Research Article

Bioinformatic Analysis of Calcium-dependent Protein Kinase Family of Medicago truncatula  

Gong Jiyi , Zhang  Yubin , Li  Fei , He  Xiaohong , Zhang  Ximin , Yi  Yin
College of Life Science, Guizhou Normal University, Guiyang, 550025, China
Author    Correspondence author
Molecular Plant Breeding, 2018, Vol. 9, No. 4   doi: 10.5376/mpb.2018.09.0004
Received: 22 Mar., 2018    Accepted: 25 Apr., 2018    Published: 11 May, 2018
© 2018 BioPublisher Publishing Platform
This article was first published in Molecular Plant Breeding (2017, 15: 3454-3462) in Chinese, and here was authorized to translate and publish the paper in English under the terms of Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:

Gong J.Y., Zhang Y.B., Li F., He X.H., Zhang X.M., and Yi Y., 2018, Bioinformatic analysis of calcium-dependent protein kinase family of Medicago truncatula, Molecular Plant Breeding, 9(4): 26-35 (doi: 10.5376/mpb.2018.09.0004)

Abstract

Calcium plays an important biological role in plants, regulating cell wall formation, pollen tube development, biotic and abiotic stress signal transduction and so on. Calcium-dependent protein kinases (CDPKs) genes are able to perceive the change of intracellular Ca2+ concentrations and convert it into specific phosphorylation events, initiating downstream signal transduction. We used bioinformatics data and analysis techniques to identify family members of the CDPKs gene in two varieties of Medicago, namely Medicago truncatula and Medicago lupulina L., and to analyze the structure, evolution and expression of these CDPKs genes, preliminarily predicted the functions of these genes. 25 CDPKs genes present in the genome of Medicago truncatula, which were divided into four evolutionary groups. The evolutionary relationship of CDPKs gene of Medicago truncatula was close to that of Arabidopsis thaliana but had a distant relation with Oryza sativa. Three CDPKs genes, Medtr1g041150, Medtr1g054865 and Medtr1g055255, clustered on chromosome 1 of Medicago truncatula, were highly similar and might be amplified from one gene. We analyzed the expression of CDPKs gene of Medicago truncatulain in different organs and under different treatments with existing gene expression data of Medicago truncatula, and found several genes related to the salt tolerance, root rot resistance and nodule growth of Medicago truncatula.

Keywords
Medicago truncatula; CDPKs gene family; Evolution; Expression pattern

Background

Ca2+ has been regarded as an important secondary messenger with conservative molecular structure in eukaryotic signal transduction for a long time. Calcium-dependent protein kinases and Ca2+ receptors/protein kinase effectors with conservative encoding structure can be found in plants. The role of CDPKs is to perceive the changes of the intracellular Ca2+ concentration and convert them into specific phosphorylation events to further activate downstream signal transduction. There have been numerous studies on the role of CDPKs in abiotic stress and immune signaling, while little is known about that in the long-term evolution and development of plants. This study identified the target proteins of CDPKs in plants, including the analysis of phosphorylation sites in target proteins, which might pave the way to elucidate the mechanism for CDPKs at molecular and biochemical level.

 

The analysis of specific phosphorylation patterns of target proteins indicated that CDPKs acted as signaling centers in the stress signal transduction and development of plants. The family of CDPKs genes included four subfamilies (Harmon et al., 2000; Cheng et al., 2002). 34 family members of CDPKs were identified in Arabidopsis thaliana, 29 family members in Oryza sativa, 20 family members in Triticum aestivum, 35 family members in Zea mays, and 20 family members in Populus trichocarpa (Harmon et al., 2000; Asano et al., 2005; Li et al., 2008; Ma et al., 2013; Zuo et al., 2013). CDPKs had conservative molecular structure, which consisted of a variable N-terminal, a Ser/Thr kinase domain, and a CDPK activation domain (CAD). CAD was composed of pseudo-substrate domain (inhibitory connecting domain) and Ca2+ binding domain (also known as calmodulin-like domain) which was highly homologous with calmodulin. Typical calcium binding domain of CDPKs contained up to 4 EF-hand motifs (Harper et al., 2004; Harper and Harmon, 2005; Boudsocq and Sheen, 2013; Liese and Romeis, 2013). Combining Ca2+ with CDPK activation domain would induce changes of the whole CDPKs conformation and cause the pseudo-substrate domain to leave the active sites and then kinase activity was activated, making CDPK functionable (Liese and Romeis, 2013).

 

CDPKs are involved in many aspects of plant life processes, from environment stress perception and signal transduction to hormone regulation at different developmental stages (Cheng et al., 2002; Ludwig et al., 2004; Asano et al., 2012; Boudsocq and Sheen, 2013). CDPKs play a significant role in plant development. The best known example is the process of pollen tube growth. In elongated pollen tube, Ca2+ presents a volatile gradient. 12 family members of the 34 members of CDPKs gene are highly expressed in Arabidopsis thaliana pollen (Myers et al., 2009). In the double mutants of AtCPK17/AtCPK34 and AtCPK11/AtCPK24, pollen tube growth were deeply flawed (Myers et al., 2009; Zhao et al., 2013). Another important feature of CDPKs is to participate in Ca2+-related signal transduction induced by abiotic stress factors such as salt, drought and low temperature, especially ABA-dependent signal transduction. There have been plenty of reports on changes of CDPKs-dependent ion channels, intracellular metabolic changes, and gene expression changes. CDPKs are considered to have the function of promoting the resistance to abiotic stress, which are able to boost the overexpression of resistance-related kinases in plants as well (Asano et al., 2012; Boudsocq and Sheen, 2013). CDPKs have been acknowledged as a key participator to convert Ca2+ concentration changes which is induced by pathogen infection signals into plant defense responses. These include ROS synthesis, gene expression changes, plant hormone synthesis and signal transduction, even apoptosis (Tena et al., 2011; Boudsocq and Sheen, 2013). After pathogen infection, CDPKs were activated rapidly and then showed lower expression after 15-120 min (Romeis et al., 2000), which indicated that CDPKs played a fundamental role in plant immune responses.

 

Medicago Sativa, as an important pasture, is major feed source for animal husbandry, which is of great significance in economics. Besides, alfalfa cultivars generally have the characteristics of strong resistance and wide adaptability. In this study, through bioinformatics data and analysis technique, we identified family members of the CDPKs gene in two varieties of Medicago, namely Medicago truncatula and Medicagolupulina L. We analyzed the structure, evolution and expression of Medicago CDPKs genes and preliminarily predicted the functions of related genes. This study identified 1 CDPKs gene related to salt-tolerance of Medicago truncatula, 7 CDPKs genes related to nodule growth of Medicago truncatula, and 9 CDPKs genes related to root rot infection of Medicago truncatula, which might lay research basis for further gene cloning and function analysis of these genes.

 

1 Results and Analysis

1.1 Identification and preliminary analysis of CDPKs genes

Genome sequencing of Medicago truncatula had been completed (JCVI, http://medicago.jcvi.org/medicago/). We took CDPKs family members of Arabidopsis thalianaas as query, and retrieved databases of genome sequences (BLASPn) and genome protein sequences (BLASTp) of Medicago truncatula with local BLAST software. Parameter settings for BLAST used the default values. In addition, we downloaded serine and threonine kinase domain (PF10494) and EF-hand domain (PF00036) from Pfam, and built Hidden Markov Model to search databases of local genomes and protein sequences of Medicago truncatula. Genes with both of the 2 domains were ranked as CDPKs candidate genes of Medicago truncatula for further analysis. As for the genes with multiple expressed sequences on a gene locus, we retained the longest amino acid sequence for further analysis.

 

We integrated the CDPKs candidate genes of Medicag otruncatula (Table 1) obtained from domain retrieval with HMMER software and local BLAST retrieval, and analyzed the domains on SMART protein domain site. Complete genes both with serine/threonine kinase domain and EF-hand domain were considered to be CDPKs genes of Medicago truncatula. 25 CDPKs genes existed in Medicago truncatula genome. There were no significant differences in size among these 25 CDPKs genes. The smallest Medtr3g098070 was composed of 495 amino-acids and the size of protein was about 55.92 KD, while the largestMedtr1g054865 consisted of 607 amino-acids and the size of protein was 66.45 KD. The isoelectric point distribution of CDPKs genes of Medicago truncatula was relatively centralized as well, and the isoelectric points of 23 genes were less than 7 (acidic). The isoelectric points of proteins corresponding to the only 2 genes, Medtr5g092810 (7.67) and Medtr5g022030 (9.1), were greater than 7. CDPKs gene of Medicago truncatula contains 4 EF-hand domains, except Medtr8g043970, which contains 3.

 

 

Table 1 Family information of Medicago truncatula CDPKs gene

 

Medicago lupulina L. is a common species of Medicago in karst areas in Guizhou Province, which has adapted to the special conditions of karst areas with high exchangeable calcium content and arid surface. We completed transcriptome sequencing for tender leaves of Medicago lupulina L., and then conducted retrieval and analysis (mentioned above) to the sequences obtained, found that there were only 4 complete genes with both serine/threonine kinase domain and EF-hand domain. Next we analyzed the genetic relationship among these four CDPKs genes of Medicago lupulina L. (Table 2) and that of Medicago truncatula.

 

 

Table 2 Family information of Medicago lupulina L. CDPKs genes

 

1.2 Phylogenetic relation analysis of CDPKs genes of Medicago

We downloaded CDPKs sequences of Oryza sativa from Phyzome website, together with CDPKs sequences of Arabidopsis thaliana, Medicago truncatula and Medicago lupulina L., and constructed the phylogenetic tree by Neighbor Joining (Figure 1). Amino acid sequences in CDPKs of Arabidopsis thaliana were divided into 4 groups (Cheng et al., 2002). The phylogenetic tree we constructed had this characteristic as well, which was divided into 4 branches. CDPKs sequences of Medicago Sativa were closely related to the gene sequences of Arabidopsis thaliana, which was inconsistent with U-box gene family. U-box gene family of Medicago truncatula was more closely related to the sequences of Oryza sativa but relatively distant from Arabidopsis thaliana (Zheng et al., 2015). One of the genes of Medicago truncatula, Medtr8g043970, together with Os06g50146.1 of Oryza sativa, was both genetically distinct from other CDPKs genes.

 

 

Figure 1 Phylogenetic relationship of CDPKs genes in Medicago truncatula, Medicago lupulina L., and Arabidopsis, Oryza. Sativa L.

 

1.3 Analysis of CDPKs domains and gene structure of Medicago truncatula

Genome sequence and cDNA sequence of Medicago truncatula exist simultaneously in JCVI, the bioinformatics database of Medicago truncatula. We extracted cDNA and gene sequence of CDPKs gene of Medicago truncatula and analyzed gene structure in GSDS (gene structure display server) (Figure 2; Figure 3). CDPKs genes of Medicago truncatula generally have 7 or 8 exons and the position of introns is more conservative. Except Medtr8g043970, CDPKs gene of Medicago truncatula included 4 EF-hand domains. Medtr8g043970 only contained 3 EF-hand calcium binding domains.

 

 

Figure 2 Structural analysis of Medicago truncatula CDPKs genes

 

 

Figure 3 Family phylogenetic tree and protein structure of Medicago truncatula CDPKs genes

 

1.4 Chromosome localization analysis of CDPKs in Medicago truncatula

The location information of CDPKs gene of Medicago truncatula could be found in JCVI. We used the location information of CDPKs gene of Medicago truncatula to identify the location of each CDPKs gene on 8 chromosomes of Medicago truncatula by Map Inspect software (Figure 4). CDPKs gene distributed unequally on chromosomes of Medicago truncatula. There was no CDPKs gene on chromosome 2 and 6. Chromosome 1 had a maximum of 6 CDPKs genes. Among them, Medtr1g041150, Medtr1g054865 and Medtr1g055255 aggregated atone place. Besides, phylogenetic analysis showed that these three genes had orthologous relationships, and gene structure analysis indicated that they were genes with conserved structure. Further experiments are needed to analyze that whether Medtr1g041150, Medtr1g054865 and Medtr1g055255 are amplified from the same gene.

 

 

Figure 4 Locations of Medicago truncatula CDPKs genes on 8 chromosomes

 

1.5 CDPKs gene expression and possible function prediction of Medicago truncatula

We found expression data of 17 genes in 25 CDPKs genes of Medicago truncatula from Medicago truncatula genechips database (http://mtgea.noble.org/v2/). The clustering analysis was processed on the expression data of these genes by Cluster software. CDPKs gene of Medicago truncatula showed obvious gene expression specificity (Figure 5). Expression status of the same gene was very different in roots, stems, leaves, flowers, petioles and leaf buds. A lot of CDPKs genes expressed in roots. Among 17 CDPKs of Medicago truncatula with expression data, 10 of them were detected highly expressed in roots. In contrast, among these 17 CDPKs genes, only Medtr5g089320 expressed in leaves. And it specially expressed in leaves and flowers. During the development of Medicago truncatula root nodules, the expression of CDPKs gene in roots dramatically changed. The expression level of Medtr3g051770 reduced with the growth of root nodules, while that of Medtr7g106710 increased. Besides, during the growth of root nodules, the expression level of Medtr5g009830, Medtr5g089320, Medtr7g068710, Medtr3g098070 and etc. all changed. Not many CDPKs genes in roots involved in the response to salt stress. The Medtr5g022030 didn’t express in roots under control condition. After salt stress, its expression level raised and maintained a high level. By contrast, root rot infection had a powerful influence on gene expression profile of CDPKs in roots of Medicago truncatula. Expression levels of Medtr7g106710, Medtr3g051770 and Medtr8g095440 increased after root rot infection, while expression levels of Medtr7g091890, Medtr4g132040, Medtr1g041150, Medtr3g098070, Medtr8g043970 and Medtr8g099095 decreased.

 

 

Figure 5 Expression patterns of CDPKs gene in Medicago truncatula

Note: 1: Flower; 2: Leaf; 3: Petiole; 4: Pod; 5: Root; 6: Stem; 7: Vegetative bud; 8: Seed coat (16~24 dap); 9: Root-denodulated; 10: Root-0 dpi (Nodule control); 11: Nodule-10 dpi (Mature nodules); 12: Nodule-14 dpi (Fixing nitrogen); 13: Nodule-28 dpi; 14: Root-0 h (200 mmol/L NaCl)-control; 15: Root-1 h (200 mmol/L NaCl); 16: Root-2 h (200 mmol/L NaCl); 17: Root-5 h (200 mmol/L NaCl); 18: Root-10 h (200 mmol/L NaCl); 19: Root-24 h (200 mmol/L NaCl); 20: Root-0 h phymatotrichum; 21: Root-72 h phymatotrichum; 22: Root-96 h phymatotrichum; 23: Root-non mycorrhizai; 24: Root-mvcorrhizai different developmental stages and treatments include Nodule (10 d, 14 d, 28 d of nodule development), root phymatotrichum (phymatotrichum pathogen infection for 72 h and 96 h), 200 mmol/L sodium chloride stress (1 h, 2 h, 5 h, 10 h, 24 h)

 

2 Discussion

Calcium plays an important biological role in plants (cell wall formation, pollen tube development, biotic and abiotic stress signal transduction, etc.). CDPKs genes able to perceive the changes of the intracellular Ca2+ concentration and convert them into specific phosphorylation events to further activate downstream signal transduction. CDPKs are intricately involved in many aspects of plant life processes, from environment stress perception and signal transduction to hormone regulation at different developmental stages (Cheng et al., 2002; Ludwig et al., 2004; Asano et al., 2012; Boudsocq and Sheen, 2013). In this study, through bioinformatics data and analysis technique, we identified family members of the CDPKs gene in two varieties of Medicago, namely Medicago truncatula and Medicago lupulina L. We analyzed the structure, evolution and expression of CDPKs genes and preliminarily predicted the functions of related genes.

 

Medicago lupulina L. is a common species of Medicago in karst areas, which has remarkable ability to adapt to the special conditions of karst areas with high calcium content and arid surface. In this study, we identified 4 complete CDPKs genes from the leaf transcriptome of Medicago lupulina L., which was far less than what it obtained from genome sequences of Medicago truncatula, 25 genes. That should be related to the tissue specific expression of CDPKs genes. We analyzed the expression of Medicago truncatula CDPKs gene and found that only 1 Medicago truncatula CDPK highly expressed in leaves (Medtr5g089320). However, that gene (Medtr5g089320) was not orthologous to the genes found in the transcriptome of Medicago lupulina L.

 

The analysis of genetic evolution and chromosome localization of 25 CDPKs genes of Medicago truncatula suggested that Medtr1g041150, Medtr1g054865 and Medtr1g055255 aggregated at the same position of chromosome 1. Besides, these 3 genes with conservative genetic structure were orthologous. The amino acid sequences of these three genes showed 97% homology, with only a few different amino acids in the hyper variable region of N-terminal. Further experiments are needed to analyze that whether Medtr1g041150, Medtr1g054865 and Medtr1g055255are amplified from the same gene. It is a pity that we have not found the expression data of these 3 genes, thus it is unable to analyze expression characteristics and gene functions of these 3 genes.

 

Existing Medicago truncatula gene chip expression data was used to analyze the expression of CDPKs gene of Medicago truncatulain in different organs and under different treatments, and multiple genes related to salt-tolerance, root rot infection and nodule growth of Medicago truncatula were found, which might lay research basis for further gene cloning and function analysis of these genes.

 

3 Materials and Methods

3.1 Data collection and CDPKs gene excavation

The sequences of Medicago truncatula used in this study were downloaded from Medicago truncatula sequence database (JCVI, http://medicago.jcvi.org/medicago/) (Tang et al., 2014). The transcriptome sequence of Medicago lupulina L. used in this study came from tender leaves of Medicago lupulina L. cultivated out of the soil with high calciumin greenhouse, which was handed over to the sequencing company for transcriptome sequencing. Serine/threonine protein kinase domain (PF10494) and EF-hand domain (PF00036) were downloaded from Pfam (Finn et al., 2006).

 

The software used for CDPKs gene excavation was local BLAST and HMMER with default arguments for sequence searching. And online software SMART and Expasy were used for the analysis to characteristics of domain and protein of sequences obtained by searching.

 

3.2 The construction of CDPKs gene phylogenetic tree

CDPKs sequences of Oryza sativa were downloaded from Phyzome. And CDPKs gene sequences of Arabidopsis thaliana were downloaded from Tair. The construction the phylogenetic tree of amino acid sequence in CDPKs gene of Oryza sativa, Arabidopsis thaliana, Medicago truncatula and Medicago lupulina L. employed MEGA 6.0 (Tamura et al., 2013). ClustW method was used for Alignment with default arguments. The phylogenetic tree was constructed by Neighbor Joining.

 

3.3 Construction of CDPKs genetic map

The location information of CDPKs gene came from bioinformatics database of Medicago truncatula (JCVI, http://medicago.jcvi.org/medicago/) (Tang et al., 2014). Gene positioning map was constructed by MapInspect.

 

3.4 Analysis of CDPKs gene expression

CDPKs gene expression data of Medicago truncatula was downloaded from Medicago truncatula Gene Expression Atlas of Noble Research Institute (http://mtgea.noble.org/v2/) (He et al., 2009). Cluster analysis of gene chip data used Cluster 3.0. Java Treeview was used for observing clustering results.

 

Authors’ contributions

GJY was the designer and executor of this study, finishing the paper writing; ZYB completed the data collecting of this study; LF was responsible for furnishing transcriptome data of Medicago lupulina L.; HXH participated in experimental design and paper modification; ZXM was in charge of the experiment, guiding paper writing and modification; YY offered invaluable guidance for paper writing. All authors read and approved the final manuscript.

 

Acknowledgments

This study was supported by the Science Foundation of Guizhou Province (2017GZ66249).

 

References

Asano T., Hayashi N., Kikuchi S., and Ohsugi R., 2012, CDPK-mediated abiotic stress signaling, Plant Signaling & Behavior, 7(7): 817-821

https://doi.org/10.4161/psb.20351

PMid:22751324 PMCid:PMC3583972

 

Asano T., Tanaka N., Yang G., Hayashi N., and Komatsu S., 2005, Genome-wide identification of the rice calcium-dependent protein kinase and its closely related kinase gene families: comprehensive analysis of the CDPKs gene familyin rice, Plant & Cell Physiology, 46(2): 356-366

https://doi.org/10.1093/pcp/pci035

PMid:15695435

 

Boudsocq M., and Sheen J., 2013, CDPKs in immune and stress signaling, Trends in Plant Science, 18(1): 30-40

https://doi.org/10.1016/j.tplants.2012.08.008

PMid:22974587 PMCid:PMC3534830

 

Cheng S.H., Willmann M.R., Chen H.C., and Sheen J., 2002, Calcium signaling through protein kinases: the Arabidopsis calcium-dependent protein kinase gene family, Plant Physiology, 129(2): 469-485

https://doi.org/10.1104/pp.005645

PMid:12068094 PMCid:PMC1540234

 

Finn R.D., Mistry J., Schuster-Böckler B., Griffiths-Jones S., Hollich V., Lassmann T., Moxon S., Marshall M., Khanna A., Durbin R., Eddy S.R., Sonnhammer E.L., and Bateman A., 2006, Pfam: clans, web tools and services, Nucleic Acids Research, 34: 247-251

https://doi.org/10.1093/nar/gkj149

PMid:16381856 PMCid:PMC1347511

 

Harmon A.C., Gribskov M., and Harper J.F., 2000, CDPKs: a kinase for every Ca2 signal? Trends in Plant Science, 5 (4):154-159

https://doi.org/10.1016/S1360-1385(00)01577-6

 

Harper J.F., and Harmon A., 2005, Plants, symbiosis and parasites: a calcium signalling connection, Nature Reviews Molecular Cell Biology, 6(7): 555-566

https://doi.org/10.1038/nrm1679

PMid:16072038

 

Harper J.F., Breton G., and Harmon A., 2004, Decoding Ca2+ signals through plant protein kinases, Annual Review of Plant Biology, 55(1): 263-288

https://doi.org/10.1146/annurev.arplant.55.031903.141627

PMid:15377221

 

He J., Benedito V.A., Wang M., Murray J.D., Zhao P.X., Tang Y., and Udvardi M.K., 2009, The Medicago truncatula gene expression atlas web server, BMC Bioinformatics, 10(1): 441

https://doi.org/10.1186/1471-2105-10-441

PMid:20028527 PMCid:PMC2804685

 

Li A.L., Zhu Y.F., Tan X.M., Wang X., Wei B., Guo H.Z., Zhang Z.L., Chen X.B., Zhao G.Y., Kong X.Y., Jia J.Z., and Mao L., 2008, Evolutionary and functional study of the CDPK gene family in wheat (Triticumaestivum L.), Plant Molecular Biology, 66(4): 429-443

https://doi.org/10.1007/s11103-007-9281-5

PMid:18185910

 

Liese A., and Romeis T., 2013, Biochemical regulation of in vivo function of plant calcium-dependent protein kinases (CDPK), Biochimica Et Biophysica Acta, 1833(7): 1582-1589

 

Ludwig A.A., Romeis T., and Jones J.D., 2004, CDPK-mediated signaling pathways: specificity and cross-talk, Journal of Experimental Botany, 55(55): 181-188

PMid:14623901

 

Ma P., Liu J., Yang X., and Ma R., 2013, Genome-wide identification of the maize calcium-dependent protein kinase gene family, Applied Biochemistry and Biotechnology, 169 (7):2111-2125

https://doi.org/10.1007/s12010-013-0125-2

PMid:23397323

 

Myers C., Romanowsky S.M., Barron Y.D., Garg S., Azuse C.L., Curran A., Davis R.M., Hatton J., Harmon A.C., and Harper J.F., 2009, Calcium-dependent protein kinases regulate polarized tip growth in pollen tubes, The Plant Journal, 59(4):528-539

https://doi.org/10.1111/j.1365-313X.2009.03894.x

 

Romeis T., Piedras P., and Jones J.D.G., 2000, Resistance gene-dependent activation of a calcium-dependent protein kinase in the plant defense response, Plant Cell, 12(5): 803-816

https://doi.org/10.1105/tpc.12.5.803

PMid:10810151 PMCid:PMC139928

 

Tamura K., Stecher G., Peterson D., Filipski A., and Kumar S., 2013, Mega6: molecular evolutionary genetics analysis version 6.0., Molecular Biology & Evolution, 30(12): 2725-2729

https://doi.org/10.1093/molbev/mst197

PMid:24132122 PMCid:PMC3840312

 

Tang H., Krishnakumar V., Bidwell S., Rosen B., Chan A., Zhou S., Gentzbittel L., Childs K.L., Yandell M., Gundlach H., Mayer K.F., Schwartz D.C., and Town C.D., 2014, An improved genome release (version mt4.0) for the model legume Medicago truncatula, BMC Genomics, 15(1): 312

https://doi.org/10.1186/1471-2164-15-312

PMid:24767513 PMCid:PMC4234490

 

Tena G., Boudsocq M., and Sheen J., 2011, Protein kinase signaling networks in plant innate immunity, Current Opinion in Plant Biology, 14(5): 519-529

https://doi.org/10.1016/j.pbi.2011.05.006

PMid:21704551 PMCid:PMC3191242

 

Zhao L.N., Shen L.K., Zhang W.Z., Zhang W., Wang Y., and Wu W.H., 2013, Ca2+-dependent protein kinase11 and 24 modulate the activity of the inward rectifying K+ channels in Arabidopsis pollen tubes, Plant Cell, 25(2): 649-661

https://doi.org/10.1105/tpc.112.103184

PMid:23449501 PMCid:PMC3608784

 

Zheng X.W., Shao L.H., and Li C., 2015, Genome-wide screening and characterization of the U-box gene family in Medicago truncatula, Caoye Xuebao (Acta Prataculturae Sinica), 24(8): 130-141

 

Zuo R., Hu R., Chai G., Xu M., Qi G., Kong Y., and Zhou G., 2013, Genome-wide identification, classification, and expression analysis of CDPK and its closely related gene families in poplar (Populus trichocarpa), Molecular Biology Reports, 40(3): 2645-2662

https://doi.org/10.1007/s11033-012-2351-z

PMid:23242656

Molecular Plant Breeding
• Volume 9
View Options
. PDF(588KB)
. FPDF
. HTML
. Online fPDF
Associated material
. Readers' comments
Other articles by authors
. Gong Jiyi
. Zhang  Yubin
. Li  Fei
. He  Xiaohong
. Zhang  Ximin
. Yi  Yin
Related articles
. Medicago truncatula
. CDPKs gene family
. Evolution
. Expression pattern
Tools
. Email to a friend
. Post a comment

ass fuck sexo anal Gruppen Pornos Blondine Pornos inzest porn hd
ankara escort
casumo
escort bodrum
erotik filmler
beykent spor salonu
free brazzers
istanbul escort