Abstract
Soil salinity is a major constraint to crop production in Australia. This has prompted the need to produce salt tolerant cereal cultivars, through the understanding of genes involved in salt tolerance mechanisms and manipulating their expression levels. Arabidopsis thaliana Calcineurin B-like Interacting Protein Kinase 16 (AtCIPK16) has been identified as a gene involved in sodium (Na+) exclusion. Analysis of AtCIPK16 alleles from Arabidopsis ecotypes suggests variances in expression are due to differences in the promoters. Experiments in Arabidopsis, barley and wheat (preliminary) have illustrated that AtCIPK16 overexpression can enhance biomass production through increased Na+ exclusion, although its full effect in barley and wheat has yet to be properly characterised in both greenhouse and field environments. The first focus of this project evaluated the salt tolerance of 35S:AtCIPK16 barley (cv. Golden Promise) grown under low and high salinity field conditions in 2013 and 2014 at Kunjin, Western Australia. Comparisons between years were difficult due to waterlogging of the 2013 high salt site and the increased variability in plot establishment in 2014. 35S:AtCIPK16 barley lines had varying responses to high salt conditions depending on the annual rainfall. Results showed Na⁺ and Cl⁻ exclusion in certain lines, although this correlated with decreased biomass and yield in high rainfall years. AtCIPK16 expression also increased Na⁺ and Cl⁻ exclusion in 2012 (a low rainfall year) which instead lead to increasing plant growth and yield. The second focus of this project aimed to fully characterised the effects of the constitutive expression of Ubi:AtCIPK16 in wheat (cv. Gladius). Despite conducting three hydroponic experiments, no definitive conclusions about the effects of AtCIPK16 expression on wheat salt tolerance could be drawn. Although, one sibling transgenic line showed increased Na⁺ and Cl⁻ exclusion from both root and shoot tissue accompanied by larger biomass under 200 mM salt stress. Despite this finding several factors hinder the analysis of data including the high number of null segregants, considerable variability between siblings of the same transformation event and minimal transgene expression. The third focus of this project aimed to investigate expression differences between two AtCIPK16 alleles from the Arabidopsis ecotypes Bay-0 and Shahdara. Since the only differences between the two alleles was a 10 base pair deletion in the Bay-0 promoter, it was hypothesised this deletion was the reason for the increased expression of AtCIPK16 in Bay-0 as it forms a TATA box (TATATAA). The aim of this project was to alter the expression of each allele by: mutating the last A to a T, removing the TATA box in Bay-0, and mutating the T after the TATATA sequence to an A in Shahdara, forming a TATA box without the deletion. Through PCR mutagenesis the required point mutations were introduced into portions of the two promoter alleles, however due to technical difficulties and time constraints the point mutations were not introduced back into the full promoter constructs driving GFP. It was therefore unable to be determined if the point mutations to the TATA box would indeed affect AtCIPK16 expression.
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Claims 10
Claim · model-written summary
Certain transgenic barley lines excluded Na⁺ and Cl⁻ under high salt.
- Taxon
- barley
.jpg?width=60)
Hordeum vulgaret0 - Material
- certain linese0
- Outcome
- Na⁺ and Cl⁻ exclusione0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
.jpg?width=500)
Verbatim
e0“Results showed Na⁺ and Cl⁻ exclusion in certain lines”
t0“barley and wheat”
Claim · model-written summary
In high rainfall years, ion exclusion in barley correlated with decreased biomass and yield.
- Taxon
- barleyHordeum vulgaret0
- Material
- 35S:AtCIPK16 barley linese1
- Outcome
- biomass and yielde0
- Direction
- decreasede0
- Qualifier
- high rainfall yearse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“although this correlated with decreased biomass and yield in high rainfall years”
t0“barley and wheat”
e1“35S:AtCIPK16 barley lines”
Claim · model-written summary
AtCIPK16 expression increased Na⁺ and Cl⁻ exclusion in barley in the low rainfall year 2012.
- Taxon
- barleyHordeum vulgaret0
- Treatment
- AtCIPK16 expressione0
- Gene
- AtCIPK16e0
- Outcome
- Na⁺ and Cl⁻ exclusione0
- Direction
- increasede0
- Qualifier
- 2012 (a low rainfall year)e0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“AtCIPK16 expression also increased Na⁺ and Cl⁻ exclusion in 2012 (a low rainfall year)”
t0“barley and wheat”
Claim · model-written summary
In the low rainfall year, AtCIPK16 expression increased barley growth and yield.
- Taxon
- barleyHordeum vulgaret0
- Treatment
- AtCIPK16 expressione0
- Gene
- AtCIPK16e0
- Outcome
- plant growth and yielde0
- Direction
- increasinge0
- Qualifier
- 2012 (a low rainfall year)e0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“AtCIPK16 expression also increased Na⁺ and Cl⁻ exclusion in 2012 (a low rainfall year) which instead lead to increasing plant growth and yield”
t0“barley and wheat”
Claim · model-written summary
Three hydroponic experiments gave no definitive conclusion on wheat salt tolerance effects.
- Taxon
- wheat
Triticum aestivume0 - Gene
- AtCIPK16e0
- Outcome
- no definitive conclusions about the effects of AtCIPK16 expression on wheat salt tolerancee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Despite conducting three hydroponic experiments, no definitive conclusions about the effects of AtCIPK16 expression on wheat salt tolerance could be drawn”
Claim · model-written summary
One wheat transgenic sibling line showed increased Na⁺ and Cl⁻ exclusion from roots and shoots under 200 mM salt.
- Taxon
- wheatTriticum aestivumt0
- Material
- one sibling transgenic linee0
- Organ / stage
- root and shoot tissuee0
- Treatment
- 200 mM salt stresse0
- Outcome
- Na⁺ and Cl⁻ exclusione0
- Direction
- increasede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“one sibling transgenic line showed increased Na⁺ and Cl⁻ exclusion from both root and shoot tissue accompanied by larger biomass under 200 mM salt stress”
t0“barley and wheat”
Claim · model-written summary
That wheat line had larger biomass under 200 mM salt stress.
- Taxon
- wheatTriticum aestivumt0
- Material
- one sibling transgenic linee0
- Treatment
- 200 mM salt stresse0
- Outcome
- biomasse0
- Direction
- largere0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“one sibling transgenic line showed increased Na⁺ and Cl⁻ exclusion from both root and shoot tissue accompanied by larger biomass under 200 mM salt stress”
t0“barley and wheat”
Claim · model-written summary
Point mutations were introduced into portions of two Arabidopsis AtCIPK16 promoter alleles.
- Taxon
- Arabidopsis
Arabidopsise1 - Outcome
- point mutations were introduced into portions of the two promoter allelese0
- Value
- two promoter allelese0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Through PCR mutagenesis the required point mutations were introduced into portions of the two promoter alleles”
e1“Arabidopsis ecotypes Bay-0 and Shahdara”
Claim · model-written summary
Technical difficulties prevented introduction of mutations into full promoter constructs driving GFP.
- Taxon
- ArabidopsisArabidopsise1
- Outcome
- point mutations were not introduced back into the full promoter constructs driving GFPe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the point mutations were not introduced back into the full promoter constructs driving GFP”
e1“Arabidopsis ecotypes Bay-0 and Shahdara”
Claim · model-written summary
The effect of TATA box point mutations on AtCIPK16 expression remained undetermined.
- Taxon
- ArabidopsisArabidopsise1
- Gene
- AtCIPK16e0
- Outcome
- unable to be determined if the point mutations to the TATA box would indeed affect AtCIPK16 expressione0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“It was therefore unable to be determined if the point mutations to the TATA box would indeed affect AtCIPK16 expression”
e1“Arabidopsis ecotypes Bay-0 and Shahdara”
The same questions in other papers
salinity · growth · Triticum aestivum (wheat)Positive and negative results
48 papers in the same direction as this paper · 327 in the opposite direction · 1 reporting no difference from an alternative
- negative
- negative
- negative
373 papers more in this group
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