April 2010, Volume 52 Issue 5, Pages 434ĘC514.

Cover Caption: MAP Kinase in Environmental Responses
The mitogen-activated protein kinase (MAP Kinase) cascade is one of the major pathways by which extracellular stimuli are transduced into intracellular responses in plants. MAP Kinases have been shown to play important roles in plant growth and development, and in responding to environmental stresses. In this issue, Wang et al. (442ĘC452) report the isolation and functional analysis of a novel MAP Kinase gene, ZmMPK3, from maize. The expression profile and kinase activity suggest that ZmMPK3 is involved in delivering various stress signals in maize leaves.


          Cell and Developmental Biology
Viability Loss Pattern under Rapid Dehydration of Antiaris toxicaria Axes and its Relation to Oxidative Damage
Author: Xia Xin, Xing-Ming Jing, Yong Liu and Song-Quan Song
Journal of Integrative Plant Biology 2010 52(5): 434-441
Published Online: February 25, 2010
DOI: 10.1111/j.1744-7909.2010.00924.x

The relation between oxidative damage and viability loss of excised embryonic axes of Antiaris toxicaria subjected to rapid drying with silica gel at 15 °C was studied. Changes of survival rate, accumulation of thiobarbituric acid-reactive substances (TBARs), activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), glutathione reductase (GR) and the permeability of cell membrane that was determined as relative electrolyte leakage (REL) were measured. The half-life moisture content (MCL50) was 0.41 g H2O/g DW (dry weight basis). During drying, the activities of SOD, CAT and APX increased until MCL50, and declined thereafter. The generation speed of ·O2, and content of H2O2 and TBARs remained steadily or even decreased at MC levels higher than MCL50, demonstrating a low oxidative level in these axes. There was no significant correlation between viability loss and accumulation of reactive oxygen species or lipid peroxidation within the dehydration process until MCL50. Whereas the increase in REL from the beginning of the drying process indicated that the cell membrane was damaged. In conclusion, under rapid drying with silica gel the viability loss of excised recalcitrant A. toxicaria axes seemed to be triggered by mechanical or physical damage, rather than metabolic damage.

Xin X, Jing XM, Liu Y, Song SQ (2010) Viability loss pattern under rapid dehydration of Antiaris toxicaria axes and its relation to oxidative damage. J. Integr. Plant Biol. 52(5), 434–441.

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          Plant-environmental Interactions
A Novel Mitogen-Activated Protein Kinase Gene in Maize (Zea mays), ZmMPK3, is Involved in Response to Diverse Environmental Cues
Author: Jinxiang Wang, Haidong Ding, Aying Zhang, Fangfang Ma, Jianmei Cao and Mingyi Jiang
Journal of Integrative Plant Biology 2010 52(5): 442-452
Published Online: March 1, 2010
DOI: 10.1111/j.1744-7909.2010.00906.x

In search for components of mitogen-activated protein kinase (MAPK) cascades in maize (Zea mays) involved in response to abscisic acid (ABA) stimulus, a novel MAPK gene, ZmMPK3, from ABA-treated maize leaves cDNA was isolated and characterized. The full length of the ZmMPK3 gene is 1 520 bp and encodes a 376 amino acid protein with a predicted molecular mass of 43.5 kD and a pI of 5.83. ZmMPK3 contains all 11 MAPK conserved subdomains and the phosphorylation motif TEY. Amino acid sequence alignment revealed that ZmMPK3 shared high identity with group-A MAPK in plants. A time course (30–360 min) experiment using a variety of signal molecules and stresses revealed that the transcripts level of ZmMPK3 accumulated markedly and rapidly when maize seedlings were subjected to exogenous signaling molecules: ABA, H2O2, jasmonic acid and salicylic acid, various abiotic stimuli such as cold, drought, ultraviolet light, salinity, heavy metal and mechanical wounding. Its transcription was also found to be tissue-specific regulated. Here, we show that ABA and H2O2 induced a significant increase in the ZmMPK3 activity using immunoprecipitation and in-gel kinase assay. Furthermore, the results showed that the ZmMPK3 protein is localized mainly to the nucleus. These results suggest that the ZmMPK3 may play an important role in response to environmental stresses.

Wang J, Ding H, Zhang A, Ma F, Cao J, Jiang M (2010) A novel mitogen-activated protein kinase gene in maize (Zea mays), ZmMPK3, is involved in response to diverse environmental cues. J. Integr. Plant Biol. 52(5), 442–452.

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Identification and Cloning of Differentially Expressed Genes Involved in the Interaction Between Potato and Phytophthora infestans using a Subtractive Hybridization and cDNA-AFLP Combinational Approach
Author: Maria Antonia Henriquez and Fouad Daayf
Journal of Integrative Plant Biology 2010 52(5): 453-467
Published Online: March 25, 2010
DOI: 10.1111/j.1744-7909.2010.00943.x

Using a subtractive hybridization (SH)/cDNA-AFLP combinational approach, differentially expressed genes involved in the potato-Phytophthora infestans interaction were identified. These included genes potentially controlling pathogenesis or avr genes in P. infestans as well as those potentially involved in potato resistance or susceptibility to this pathogen. Forty-one differentially expressed transcript-derived fragments (TDFs), resulting from the interaction, were cloned and sequenced. Two TDFs, suggested as potential pathogenicity factors, have sequence similarity to N-succinyl diaminopimelate aminotransferase and a transcriptional regulator, TetR family gene, respectively. Two other TDFs, suggested as potential avr genes, have sequence similarity to an EST sequence from Avr4/Cf-4/Avr9/Cf-9 and a P. infestans avirulence-associated gene, respectively. Genes' expression and origin were confirmed using Southern blots, Northern blots and qRT-PCR. I.e., potential resistance gene DL81 was induced at 12 hpi in the moderately resistant cultivar, whereas it was down-regulated as early as 6 hpi in the susceptible cultivar. On the other hand, DL21 was induced at 6 hpi (3.38-fold) in response to the highly aggressive isolate (US8) and strongly up-regulated thereafter (25.13-fold at 120 hpi.), whereas it was only slightly up-regulated in response to the weakly aggressive isolate US11 (3.82-fold at 96 hpi), suggesting its potential involvement as a susceptibility gene.

Henriquez MA, Daayf F (2010) Identification and cloning of differentially expressed genes involved in the interaction between potato and Phytophthora infestans using a subtractive hybridization and cDNA-AFLP combinational approach. J. Integr. Plant Biol. 52(5), 453–467.

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Growth Response to Ionic and Osmotic Stress of NaCl in Salt-tolerant and Salt-sensitive Maize
Author: Ke-Fu Zhao, Jie Song, Hai Fan, San Zhou and Meng Zhao
Journal of Integrative Plant Biology 2010 52(5): 468-475
Published Online: April 22, 2010
DOI: 10.1111/j.1744-7909.2010.00947.x

Salt-tolerant maize (STM) and salt-sensitive maize (SSM) were treated with 100 mM NaCl for 1, 3 and 6 d and the contents of Na+ and Cl (cps) of different organelles of leaf cells determined by X-ray microanalysis. The results showed that Na+ and Cl entered the cytoplasm, vacuole, chloroplast and apoplast simultaneously. When STM and SSM were treated in 100 mM NaCl at atmospheric pressure (−P) and with pressure equivalent to the osmotic pressure of the NaCl (+P), the dry weights of STM (+P) and SSM (+P) plants were greater than that of STM (−P) and SSM (−P) plants, showing that the inhibitory effect of salt on plant growth was at least partially due to the osmotic effect of the NaCl. When STM and SSM were treated with NaCl and iso-osmotic polyethlene glycol, the dry weights of plants given the iso-osmotic polyethlene glycol treatment were lower for both maize lines than that of the NaCl-treated plants. Our data show that under NaCl stress, both STM and SSM seedlings simultaneously suffered from osmotic and ion stresses.

Zhao KF, Song J, Fan H, Zhou S, Zhao M (2010) Growth response to ionic and osmotic stress of NaCl in salt-tolerant and salt-sensitive maize. J. Integr. Plant Biol. 52(5), 468–475.

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Inducible Positive Mutant Screening System to Unveil the Signaling Pathway of Late Blight Resistance
Author: Zhiqi Jia, Yanhong Cui, Ying Li, Xiaoxuan Wang, Yongchen Du and Sanwen Huang
Journal of Integrative Plant Biology 2010 52(5): 476-484
Published Online: February 5, 2010
DOI: 10.1111/j.1744-7909.2010.00915.x

Late blight is the most devastating potato disease and it also causes serious yield loss in tomato. Several disease resistance genes (R genes) to late blight have been cloned from potato in the past decade. However, the resistance mechanisms remain elusive. Tomato and potato belong to the botanical family Solanaceace and share remarkably conserved genome structure. Since tomato is a model system in genetic and plant pathology research, we used tomato to develop a powerful mutant screening system that will greatly facilitate the analysis of the signaling pathway of resistance to Phytophthora infestans. First we proved that the R3a transgenic tomatoes developed specific hypersensitive cell death response (HR) to P. infestans strains carrying the corresponding avirulence gene Avr3a, indicating that the signaling pathway from the R3a-Avr3a recognition to HR is conserved between potato and tomato. Second, we generated transgenic tomatoes carrying both R3a and Avr3a genes, with the latter under the control of a glucocorticiod-inducible promoter. Dexamethasone induced expression of Avr3a and resulted in localized HR. This versatile system can be used to construct a mutant library to screen surviving mutants whose resistance signal transduction was interrupted, providing the basis to identify key genes involved in the resistance to late blight in Solanaceae.

Jia Z, Cui Y, Li Y, Wang X, Du Y, Huang S (2010) Inducible positive mutant screening system to unveil the signaling pathway of potato late blight resistance. J. Integr. Plant Biol. 52(5), 476–484.

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Molecular Characterization of Three Ethylene Responsive Element Binding Factor Genes from Cotton  
Author: Long-Guo Jin, Hui Li and Jin-Yuan Liu
Journal of Integrative Plant Biology 2010 52(5): 485-495
Published Online: March 7, 2010
DOI: 10.1111/j.1744-7909.2010.00914.x


Ethylene-responsive factors (ERFs) are important regulators of plant gene expression. In this study, three novel ERF genes, GhERF2, GhERF3 and GhERF6, were isolated from cotton (Gossypium hirstum) using rapid amplification of cDNA ends-polymerase chain reaction. Transient expression analysis using GhERF-green fluorescent protein fusions showed that these three proteins were targeted to the nucleus. Fusion proteins consisting of GhERF2, GhERF3 or GhERF6 coupled to the GAL4 DNA binding domain strongly activated transcription in yeast. Furthermore, GhERF6 was shown to be able to bind specifically to GCC boxes using a particle bombardment assay in tobacco cells. Semi-quantitative reverse transcription-polymerase chain reaction revealed that GhERF2 and GhERF3 are constitutively expressed in all organs, while GhERF6 is only constitutively expressed in vegetative organs. When plants were treated with ethylene, abscisic acid, salt, cold and drought, the transcripts of GhERF2, GhERF3 and GhERF6 were rapidly induced to high levels. Promoter analysis also indicated that the 5' upstream regions of the three genes possess elements induced by these physiological and environmental factors. Collectively, our data suggest that GhERF2, GhERF3 and GhERF6 might function as positive trans-acting factors in the plant responses to ethylene, abscisic acid and other stresses and provide useful clues for further research into the mechanism of them in regulating cotton multiple stress responses.

Jin LG, Li H, Liu JY (2010) Molecular characterization of three ethylene responsive element binding factor genes from cotton. J. Integr. Plant Biol. 52(5), 485–495.

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          Molecular Physiology
Reverse Genetic Identification of CRN1 and its Distinctive Role in Chlorophyll Degradation in Arabidopsis  
Author: Guodong Ren, Qian Zhou, Shouxin Wu, Yufan Zhang, Lingang Zhang, Jirong Huang, Zhenfei Sun and Benke Kuai
Journal of Integrative Plant Biology 2010 52(5): 496-504
Published Online: March 23, 2010
DOI: 10.1111/j.1744-7909.2010.00945.x

Recent identification of NYE1/SGR1 brought up a new era for the exploration of the regulatory mechanism of Chlorophyll (Chl) degradation. Cluster analysis of senescence associated genes with putative chloroplast targeting sequences revealed several genes sharing a similar expression pattern with NYE1. Further characterization of available T-DNA insertion lines led to the discovery of a novel stay-green gene CRN1 (Co-regulated with NYE1). Chl breakdown was significantly restrained in crn1-1 under diversified senescence scenarios, which is comparable with that in acd1-20, but much more severe than that in nye1-1. Notably, various Chl binding proteins, especially trimeric LHCP II, were markedly retained in crn1-1 four days after dark-treatment, possibly due to a lesion in disassociation of protein-pigment complex. Nevertheless, the photochemical efficiency of PSII in crn1-1 declined, even more rapidly, two days after dark-treatment, compared to those in Col-0 and nye1-1. Our results suggest that CRN1 plays a crucial role in Chl degradation, and that loss of its function produces various side-effects, including those on the breakdown of Ch-protein complex and the maintenance of the residual photosynthetic capability during leaf senescence.

Ren G, Zhou Q, Wu S, Zhang Y, Zhang L, Huang J, Sun Z, Kuai B (2010) Reverse genetic identification of CRN1 and its distinctive role in chlorophyll degradation in Arabidopsis. J. Integr. Plant Biol. 52(5), 496–504.

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Identification of Rice Al-responsive Genes by Semi-quantitative Polymerase Chain Reaction using Sulfite Reductase as a Novel Endogenous Control
Author: Jianjun Zhang, Ying Yin, Yuqi Wang and Xinxiang Peng
Journal of Integrative Plant Biology 2010 52(5): 505ĘC514
Published Online: March 16, 2010
DOI: 10.1111/j.1744-7909.2010.00931.x

Based on the evidence that Al resistance is an inducible process and rice is an Al-resistant crop, identification of Al-responsive genes from rice may help to further clone Al-resistant genes in plants. Semi-quantitative and real-time polymerase chain reaction (PCR) is widely applied in gene transcriptional analyses, particularly for those genes with low transcript abundance. Normalization with proper endogenous control (EC) genes is critical for these two approaches in terms of reliability and precision. We first noticed that the expression of several commonly-used EC genes was depressed under Al stress, while sulfite reductase gene (SR) was stable throughout the Al treatment. The reliability of SR as an EC gene was further tested by analyzing the expression of a number of genes in response to Al challenge. Except for the consistent results obtained for the four previously-identified genes, nine additional genes were newly defined as Al-responsive in this study. Collectively, our results suggest that SR can be used as a novel EC gene for semi-quantitative and real-time PCR analysis of Al responsive genes, and that activated transport of silicon and stimulated metabolism of carotenoid and terpenoid could be involved in Al resistance in rice plants.

Zhang J, Yin Y, Wang Y, Peng X (2010) Identification of rice Al-responsive genes by semi-quantitative polymerase chain reaction using sulfite reductase as a novel endogenous control. J. Integr. Plant Biol. 52(5), 505–514.

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