To have everything in one thread - I found this today:
While looking at what might be associated patents, I find this interesting and think it clears up another mystery, even though this belongs in the food sector:
From a arm lesion, the petri dish had started drying out, this is 2 months after culture:
tinypic.com/r/ve0hl5/7It looks like what we called a 'magnolia'... well - it might be a petunia? In a separate instance, Toni had a sample that was identified as dicotyledon, I do believe?
www.ancorainviaggio.it/petunia3.jpgPLANTS EXPRESSING ENVIRONMENTAL STRESS TOLERANCES HAVING PETUNIA CBF GENES THEREIN
www.freepatentsonline.com/y2009/0307793.html"18. The vector of claim 16 wherein the host is a dicotyledon.
FIELD OF INVENTION
This invention relates to stress tolerance in plants. In addition, this invention relates to transcription factors and gene expression involved in stress tolerance in plants.
Embodiments of the present invention also provide a nucleic acid which is hybridizable to a PetCBF nucleic acid, or to a nucleic acid encoding a PetCBF variant, under conditions of high stringency.
In another preferred embodiment, a binary vector system, a plasmid containing the left and right borders of T-DNA is engineered to contain an exogenous gene of interest. If needed, an antibiotic resistance gene may be introduced in order to provide a basis of selecting transformed plant cells. Other genes conferring a selectable phenotype in the presence of an exogenous agent other than an antibiotic may also be utilized. The modified plasmid may be transformed into E. coli and then transferred by conjugation to an A. tumefaciens strain containing a helper plasmid. The helper plasmid provides the vir functions. On infection of a plant, the activated vir functions recognize the left border sequence of the modified plasmid and transfer all DNA between left and right borders to a plant chromosome.
FIG. 2 illustrates PetCBF4 gene expression as described above.
Table 2—Primer used for isolation of CBF genes and for generation of probes for Northern hybridization
SEQ
ID
Primer NO: Sequence
PetF4 1 AATCCNAARAARCCNGCNG
PetR5 2 ATCAGCRAARTTNARRCANGC
3RACE1 3 GGGAGGAAGAAGTTTCAAGAAACTCGACATCC
3RACE2 4 GGGAGGAAGAAGTTTCAAGAAACACGACATCC
3RACE3 5 GCCGGGAGGAAGAAGTTTAGAGAAACACGACA
5RACE1 6 GGATGTCGAGTTTCTTGAAACTTCTTCCTCCC
5RACE2 7 GGATGTCGTGTTTCTTGAAACTTCTTCCTCCC
5RACE3 8 TGTCGTGTTTCTCTAAACTTCTTCCTCCCGGC
ParF1 9 CCTCAAACTGAAATAACATTCAGTACTAGTACT
ParF3 10 CCTCTAACTGAAACAACATCCAATACAACC
ParF2 11 TAACATTCAGTACTAGTACTATACACTTACTA
ParF4 12 CAAAAACCTCAAACTGAACAACATTC
ParR1 13 GGCAAACTACACGATGTTCTTGTCTCTCATC
ParR2 14 TGGATCTTTCATTCAATACAAGGGCTTGG
ParR3 15 CAGTCAAATGTCGTGGTTTGAAAAAACCG
ParR4 16 CCGCGCCAAGTCAAACACAGACACTC
CBF1ProbeF 17 CAAGAAGTCACTCCGGCTTT
CBF1ProbeR 18 TTGCATTCAAAAGTGGCAAA
CBF2ProbeF 19 AATTCTGTTAGTACTTCTTTGGGATAG
CBF2ProbeR 20 TTAGCTGCTCACTTGGATCTTTC
CBF3ProbeF 21 TTGGTACCTAATATTTGGACGGTA
CBF3ProbeR 22 TCAAATGTCGTGGTTTGAAAAA
CBF4ProbeF 23 ATATTTGGGCGGTACGTCTG
CBF4ProbeR 24 AGATTTGTTTTGGACCACATGA
ActinF 25 GAAGCGCCTCTGAACCCAAA
ActinR 26 CCGCAGCTTCCATTCCAATC
Biolistic Mediated Transformation
Plasmid DNA containing: PetCBF1 [SEQ ID NO: 27], PetCBF2 [SEQ ID NO: 28], PetCBF3 [SEQ ID NO: 29], and PetCBF4 [SEQ ID NO: 30] was isolated using a HiSpeed Plasmid Midi Kit (Qiagen Sciences, Maryland, USA) and the DNA concentration was adjusted to 900 ng/μl. Gold particles of 0.6 μm in diameter (Bio-Rad Laboratories, Hercules, Calif., USA) were coated with 10 μl plasmid DNA. The coated gold particles were then mixed with 50 μl of 2.5 M CaCl 2 , 20 μl of 0.1 M sperimidine and were vortexed for 20 minutes at 4° C. The coated particles were washed 3 times with 200 μl ethanol and centrifuged for 1 minute after each wash. Finally, the particles were re-suspended in 35 μl ethanol and kept on ice. 8-9 μl of the suspension were spread on each macro-carrier and allowed to dry prior to bombardment.
Regeneration and Maize Genotypes
Six different hybrids and an inbred of corn were tested for suitability with Agrobacterium mediated transformation. The hybrids LH 74x A 641, LH 262x LH 252, LH 198x LH 227, FR 1064x FR 1064 (SDMS) x LH 185, and LH 176x LH 177 DMS were obtained..."