spruce seeds.
- Taxon
- Norway sprucePicea abiest0
- Organ / stage
- mature seedse2
- Outcome
- abundante2
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Aquaporin homologues in Norway spruce (Picea abies)”
e2“One isoform was abundant in mature seeds”
Claim · model-written summary
Another aquaporin isoform was detected in extracts of young Norway spruce seedlings, particularly in the roots.
- Taxon
- Norway sprucePicea abiest0
- Organ / stage
- young seedlingse3
- Outcome
- detectede3
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Aquaporin homologues in Norway spruce (Picea abies)”
e3“another was detected in extracts of young seedlings, particularly in the roots”
Claim · model-written summary
Full-length gene mipfg was identified from Norway spruce seeds.
- Taxon
- Norway sprucePicea abiest0
- Organ / stage
- seedse4
- Gene
- mipfge4
- Outcome
- identification of full-length genese4
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Aquaporin homologues in Norway spruce (Picea abies)”
e4“resulted in the identification of full-length genes in both seeds (mipfg) and roots (mipr)”
Claim · model-written summary
Full-length gene mipr was identified from Norway spruce roots.
- Taxon
- Norway sprucePicea abiest0
- Organ / stage
- rootse5
- Gene
- mipre5
- Outcome
- identification of full-length genese5
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Aquaporin homologues in Norway spruce (Picea abies)”
e5“resulted in the identification of full-length genes in both seeds (mipfg) and roots (mipr)”
Claim · model-written summary
The two identified genes showed high homology to the tonoplast sub-class of aquaporins.
- Taxon
- Norway sprucePicea abiest0
- Outcome
- high homology to the tonoplast sub-class of aquaporinse6
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Aquaporin homologues in Norway spruce (Picea abies)”
e6“These two genes showed high homology to the tonoplast sub-class of aquaporins”
Claim · model-written summary
In mature seeds, aquaporin labelling was high in the tonoplast of protein storage vesicles.
- Taxon
- Norway sprucePicea abiest0
- Organ / stage
- mature seedse7
- Outcome
- high labelling in the tonoplaste7
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Aquaporin homologues in Norway spruce (Picea abies)”
e7“In addition, in mature seeds, immunolocalization experiments gave a high labelling in the tonoplast of protein storage vesicles”
Claim · model-written summary
Before protein storage vesicle formation, labelling was seen in the plasmamembrane of jacket cells lining the archegonia in young female gametophytes.
- Taxon
- Norway sprucePicea abiest0
- Organ / stage
- jacket cells lining the archegonia in young female gametophytese1
- Outcome
- labelling was seene1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Aquaporin homologues in Norway spruce (Picea abies)”
e1“Contrary, in the early stages of seeds, before PSV formation, labelling was seen in the plasmamembrane of the jacket cells lining the archegonia in young female gametophytes”
Claim · model-written summary
In situ hybridization gave strong expression signals in jacket cells and in maturing seeds of Norway spruce.
- Taxon
- Norway sprucePicea abiest0
- Organ / stage
- jacket cellse1
- Outcome
- strong expression signalse1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Aquaporin homologues in Norway spruce (Picea abies)”
e1“In situ hybridization experiments agreed with these results, as strong expression signals were present in the jacket cells as well as in the maturing seeds”
Claim · model-written summary
In maturing seeds, the hybridization signal was found in the entire female gametophyte.
- Taxon
- Norway sprucePicea abiest0
- Organ / stage
- entire female gametophytee1
- Outcome
- hybridization signal was founde1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Aquaporin homologues in Norway spruce (Picea abies)”
e1“In the latter the hybridization signal was found in the entire female gametophyte”
Claim · model-written summary
A strong signal was seen in the spongy tissue surrounding the female gametophyte of early ovules.
- Taxon
- Norway sprucePicea abiest0
- Organ / stage
- spongy tissue surrounding the female gametophyte of early ovulese1
- Outcome
- strong signale1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Aquaporin homologues in Norway spruce (Picea abies)”
e1“A strong signal was also seen in the spongy tissue surrounding the female gametophyte of early ovules”
Claim · model-written summary
The in situ high signal corresponded well with results from Northern analyses.
- Taxon
- Norway sprucePicea abiest0
- Outcome
- high signale1
- Comparator
- results from Northern analysese1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Aquaporin homologues in Norway spruce (Picea abies)”
e1“The high signal corresponded well with results from Northern analyses”
Claim · model-written summary
A strong hybridization signal was seen in root tips of plants of various ages, including lateral roots and adventitious roots.
- Taxon
- Norway sprucePicea abiest0
- Organ / stage
- root tipse1
- Outcome
- strong hybridization signale1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Aquaporin homologues in Norway spruce (Picea abies)”
e1“Similarly a strong hybridization signal was seen in root tips from plants of various ages, including lateral roots and adventitious roots formed on hypocotyl cuttings”
Claim · model-written summary
Columella cells of the root cap and guard cells of stomata in needles stained intensely.
- Taxon
- Norway sprucePicea abiest0
- Organ / stage
- columella cells of the root cape1
- Outcome
- stained intenselye1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Aquaporin homologues in Norway spruce (Picea abies)”
e1“The columella cells of the root cap stained intensely, as did guard cells of stomata in needles”
Claim · model-written summary
A gene transcript identical to mipfg was identified in somatic embryos of Norway spruce.
- Taxon
- Norway sprucePicea abiese1
- Organ / stage
- somatic embryose1
- Gene
- mipfge1
- Outcome
- gene transcripte1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“A gene transcript identical to mipfg was also identified in somatic embryos of Norway spruce”
Claim · model-written summary
In situ hybridization staining patterns in plantlets from somatic embryos were similar to the signals in seedlings grown from seeds.
- Taxon
- Norway sprucePicea abiest0
- Organ / stage
- plantlets derived from somatic embryose1
- Outcome
- staining patternse1
- Comparator
- staining signals seen in seedlings grown from seedse1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Aquaporin homologues in Norway spruce (Picea abies)”
e1“The in situ hybridization staining patterns in plantlets derived from somatic embryos were similar to the staining signals seen in seedlings grown from seeds”
Claim · model-written summary
Aquaporin function is indicated to have important roles in regulating transmembrane water and solute transport in Norway spruce.
- Taxon
- Norway sprucePicea abiest0
- Outcome
- regulating transmembrane water and solute transporte1
- Qualifier
- in response to stress reactionse1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“Aquaporin homologues in Norway spruce (Picea abies)”
e1“The results indicate important roles of aquaporin function in Norway spruce, in regulating transmembrane water and solute transport and in response to stress reactions”