Os02g0677300 - Revision history

Zhennan at 07:37, 23 March 2017

2017-03-23T07:37:08Z

192.168.72.52 at 06:46, 14 May 2015

2015-05-14T06:46:08Z

Angela: /* Evolution */

2014-06-06T14:19:25Z

‎Evolution

Angela: /* Function */

2014-06-06T14:02:04Z

‎Function

Fengdongmoqing: /* References */

2014-06-04T02:26:31Z

‎References

Fengdongmoqing: /* Expression */

2014-06-04T02:26:08Z

‎Expression

Fengdongmoqing: /* Expression */

2014-06-04T02:21:42Z

‎Expression

Fengdongmoqing: /* Expression */

2014-06-04T02:16:15Z

‎Expression

Fengdongmoqing: /* Expression */

2014-06-04T02:14:14Z

‎Expression

Angela: /* Function */

2014-05-31T16:59:42Z

‎Function

@@ Line 1: / Line 1: @@
-Please input one-sentence summary here.
+The rice '''''Os02g0677300''''' was reported as '''''OsERF#025''''' in 2006 <ref name="ref1" /> by researchers from Japan.
 ==Annotated Information==
 ===Function===
-Nine CBF/DREB1homologous genes in rice were obtained by BLAST search in the NCBI database, which share conserved amino acid sequences with DREB1 protein in Arabidopsis. Three CBFgenes organized in tandem, named OsCBF1, OsCBF2and OsCBF3, showed a transient induction in the process of cold acclimation, much stronger in indica rice 93-11 compared with japonica rice Nipponbare.The candidate downstream genes OsLIP5and OsLIP9were induced in 93-11 but notin Nipponbare. The differential expression of CBF regulon might be caused by polymorphisms within promoter sequences between these two rice varieties.One of the nine homologous CBF/DREB1 genes and the center of resistance to cold way.The encoding products of CBF genes belongs to the transcriptional activator of AP2/ERF family, which can specifically binding to the promoter that contain CRT/DRE cis-acting elements Kai and start the expression of the downstream genes to improve the cold tolerance of plants.Gene expression profiling analysis shows that the CBF genes of rice can upstream the express of stress tolerance genes to enhance the cold tolerance of transgenic rice,like OsP5CS,OsLIP5,Os-LIP9,OsRAmy3D and so on.
+* Genes in the ERF family encode transcriptional regulators with a variety of functions involved in the developmental and physiological processes in plants.
+* It has been demonstrated that the AP2/ERF proteins have important functions in the transcriptional regulation of a variety of biological processes related to growth and development, as well as various responses to environmental stimuli.
-===Expression===
+* Genes in the AP2 family have been shown to participate in the regulation of developmental processes, e.g. flower development (Elliott et al., 1996), spikelet meristem determinacy (Chuck et al., 1998), leaf epidermal cell identity (Moose and Sisco, 1996), and embryo development (Boutilier et al., 2002).<ref name="ref2" />
 ===Evolution===
-Please input evolution information here.
+* The ERF family is a large gene family of transcription factors and is part of the AP2/ERF superfamily, which also contains the AP2 and RAV families
 You can also add sub-section(s) at will.
 ==Labs working on this gene==
-Please input related labs here.
+* Molecular and Cellular Breeding Research Group, Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki 305–8566, Japan (T.N., K.S., H.S.);
 ==References==
-.Canella D, Gilmour S J, Kuhn L A, Thomashow M F. 2010. DNA binding by the ArabidopsisCBF1 transcription factor requires the PKKP/RAGRxKFxETRHP signature sequence.  Biochim
+<references>
-Biophys Acta, 1799: 454–462.
+* <ref name="ref1">
-.Wang Y, Hua J. 2009. A moderate decrease in temperature induces COR15aexpression through the CBF signaling cascade and enhances freezing tolerance. Plant J, 60: 340–349.
+Nakano T, Suzuki K, Fujimura T, Shinshi H. Genome-wide analysis of the ERF
-.Chinnusamy V, Ohta M, Kanrar S, Lee B H, Hong X, Agarwal M, Zhu J K. 2003. ICE1: A regulator of cold-induced transcriptome and freezing tolerance in Arabidopsis. Genes Dev, 17: 1043–1054.
+gene family in Arabidopsis and rice. Plant Physiol. 2006 Feb;140(2):411-32.
-.Miura K, Jin J B, Lee J, Yoo C Y, Stirm V, Miura T, Ashworth E N, Bressan R A, Yun D J, Hasegawa P M. 2007. SIZ1-mediated sumoylation of ICE1 controls CBF3/DREB1Aexpression and freezing tolerance in Arabidopsis. Plant Cell, 19: 1403–1414.
+PubMed PMID: 16407444; PubMed Central PMCID: PMC1361313.
-.Jaglo K R, Kleff S, Amundsen K L, Zhang X, Haake V, Zhang J Z, Deits T, Thomashow M F. 2001. Components of the Arabidopsis=C-repeat/dehydration-responsive element binding factor cold-response pathway are conserved in Brassica napusand other plant species. Plant Physiol, 127: 910–917.
+</ref>
-.Zhang X, Fowler S G, Cheng H, Lou Y, Rhee S Y, Stockinger E J, Thomashow M F. 2004. Freezing-sensitive tomato has a functional CBF cold response pathway, but a CBF regulon that differs from that of freezing-tolerant Arabidopsis. Plant J, 39: 905–919.
+* <ref name="ref2">
-.Kume S, Kobayashi F, Ishibashi M, OhnoR, NakamuraC, Takumi S. 2005. Differential and coordinated expression of Cbfand Cor/Leagenes during long-term cold acclimation in two wheat cultivars showing distinct levels of freezing tolerance. GenesGenet Syst, 80: 185–197.
+Chuck G, Muszynski M, Kellogg E, Hake S, Schmidt RJ. The control of spikelet
-.Dubouzet J G, Sakuma Y, Ito Y, Kasuga M, Dubouzet E G, Miura S, Seki M, Shinozaki K, Yamaguchi-Shinozaki K. 2003. OsDREBgenes in rice, Oryza sativa L., encode transcription activators that function in drought-, high-salt- and cold-responsive gene expression. Plant J, 33: 751–763.
+meristem identity by the branched silkless1 gene in maize. Science. 2002 Nov
-.Ito Y, Katsura K, Maruyama K, Taji T, Kobayashi M, Seki M, Shinozaki K, Yamaguchi-Shinozaki K. 2006. Functional analysis of rice DREB1/CBF-type transcription factors involved in cold-responsive gene expression in transgenic rice. Plant Cell Physiol, 47: 141–153.
+;298(5596):1238-41. PubMed PMID: 12424380.
+</ref>
 ==Structured Information==
       [[Category:Genes]][[Category:Oryza Sativa Japonica Group]][[Category:Japonica Chromosome 2]]

← Older revision		Revision as of 06:46, 14 May 2015
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	==Structured Information==		==Structured Information==
−	~~{{JaponicaGene\|~~	+	[[Category:Genes]][[Category:Oryza Sativa Japonica Group]][[Category:Japonica Chromosome 2]]
−	~~GeneName = Os02g0677300\|~~
−	~~Description = Similar to CRT/DRE binding factor 1\|~~
−	~~Version = NM_001054262.1 GI:115447894 GeneID:4330306\|~~
−	~~Length = 1272 bp\|~~
−	~~Definition = Oryza sativa Japonica Group Os02g0677300, complete gene.\|~~
−	~~Source = Oryza sativa Japonica Group~~
−
−	~~ORGANISM Oryza sativa Japonica Group~~
−	~~Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;~~
−	~~Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP~~
−	~~clade; Ehrhartoideae; Oryzeae; Oryza.~~
−	\|
−	~~Chromosome = [[:category:Japonica Chromosome 2\|Chromosome 2]]\|~~
−	~~AP = Chromosome 2:28539796..28541067\|~~
−	~~CDS = 28539841..28540515\|~~
−	~~GCID = <gbrowseImage1>~~
−	~~name=NC_008395:28539796..28541067~~
−	~~source=RiceChromosome02~~
−	~~preset=GeneLocation~~
−	~~</gbrowseImage1>\|~~
−	~~GSID = <gbrowseImage2>~~
−	~~name=NC_008395:28539796..28541067~~
−	~~source=RiceChromosome02~~
−	~~preset=GeneLocation~~
−	~~</gbrowseImage2>\|~~
−	CDNA = <cdnaseq>atggacgtttctgctgcgctcagcagcgactactcgtcggggacgccgtcgccggtggcggccgacgccgacgacggctcctccgcctacatgacggtgtcgtcggcgccgcccaagcggcgagcggggcggaccaagttcaaggagacgcggcaccccgtgttcaagggcgtgcgccggaggaaccccgggaggtgggtgtgcgaggtgcgcgagccgcacggcaagcagcggatatggctcgggacgttcgagacagcagagatggcggcgcgcgcgcacgacgtcgccgcgctcgcgctccgcggccgcgccgcctgcctcaacttcgccgactcgccgaggcgcctccgcgtcccgcccatcggcgcaagccacgacgacatacggagggcggcggctgaggcggccgaggcattccggccgccaccagatgagagcaatgcggccaccgaggtggcagccgccgcatcgggcgccactaattcgaacgccgaacagttcgcctcccacccgtactacgaggtcatggacgatgggctggacttggggatgcagggctatctcgacatggcgcaagggatgctcattgacccgcctccaatggccggtgatcctgccgtaggtagcggcgaagacgacaacgatggcgaggtccagctatggagctactga</cdnaseq>\|
−	AA = <aaseq>MDVSAALSSDYSSGTPSPVAADADDGSSAYMTVSSAPPKRRAGR TKFKETRHPVFKGVRRRNPGRWVCEVREPHGKQRIWLGTFETAEMAARAHDVAALALR GRAACLNFADSPRRLRVPPIGASHDDIRRAAAEAAEAFRPPPDESNAATEVAAAASGA TNSNAEQFASHPYYEVMDDGLDLGMQGYLDMAQGMLIDPPPMAGDPAVGSGEDDNDGE VQLWSY</aaseq>\|
−	DNA = <dnaseqindica>46..720#agaattcaaaccggatcaacctcgctcgcttactcgtgtttaggcatggacgtttctgctgcgctcagcagcgactactcgtcggggacgccgtcgccggtggcggccgacgccgacgacggctcctccgcctacatgacggtgtcgtcggcgccgcccaagcggcgagcggggcggaccaagttcaaggagacgcggcaccccgtgttcaagggcgtgcgccggaggaaccccgggaggtgggtgtgcgaggtgcgcgagccgcacggcaagcagcggatatggctcgggacgttcgagacagcagagatggcggcgcgcgcgcacgacgtcgccgcgctcgcgctccgcggccgcgccgcctgcctcaacttcgccgactcgccgaggcgcctccgcgtcccgcccatcggcgcaagccacgacgacatacggagggcggcggctgaggcggccgaggcattccggccgccaccagatgagagcaatgcggccaccgaggtggcagccgccgcatcgggcgccactaattcgaacgccgaacagttcgcctcccacccgtactacgaggtcatggacgatgggctggacttggggatgcagggctatctcgacatggcgcaagggatgctcattgacccgcctccaatggccggtgatcctgccgtaggtagcggcgaagacgacaacgatggcgaggtccagctatggagctactgatcctgcgcgtttgaactcaacttggtttggcgcgaagagatcgcatgtacagcttaagggagtcgagtacaagtacctcaggtgtactccactcgttgcccctttcccttccctttcgtttttcttgagcttatctgcagggtaatgttatgtattgctgctcttctgatgaaatgtgatcggaagaagcggaaggccagatcgagcttatgggttctgaagacggtgaaggcttgtcgagtgttgtgagcatatattcagaaagtcaggcactgtgaaagtatgaatcagatcagccttgttacgaatgagagtgatcgaccttgttcagtgtttataattgaaccacttgtgtgtaataagcagcaaagccatgttgcttgcttgatctgactcttgggaatggtatatttctcaaagaatgcaggattgactactcagaatttgacattttgcagtgaaatgataggattgttaaattaacattggaggagaggcatgtgtatatatgttaagaaacattagtaatgatgagcctatgatacttcgatc</dnaseqindica>\|
−	~~Link = [http://www.ncbi.nlm.nih.gov/nuccore/NM_001054262.1 RefSeq:Os02g0677300]\|~~
−	}}
−	[[Category:Genes]]
−	~~[[Category:Japonica mRNA]]~~
−	[[Category:Oryza Sativa Japonica Group]]
−	~~[[Category:Japonica Genes]]~~
−	[[Category:Japonica Chromosome 2]]
−	~~[[Category:Chromosome 2]]~~
−	1.PAN, X.-W.; LI, Y.-c.; LI, X.-x.; LIU, W.-q.; MING, J.; LU, T.-t.; TAN, J.; SHENG, X.-n., Differential Regulatory Mechanisms of CBF Regulon Between Nipponbare (Japonica) and 93-11 (Indica) During Cold Acclimation. Rice Science 2013, 20 (3), 165-172.

@@ Line 14: / Line 14: @@
 You can also add sub-section(s) at will.
-CBF/DREB1 (dehydrationresponsiveelement-binding protein) proteins belong to a subfamily of AP2/ERF(APETALA2/ethyleneresponsivefactor) transcription factor and contain a highly conserved DNA-binding domain which interacts with CRT/DRE (C-repeat/drought-responsive element)motif and activates downstream genes.This genes share  extensive homology, not only in the AP2/ERF region but also in the C-terminal regions and signatures bordering the AP2/ERF domain[1].we found that all the CBF/DREB1 homologous proteins in rice had a conserved valine in the V14 position and a conserved glutamic acid in V19 position, which play important roles in DNA-binding specificity.At the same time,V14 is a great important for the transcriptional activity of CBF protein.
+CBF/DREB1 (dehydrationresponsiveelement-binding protein) proteins belong to a subfamily of AP2/ERF(APETALA2/ethyleneresponsivefactor) transcription factor and contain a highly conserved DNA-binding domain which interacts with CRT/DRE (C-repeat/drought-responsive element)motif and activates downstream genes.This genes share  extensive homology, not only in the AP2/ERF region but also in the C-terminal regions and signatures bordering the AP2/ERF domain[1].we found that all the CBF/DREB1 homologous proteins in rice had a conserved valine in the V14 position and a conserved glutamic acid in V19 position, which play important roles in DNA-binding specificity.At the same time,V14 is a great important for the transcriptional activity of CBF protein.According to analyze three promoter sequence ofrice CBF and compare the difference sequence between Nipponbare and 93-11,we find that coding region sequence is very conservative among species ,while the promoter region are difference. Somebody  speculate that this may be related to the evolutionary process of selection pressure.
 ==Labs working on this gene==

@@ Line 3: / Line 3: @@
 ==Annotated Information==
 ===Function===
-Nine CBF/DREB1homologous genes in rice were obtained by BLAST search in the NCBI database, which share conserved amino acid sequences with DREB1 protein in Arabidopsis. Three CBFgenes organized in tandem, named OsCBF1, OsCBF2and OsCBF3, showed a transient induction in the process of cold acclimation, much stronger in indica rice 93-11 compared with japonica rice Nipponbare.The candidate downstream genes OsLIP5and OsLIP9were induced in 93-11 but notin Nipponbare. The differential expression of CBF regulon might be caused by polymorphisms within promoter sequences between these two rice varieties.One of the nine homologous CBF/DREB1 genes and the center of resistance to cold way.Gene expression profiling analysis shows that the CBF genes of rice can upstream the express of stress tolerance genes to enhance the cold tolerance of transgenic rice,like OsP5CS,OsLIP5,Os-LIP9,OsRAmy3D and so on.
+Nine CBF/DREB1homologous genes in rice were obtained by BLAST search in the NCBI database, which share conserved amino acid sequences with DREB1 protein in Arabidopsis. Three CBFgenes organized in tandem, named OsCBF1, OsCBF2and OsCBF3, showed a transient induction in the process of cold acclimation, much stronger in indica rice 93-11 compared with japonica rice Nipponbare.The candidate downstream genes OsLIP5and OsLIP9were induced in 93-11 but notin Nipponbare. The differential expression of CBF regulon might be caused by polymorphisms within promoter sequences between these two rice varieties.One of the nine homologous CBF/DREB1 genes and the center of resistance to cold way.The encoding products of CBF genes belongs to the transcriptional activator of AP2/ERF family, which can specifically binding to the promoter that contain CRT/DRE cis-acting elements Kai and start the expression of the downstream genes to improve the cold tolerance of plants.Gene expression profiling analysis shows that the CBF genes of rice can upstream the express of stress tolerance genes to enhance the cold tolerance of transgenic rice,like OsP5CS,OsLIP5,Os-LIP9,OsRAmy3D and so on.
 ===Expression===

@@ Line 28: / Line 28: @@
 .Zhang X, Fowler S G, Cheng H, Lou Y, Rhee S Y, Stockinger E J, Thomashow M F. 2004. Freezing-sensitive tomato has a functional CBF cold response pathway, but a CBF regulon that differs from that of freezing-tolerant Arabidopsis. Plant J, 39: 905–919.
 .Kume S, Kobayashi F, Ishibashi M, OhnoR, NakamuraC, Takumi S. 2005. Differential and coordinated expression of Cbfand Cor/Leagenes during long-term cold acclimation in two wheat cultivars showing distinct levels of freezing tolerance. GenesGenet Syst, 80: 185–197.
-.Kato-Noguchi H. 2008. Low temperature acclimation mediated by ethanol production is essential for chilling tolerance in rice roots. Plant Signal Behav, 3: 202–203.
+.Dubouzet J G, Sakuma Y, Ito Y, Kasuga M, Dubouzet E G, Miura S, Seki M, Shinozaki K, Yamaguchi-Shinozaki K. 2003. OsDREBgenes in rice, Oryza sativa L., encode transcription activators that function in drought-, high-salt- and cold-responsive gene expression. Plant J, 33: 751–763.
-.Dubouzet J G, Sakuma Y, Ito Y, Kasuga M, Dubouzet E G, Miura S, Seki M, Shinozaki K, Yamaguchi-Shinozaki K. 2003. OsDREBgenes in rice, Oryza sativa L., encode transcription activators that function in drought-, high-salt- and cold-responsive gene expression. Plant J, 33: 751–763.
+.Ito Y, Katsura K, Maruyama K, Taji T, Kobayashi M, Seki M, Shinozaki K, Yamaguchi-Shinozaki K. 2006. Functional analysis of rice DREB1/CBF-type transcription factors involved in cold-responsive gene expression in transgenic rice. Plant Cell Physiol, 47: 141–153.
 ==Structured Information==

@@ Line 7: / Line 7: @@
 ===Expression===
 Please input expression information here.
-CBF (C-repeat-binding factor) cold response pathway has proven to play important roles in cold acclimation<ref name="ref1"/>. CBF/DREB1 (dehydration- responsive element-binding protein) proteins belong to a subfamily of AP2/ERF  (APETALA2/ethylene- responsive factor) transcription factor and contain a highly conserved DNA-binding domain which interacts with CRT/DRE (C-repeat/drought-responsive element) motif and activates downstream genes <ref name="ref2"/>. The expression of CBF/DREB1genes is regulated by an upstream transcription factor ICE1 (inducer of CBFexpression 1) <ref name="ref3"/>. After activated by low temperature, the ICE1 protein binds specifically to the MYC (myelocytomatosis oncogene) recognition sequences present in CBF/DREB1promoters and stimulates the transcriptions of CBF/DREB1genes <ref name="ref4"/>. CBF cold response pathway is conserved not only intemperate plants like Arabidopsis, wheat and Brassica napus, but also in tropical plants like rice and tomato<ref name="ref5"/>, <ref name="ref6"/>, <ref name="ref7"/>, <ref name="ref8"/>. Several CBF/DREB1homologous genes identified from rice have proven to improve cold tolerance of transgenic Arabidopsis and rice <ref name="ref9"/>,<ref name="ref10"/>.Using the microarray analysis, several candidate target genes of CBF/DREB1 protein were identified in rice, such as OsP5CS, OsLIP5, OsLIP9,and OsRAmy3D<ref name="ref10"/>.The induction of these target genes improves the cold tolerance of rice plants through mediating various physiological and biochemical processes.
+CBF (C-repeat-binding factor) cold response pathway has proven to play important roles in cold acclimation<ref name="ref1"/>. CBF/DREB1 (dehydration- responsive element-binding protein) proteins belong to a subfamily of AP2/ERF  (APETALA2/ethylene- responsive factor) transcription factor and contain a highly conserved DNA-binding domain which interacts with CRT/DRE (C-repeat/drought-responsive element) motif and activates downstream genes <ref name="ref2"/>. The expression of CBF/DREB1genes is regulated by an upstream transcription factor ICE1 (inducer of CBFexpression 1) <ref name="ref3"/>. After activated by low temperature, the ICE1 protein binds specifically to the MYC (myelocytomatosis oncogene) recognition sequences present in CBF/DREB1promoters and stimulates the transcriptions of CBF/DREB1genes <ref name="ref4"/>. CBF cold response pathway is conserved not only intemperate plants like Arabidopsis, wheat and Brassica napus, but also in tropical plants like rice and tomato<ref name="ref5"/>, <ref name="ref6"/>, <ref name="ref7"/>. Several CBF/DREB1homologous genes identified from rice have proven to improve cold tolerance of transgenic Arabidopsis and rice<ref name="ref8"/>, <ref name="ref9"/>.Using the microarray analysis, several candidate target genes of CBF/DREB1 protein were identified in rice, such as OsP5CS, OsLIP5, OsLIP9,and OsRAmy3D<ref name="ref9"/>.The induction of these target genes improves the cold tolerance of rice plants through mediating various physiological and biochemical processes.
-CBF cold response pathway has proven to play critical roles in cold acclimation in many subspecies <ref name="ref5"/>. In order to know the molecular basis of rice cold acclimation, we examined the expression of CBF/DREB1genes in the process of cold acclimation at 10 °C. As shown in Fig. 4[[File: expression.jpg‎‎|left|thumb|150px|Table : Time-course expression of rice genes and candidate downstream genes in the process of acclimation.]], the three CBF/DREB1genes (OsCBF1,  OsCBF2and OsCBF3) exhibited similar expression profiles during cold acclimation in Nipponbare and 93-11. The transcriptions increased at 1 h and subsequently reachedtheir peaks at 2 h and then decreased. Interestingly, the gene induction was much stronger in 93-11 than in Nipponbare. The CBF proteins can bind to the CRT/DRE regulatory element and activate the expression of downstream target genes<ref name="ref9"/>. Therefore, we further analyzed the expression of several candidate down- stream genes. Cold-induced genes OsLIP5and OsLIP9showed no obvious changes in Nipponbare during cold acclimation. Differently, the expression of these two genes was induced after acclimation for 2 h and then decreased in 93-11. Another candidate target gene OsP5CS, encoding a central enzyme in the proline biosynthesis, exhibited induction both in Nipponbare and 93-11. However, the induction was stronger in 93-11 than in Nipponbare. Based on these results, we proposed that differential induction of CBF/DREB1genes during cold acclimation might result in differential expression of downstream genes and was responsible for significant EL decrease in 93-11 compared to that in Nipponbare.
+CBF cold response pathway has proven to play critical roles in cold acclimation in many subspecies <ref name="ref5"/>. In order to know the molecular basis of rice cold acclimation, we examined the expression of CBF/DREB1genes in the process of cold acclimation at 10 °C. As shown in Fig. 4[[File: expression.jpg‎‎|left|thumb|350px|Table : Time-course expression of rice genes and candidate downstream genes in the process of acclimation.]], the three CBF/DREB1genes (OsCBF1,  OsCBF2and OsCBF3) exhibited similar expression profiles during cold acclimation in Nipponbare and 93-11. The transcriptions increased at 1 h and subsequently reachedtheir peaks at 2 h and then decreased. Interestingly, the gene induction was much stronger in 93-11 than in Nipponbare. The CBF proteins can bind to the CRT/DRE regulatory element and activate the expression of downstream target genes<ref name="ref8"/>. Therefore, we further analyzed the expression of several candidate down- stream genes. Cold-induced genes OsLIP5and OsLIP9showed no obvious changes in Nipponbare during cold acclimation. Differently, the expression of these two genes was induced after acclimation for 2 h and then decreased in 93-11. Another candidate target gene OsP5CS, encoding a central enzyme in the proline biosynthesis, exhibited induction both in Nipponbare and 93-11. However, the induction was stronger in 93-11 than in Nipponbare. Based on these results, we proposed that differential induction of CBF/DREB1genes during cold acclimation might result in differential expression of downstream genes and was responsible for significant EL decrease in 93-11 compared to that in Nipponbare.
 ===Evolution===